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TRANSCRIPT
Curcuma cf comosa Roxb
A study on wound healing effect of a herbal plant,
Curcuma cf comosa Roxb
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~1'L.!~2.J1~'tliJt')'Vlfln'l::;~'Wn1'1~1tl"lJ'El'IU1!11LLCJ~'Vi'..:J1'L.!~~'ElVI'Vl !11~'El'l (in vitro) LL~::;1ul!PlrJ'Vl VI~'El'l (in
vivo) Ld'El~ 00-1'1..11 LU'Wl'h fu l"l12.J~il~'l'WCJ~2.J"1J'El..:J~1'1~n t11 ~l'lu L'Elfi~'El::;sn L!Pl'Vl"ll'El'l'i1'L.!~2.J1~'El ~u'i1il
t')'Vl'ifn'l::;~'L.!n1'1~1ti"1J'El'IU1!11LLCJ~1'Wl1Plrf'Vlt11~'El'l (in vivo) .,
~'IU'W Yl"l!~2.J'L.!L~'l'i1'W~2.J1~'tl (Curcuma cf. comosa Roxb.) ~..:~ilfll'nwnv.lum'lU1LU • ~ ~
~ ~u1 Luum 1 mJ1 'L.!'El'L.!11"l!Pl ~1'tl'El1"'1 ~..:~ L~1~ 1 ~iJ m '11 ~tJ'l::; LU"l!U"'11n~ 3-!'WL v. '1Yl'L.!U1'L.!UL~ LU'L.!U1
fn'l!t1 LLCJ~ 'VI t11 LL 'Vl'L.!m LLe.~u11"'1"'1U'L.! LL~ ::;'tl1"'1U1LU~n1'1CJ~ 1?1 Luw • .nl"l!.ri'ru"VIL:n..:~~1W'llu1~ 'L.!'El'U11"l (;! . ~
'ii\Jini .. W. .. ~.~k .. ~.o~%,111 -tci1Jtl:!UUU.'!-f.YJ .. b..~}fg ... ::·~....! )...!}~~ lWt~BWfl .. ~ ... ~.J~!:~· ... ?~60;
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n1 ~ijr.ft~Uilf r;]Cp.J~~i'l.:J f'fL ~'fl ~ m~n~VIim~~'Wn1 ~1.-1'1 tl"ll'fl.:JU'l"' LLI:J~i.:~ L 'Wt-1~'fl "'VI"'~'fl.:J (in
vitro) LL~~lul~J~rfVI"'~'fl.:J (in vivo) "ll'fl.:Ji'l'l~i'ln(P)t-1tl'lU~'lm'flfi~'fl~~LIJIVI"ll'fl.:J~'l'Wt-13.J'l~'fl (Curcuma
ct. comosa Roxb.) LL~~VI(P)i'l'flU~VIim~~'Wn1~1.-1'ltl"ll'fl.:JU'l"'LLI:J~L'Wi!JirfVI"'~'fl.:J"ll'fl.:JI;'hfuF~1J.J~il ~'l'We.li'l3.J"ll'fl.:Ji'l'l~i'l fi"' ~'l tJ L'flfi~'fl~~ LIJIVI"ll'fl.:J~ 'l'W\-13-J'l~'fl
m~~n'l!t'l~VIim~~'Wn'lffi'ltl"ll'fl.:JU'l(P) LLI:J~ l 'Wt-1~'fl(P)YJ(P)~'fl.:J (in vitro) v'h L(P)tJVI (P)i'l'flU ~VIim~ L~J.J~'l'W'J'WL"l!~~L V'l'l~Lit~.:~ Lil'fl L~'flL~&J'JWW.:J"ll'fl.:!t-1'W"l!il(P) l929 LL~~ L"11~~3.J~L1.:~thnJ.J (P)~n"ll'fl-:!3-J'WMcl'
~ ~ . ...
olll:lllo I.ICI!t.cl • • Clit." "l!'W(P) Hela cell (P)'Jtl'Jfi WST-1 cell proliferation assay LL~~VI(P)~'fllJ~VIfim~f?!'Wn1~1.-1'ltl"ll'fl~
o II .c:~~o.~ ....,. 1.1
lJ'l(P)LLI:J~'"l'lnLLUU'"l'l~'fl.:Jn'l~t-1'ltl"ll'fl~U'l(P)LLe.J~ (P)'Jtl'Jfi Scratch assay "ll'fl.:Ji'l'l~~n"'t-1mU"''ltl I I
Ci1t. C11t. I II .o:::ai 1.1 1.1 Cilt..... I OIV I II Qj
L'flfi~'fl~"l! LIJIVI"ll'fl.:J'l'l'Wt-13-J'l'l'flVI fil'l'l3.JL "113-J"li'W 25-400 f.JQ/ml I:J~n'l~'l'"lt!V'IU'l'l i'l'l~i'lnm'l'W\-13-J'l'l'flVI r 1 ., .,
1.1 1-' -=1 Cllt.CIIt. 0 cr" oiiC:II cv Cit. I <ICill
fil'l'l3.JL"113.J"li'W 400 f.JQ/ml 3.J~VIfiLV'I3.J'"l'l'W'J'WL"l!~~LV'I'l~L~tJ.:JVI.:J"l!'W(P) L929 LL~~ Hela cell 'fltl'l-:!3-J
l!t~t1'l~n 'Ld'fl Lmt~uL~t~uriul?i''lmUFIJ.J~LlJil~'l~i'ln"'i'lJ.J'WLV'I~ L(P)t~ilf'l'l Proliferation index (P.I.) •~;~ . . tr I I li
L'Vhnu 1.09 LL~~ 1.19 IJI'l3.J~'l~U LL~~i:J~VIfi~~~"' L'Wn'ln~J.J~'l'W'l'WL"l!~~vl~"l!il(P) L929 LL~~ Hela
cell lif=JJ"'riuL'f'l~~"'~L'l~'l 24 -i'ltJ.J~ m ~~nM'l ~VIim~~um~vt'ltl"ll'fl~U'l"' LLI:J~ 1ul~JJ1VI "'~'fl~ (in vivo) "ll'fl~i'l'l~i'ln "'vtmu ~'J tJ
I I oC1to C1it. I !.1 4::111 !,/ !.1 0 CV q Q.o:::al IV !11
L'flfi'~'fl~"l!Lr;]VI"ll'fl~'l'l'Wt-13.J'l'l'e1VIfil'l'l3.J L"113.J"ll'W 0.1% w/v LL~~ 0.2% w/v LL~~IJI'l~Ufi1~3.JYI3.Ji'l'l~i'ln(P)(P)'Jtl
L'flfl~'e1~6JiLIJIVI"ll'fl~~'l'Wt-13.J'l~'fl~fii'J'l3.JL~3.J~'W 5% w/w LL~~ 10% w/w vi'lL'Wt-1t:YJ(P)~'fl~i'l'ltlvr'W~
C57BU6 'fl'ltj 8 lu"'1'1f ~'l'W'l'W 6 n~J.J n~J.J~:; 6 l?i''l L(P)t~i'l1'l~U'l(P)LLI:J~~'lt~mn'"l'l~lifL11ul~1u~ ... I
vl~i'l'fl~~.,~~'Jtl metal ear punch "li'W1(P)L~'We.hfl!'W!fmn-3 2 ij~~L:W(;J1 VI1~11VIf'l~'fl1JVJL1JVJU1L'dnJ .. .. ...
U'l(P)LLe.J~Vjn1'u 1'u~~ 1 F~f~ LijuL'l~'l 30 1'u t"'t~liflu~~'l.:Joii'lt~"ll'fl.:Jt-1t:'V!nl>i'lVI'l~'ltJL>i'lF~'lU'")J.J n~J.J
~ 1-3 Vl'l~'lm'flfi~'fl~=llLIJIVI LL~~t-1t:n~J.J~ 4-6 Vl'l~'ltl cream base ~'Julu~~'l~"ll'l'lVI'l~ht~i'l'l~i'lri"'
I I I CV !.1 Clit. Cllt. I !.1 cf !1,.1 !.1 1 .rc:i 0 .., Q .q_q Q..o"
i'l'l~i'ln"'(P)'lUL'flfi'~'fl~"l!LIJIVI"ll'fl~'l1'WV13.J'l'l'flVIFI'l'l3.JL"113.J"ll'W 5% w/w (n~J.JVI 4)1 IJI'l~Ufii~3.JV13.Ji'l'l~~n(P) • ~'lUL'flfl~'fl~~LIJIVI"ll'fl~~'l'Wt-13-J'l~'fl~fil'l'lJ.JL~J.J~'W 10% w/w (n~J.J~ 5) LL~~ Hirudoid®(n~J.J~ 6) • •
I
healing) e.~~m~ij{tJV'IU~'l i'l'l~i'ln(P)~'lm'flfi~'fl~6JiLIJIVI"ll'fl\l~'l'Wt-13.J'l~'fl~fil'l13.JL~J.J~'W o. 2% w/v il~'fltl I
.::::.1~ ..::;,! I fV I oO:::il IV 0 IV .q.q
~~"ll'fl·m'lJ'Vl'ltl"ll'fl'llJ'l(P)LLe.J~(P)YJ~(P) (P)n'J'l!JI'lfil'llJ'")J.JLL~~ Oxoferin® 'fltl'l\IJ.J'Wtli'l'lfiltlJVI'l'li'lrliJI (p <
0.05) L(P)t~vh1~u'l"'LLe.J~u"'i'li1VJ1~1u1'u~ 30 (~'fltJ~:; 100) ~'l'W~'lfuF~1J.J~ili'l'l~~ri"'~'lm'flfi~'fl~6Ji
Ill
L(;)'Vl'lJ'tl~~TW'I.-U.J17J'tl~rl'l1:WL ;:W;'W 5% LL~:; 10% w/w iJ1'tltJ~:;'lJ'tl~n1~VI1tl'll'tl~1J1 I?ILLe.J~ ln~ L~tl~rl'W
Pi'fl1'fltJ~:; 66.67 LL~:; 62.12 lurfu~ 30 rn1:wfhc;i'u :i~~n~11K'lr!'llll"l:W LL~:; Hirudoid® 'tlth~iJ •
tl't~~11"i'qJ'V11~~ti ~ (p < 0.05) Lrl'tl~:W(;)1'l"'l=i'uL rl''tl'V11~,~n1ti~Jl1 r~f'11~ rnf'll'fl~ Lif'tl L~'fllllL'lru I I tr t ., 1 coil -=1 4Cilt. "' ell Cl dl dl 1
1J11?1LLe.J~'Vl 1J~'ll'tl~VI~'Vl'Vll?l~'tl1J~'Vlnn1:;~'Wn1~VI1tl'll'tl~1J11?1LLe.J~ L ~'tl~~tl1n~Jl1~'ll'tl~ L'W'tl Ltl'tl 'W
tJ'W~ 0, 2, 5, 10 LL~:; 20 LI"Plmh:;d1'W"'l1nrl'l1:W,.'W1LLU'W'll'tl~ polymorphonuclear (PMN),
"' ' fibroblast, collagen fiber LL~:;VI~'tli?IL~'tll?l LL~:;m~~11~'li''WVIU~rl1~11LoW'tlUIPI1.hnLLe.J~ e.J~m~~{tl
' ... .... 1,1 .c:::llio 4Cilt. I II -=f 4Cilt. .:::5I
~1~~nl?ll?l'ltiL'tln~'tl:;•l!Lrn'Vl'll'tl~'l1'WVI:W1'l'tl :wmn~:W'li'W'll'tl~ PMN, fibroblast, collagen fiber LL~:;
Vl~'tli?IL~'tll?l:W1nn~1 Oxoferin®, Hirudoid® LL~:;n~:Wrl'll.Jfl:W ~1'W~{tlif~l1.JL~~1 ~11~nl?l~'ltl
L'tlfl~'tl :;en L[;J 'V1 'll'tl~ ~1 'WVI :W 17J'tl iJ ~ 'V1 in 1:; ~'W n 11VI1 tl 'll 'tl~ 1.J 1 1?1 LLe.J ~ i~ l 'W VI~ 'tll?l 'V1 1?1~ 'tl~ LL~:; l 'W
~(;)tl'Vll?l~'tl~ 9-:JiJPi'nt~.n1~1um1ih11.J~oo.J'W1LU'WmlVI3-llu'tlu1r~rn
fi1ft1trty : ~'Vlin1:;~'Wn1~VI1tl'll'tl~l.J11?1 LLe.J~ l'WVI~'tll?l'Vll?l~'tl~ ~'Vlim:;~'Wn1~VI1tl'll'tl~1J11?1LLe.J~ lu
~ (;)tl'Vll?l~'tl~ ~11~ rl 1?1VIti11.J ~'l tiL'tlfl~'tl:;en L(;)'Vl'lJ'tl~~1'WVI:W17J'tl ~~~j, fu 1"1 ~:w~SJ~'J'We.J~:W
IV
ABSTRACT
The objectives of these researches were to investigate for in vitro and in vivo wound
healing effects of ethyl acetate crude extract of Curcuma cf. comosa Roxb. and in vivo
wound healing effects of formulated cream of Curcuma cf. comosa Roxb.
In vitro wound healing effects of ethyl acetate crude extract of Curcuma cf. comosa
Roxb. at concentration of 25-400 j.Jg/ml were studied from cell proliferative activities in both
mouse subcutaneous connective tissue cell cultures (L929) and human cervical carcinoma
cell cultures (Hela) by WST-1 cell proliferation assay and scratch assay that using as
wound healing models. The results showed that crude extract at concentration of 400 j..lg/ml
had significantly increased amount of cell cultures both L929 and Hela with proliferation
index (P.I.) of 1.09 and 1.19, respectively and also had strong activity to stimulate cell
growth to close the space within 24 hours.
In vivo wound healing effects of ethyl acetate crude extract of Curcuma cf. comosa
Roxb. at concentration of 0.1% and 0.2% w/v, formulated creams composed of 5% and 10%
w/w ethyl acetate crude extract of Curcuma cf. comosa Roxb. were studied. The 8 weeks
old of C57BL/6 mice were divided into 6 groups (n=6). Both ears of mice were punched to
make a hole with 2-millimeters of diameter by metal ear punch. Samples were applied to
wounds once a day for 30 days. Left ears of group 1-3 were applied with the control of ethyl
acetate solvent. Right ears of group 1 and 2 were applied with 0.1% w/v and 0.2% w/v of
ethyl acetate extracts of Curcuma cf. comosa Roxb., respectively. Right ears of group 3
were applied with Oxoferin®. While left ears of group 4-6 were applied with the control of
cream base. Right ears of group 4 and 5 were applied with 5% w/w and 10% w/w of
formulated creams of Curcuma cf. comosa Roxb., respectively. Right ears of group 6 were
applied with Hirudoid®. Wound healing effects were determined by taking photographs and
measuring the diameter of the wounds that be calculated into % of wound healing. The
results showed that 0.2% w/v of ethyl acetate extract had % of wound healing significantly
more than control and Oxoferin® (p < 0.05). The wound was completely healed at the day of
30 (100%). Whereas 5% and 10% w/w of curcuma creams had indifferent of %wound
healing about 66.67% and 62.12% at the day of 30, respectively. But both formulated
-
v
creams had significantly wound healing effect better than cream base and Hirudoid® (p <
0.05). The pathological effects of histology of these ears were also examined at day o, 2, 5,
10, 20 to evaluate the amount of polymorphonuclear (PMN), fibroblast, collagen fiber and
vessels and the formation of epithelium healing the wound. The histological effects
expressed that tissues around the wound area of treatment groups were found PMNs,
fibroblasts, collagen fibers and vessels greater than control groups. In conclusion, ethyl
acetate extract of Curcuma cf. comosa Roxb. had potential both in vitro and in vivo wound
healing effects to be developed to a new drug in the future.
Keywords : In vitro wound healing effect, In vivo wound healing effect, ethyl acetate crude
extract of Curcuma cf. comosa Roxb., formulated creams of ethyl acetate crude
extract of Curcuma cf. comosa Roxb.
VI
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... 1111..11
.. 1 ~'VIflnj'~~'l.!n1j''W1U"ll'el~tJ1j;JLL~~ (%wound healing) 1-uilwf'VIj;J~fl-1 57
' - "' I II Cll 1.1 1.1
"ll'el-3~11~Mrmnu!Pl'dU ethyl acetate "llfl..:J'J1'WVI2-J1'J'el'VIf'l'J1l-JL"lll-J"ll'U
0.1% w/v LL~~ 0.2% w/v LdflLmtltJL~tltJntJ Oxoferin® L~'tlL'J~1wi1..:11 nu .. 2 '1VIfim~~'Wn1~V11U'll'ti..:JtJ1!PILL~~ (%wound healing) 1u~~ntfVJ~~fl..:J 61
' ' D &.1 ...::!~ .£!11..:i - S.. I tJ Q 1.1 1,.1
"llfl-11n11tJml-l'VIl-1~17~n!Plmu ethyl acetate "llfl~'J1'WVI2-J1'J'el'VIf'l'l12-JL"lll.l"ll'U
5% w/w, 10% w/w Ld'tlLtnUULfiuunu Hirudoid® Ld'flL'Jt'il1wi1-31 n'W
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1
2
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4
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7
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9
10
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16
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cv ~1'i'l.HU111~ ..,
~'\.l'l1tJVIl.J1~'fl Curcuma cf comosa Roxb.
L1J'll'fl~~1'1JVIl-J1~'fl Curcuma cf. comosa Roxb.
~'IJLL~:;t?l'flfi'll'fl~~1'1JVIl-J1~'fl Curcuma cf. comosa Roxb.
t?l'flfi'll'fl~~1'1JVIl-J11'fl Curcuma cf. comosa Roxb.
~'IJLL~:;LVI~1'll'fl~~1'1JVIl-J11'fl Curcuma cf. comosa Roxb.
LVI~1'll'fl~~1'1JVI:IJ11'fl Curcuma cf. comosa Roxb.
"' Brick and mortar model 'll'fl~Ej'.JVIU~"li'IJ stratum corneum
m:;1J'.l'IJn1~V11tl'll'fl~1J1t?lLLtJ~ (wound healing)
Oxoferin® (Tetrachlorodecaoxide)
Hirudoid® (Heparinoid)
n1~L~1:; L1JVI'll'fl\IVI'IJL~'f!Lil Lii t?lLLCJ~ .. v . "'
n1n'l1~ 1 ~'Vl 1?1~ 'fl1JVl1J1 L '.l n.\1J 1 1?1 LLC.Jfl 'll'fl\IL1JVI'Yl\l~'fl~olh~'ll'fl\IVI'IJ'Vl 1?\~'fl\1 .. .. ~'VljfL~l-J~1'1J'.l'IJL"llfl~Ltl'f!L~'flL~Ej'.lVIU~'ll'fl\IVI~"l!Ut?l L929 (mouse
subcutaneous connective tissue) 'll'fl~~1~~nt?lrJl'.ltJ ethyl acetate I
'll'fl\1~1'1JVIl-J11'fl (Curcuma cf. comosa Roxb.) Vlrl'.l1l-JL~:IJ~'1Jt;h~1rl'IJ .. I
fl'VlflLYil-J~1'1J'.l'IJL"llfl~:IJ:;L~\IU1n:IJI?lfln'll'fl~:IJ'IJMU"l!Ut?l Hela cell (human ., .. . cervical carcinoma) "ll'fl\li:'l11i:'lrll?l~btl ethyl acetate "ll'fl\lrJ1'1JVIl-J1~'fl
I q !1!11 cv
(Curcuma cf. comosa Roxb.) 'Vlrl'.l1l-JL'li:IJ'li'IJlPl1~1n'IJ
~'Vlim:;~um~VI1t.l'll'fl~1J1t?lLLCJflL'IJVI~'flt?l'Vlt?l~'fl~ ~'.ltl~fl Scratch assay
Ln'flL:iJil~1~~nt?l~:l-j'IJLoY~~L'IJL"l!~~L'Y'l1:;Liti~"J!ilt?l L929 ~L'.l~1t;i1~1 rl'IJ
~'Vlim:;~um~VI1 t.l'll'fl\11J1 1?1 LLCJ~ 1 'WVIfl'fl t?l'Vl t?l~'fl\1 ~'.ltl~fi Scratch assay
'll'fl~~1~~nt?l~'.lt.l ethyl acetate 'll'fl\1~1'1JVI:W11'fl (Curcuma cf comosa Roxb.)
~rl'.l1:1JL~:IJ~'IJ 25 IJQ/miL'WL"l!~~LYn:;L~"'t.l\l"l!Ut?l L929 ~L'.l~111h~1 rl'W
., 'WIJ1
4
5
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12
14
22
23
32
32
36
37
39
40
~'Vlim:;~um~VI1tl'll'fl\11J1 t?lLLCJflL'IJVI~'flt?l'Vl t?l~'fl~ ~'.ltJ~fi Scratch assay 41
'll'fl~~1~~nt?l~'.lt.l ethyl acetate 'll'fl\1~1'WVI:IJ1~'fl (Curcuma cf. comosa Roxb.) I " I
Vlrl'.l1:1JL;]:IJ'll'W 50 IJQ/miL'IJL"l!~~LYn:;L~t.l\l"l!Ut?l L929 VlL'.l~1t;i1\11 rl'IJ
IX
18
19
20
21
22
~'Vlim:::~um~VI'U..I"ll'il..:J1J11?\LLI:J~L't..!VI~'ill?l'Vll?l~'il..:J rJI'lu~ Scratch assay
"ll'il~~1~i'Hil?l~hu ethyl acetate "ll'il~tl1't..!VI3J1~'il (Curcuma cf. comosa Roxb.) ' "' '
Vll"l'l13JL~3J~'t..! 100 ~g/miL't..!L"ll~£fLvn:::L~U~"l!UI?l L929 Vl L'l~1r;l1~1 riu
~'Vlim:::~um~V11U"ll'il~1J11?\LLI:J~L't..!VI~'ill?l'Vll?l~'il~ rJI'lu~ Scratch assay
"ll'il~~1~~ril?lrJ!'lU ethyl acetate "ll'il~t)1't..!VI3J1~'il (Curcuma cf. comosa Roxb.)
~l"l'l13JL~3JiJU 150 ~g/miL't..!L"ll~£fL'Yl1:::Lifu~"l!UI?l L929 ~L'ltnr;l1..:!1 riu
~'Vlim:::~um~V11U"ll'il..:J1J11?\LLI:J~L't..!VI~'ill?l'Vll?l~'il~ rJI'lu~~ Scratch assay
"ll'il~~1~~ril?lrJ!'lU ethyl acetate "ll'il~tl1't..!VI3J1~'il (Curcuma cf. comosa Roxb.)
' "' ' Vll"l'l13JL~3J~'t..! 200 ~g/miL't..!L6Jl~~L'Yl1:::L~U~"l!UI?\ L929 VJL'l~1r;l1~1 riu
~'Vlim:::~um~V11U"ll'il..:J1J11?\LLI:J~luvr~'ill?l'Vll?l~'il~ rJI'lu~ Scratch assay
"ll'il~~1~~nl?l~l'lu ethyl acetate "ll'il~~1't..!VI3J1~'il (Curcuma cf. comosa Roxb.)
~l"l'l13JL~3J~'t..! 400 ~g/miL't..!L"ll~~L'Yl1:::Lifu~"l!UI?l L929 ~L'l~1r;l1~1 nu
~'Vlim:::~um~VI1 U"ll'il~1J11?l LLI:J~ luvr~'ill?l'Vll?l~'il~ rJI'l u~~ Scratch assay
"ll'il~ Doxorubicin L't..!L"ll~£fL'Yl1:::Lifu~"llill?l L929 ~L'l~1r;l1~1 riu
., 'III'W1
42
43
44
45
46
23 ~'Vlim:::~um~VI1U"ll'il~1J11?\LLI:J~L't..!VI~'ill?l'Vll?l~'il~ rJI'lu~~ Scratch assay 47
L~'ilhJiJ~1~~nl?l~~'t..!L'Yl~L't..!L6J!~£fL'Yl1:::Lifu..:~"J!ill?l Hela cell ~L'l~1r;i1..:!1 nu
24 ~'Vlim:::~l..!n11'VI1E.I"ll'il-:J1J11?\LL!:.i~b'wvr~'ill?l'Vll?l~'il'l ~'lEJfjfi Scratch assay 48
"ll'il'l~1~~nl?l~'ltl ethyl acetate "ll'il'lt)1'WVI3J11'il (Curcuma cf. comosa Roxb.)
' "' ' Vll"l'l13JL~3J~'t..! 25 ~g/miL't..!L"ll~£fL'Yl1:::L~H>I"l!UI?l Hela cell VJL'l~1r;i1~1 nu
25 ~'Vlim:::~um~V11U"ll'il'11J11?\LL!:.i~luvr~'ill?l'Vll?l~'il~ rJI'lu~ Scratch assay 49
"ll'il~~1~~nl?lrJI'lu ethyl acetate "ll'il'ltl1't..!VI3J1~'il (Curcuma cf. comosa Roxb.) ' "' '
Vll"l'l13JL~3J~'t..! 50 ~g/miL't..!L6J!~£fL'Yl1:::L~U'I"l!UI?l Hela cell VJL'l~1r;i1..:~1 nu
26 ~Vlim:::~um~VI1U"ll'il'I1J11?\LLI:J~luvr~'ill?l'Vll?l~'il'l rJI'lu~~ Scratch assay 50
"ll'il'l~1~~nl?lrJI'lu ethyl acetate "ll'il'ltl1't..!VI3J1~'il (Curcuma cf. comosa Roxb.) ' "' '
Vll"l'l13JL~3J~'t..! 100 ~g/miL't..!L"ll~£fLvn:::L~U'I"l!UI?l Hela cell VJL'l~1r;l1..:~1 nu
X
27
28
29
30
31
32
33
34
35
~'Vlim~~'Wn11~1£J"ll'EI~1J1!?1LLt:.~~'l'W'IH"l'fll?l'Vll?l~'EI~ ~'l£.1~ Scratch assay
"ll'fl~~11~nl'l~'l£J ethyl acetate "ll'fl~~1'W~:J.J1~'fl (Curcuma cf. comosa Roxb.) ' ... '
Vll"l'l1:J.JL-ii:J.J<t'W 150 IJg/ml L'WL'11~iL'Y'I1!::L~U\I"llill'l Hela cell VJL'J~1~1\11 n'W
~'Vlim~~'Wn11~1£J"ll'E1~1J1!?1LLt:-~~1'W~~'Eil'l'Vll?l~'fl..:J ~'lu~fi Scratch assay
"ll'fl..:J~11~nl'l~'lU ethyl acetate "ll'EI~rl1'W~:J.J1~'fl (Cwcuma cf. comosa Roxb.)
~l"l'l1:J.JL<t:J.J<t'W 200 1Jg/mi1'WL'11~iL'Y'I1:::Liu..:~"llill'l Hela cell ~L'J~1~1..:!1 n'W
~'Vlim:::~'Wn11~1U"ll'fl~1J1!?1LLt:.~~'l'W~~'fll'l'Vll'l~'fl..:J ~'lu% Scratch assay
"ll'fl..:J~11~nl'l~'JU ethyl acetate "ll'fl..:J~1'W~:J.J1~'fl (Curcuma cf. comosa Roxb.) ' ... '
Vll"l'l1:J.JL<t:J.J<t'W 400 1Jg/miL'WL'l!~iL'Y'I1!::L~U..:J"llill'l Hela cell VlL'J~1~1..:!1 n'W
~'Vlim:::~'Wn11~1U"ll'fl..:J1J1!?1LLt:-~~1u~~'fll'l'Vll'l~'fl~ ~'lu% Scratch assay
"ll'fl..:J Doxorubicin L'WL'11~iL'Y'I1:::Liu~"llill'l Hela cell ~L'J~1~1~1 n'W
~mtru:::n1U'W'fln"ll'EI..:J1J1!?1LLt:.~~~u1L'Jru1u~"ll'fl~~'W'Vll'l~'fl~ 1u1'u~ o (A) LL~~ .. 'II
1'u~ 30 (B) L~'fl'V111u~oii1..:J"ll'J1~'lU~11~nl?l~'JU ethyl acetate "ll'fl~~1'W~:J.J1~'fl
l"l'l1:J.JL<t:woiiu 0.1% w/v LL~!::'V111u~<t1~~1U~'lU~'J'Vh~:::~1U ethyl acetate
~nMru:::1l1£J'W'fln"ll'fl..:J1J1!?1LLt:.~~~u1L'Jruluwn'fl~~'W'Vll?l~'fl..:J 'lurfu~ o (A) LL~::: .. .. 1'u~ 30 (B) L~'fl'V111u~oii1-:J"ll'J1~'JU~11~nl?l~'Ju ethyl acetate "ll'fl-:J~1'W~:J.J1~'fl fl'l1:J.JL;J:w;Ju 0.2% w/v LL~!::'V11'lu~;J1-:~.j!1u~l'Jut;i''lvh~~~1£J ethyl acetate
~nMru:::n1U'W'fln"ll'fl-:J1J1!?1LLt:.~~~u1L'1ru1u~"ll'fl-:Jwt.J'Vll?l~'fl-:J lurfu~ o (A) LL~::: ' ' "" ... "" 1 ... ... ® 1 ... ... ... 'J'W'Vl 30 (B) L:J.J'fl'V11 1JW1J1-:J"ll'J1l?l'JU Oxoferin LL~!::'V11 1J~"ll1-:J'1!1Ul?l'JU .. ..
!K'J't11~:::~1U ethyl acetate
m1'Y'IL~'WLL~l'l-:J~'Vlim:::~'Wn11~1U'll'fl-:J1J1!?1LLt:.~~ (%wound healing) lu~~rf ' IV i.l I 1.1' q t,r !I
'Vll'l~'fl\l"ll'fl\1~11~n l'l~U11Jl'l'JU ethyl acetate "ll'fl-:J'J1'W~:J.J1'J'fl'Vlfi'11:J.J L"ll:W"ll'W ' ' 4 -~ q tV ® dl I Q..o'
0.1% w/v LL~::: 0.2% w/v L:J.J'flLuJ'U1JL'VlU1Jn1J Oxoferin L:W'flL'J~1~1-l1 nu .. m1'V'ILLvl-:JLL~l'l\I~'Vlfim:::~'Wn11~1U'll'fl-:J1J1!?1LLt:.~~ (%wound healing) lu~~rf
' CV 9.1 I !.1 ..::1 i,l !.1
'Vll'l~'fl\l"ll'fl\1~1J>~n l?l~U11Jl'l'JU ethyl acetate "ll'fl-:J'J1'W~:J.J1'J'fl'Vlfi'11:W L'll:J.J"ll'W ' ' .dl -~ .oC::li CV @ .dJ I CV
0.1% w/v LL~::: 0.2% w/v L:J.J'flLuJ'U1JL'VlU1Jn1J Oxoferin L:J.J'flL'J~1~1-l1 n'W
XI
... ~1.11
51
52
53
54
55
56
56
58
59
36
37
38
39
imjnJ~fi1U'Uf.ln~f.l,UJ1~LLCJfl~1Jh'lruLU~~f.l"'VI~VI~flf.l"' luiu~ 0 (A) LLfl~ 1u~ 30 (B) L~f.lV11L1JVI;1"1~'l1~'lf.ll;'hful"i12-l~ij~11t'Hl~~'lU ethyl acetate .. ~f.l-3~1'UVI2J1~f.l~fll'l12JL;2J;'U 5% w/w LLfl~VI1Lll\1;1-3sif1U~'lU cream base
imjru~fi1U'Uf.ln"'lf.l-31J1~LLC.Jfl~1J1L'lt1JL1J\1"11f.l-3VI~VI~flf.l"' L'UrJ'U~ 0 (A) LLfl~ 1u~ 30 (B) L~f.lV11LU\1;1"1"11'l1~'lU~1fUfli12J~ij~11~ri~~'lU ethyl acetate
"..lf.l-3~1'UVI2J1~'El~fll'l12JL~2J;u 10% w/w LLfl~V11LU\1;1-3.Jl1u~'lU cream base
in]jnJ~ll1U'Uf.ln"11f.l-31J1~LLCJfl~1J1L'lnJL1J\1"11f.l-3VI~VI~flf.l-3L'UrJ'U~ 0 (A) LLfl~
iu~ 30 (B) L~f.lV11L1JV1~1-3"11'l1~'lU Hirudoid® LLfl~V11L1JV1;1-3.Jl1U~'lU .. .. cream base
m1~L~'ULL~~-3~Vtim~~'Un11VI1U"'If.l-31J1~LLC.Jfl (%wound healing) lui[;lrf
' ' 0 U .o=f c=i.cll Goo" ~ I 1.1 ..::::1
Vltnflf.l-3"11'El-3 [;I'WlJ fll12-l'Vl2J~11~ n tn tn'l u ethyl acetate "11f.l-3'l1'UV12J1'lf.lVI fll'l12J
L;2J;'U 5% w/w LLfl::; 10% w/w Ldf.lLmUULVlU1Jrl1J Hirudoid® Ldf.lL'lr11c;i1-31 rl'U .,
.... VI'U1
60
60
60
62
40 m1~LLvl-3LL~tn-3~'Vlfim::;~Un11V11U"11f.l-31J1~LLCJfl (%wound healing) lui[;lrf 63
' ' 0 V <11:::11 <C:il-=f - !.1 1 !.1 .c:lil 'Vl~flf.l\1"11f.l-3[;1111Jfli12J'Vl2J~11~n~~'lfl ethyl acetate "11f.l-3'l1'UV12J1'lf.l'Vlfll'l12J
L~2J~'U 5% w/w LLfl::; 10% w/w Ldf.lLmU1JLYlU1Jrl1J Hirudoid® Ldf.lL'lr11c;i1-31 rl'U
41 CJfln11[;11'l"'liuLtlf.l"'lf.l\l L tlf.l Lmf.lu1L 'l t1J1J1 ~ LLC.Jfl~L1J\1~f.l-3VI~ L 'UrJ'U~ 0 64
42 CJfln1~m'l"'l:fuLtlf.l"lJ'El-3 Ltl'fl L~'fl1J1L'l t1J1J1tn LLC.Jfl~L1JV1"11'fl-3VI'W L 'UrJ'W~ 2 Ldf.l'Vl1 65 .. ..
' L~2J;'U 0.1% w/v, (B) ~1~~ri~VIti11J~'lfl ethyl acetate "11f.l-3~1'WVI2J11'flVlfll'l12J
L~2J~'W 0.2% w/v, (C) Oxoferin® LLr~::; (D) ethyl acetate
43 " !.f ""' I I I I
C.Jfln1 :i'[;l1'l"'l='n'UL il'fl"llf.l-3 L il'fl LU'fl1J1L 'l n.l1J1 ~ LLCJr!VlL1JV1"11f.l\l VI'U l 'UrJ 'UVl 5 Lij'fl'Vl1 'IJ ..
66
' ~'lU~11'Vltn~'El1J (A) ~11~ri~VIti11J~l'lu ethyl acetate "'I'El-3~1'UV12J1~'flVlfll'l12J
' L~2J~'U 0.1% w/v, (B) ~1:i'~rl~VIU11J~'lfl ethyl acetate "11'fl-3~1'UVI2J11'flYlfll'l12J
L~2J~'W 0.2% w/v, (C) Oxoferin® LLtl::; (D) ethyl acetate
XII
XIII
... 'VI'U1
44 S,. S, "' 1 I I I
~~n'l~(;]~'J~=ii'WLU'a"ll'fNILU'a L~'fl1J1L 'l CW1J1 (;I LL~~Vil1J~"11tl'I~~L'WtJ'WVi 1 0 Li4'fl'VI1 68 ' 1.1 CV 1.1 I 1.1 _q
(;l'Jti~1~'VI(;l~fl1J (A) ~1~~n(;l~ti11J(;'I'Jtl ethyl acetate "11'tl'l'l1'W~:J.J1'ltl'VIFI'l1:J.J ' 1.1 J,l - 1,1 I 1.1 c:;rl
L"ll:J.J"11'W 0.1% w/v, (B) ~1~~n(;l~ti11J(;l'ltl ethyl acetate "11'tl'l'l1'W~:J.J1'l'a'YIFI'l1:J.J
L~:J.J~'W 0.2% w/v, (C) Oxoferin® LL~::: (D) ethyl acetate
45 69 ' 1,1 U 1,1 I So/c:i
(;l'Jti~1~'VI(;l~fl1J (A) ~1~~n(;l~ti11Jmtr ethyl acetate "11'a'l'l1'W~:J.J1'l'a'VIFI'l1:J.J ' 1.1 1.1 - !.1 I 1.1 ~
L"ll:J.J"ll'W 0.1% w/v, (B) ~1~~n(;l~ti11J(;l'ltl ethyl acetate "ll'a'l'l1'W~:J.J1'l'tl'VIFI'l1:J.J
L~:J.J~'W 0.2% w/v, (C) Oxoferin® LL~::: (D) ethyl acetate ll 1.1 II I I I I
46 ~~n'l~(;]~'l"i=ii'WLU'tl"ll'tl'ILU'tlLU'tl1J1L'lru1J1(;1LL~~Vi1u~"11'tl'I~'WL"WiuVi 2 Li4fl'VI1 .. .. 70 ' "' 0 11V ..:::.1 G:lil.c::i fV !.1 I 1.1
(;l'lti~1~'VI(;l~fl1J (A) (;]1~1JM:J.J'VI:J.J~1~~n(;l(;l'Jtl ethyl acetate "11'tl'l'l1'W~:J.J1'ltl
' I !,1 q !,1 J,.l • • ® 'l1'W~:J.J1'J'a'VIFI'J1:J.JL"ll:J.J"ll'W 10% w/w, (C) H1rudo1d LL~::: (D) cream base
47 ~~n'l~(;]~'l"i~'WLif'a"ll'a\1 Lif't!L~'a1J1L 'ln.!1J1 (;'ILL~~~L1J~"ll'a\IWI~L'WtJ'W~ 5 L~'a'VI1 71
' ' ViFI'l1:J.JL~:J.J~'W 5% w/w, (B) ~1fuFI1:J.JVii1~1~~rl(;l~'Jtl ethyl acetate "ll'a\1
' I !JQ !Ill , ,@
'J1'W~:J.J1'J'eJ'VlFI'l1:J.JL"ll:J.J"11'W 10% w/w, (C) H1rudo1d LL~::: (D) cream base
48 73 '
~'Jti~1~'VI'"l~'a1J (A) ~1f1JFI1:J.JVii1~1~~rl(;l~'ltl ethyl acetate "ll'e:l\1~1'W~:J.J11'a ' '
.11::11 "' "' 0 .... q .:::l.c:i .... "' 'V1FI'l1:J.JL"ll:J.J"11'W 5% w/w, (B) (;]1~1JFI~:J.J'VI:J.J~1~~n'"l(;l'ltl ethyl acetate "ll'a\1
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'l1'W~:J.J1'J'tl'VIFI'l1:J.JL"ll:J.J"ll'W 10% w/w, (C) Hirudoid LL~::: (D) cream base
49 ~~ n'l ~(;] ~'l~~'WL if'a"ll'a\1 L llfl L~'a1J1L 'l CW1J1 '"1 LL~~~L1J~"ll'a'I~'W L 'WtJ 'W~ 20 L~'tl'VI1 .. " 74 ' !I 0 IV .c:i _qq CV i,l 1 1.1
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'VlFI'l1:J.JL"ll:J.J"11'W 5% w/w, (B) (;]1~1JFI~:J.J'VI:J.J~1~~n'"l(;l'ltl ethyl acetate "11'a\l
' I 1.1 ...::.1 S,.. i,.l ® 'l1'W~:J.J1'l'a'VIFI'l1:J.JL"11:J.J"11'W 10% w/w, (C) Hirudoid LL~::: (D) cream base
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n~~ t L~n~ ~tl 1:-Bif.t.LULBL~U.f~ ,n.L[L9":t LttL!b~t Lr:-L~ ~!}!I.t.M tnii~!I'.~:::LM.~, 11~1 _!1.!I.Il1:::tf1~ ~,~t .. . ~tLttllLUIT.Bl.)lB~~~L!I'.ttt.t.Yt.ULilM.11A.LB1L!\tl.Jil~~t.LU~~tlf.~?Ut.LYL~t.LUUL~~Utt-L~~IA.IT.tl.!l'.tl.~
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t.LunL~ntrt.Lf:Lt.Lu.Lrnrn~~LI 11LU!.'LfM.LVIT.~B~~~~~~ Lr:-L~~ray~t.1uL~~Bt.t.,~LB1t.LU~~~~B I
t1. trt.~ tl.,(t~~~np;. 1 1:-~ M.LV!aU _lel t!~ t Ltl.I'M ,M.t. LUIT._f~t~ttt I:-,!1_!31A.M. 11t.L UI:-LIA.t.LUL!aiA.~~I:-1~ Bts1 I ... I
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1J1 1'1 LLf.Jf'l (wound) Vl~1tltl-1 ~'fltl'iln"ll1 I'I"J..'fl-.1 f.J'JVI'U..:JVI~'flL'U'fl Ltl'fl LLf'l~~'J'U'Vlf'ln n'l1"11'Uf.J'JVI'U-1
" ' ' Qn.Yh~1 tl li11~chvru..:J LLtln'"l1 n rl'U Ll'l 1:.1~1 L Vl'111'1ri [;11~ LLf'l~li11ih U'el LU'fl i.~f1Jtl'U[;111tl ~..:)
" nr:::U'J'Un11VI1 1:.1"1.1 'fl-1 LLf.J f'l~::: Ln I'I~'UL~ L'fl-1 [;11 ~fi11~"111 ~ LLr;iVI1 nu1 1'1 LLf.J f'l ;j Fl'l 1 ~ ('ULLN ~ 1 n
nr:::U'l'Un11~..:Jn~1'l~~~'fl..:J1-iiL'l~1'U1Un~1~~vr1tJd~uun~ ~-1~1d1u~~~'fl-11-iimfm!t11J11'1LLr.J~i'l~
~'Jtl ~..:Ji.~;jn-wfii"' fl'ULLf'l :::r.J~ [;lt11fn 1i1 LLf.Jf'lVIf'l1 ti"llill'l vT-.11:.11 ~;j~'JUU~~n'fl1J"JJ'il-1ti1U!j~'l'U:::1'J ~ n-.1
..J.a~ .J 1 I .:61 IV .J ti1'Vl~~'J'Uf.J~~'fl'U1'UlULL1JUm-11 L"ll'U Solcoseryl® L'i1u protein free haemolysate ~1nLf'l'fll'l'l'l "11-1
"li'Jmf-1"111J'J'Un1~~~1'ULLf.Jf'l vi11~LLf.Jf'lVI1tiL~'J LLf'l:::ilm1~~1-1~'JVI'I!-11~~~\j~nf " ' ~1n~'fl~f'l!?]1nmfiuu1u"1.1tl-1Lvm ~u~1il~~ui.~1VIf'l1tJ"llili'IVI;j~n~Firufn1i1LLr.Jf'l L"liu .. . .
~1'UV11-1~~~L~ (Aloe vera) U'JUn (Centella asiatica) LLf'l:::"llifu"liu (Curcuma tonga Linn) LU'U~'U L"'ti~1'UVI1-1~ 1~ L ~~~1-ii~'l'ULiJ'fln 1 um1fn1i1 LLr.Jf'l i. rJi. vriJ J1~'il'Uf'l'ln 1uu'lu n 1-ii1 umrfn1i1 LLf.Jf'l
o" 1.1 ca.".., 1.1 ....,. J'.,.q~ L ctl -.J.J rJn"111 LLf'l~LVI-11"1J~'U"ll'U1"111 'Un11m1i1LLC.Jf'l1 'Um'~L ~1~ 'U'fln~1n'Uti-1~~"11~~'U Vi1'flnVI~1tl"ll'UI'I'Vl L"ll'fl
~1i1~11~Firu1um1fn1i1LLr.Jf'l LLr;iu..:Jhii.~ilm1~n1i1'Vl1..:J~'Vlmfll1~[;lf'flu1..:JLi1u1:::uu • ~1'UVI~1~'fl (Curcuma cf. comosa Roxb) LU'U~~ui.~r~~ui.~1uiu~fl1FI
11V ctl .:61 I .dl CV .c::l J 1.1 0 'l"' [;l:::'J'U'il'flnL'ilti-1LVI'U'fl[;l'fl'Uf'l1~ ~1nn11flln1i1f'ln1in.l~ll1ti'U'eln"ll'fl~~"ll (morphology) LU'fl~[;l'U 'Vl1 ~VI
'Vl111J~1~1'U"llill'lfuuufi"111u[;11~Qf'l Curcuma 'l..:Jff Zingiberaceae LVI~1iiin1iru~nf'l~1 "1l'U11'1 L[;l
LVhn1,j'U~1vr'JJ (L~'Uc.i1~'U~nf'l1~U~~~1n.l 3-10 "11~.) ~"1.11'l'U'lf'l iin~'UL~nU'eltl vT..:J~'l'ULVI~1LLf'l~1u ilin1iru~FI~1t.J~1Uin~l'l~n (Curcuma comosa Roxb.) LLr;i~1nn1~[;l~'J~L'flnin1iru11'1t.Jn11
fJLI"ln~'lf DNA "ll'fl-.1 18s ribosome LLf'l~ Trnk Ll'lt.l Professor Katsuko Komatsu LL1A~ Toyama
' " Medical and Pharmacuetical University u~~L'VlfllqJ~'U ~u~1fi"ll~~ui.~nill'luiiil'll"l~L'flL'Vl~
LL[;1nr;i1-1~1n~1uin~!'!~n (Curcuma comosa Roxb.) 'flth-:~ihm 2 ~1LLVIU-1 (;J'fl2.!~U..:JL3JL~~~~v~) I . II' ll I I
~-1il Fl'l1 ~ LU'U 1tJ1~~1~ 1-u-Yi uLil'fl..:JUm~ Li~u~ 1t.1'rl'ufil vr3JV11n~ L~t.J~ nu~1ui n~ !'If'~ n ~..:J1-ii~'fl'Vl1~ . .. " " II"'..:;!! 1.1 I I ..::::.1
~~n1ifii1~[;11L1J'fl~[;l'Un'fl'U'l1 Curcuma cf. comosa (cf. = look alike) 'U'eln~1nu Teerawatanasuk
LLf'l~Fin.l~ (2004) 1~~n1i1~'Vli~1'U~~L;~"1.1'fl'lfi"ll~~'UL~1 Curcuma cf. comosa Roxb (~1'UVI~1~'fl) ~1JrJ1 ~1r~nl'l~'l'U ethyl acetate ~1nLVI~1ii~'Vli anti-invasive LL~~ anti-angiogenesis ~'l'U~'Vli m~~'Un1~VI1t.l"ll'fl'I1J11'1LLf.J~U'Ihliim1~n1i1~~t.l
" r
~..:JU'U l"ln.l~~~~t.l~'I~'UL~~n1i1~'Vlfim:::~'Un11VI1t.l"ll'fl'I1J11'1LLf.Jf'l"JJ'fl~~11~rll'l Curcuma cf.
comosa Roxb vT-11 'UVIf'l'fl I'! 'VI l'lf'l'fl-1 (in vitro) LLf'l ~1 'U~[;11"'Vl l'l~'fl'l (in vivo) ~'l~vT'I'Vl !'!~'flU
U1~~'Vlflll1~"1J'fl'l !?]1 fu Fl1~~i1~1~~ n I'I"JJ'fl'I~1'UVI~1~'fl1 'Un11n1~~'Un11V11tl"1.1'fl-11J1"' LLf.Jf'l1 'U • ~[;lf'Vll'lf'l'fl-1 L~'fl Luuur~ Lt.J"llu1um r'rl'W-~u1 r.J~ [;lrl rusn~~ui. ~nLf'l~~~1~ Fl rul"h 1 ~nufi"ll~~ui. ~11 'Vlt.J • • • 1u'fl'U1FI[;l
3
~~~u~~~~A~~~n1~1~~ • 1. L~'ilMn'l11~'Vlifm:;~Un11'\.ntl'll'il-'11J1(;)LLc.J~LUV1~'il(;)'VI(;)~'tN (in vitro) 'll'il~~11~rl(;)~1UV13-11~'il
2. L~'ilMn'l!t1~'VIim:;~um1V11tl'll'il-'11J1(;)LLc.J~1ulrwf'VI(;)~'il..'l (in vivo) 'll'il~~11~rl(;)11UV13-11~'il 3. L~'il'VI(;)~'illJ~'VIim:;~um~,..,1tl'll'il~lJ1(;)LLc.J~1ulrwr'VI(;)~'tl~'ll'il~r;·h1'ufl13-l~i1~11~ri(;)'ll'tl-'111u
u~~ Lti~UviFI1~';h~~ l~~u
1. HHi'il3-I~'V11~~'VIti1PI1~1?lfL~'ilUUUU~1~YiflnJ'll'il~~1UV13-11~'il (Curcuma ct. comosa Roxb) 1u .. . ...
n1~m'!;f'lUn11V11tl'll'il~lJ1(;)LLc.J~
2. L~'tll'lft~r;i1fufl13-l~i'.i~'Juc.J~3-I'll'il~~1~~ri(;)rj1uV13-11~'tl~il~'Vlim:;~um~V11tl'll'il..'llJ1(;)LLc.J~ L~'il
Ll~ULL'W'J'V11~L'Wn11~00-IU1c.J~I?ltlnJ"tf~~UL'Yi~~l-i!1um~fn'I11LLc.J~r;i'tlhJ
3. L~'il~,:JL~13-Im~1-i!~~UL'Yi~LL'Vl'Wn1~L-ifmLLc.JUU'"l,1.TU
4. L~'tldju;J'tl~~~1~~-'IL'Wn11r;i'tltl'tl(;)~1U~4fti~L~mriu~"JJ~~UL'Yi~~1UV13-11~'il
~'ilUL~I?l~~·:Jn1~1~t1
1. m~Mn'l!t1~'VIim:;~um~V11tl'll'il~lJ1(;)LLc.J~1uV1~'tl(;)'VI(;)~'tl~ (in vitro)
1.1 n1~'Vl(;)~'fllJ~'VliL~:J..J91U'l'WL"ll~~ (Cell Proliferation activity) 'll'il~~1~~rl(;)11UV1:J..J1~'il
(Curcuma cf comosa Roxb.) ~'Jtl~fi WST-1 cell proliferation assay .. 1.2 m~'VI(;)~'tllJ~'Vlfin~:;~um~V11tl'll'il~lJ1(;)LLc.J~ L(;)tJL-ilLLUU91~'il~n1~V11tl'll'il..:JlJ1(;)LLc.J~ (in vitro
• "' .o=:tr.q wound healing model) (;'}')ti':Jfi Scratch assay
2. m~Mn'l!t1~'Vlim:;~um~V11tl'll'il~lJ1(;)LLc.J~1ull?lrf'VI(;)~'il..:J (in vivo) .. ~ !J U I il
2.1 n1~'Vl(;)~'tllJ~'VIfim'!;f'l'Wn1~V11tl'll'il~lJ1 (;'} LLt:.J~'ll'il-'1~1 ~~n (;)'d1UV13-11'd'il
2.2
2.3
,- I .!Clio il 0 G.' .Qj ez:lc:J CV I !I
n11'Vl (;)~'tllJ ~'VI fim'!;f'l'Wn 11V11tl'll'il~1J1 (;'} LL(:.J ~'ll'il~ 1?11~1J fl~3-I'V13-1~1 ~~ n (;)'lJ'il~'d1UV13-11'd'il " l.t 11 I
<C:It. 40!1 .!Clio tr ..::!1 ~
n1~1?l~'J'"l"ll'WL U'il'Vl1..:1,~ n1 ti'Jll1 rl Pl1~ l?l~'ll'il..:J L U'il Ltl'il (Histology)
..,:. 'U 'Vl'Vl 2
~'fl~'JLu
~'fl~'VImfli1?H?l~ Curcuma cf. comosa Roxb.
Zing iberaceae
v ~
~nHCU:::'V11-:JYHlnHfli1~1Pl'i
blh.!~"llb 'Wl?lJ:::Q,~ Curcuma 'l..:J ~ Zingiberaceae ~'I bU'W'J..:J~~iJmdb-ii'tl~.h..:~ bb'V'l~VI~1tlb'W " " ~1~tJti1~'Wlh'W b'li'W "ll~'Wi'W (Curcuma tonga Linn.) ~1'W'W1..:Jfi1 (Curcuma aromatica Salisb.)
"' "ll~'Wil'flt.J (Curcuma zedoaria) bb~:::~1'Wim.JI'l~n (Curcuma xanthorrhiza Roxb., Curcuma "
comosa Roxb .)
~1'WVI~1~'flbU'W~'ll~~~n91'V'l'ln~..:~~1 ~..:11.h:::~1ru 2 b~m ~1~'WiJim1ru:::!Pl~1tlrltJ~1'Win ' "
~tJ-¥11 ~'W~1'WVIm~'fl Curcuma cf. comosa Roxb.
5
11HU'W1Ub~tn ~t.h'IL'l,.HU'Wm'~'"ln1n~nn in~ru~1udJ'WnJweJn "ll'flU1U~tJU rh'W1Um'd ' ~
'iuV\ 2 1U"ll'fl..:J~1'W'>U-n1'fl Curcuma cf. comosa Roxb. "'
6
-atl~ 4 (?)'EJn'll'El'l~1'klV!:J.J1~'El Curcuma cf. comosa Roxb . ..
';lU~ 5 ~'klbbiN~bV!~1'll'El~~1'klVI:J.J1~'El Curcuma cf. comosa Roxb . ..
7
' t)'Vl fivn..:~ blli"li~'Vl m
~1tJ~1kln1~~mn~9tJ~'ll~~kl1'Y'l~~1mn.n~'r:lb~mnu~'Vli'Vl1~~'1Il1'Y'l~b'U'Y'l1~n1~~mj-1~9tJ
t)'Vli l?i'1k!'fl'fln;nL ~i'u (Antioxidant activity)
thVi'Y'ltf '1~~v1n~ LL~~f1ru~ (2548) 1~~n'l11~'Vli~1kl'il'iln~lhl9likl"ll'r:l'l~1~~nl9l~1kl ..
Vlj.j1~'il~'";itJ!?hvh~~~1tJ'll'Wl9ll?i1~1 1~bbrl methanol. ethyi acetate LL~~ hexane bl9ltJ'Vll9l~'r:lU~Ylfi
8
~uJ..:~mnnl'l lipid peroxidation L'W~~'Ell'l'Yll'l~'El'l (in vitro) ~'JtJ1fi thiobarbituric acid (TBA) bl'ltJ
" 1-D~1~ curcumin dj'W~1~l.J1(;1~~1'W 'Y'ltJ~1 ~1~~nl'l~'JtJ ethyl acetate ilm1l.J~1l.J1Hl~U~~mnnl'l d.9
lipid peroxidation ~~~(;] t~milrh IC50 whnu 29.49 IJQ/ml ~'El~'tl~l.J1~'El ~1~~nl'l~'JtJ methanol
' " LL~::: hexane ml.Jii1r?1u L~'Elll1~1~~nl'l~tn1J'll'fl~~1'W~J.,J1~!l~".ltJ ethyl acetate 1u~nV'lLLtJni'W1~
bU'W~1~~~u1~'Yli (partial purification) dj'W~'J'W~rll'l fraction L'i1..:11 bb'tl:::1.l1l.J1~n1~1~'Yli~1'W " bl'ltJ~m~nm~~u~~'El'Wl.J~;aM:::~'JtJ1fi
'"' DPPH assay lipid
peroxidation ~'tlm~1r.i'tJ'Y'ltJ~1 ~'YliuuJ~u.fjrntn'El'Eln~LV'li'W'll'El~~1~~n~'~~1'W~l.J1~'El~'JtJ r?i''J~1~:::~1tJ ethyl acetate l.J1'"l1n~1~~1rlb1JL'W~'J'W~rll'l 4 "ll\11'1 HiLLri CE-1-18, CE-1-21,
CE-1-23 bb'tl::: CE-1-24
'W'Eln'"l1nd' n'Wn'J~nw ~(;l{m LL'tl:::l"lru::: (2550) 1~~m.'.f1~'Yli~1'W'Elt.p~'tl;a~~:::'ll'El'1~1~~nl'l
~1'W~l.J1~'El1 til [;]rJ'Yl Vl'tl'El~ bV'ltJ ~m~n ~'Yliu'El~ n"Wm~Ln ~ fl'J'll.J ~ ~un ~'ll'fl~~'tl'fl Vl L ~'fl Vl LLV'l~ 1 ~ qj1 'W~~ 'Y'ln'JJ1'J~nn L~timll1 Hhi1'WLtJ1~".l1'W~'JtJ streptozotocin LmtJu LVJtJunum'tli'IJ1(;11'tlL'WL~'El ~ ' "' ' ..
(gliburide) LL'tl:::~1~V1iJ~'Ylfi~1'W'El'4~'tl;a~ ~::: (vitamin E) t~mmtJu LVltJtJ rhm~~~!K'J~~~~'ll'El~
Wi'l!l~b~'El VlbbV'l'IL mk)~ (;l'E:JtJ~'W'El~ L'i'El phenylephrine ~'tl m~1r.i'tJ'Y'ltJ~1 rh m~~ ~ !K'J~'I~~'ll'El~~'tl'El ~
L~'Eli'ILLI'l~1mu'll!l~~'WLU1~'J1'W~1~fu~1~~n~~1'W~l.J11'El ilrh~~n~11'W~'Wun~ LL'tl:::1'W~'WLU1~'J1ti IV '\1 'IJ 'IJ 'U
~1~fu gliburide bb'tl::: vitamin E bb'tl:::m~1~~1~~n~~1ti~l.J11'El~~L'i'Eln'W 6 lu~1,; ~1l.J1~bi'tl~rh
n1 ~~ 1'\ r?i''J ~'I~ ~'JJ 'El'l ~'tl'Ell'l b~'Ell'l bbl'l'lb ~ rul~'El ti1..:1 iJ-JtJ~1 rl ru b~'El LmtJtJ b Vl tJtJml.J1 ru~1~~ rl ~~l~ftJ 'IJ q cv Q,l
'Y'lU~1 ~'iltJ1~'J1ti~L~fu~1~~nV1~1ti~l.J1~!l 600 mg/kg L'i'Eli'W ~~L'i'ElrltibUtib".l'tl1 6 lu~1,;
~1l.J1~bl'tl ~ rhm~~~!K'J~~ ~ ~'JJ'El~~'tl'El ~ b~'El~ bb~~1~ru'tl~l~ bbL'i~'WbtJ1~'J1'W~1~ftJ~1~~n ~~1'W 'U q Q.l 'U
~l.J1~tl 300 mg/kg L'i'Eli'W hJ~1l.J1~bl'tll'lfhm~~~'~r?i''J~~~~'ll'El~~'tl'El~b~'El~bbV'l'ILmk} '"l'ln~'tl
m~~n~1L1f'El~~uLL~~~HL~u~1 ~1~~n~~1u~l.J1~'El1'W'll'W1Vl 600 mglkg L'i'Eliu il~'Yli1um~ u!l..:~num ~~1~1u~ !?lun ~'ll'El~~'tl'El ~ L~'El~ 1 ti~'WLU1~'J1u
"' fi~'"'ni 'J'I~'J1ti LL'tl:::flru::: (2550) 1~~n~1~'Yli'll'El~~1~~n~~1'W~l.J1~'El1'Wm~unu'El~fl1".l:::
Qn!l'Eln~1Vl'lfLn'W~ l.J ~ 'tlL ti~l.J'El~'ll'El~~'i'l"in 'll 'l".l~ Q n L ~timll11 ~ LU'WbtJ'l~".l'lti~".ltJ streprozotocin
bl'ltlfi1~u~1~l[;]rJ'Yll'l'tl'El..:l~iJ~:::r?luJ1(;11'tlL'WL~!ll'l'lJru:::'El~'El1~1~ (FBS) l.J1nn~1 150 mg/dl bUt..!
mil.JLU1~".l1'W '"l1nt.f'WU'El'W~1~'Yl~~'fltJbb'tl:::~1~~n~~1'W~l.J1~'El (;l~".l'"lrJ~ FBS 'Yln 2luV11,; b~'Elfl~tJ ' '
6 lu~1,; Lf1u~l.J'El~b~'El~Lm1:::,;~1't!dl.J1ru malodialdehyde (MDA) L'W~l.J'El~ ~".l'Wnl~'tl~~:::r?lu
J1(;11'tlL'WL~'ElV'lLLuuL~tJu'Y'li'W vi'lm~~n~1~L".l'tl1 o.5, 1, 3 LL'tl::: 6 i'".l'fl.J,mi~m~u'Elti~1~'YlV'l~'Elu ~~m~~r.i'tJ'Y'ltJ~1 illl.J1ru MDA L'W~l.J'El~'ll'El'l~uun~LL'tl:::~'WLtJ1~'J1ti~1~fu~1~'Yl~~'EltJ LL'tl:::~1~
"' "'
9
n~~un~tJih~ih!tJ~1Ab\J ~~LU1~'J1kl~1~fu glibenclamide ~f11 FBS ~1~1~~LU1~'J1k!n~~ fl'JUfl~'flEh~~utJ~1A'ru1w~'u(;n~~ 2 u~hJ~e.J~1kliu(;l1~~ 4 LL~:; lu(;l1~~ 6 m~1ih!l1~~·H1(;1~1kl , ~
~~1~t1 ~1t1 vitamin E 1~~e.J~~t1 FBS mh~~utJ~1Ab\JlkliUW1~~ 2 ti~ 6 e.J~n1~~mn~Yli~V1 ~ I I ~
~:;~UU11?11~lklb~'fl(;lbbUUb~tJU'Y'likl'Y'lU~1 ~klbU1~'J1klYl1~fu corn oil ~f11 FBS LYl~~kllk!'Ji'J~ 1 'lJ
1 li I 11
"Ji''JL~-.:JLLm m~1~ glibenclamide LL~:;~1~~nV1~1kl~~1~tl ~e.J~tTutT~mnYl~~kl"lltl~ FBS lkl~~ ' " LU1~'J1kllkl'li'J-.:J 1 "Ji''J hN LL~n e.J~ m~~r.i'tJuMu1~~1m~1~fu~1~~n (;1~1kl~~1~t11~~ e.J~ lkln1~ ,
kltlnr.nnrl' ~~r.r tJ tT~ 1~~ n111 ~'Vli ~1\.ltl~~~ ~~ ~:;"lJ'f}-.1~1 ~~ n (;1~1kl~~1~tll kln1 ~~ (;)Jl1'J :;m ~
Ln(;l oxidative stress 1kl~kl'Y'ln"ll1'J~nnmtimu11ihuk!LU1~'J1kl~'JtJ streptozocin L~tJ1•ihuk! "' , 'lJ
LLkl'J'Vl1-.:JLklm ~fm11 LL~ :;~ (;)Jl1'J :;LL 'Vlm.JitJkl~ Ln (;191 n hfl LU1~'J1kl L(;ltJ vu'Y'l1 :;l:~fl LL 'Vlm.Ji'flkl'Vl1-.:J11?1
m~Yl(;l~tJ-.:Jvht(;ltJ~(;ll?l1:Wn1~~1~1kl"ll'fl~11?1Yln 2 lu(;l1~ LUklL'J~1 6 lu(;l1~ ~'JtJm~rf(;lf11 BUN '
' ' " LL~:; serum creatinine LL~:;i-.:JLnl?ln1~LU~tJklLLU~-.l"ll!l-.l malondialdehyde (MDA) ~~LU\.l~'J~rf(;l"ll!l-.1
" ' ' mnn (;)Utjmm lipid peroxidation lklLi1'fl L~t11m'fl~il?lrf'VlV1~t1~ 91ne.J~m~~r.i'tJY!~m11i(;l'i'l'Jkl
~:;~~1-.:Jtl1~un11?1LL~:;J1~un~'J m~rfm.fi:w1ru BUN LL~:; serum creatinine 'Y'lU~1 m~fmn~'JtJ " m~1~~1~~rl(;l~1kl~:W1~!l"llkl1(;1 300 mg/kg ~'J:wnum~(;l~:;~uU11?11~ Glibenclamide ~e.J~lkln1~
u!l~nklm~~1-.:J1kl"ll'fl-.:J11?11kl~~Lu1~'J1kl LL~~1 ~~n (;1~1kl~:w1~!l1~~1~1 ~n~ (;lmnn V1 MDA level
"ll'fl~mnnVluBmm lipid peroxidation Lkl~~LU1~'J1kl1~
C]'Vli~1kl'1~:a~ (Antimicrobial activity)
ruJ1~ t:.J~--11~~ ll~!:: rPlrt.J !:: (2549) 1~Mmj1~'Vli~h'WLL1JflYlLhm'l'l:::L~"'flnlJ1'1'llil~'ll'fl'1~1~ " ..... !J ' !J !.1 cv 0 .c:::t. ~ ..... !J 0
~n(;l~tJ1U91nL~~1"lltl~'J1kl~:W1'J'fl mtJ l?l'JY11~:;~1tJ 4 "Jlkl(;l fl'fl ~1~~n (;l(;l'JtJL:Wn1kl'flrl :\.!1 ( 1:1), ~1~
" " ~rl(;l~'Jm:Wn1kl!l~ :u1 (9 :1 ), ~1~~rl(;l~'Jme:Jm"llkl u~:;~1~~rl(;l~'Jm'flfi~!l:;;nLI?1Yl ~t1L~m~klY11u '
0 ~ ""
'Vl(;l~'flU91kl'Jkl 9 'Jl\.!(;1 fltl Staphylococcus aureus, Streptococcus pyogenes, Streptococcus
pneumonia, Bacillus cereus, Campylobacter jejuni, Shigella sonnei, Escherichia coli,
Candida albicans LL~:; Aspergillus niger ~'JtJ~fi agar disc diffusion 'Y'lU~1~1~~rl(;)Y!n"Jlti(;l"lJ'fl~ 1 r 11 11
~1kl~~1~t1Yl100 iJrl~nf:w /iJ~~~I?l~ ~~YlfltJUtJ-.:JL~tl~1 A. niger 1~ LL~~fl'J1:W~1:W1~bllkln1~~1kl tl " tr "
L~!lLLU flYl L'fmLI?ln ~1~ rlkl b(;ltJ'Y'lU~11."l1~~n (;) L:Wn1kl!l~ :U1 (9: 1) ~~Ylfi~1klb~tl LLU flYl L'ftJ l."l1~1~bl " " v v .c:Y '1 !.1 .c:::t. .:!I .c:Y
tJUtJ--lb'lltl L(;l 5 "Jl\.!(;1 fl!l S. aureus, S. pneumoniae, B. cereus, C. jejuni, S. sonnei, ~tl-.1~~~1fl'fl
" " l."l1~\."lrl(;)L!Jflrl!l:;;nLI?l'Vl ~1:W1~mJUtJ'lL~tlLLUflYll'ftJ1~ 4 "Jlti(;l ~!l B. cereus, C. jejuni, S. sonnei,
r " " LLrl:; S. pyogenes ~1~1."lrl(;lbe:Jnb"llkl~~YlfitTutT'lb~'fl1~ 3 "Jlti(;l ~'fl S.pneumoniae, S. sonnei bb~:::
10
B. cereus LL'tl:;~1~~n~LJ-Jfi1'W'El'tl: J1 (1:1) ~~'Vli~1'WLLU~Vlrft.JL~ 1 -nU~ ~'El S. pyogenes b~'El tl1~1~~n ~LJ-Jn1'W'El'l'l :J1 (9: 1) LL'I'l:;~1~~n ~ L'Elfi'l'l 'E):;~L[;l'VlhJ,.nrh M 1c Tiu~1rh M 1c ~'El'l~1~~n ~
" " LJ-Jfi1'W'El'tl:U1 (9:1) (;]'ElL~'El B. cereus, S. aureus, C. je)uni, S. sonnei Lvhnu 50, 60, 100, LL'tl:;
" .Q C. V Q ~Q 0 CV I V Q Q I c:!il
100 J-J'C'l'C'lnJJ-J/J-J'C'l'C'l'C'l [;]~ ln1J-J'C'l1~U LL'C'l::;~1 MIC ~'El'l~1~~n r1 L'Elfi'C'l'EJ::;'ll L[;l'Vl (;]'EJ L"1l'El B. cereus,
C. jejuni, S. sonnei LL'tl:; S. pyogenes Lvhnu 10, 40, 60 LL'tl::; 10 il'l'l~nfJ-J/il'l'l~~[;l~ mJ-J~1c;i'u
nJ'El'ln1~"'l,_r fi'W~iJ~~1'C'l LL'C'l:;~ru:; (2550) L~~n1i1~'Vli~1'W'1'1'l~Yi~'El'l~1~~rl~~'lU1~'Vli~ ~n~~'lt.ib'Elfl'tl!l:;~L[;l[;l~'EJ'l~"ll~J-J'WLYi~~1'Wt-~J-J1~'El 91'W'l'W 12 ~'l'W~n~ ~'El CE-7, CE-14, CE-15,
' ' ""' ......
CE-16, CE-17, CE-18, CE-19, CE-20, CE-22, CE-23, CE-24 Lb'tl:; CE-27 'Vl~'l1J-JL~J-J~'W 50
aureus,
Streptococcus pyogenes, Streptococcus pneumoniae, Bacillus cereus, Campyrobacter
jejuni, Shigella sonnei LL'tl::; Aspergillus niger L~t.J1fl'Vl~~'ElU~'lt.Jt-linmnLYli~hu1u e.J'tln1~19t.J
YiU~1 ~'l'W~rlr1~~~'Vli~1'W'1'1'l~Yi91nJ-J1nLUiJ'Elt.J ~'El CE-20, CE-18, CE-19, CE-22, CE-23,
CE-24 LL'tl:; CE-27 [;11J-J~1c;i'u L~t.J~'l'W~n~ CE-20 ~1J-J1~tl~ui'lL~'ElL~ 6 -nU~~'El S. pyrogenes,
" S. pneumoniae, B. cereus, C. jejuni, S. sonnei L~'tl:; A. niger ~'l'W~n~ CE-18 ~1J-J1~tl~U~'l
L~'f1L~ 4 -nUr11 ~'El S. pneumoniae, C. jejuni, S. sonnei LL'tl::; A. niger ~'l'W~rlr11 CE-19, CE-22
LL'tl:; CE-23 ~1J-J1~tl~uJ'lL~'ElL~ 3 'l!U~ ~'El S. pneumoniae, S. pyogenes bb'tl:; S. sonnei ~'l'W " " ~n~ CE-24 ~1J-J1~tl~U~'lL~'ElL~ 2 'l!tir11~'El S. pyogenes LL'tl:; S. sonnei LL'tl::;~'J'W~rlr11 CE-27
" ... ~1J-J1~tl~U~'lb~'Ell~ 1 -nUr11 ~'El S. pyogenes
..t .. t)'Vlfi!Pl'l\Jn'l'iti'J IPl (Analgesic activity)
1'Vlm ~1m LL'C'l:;~ru::; (2548) 1~~mj'1~'Vli~1um~u'lr1l~'El'l~1~~nr1l~'lt.JL'i''l't11'C'l::;'l'l1mJ-Jn1
'W'El'tl, L'Elfl'tl'El::;~L[;l'Vl bL'C'l:;Le:JnL"1l'W~'El'l~1'Wt-IJ-J1~'El i'lL'W'\.ol'tl'Elr1l'Vlr1l'tl'El'l (in vitro) LL'tl:;L'Wl[;lrJ'Vlr1l'tl'El'l (in
vivo) Lmt.JULVlt.Junu ibuprofen LL'tl::; curcuminoid m~'Vlr1l~'ElUL'Wl[;lrf'Vlr1l'tl'El'l't11L~mJ'El'W~1~~nr11 ...
~'W1r11(;]1'l11-timim~nu9mv1'lLYl~~LL'tl::;L~muuL'l'l'l1 60 'W1Vl ri'f1'WU1LU'Vlr11~'ElU~'lt.J1fi hot plate ' ... i.J IV i.l .Q.c::::l .::!ij 0 C:V
test bb'tl'l'l~ reaction time LL'tl:;'Vlr1l~'ElUr1l'lt.l'lfi acetic acid induce writhing LYi'El'\..1191'\..m.m~'l~'El'l
n1~U r11L'i''l1l1t.JLoW 30 '1...11Vl e.J'C'ln1~19t.JYiU~1~1~~rl~~1'W'\..IJ-J11'ElL'WL'i''J't11'C'l::;'C'l1t.Ji'l~1J-J"1ltlr11 L~J-Jrh
reaction time
" ' cyclooxygenase (COX) e.J'C'ln1~19t.JYiU~1~'l'W~ nr11~1'Wt-~m1'ElL'WL'i''l't11'tl::;'tl1t.lvl'l~1J-J"1ltir11 V1 ~'l1J-J
11
#' "
L~)-J~'W 50 IJg/ml i1~'Vlfi~u~'IL'fl'WL"l!a.J COX-I LL~:; COX-II L~tJ~11~rll?l~1'W~)-)1~'fl~'Jm)-Jfi1'W'fl~i1
fl'VliLLrlU'J~~'I~~ 1'fl'l~'l)-)1~'fl ~11~rl~~1'W~)-)1~'fl~'JtJ Le:!nL'll'W LL~:::L'flfl~'fl:::~L(;]'Vl (;]1)-)~1tiJU "I "' '
~.., a t]'Vlfi(;]1'W~~L'i.:J (Anticancer)
#' ,. "
~11~n~~'J'W ethyl acetate '"11nL~~1'll'fl'I~1'W~)-)1~'fli1~'Vlfi~1'W)-J:;L~'I L~tJi1~'Vlfl~U~'In11 '
L~)-)91'W'J'W (anti-proliferative effect) 'll'fl'IL'1l~~)-J:;L;'I{;]U'JJ'fl'IWI~ (murine hepatocellular
carcinoma) "l!oW~ CB0140C 12 LL~:::i1~'Vli~uJ'In11~n~n (anti-invasion) ~'Vli~1'Wn1d'flyjt.Jyj (anti-
" migration) 'll'fl'IL'1l~~)-J:;L;'I1'W~~'fl~'Vl~~'fl'l (in vitro) LL~:::~u~'lmnm:;~~'ll'fl'IL'1l~~)-J:;L;'I[;]'fl
extracellular matrix proteins 'W'fln'"l1nif~'li1~'Vli anti-angiogenic activity L~tJ~uJ'I hepatic ' "
sinusoidal endothelial (HSE) cells 1'Wn11~~1'1 tube-like structures L~'flL~tl'IL'1l~~L'W Matrigel-
coated substrate (Teerawatanasuk eta/., 2004)
2. n1'iVI1tl".ll'eN'IJ11?1LLe.J'Cl (Wound healing)
2.1 Ll'l'i.:~~-i1'1".ll'fl.:J~':lVIu.:~
~'J~'IX'ILLU'Im)-)Lfi1'1~~1'1L~dj'W 3 .ff'W ~'fl ~'IX'Ifhyj~1 (epidermis}, ~'IX'ILLVJ (dermis)
LL~:::Ltf'flLm'fl1"JJ6Y'W1~Eh~'IX'I (subcutaneous fatty tissue) (Strodtbeck, 2001) "' I 'J,I l,l
i'"WYi 1 VIU·uhw-i1 (epidermis) Lij'W~':m'IX'I'll'W'W'fln~~ LLU'I'fl'flnLU'W 5 'll'W 1~LLri , " " '
1. Stratum corneum (SC) 'fl~'ll'W'W'fln'i'J,~ i1 fi"J1)-J~'W1U1:::)-)1tl.J 10-20 iJ m LU'W'll"WV1i1
fl'l1)-)~1rfru~~~1'Wn11n~n'Wn11U1~'1tJ1~1'W~'l~'IX'I U1:::n'flU~'lt.Jn~)-J"JJ'fl'l keratinized cells ~ ' ,
ttli1'1LLU'W~hii:J~1(?1 ~~m'h corneocytes i:J'lJ'Wl(?]L~W·h~'WtJnm'l1..h:;)-)1tl.J 30 IJm ~'W1
U1:::mru 1-2 IJm Ln'fli'W sc LUtJmf1 corneocytes ~1)-)11Cl~~-nuu"11~n'l 60-70 %w/w "JJ'fl'l
" ' , , I Q .<Cll I f"
SC keratm1zed cells L~~1'W'"l:::QnLL'Vl'W'Vl(;]~'fl~L'l~1'"11nn1nLU'IL'1l~~'ll'fl'l viable epidermis
' " 1'WVl'i'J,~'"l:::L~L'1l~~~n1j'ru:;LLU'W1 ~'fl'WvlUrl'W U1:::n'fJU~'ltJ fibrous keratin (fibers L~~1ili1th:::'1L1j'W
~u Ln'fl pH "ll'fl'l physiological condition = 7.4} LL~::: keratohyalin ~'fl)-)1'flU~'JtJ extracellular lipid matrix Lfl1'1~~1'1in1j'ru:;ifm'"l~t.Jn~1LU'W protein bricks ~t.J'I{;]'l~'fl'Wml1'W
'IJ
lipid mortar (nJ~ 7) 'IJ
Lfl1'1~~1~"ll'fl~ epidermal intercellular lipid matrix rl'fl'W~1~'llu~m.\U1:::n'fJU~'JtJ~'l'W
~LU'W hydrophobic LL~:; hydrophillic ~~Url'W ~'l'W1~Cl.J6Tn~t.J~{;]'JLU'W bilayers ~':l'W"ll'fl'l hydrophobic 1'W SC U1:::n'flU~'JtJ hydrocarbon chain "JJ'fl'l lipid ;~LU'WU1L'ltl.J~~11[;]1~1
1'lmf~J1~1'WL~mn lipid 1'W.ff'W epidermis '"l:::ud:::n'flU~'ltJ ceramide type Ill 33%,
12
ceramide type IV 22%, cholesterol 25%, cholesterol sulfate 5%, LL~~ several fatty acids ...
I V ..::! CV ~ Q.J'
15% !119 LL(;ln (;11-.:ln'\..!(;11~ fl'11~~n'll!Wll'We.J'1~'W..:l
Intercellular route Transcellular route
Lipid Aqueous Cholesterol/ Triglyceride Minimal Keratin
Cholesterol sulfate lip rl
... 'iU~ 7 Brick and mortar model 'll!l..:l~""l~i!..:J'll'W stratum corneum <u
' Yl~1: Strodtbeck (2001)
~u1L'1ru~'1~i!..:J9~il pH 1.h~~1ru 5 ~..:! pKa 'll!l..:J sc lipid LYhnu 8 head groups 'll!l..:l
lipid 9~LL(;lnlPl'1 (ionized) HiL~niJmJ 1uiu SC ~~n~..:JlU pH 9~L~~~"'WLU'W 7.4 ~..:Jv'h1~ head
" ' " groups 'll!l..:l lipid Lb'iWi..:JU~~'"l~u n1~LL(;lnlPl'1'll!l..:l head groups 'i19~Ly;~~'W91n 10% (~'d'W'W!ln'lltl..:l
SC) LU'W 90% (~""l'W~h'WL'W'll!l..:l SC) A'11~~1'WYI1'W'll!l..:l SC 9~L~~~'Wm~A'11~t.l1'1'lltl..:l chain 'lltl\l
sphingo-lipids
membrane LL~..:l
lipid
3. Stratum granulosum
Lmhd''"l:::LU'W quasi-crystalline phase
n1 tJ 1 u L'1l~ ~Ln (;) LU'WLLn ~'W~ LL~""l n niu!l!l n ~1'W!l n L'1l ~ ~ " "
13
4. Stratum spinosum LilwL"ll~~~~in')j'ru:;mnJ '\..ihn;l~1m~~ti~LL~'Jfl'flt11 LU~mJ LU'WL"ll~~m.J'W il'1Lfl~m~'"l:;~~~'J L"ll~~L~'fl~lii'fln'W ~m~~~1-.1LU~~'W keratohyalin
" 5. Stratum germinativum (Basal layer) LU'WL"ll~~luhi~tl-.l~'JLU'WLLmi'WL~ti'J LL~:;~L"ll~~
.ffu~ 2 \nJ·.:HL...i' (dermis) ~fi'J1~~'W1LL(;lnli]1-.lfl'W(;11~~1LL~ki--l"ll'fl-.lf1--ln1tl U~:;nmJ~1'Jti " L~'W1t1LU~~'W ~'f) collagen (75% "ll'fl--lll1m!'mL~-.l) vh1~~fi'J1~L~Uti'J ~elastin 4% vh1~~fi'J1~
0 .drij I Q ~ 0 Go' .:!1
ti~~~'WLL~:;~ polysaccharide 20% Yl~1flt1Jfl'fl chondrotin sulfate, dermatan sulfate LL~:;
" " " heparin 1 uiuw'"l:;~ L~'WL~'fl ~~'fltl vi'flll1 L~~'fl-.1 U~1 t1u~:;~1Y1 LL~~lii'fl~ lii1--11
.ffu~ 3 Ltl·m~'ilL"luTu1.rPi'~'lVI~\I (subcutaneous fatty tissue) ~fi'J1~~~~~u U'fl-.lrl'ULLN
n ~:; LL VI n LL ~ :; L U 'U 'U 'W 'J 'W U 'fl \1 n 'W fl 'J 1 ~ ~ 'fl 'W U ~ :; n 'fl U ~ 'J tl L "1l ~ ~1 "lJ ~ 'W L U 'W ~ 'J 'W 1 ~ n.j LL ~ :; iJ
collagen LLYJm'fl~u~:;u~1tl
2.2 n"i::aJ'l'Un1"iVI1!.1"1J'il\11J1l?ILLc.J~
1J1l?ILLc.J~ ~~1tlel\l n1~U1~L'~U~Ln~:n"u1uutL'Jru1~utL'JCW~~-.l"llll--lf1--ln1tl vh1~~'J~U-.l ~ 1'flL~'flU LL~:;~'J'W~~n~-.llU n11.ffuEh~u\l nnv'h ~1tl ~'J~U\Ill199:; LLtl n1l'fln'"l1 n rl'W ~1'fl hi LLtln n1~ ' ~
U1 ~LLe.J~~1~1~blr.i1 LL'Wn1~ 2 U~:;LJlYIU'Wyf'W"'l1'W'll'fl-.lm:;u'J'Wn1~'li'fl~LL"ll~ (;11~fi~~~'Jl1~ 1~ LLri LLe.J~ d.S
L~tlu~i'W (acute wound) ~'fl m~~Lif'flL~'fl1~fuu1~L'~U'il'WL~'fl-.l'"l1nm~~~~1mh~~~~tlm~~ ~'~
I !.1 I I I 1,.1 I .cY ~ .dril 4 .o::;:,j1 \J V t;; Cit, ~ 1 iJ .c\j .c\1 .dj
(chronic wound) fill m~YIL'W'flLtlllYI muu1~L'"lU Ln~m:;U'J'Wn1~m')j'11Jtl1-.l'll11 L'Wll-.l'"l1nL'WllLtlll
1~fu~-.l~vh 1 ~Ln ~u1 ~ LLe..~~.f'11 ~1'fl Ln ~~m'jj~Jl1~:n"u~ Lilli L~'fl ~1'Wrl1--l ~-1m mu.ffu!?l'fl'Wm~~~tl 0 0
.o::;:,l.::!i .o::;:,j .c:::;:,j I V V Cit, .o::;:,j I Q
'JJ'fl-.1U1 ~ LLI:-J~ U1 ~LLI:-J~ll19~'ll'flL~tlnYJm-.ln'W!?l1~~n')j'Cl.J:;'ll'fl\ln1nn ~ fi'J1~ L~tl~1tl L'll'W LLI:-J~'Un"ll1 ~
(laceration) LU'WUI~LL~:-J~~arn Ln ~f'J~numm~u1 ~ v~u~~-.le.J~ 1~Ln ~fl'11~ L~tl~1tllii'fl Lif'flL~'fl LL~:-J~
2549)
m~~1tl"ll'fl'~ LLe.J~ Luum:;u'Jum~YJ1--l~·h1YJm ~--1 Ln~:TiuLu'W~1 ~u lii'flL ~'fl-.lnu LL~:;aJ'niJ
fi'J1~L~~'fl~~"1n'W1'WLLlii~:;~:;tl:; (Beanes eta!., 2003) u~:;n'flU~'Jtl 4 ~:;tl:; (dU~ 8) ~-.1if' "
Skin surface
Platelet
Macrophage
a Injury
d Late inflammation (48 h)
e Proliferation (72 h)
Epidermis and dermis of skin
The phases of cutaneous wound healing
Fibrin
b Coagulation
c Early lnnammatlon (24 h)
I Remodelling (weeks to months)
Expert Reviews in Molecular McdicinoQ2003 Cambridge University Press
'itl~ 8 m:::U'J'Wn1~'\.!f1t.J'llt!'llJ1!?lbbt:.J~ (wound healing) ... .d 'Vl~1: Beanes eta!. (2003)
' ~
14
L~mn !?lm ~u1 !?1 v~u '1.-H'l t1 !?1 L~tl !?lu1vw.J \J'W9 :::r1 n i11~1 u 9:::ih~ t1 !?1 Lt!f~t~t~ mnu1vnw "
" lJ1 !?1 bbt:.J~ 91n\J'W9:::Ln !?1 m:::U'J'Wm~ih~ b~t! !?1 L!?lmn!?ln1~'\.!f!?l i?l'J'llt!'l'\.!f~t! !?1 b~tl !?1 (vasoconstriction)
btit~'l91nm~wK'l chemical mediators L"li'W serotonin t!t~nm91nL"ll~~'llt!'lbift~b~t!lJ1L'Jru~1~¥u ~
lJ1 !?1 L9u bb~::: endothelial cells (Stadelmann et a!., 1998) n1~'\.!f!?l i?l'J'llt!'l'\.!f~t! !?1 b~tl !?19 :::Ln !?l~'W
'l.!fi'l91mn!?llJ1!?lbbt:.Jm~t.J'l 5-10 'W1~ ~t!91mf'W9:::i'.Jm~~'J~n~~'llt!'lLn~!?lb~t!!?l L!?lt.J9:::~um~!?l '
L~t~ !?lu 1L 'J ru~ Ln !?1U1 !?1 Lbt:.J~ 1 wh'W'J'W~1 nn1u 1 'WL 'J~1 'i''J !?1 L~'J LL~'J m ~ !?1 L~ t~ !?19 :::'\.!fi'l~ 1 ~'11' u tl'l1vnrsnu!?l
~1'l1l~bbrl epidermal growth factor (EGF), transforming growth factor ~1 , ~ 2 bb~::: platelet
derived growth factor bb~~::;'ll\lrn9::;r1TI.!fih~~1'lrl'Wt!t!nhJ 'Wt!n91nif Ln~!?lb~tl!?l~'l'\.!fi'l pro-
~"n(:A
\LM k~l:J . H9j
15~4q~ f'
inflammatory cytokines 1~ LLri serotonin, bradykinin, prostaglandins, prostacyclins, ~!)if! thromboxane LL~::: histamine ~~m~J..r0nn~i~'el'elmJ1L~'elm:::~'WL~L'll~~1'Wm:::u'J'l.Jn1~'Elnv~u . ... . LLti~~'J L~J..Jr.i1WJ'W LL~:::Lfl~t:J'W~J..J1U~U1 ~LL~~ m:::~'WL~~~'el ~ L~t:J~'llmu ~'J LL~:::L~J..Jm1J..J~1J..J1Hl •
1 'Wn1~~J..J~1'WL ~1'el'eln'll'el~~~~1~J..J1n('W L'W'llrw:::L~m n'Wn di l'lm:::U'J'Wninl.J~u'W fibrinogen LU'\..J
fibrin Lni'ILU'W fibrin clot 'eli'IU1L'JrtJ~Lni'IU11'1LL~~l~.j''Jm1'J L~'elLih~'ell'l~tJI'll~~ LL~:::U'el~rl'WL('elhfl ' . S, I I I I
"" ~ ."1 ' ~ ' ~ ~ "" ."1 ..,..,. 'W'elnr.nn'W fibrin LL~::: fibronectin 9:::~1'Wn'WLu'W~1~LmLYi'el'll'JU b'WnldLfl~'el'WYl LL~:::Lu'WYlUI'!Lnl:::'ll'el'l
' ' tr" I ~ ~ l;' V I ,
L'll~~m~1 Y1Lnt.J'J'llt:J~num:::u'J'Wn1'.i'~1t.J'll'el~U11'1LL~~ L'll'W neutrophlls, macrophages LL~:::
fibroblasts LU'W~'W ~L~1J..J1U1L'"lrtJU1~LL~~ ~'JJ..Ji..:tLU'W~~I'!Lnl:::'ll'el~Ln~I'IL~'ell'l~'JU (Midwood et
a/.,2004) L~'elL~'el~~tJI'ILWtlLL~'J 9:::Ln~m:::U'J'Wn1'.i'~~~t.J fibrin clot 'el'elnltJ ll'lt.Jn1~Yi1~1'W'll'fl~~~'.i' '
plasminogen activator LU~t.J'W plasminogen LU'W plasmin ~~LU'WL'fl'WL'll~rit:Jt.J fibrin 91mX'WL'll~;
lPl1'11 ~ L~ tJ'J~'el'l rllJ n ~:::U'J'Wnl ~!i'n L~U9::: Lfl~'el'W~'J L~1J..J1~U1L'J rtJU11'1 LL~~ L~'el L ~~~'.i':::U :::l'i'ellU
(Singer and Clark, 1999; Strodtbeck, 2001; Bale and Jones, 2006)
'i~tJ~Yi 2: 'i~t.l~n1'i-am~u (inflammatory phase)
nl'.i''Ei'nL~UU'.i':::n'elU~'JUoKn~rw:::m'.i'U'JI'llJ'JJ..J LLWI~ LL~:::~t:J'W~U1L'JCWLL~~ ~:::u:::ll9:::Lfil'ln1'.i' ...
QJ .dil • ~ cv q
'lJtJ1tJ ~'J'lJ'fl~VI~'fl ~ L~'el ~ (vasodilation) u~:::Ln ~m:::U'J'Wnl '.i''elnL~U91 n ~Yln'lJ'el~ prostaglandins
LL~::: histamines ~~VIi~J..J191n mast cell mr~~~'eli'IL~'eli'IU'elJ..JL~~n1'.i'~J..J~1'W'lJ'el'l'lJ'el'ILVI~'Jl~J..J1n "' .,.
'll'W (capillary permeability)
' .dJ Clo I (T" Clio .c:Y 1./ I .0:::::. .r::::;,l..:::lil QJ
91 n~~'fl~ L~'fl ~'Vl1..!11J'.i'L'Jru '.i't:Jt.J!?ltl'lltl-.1 L'll~~tj~'JVI~'fl~ L'6'l'fl~ L'li1~1J H'J ruYl3-J n1'.i''flm~u (Bale and
Jones, 2006) L'll~;~lflb!.J~L~tJ'J~'el~L'W~:::u:::'lJ'el~nt~!lm~u~ 2 'llU~ ~'el neutrophil LL~:::
macrophage ll'lt.J~ neutrophil "'l:::Lfl~'el'W~m LU'W'fl'W~U LLm J..J1tl~U1L'"l rwu1 ~ LL~~1l1t.JL'W ~:::u:::L'J~1 1 .f'J'i:J..J~VIi~Lnl'lui~LL~~ LL~:::~w.J'W1n~1t.JLU'W predominant cells L~J..J91'W'J'WJ..J1n~~~~u1L'JCW
' U11'1LL~~1l1t.JL'W 2 tJ'WLLm neutrophil LL~::: monocyte (~~lPl'elll.J9:::~w.J'W1LU'W macrophage) 9:::Qn
~~~I'IL~1~U11'1LL~~'i:l'!u chemotactic factors ~mtJ'llUI'l L-ri'W platelet-derived growth factor
(PDGF) LL~::: transforming growth factor-~ (TGF-~) (Beanes et a/., 2003) neutrophil 9:::n1r.i'l'l
~~ LLu~nu~'elJ..JLL~:::LLUAYJ ctu ll'lt.Jn~:::u'J'Wn ~~ phagocytosis 'W'eln91nllu~ ~1J..J1 ~n~.h LLU AYJ ctu1~ ~'Ju'rJfim~u~'eltJ'el'l..p;j~;sM:::L~1hl\l.hLLUflYJL1u ~un~1 m:::U'J'Wnt'.i' respiratory burst ~'JJ..Ji..:t~
m'il~f11 fl'J 1J..J~ :::'ell 1?1U1 1?1 LL~~ li?lt.Jni~VIi..:t L'el'Wl'll~n~J..J proteases L~'elJ..J1U'elmf'I~L ti't:~ L~'elu1L'"l rw
U1 i?lLL~~ neutrophil ~~'r) ~l'Wm nLL't1:::9 :::'t1 WJr.i1'W'J'W~'I'elU1'1'.i''J 1?1L;'d1l1t..IL 'W 3 tJ'WVIi~n1~U1 1?1 L~U
16
lri'1l neutrophil '\.-ll-l!PI'i-lih~mJ''¥11-11'W n9:;l~1~LUJLlnJl-ln'1J'I?I1tJ'll'fNl'll~~ (apoptosis) ll~:;tJn9u
n'WL!Pitl macrophage l'll~~biiiP'Il~'1l!PI'1J1'J"lloW!Pi~'W1 9:;l-J1U-1U1l'Hl.Jlle-J~~n Uilbrl helper T cell 9:;
wi-:J~1J'"Ji'tJ'i:I?ILrn1md1u1~ T cell llU~l?l'Jl~lJ91'W'l'Wll~:;b~lJn1J'reJm~u 'Y11L~'i-1~'1l!Pib~!l!Pi'lltl1tl l?l'J • I !.t I !J Q~ c:r, cf1 1,.1 I- ~ .:.1 IV
b'r'llJ'll'W b'r'llJA'l1lJ~1lJ1J'rln1J''lll-lm'Wb'll1'1l'1ln'll'1l~~~J'e-J1'We-J'W~'i-l~!li9lb~'1l!PI 'W'fln~Dlln'W T cell tJ~
m:;~umJ''Y11-11'W'll'1l~ macrophage LIP'Itl macrophage 9:;l~1lJ1bbYl'W~ neutrophil 1uJ':;tJ:;b'J~1 2 " I I
IV CV .0:::. V .c::Y CV .c::t. .:;:,lq I .dij .:::!rl q
'J'W'\.-1~~91mn~P~U11Plbbe-J~ UY1U1Yl'i-l~n'll'1l~ macrophage A'fl IDlUn'WblUAYlbJ'tJ bb~:;tJ'1ltJ~~~m'W'1lbtJ!lYl
b~tJ'i-11tJL!Pitln1J''i-IJ~b'1l'WL'lll-fn~lJ proteolytic enzymes (Deodhar and Rana,1997) 'W'1ln91ntl
macrophage U~'\.-IJ~~~J'~h~1l'li'W growth factors bl~:;"li'tJLI?ILAt1~91bU'W~'1lm:;U'J'Wn1J''i-11tJ'll'fl~ bbe-J~ b'li'W platelet - derived growth factor (PDGF) bl~::: transforming growth factor-~ (TGF-~)
t.uu~u 'i:IPlm'Yl'r'l1:;1u'li'l~iu~ 3 ~.~.~:;iu~ 4 'i-l~~ln~P~u11Pllw~ ~1JL'i-1~1tl9:;~~1Pl~P~b'll~~~l~tJ'l~'fl~1u '"
J':;tJ:;mnt.ti~l?l'Jb~lJ91'W'l'W (proliferative phase) L~bbrl fibroblasts b~1lJ1U~U1l'lruU11Plbbe-J~
'W!ln91 ntl macrophage u~ tJnm:;~'W~'JtJ~ll1'J:;~~'1l'1ln~ b9'W~1~ bn IPI;'WU1b'lbWU1 (.91 bbe-J~ HI 'i-Ii~ ~1J'LUnJ':;~u1~ bn ~P~nJ':;U'J'WniJ'~~~-:J'i-1~'1li9l b~!liPl1mJ (angiogenesis) nJ':;~'Wl'll~~1 unJ':;U'"l'WniJ'
re-epithelization ~b~:;m:;~'WniJ'~~~~ granulation tissue bb~:;~IJ' extracellular matrix 91nn1J'~ macrophage '\.-IJ~~~JL'\.-1~1\f ~~ e-J~ 1~m:::U'J'Wn'1J''i-11tl'll!l~ lle-J~ l ~~·;;•iJ:;tJ:;n~P~LU 'I.-lin~~ lbU~nU~'1llJ
" tJn'V11~1tJLUL~'i-llJ !PI '1.-11-'fll 'i-1~'1l91u'Juu'1ltJIDl'Whhuu~ul?lntJ 91U'l'W'll'1l~ neutrophil lb~:;
macrophage ~u1b'1ruU11P1 bbe-J~9:;~ IPl~'l J':;tJ:;mJ'~m~u n~Dl:;~"u~ !PI~'! bb~'i-11n~~ bbU~nU~'1ll-JiJ '
A'Jil-Jl'WbbJ''Ibn'Wnrh neutrophil bb~::: macrophage 9:;'¥11~1tJL~ b'll~~vf-1 2 "lloWIPI nu~A~bbU'Il?l'Jb~lJ
9iu'Ju~n m:;u'JumJ'~m~untDl:;~~ ~.uu~!l1u
i.l I I I
- Q 4 - tr .::!1 q -c:r, .c::Y!.I 'i-1~'191nLni9lU11Plbbe-J~'ll'W 2-3 'l'W b'll~~ fibroblasts 9:;bA~'1l'WYllJ'ltJ'IUJb'JruU11Plbbe-J~ b'r'l'1lb'll1
~ J':;tJ:;m nbti-:J b'll~ ~ ~m n 1Dl1ru'll'fl-:~ ~. d''1l ~.m!l ~. ~'fl'li!ll-l bb '1l l-JUIIPI bbe-J ~ 1 u'll ru:;~ J':;tJ :;n 1 J'~ n ~.~u~Dl :;~.,u~ !PI " ~ ' ~~ nJ':;U'J'Wn1J'~1~1 ~bn!PI;'WL'WJ':;tJ:;tl1~bbri n1J'~~1'1'1.-1~'1l!Pib~'fl!PILmJ (angiogenesis) ~~~-:~
" I I
btlmm!l1mJ (granulation tissue formation) ~~UJ':;n'1lU~'ltJ fibrin, fibronectin , collagens,
proteoglycans, glycoproteins bb~:; glycosaminoglycans (GAGs) b'li'W hyaluronan bU'W~'W ~niJ'
~:;~l-JA'1l~~1b9'W (collagen deposition) ~~1-:~bd''fllm!l~e:1'l1'1.-1~ (epithelization) bb~:;mJ''I.-I!Pil?l'J'lJ'1l~
bbr.J~ (wound contraction)
17
.d cv~ s.- JQ . 'i~ti~'Yl 4 : 'i~tJ~n'l'iU'itJbFI'i~~'i'l~L'lJ'f.lLtl'f.l (Remodeling phase)
J' 1:;\ J ,-.dlc. .. 9 :i'~t.J~'W granulation tissue '"l~L'"l:i'fl.JLU'W connective tissue 'Vl'i:'l~\j:i'n..l '\.-l:i''flLni1\Lu'W:i''flt.J
LLe..J'mU'W (scar) L'fl'lJL•nf~~U'VlU1'Vl~1rlfl.JL'W:i'~t.J~ci'~'fl n~~ matrix metalloproteinases (MMPs) ~'I '
'\.-l~'l'fl'flnm'"l1n macrophages, epidermal cell, endothelial cells LL~~ fibroblasts (Singer and
Clark, 1999)
collagenases collagen I 11 .J 1.1 I
connective tissue Yi'Cl~1'1LmJ L'W:i'~t.J~U fibronectin LL~~ hyaluronan '"l~tJnLL'Vl'W'Vli1l'dt.ln'Cl~'ll'fl'l
collagen bundles ~~"ll'W1 11\Lm\J LL~~LLoJJ'I LbN ~'I e..J~ L~rl'd1~ ~'l'll'fl..:J LLe..J~ (tensile strength) L~~J . .n n
('W n1:i''i:'l~1'1'\.-l~'fli11L~'fli11LmJm~i11~..:J rl'd1~'\.-l'W1LLU'U'll'fl..:JL'll~~t?i'l'l1 L"li'W macrophages ,
keratinocytes LL~~ fibroblasts ~11l~'l'"l1nn1:i'l?l1ti~htim~u'd'Wn1:i' apoptosis L'W:i'~tJ~mnH'u
br!N'i:'l~1..:JLif'flL~'flLU'lJ:i'~tl~n1:i''i:'l~1..:J'i:'l~V)m~'\.-1~1..:Jn1:i'~..:JLrl'.i'1~vf collagen L'\.-1:1-i LL~~m:i'vh~'ltl ' ' "
collagen Lrh m~U'd'Wn1:i''li'fl~LL'll~U1111LLe..J~'"l~'\.-l~l?l~'ILil'fl connective tissue Yi'Cl~1..:J~'WL'\.-l:IJ
L'"l1f),JL~Ubl?lL~~~ (Strodtbeck, 2001)
2.3 U~~tl~ij~'Ylfl'W~ l?i'fln'l'i'VI'ltl"ll'f.l~tl'l WILUJ~
n1 :i''\.-11tl'll'fl'l U1 111 Lle..J~ LbiJ'"l ~'\.-11tl l~ L 'fl'll?l1~n:i':i'~'ll1 ~ Lbt?i n~u99 t1U1..:JU'"l9 tJ~'Ji'd tl~'l L'Cl1~'\.-11-'fl "1111l'll'd1'1n:i'~U'd'Wn1:i''\.-11tl"ll'fl..:JU1111Lbe..J~ ~..:J'i:'l1m:i'rtrol1LL'Wnl~~..:J;i (Harari , 1993)
2.3.1 tr~~tiL'U'W'l~Yi
2.3.1.1 "li'U'l Wl"ll'f.l~tl'l WILUJ~ '\.-11nLLe..J~~'ll'W1111L'\.-lfkJ LL~~~ n m~U'd'Wn1:i''\.-11tl'll'fl'l LLe..J~'"l~-il1n~1 ' " qq r;:; ~
lle.Jfl'Vl:W"ll'W'l (?) lfl nLL'i'l ~191'W
2.3.1.2 r?l'lLL 'VI1l~"ll'f.l~ LLt:J~ '\.-11mn (?IU1 (?ILL e..~~ 1"Wu1L'dru~~mnr~~m.J1 '\.-l'd'fltlL'i:'l~'fl '"~~~-.:~e..~~ 1~ "
L~'WLtl collagen Lb~~'\.-l~'fl(?IL~'fl(?l~'i:'l~1'1L'\.-l:IJ~n'll1(?1l~~1tl 'v11L~LLt:J~'\.-11t1-il1
2.3.1 .3 im~tU:::"ll'f.l~LLt:J~~ijn'l'i~WIL:{'fl '\.-11-'fl~~'ILLU~m.J~'fl~'fltlL'WLLt:J~ L'li'U LLe..J~~nL'i:'lULU'U "
2.3.1.4
2.3.1.5 n'l'iL 'VI~ L1tJU"ll'fl~L~'f.l WttJ~L'".ltULLt:J~ L'U1l1'd~'ll1 (?IL~'fl(?l m~u'dtJn1:i''i:'l~1tJ~~ b~'fl (719~
~(71~'1 'v111~m'fln1'i:'lln(?I~~L~'fl(?IL'Um~'fl(?IL~'fll?l'i:'l'l L~'fll?l9~~1L~"tl'll~i1'flti~'l :i''d~vf-.:~ '"
18
2.3.1.7 'iiCUVIll~"ll'il'l LL!:J~ {]l'PI bWVIll~ ~1 nrJ1'''~bWVIlliJi1'l n1t.l '"l:;vi11 ~ LL!:J~VI1 t.l~1 ~'l ' cu q 'U ~ \1
Lti'fl..:J'"l1m:;~..:J t:J ~ 1 ~m:;u'"J't.ln1~'6'1~1'l L~'fll-!~'"l 1 mJ LL~:;m~~'l Lr\~1:::\f collagen Ln ~ 1.~
~1~'l LL~:;~..:Jt:J~1~m~Yi1'l1'W'll'fl'l neutrophil, fibroblast LL~:; epithelial cell U'flt.l~'l
2.3.2 tr~r.rt~~'llt.l
2.3.2.1 'fl1~ 1'WitJ~'l!l1tJ'"l:::i:Jm~srimm'11d-.IU1~LLt:J~L~~1n11it.JVI~d-.1'6'11'"l
i?1'flU'i'l'W'fl'll1i'flm~~m'6'1uu'f!t.J i:Jm~~..:JLr\~1:::\f collagen LL~:::m~sri'fld-JLL'11d-JL~'fll-!~'"l ~~~'l n1~'6'1~1'lVI~'fl~ L~'fl ~ 1 m.JLn ~ L~~1 L'Yi~1:::t:J't!..:JVI~'fl~L~'fl ~'"l:::bL"Ji'l LL~:::VI'W1 ~'"J Yi1
1 ~m~ LVI~ L~t.I'WL~'fl ~ L~1~ LLt:J~U'fltJ
2.3.2.2 ll1'l~Lll"lit.nn1'1' LU'WU'"lrolt.J~1~'1J1'Wm~~'lL'6'11d-.lm~VI1t.J'll'fl'lLL~~ Lsri'W hh~'W LU'W
'fl'l ri'lh:;n'f!U~1~'1J1'Wm~'6'1~1'l Lif'flL~'fl1mJ LU'W~'W ~..:JtT'Wm~'ll1~'6'11~'fl1VI1~~r.i1 LU'W " I
11i'fl m~'6'1~1'l L u'f! Lm'fl9..:J.Yi11 ~ LL!:J~V11 t.l~1
2.3.2.3 ~ll1'l~n1'1'L~uthtJ L'li'W r\'W~ LU'WL~rJLU1WJ1'W9:;i:Jm~VI1t.l'll'fl'l LLt:J~~1n11un ~
2.3.2.4 tJ1 m~l.~fu'fl~m'"li:J;sYlfiri~l1i'flm~VI1t.J'll'fl'lU1~LLt:J~ L'li'W t.11U1'ln~d-J'"l:;mm'Wm~
~'lLr1~1:;Vf1u~~'W LU'W~'W
3.1 YJ"li~n~ Curcuma • ~ I
m~Mm~1~Ylfl wound healing 'll'fl'l Curcuma longa YlLi?11t.Jd-.11'W ethanolic solution1'W
~i?11Yl~~'fl'l wistar rat ~Qn'6'1~1'lU1~LLt:J~~'"lt.J';jfi light ether anesthesia ~'lLU'W semi-aseptic
~i?11Yl(;l~'fl'lQnm.i..:JLU'W 3 n~d-.1 L~LLri control (n~d-.1~ 1 ), Curcuma longa extract (n~d-.1~ 2) LL~:;
Framycetine sulphate Cream (n~d-.1~ 3) (Reference Standard) vf'l1'W excision bb~:; incision
wound models Yi1JrJ11'W excision wound i:J LU!lfLoTI'WI1i'n1~VI1tJ'll'fl'lU1 ~ bbt:J~ 1'W n~d-.1~ 2 LL~:; 3 '
d-.11nn11L~'flLmt.JuLY1t.Junu n~d-.1~ 1 mh..:Ji:J't!t.J~1f1'1JYl1'l'i'ltl~ (p<0.001) 1'W~'"l'W'll'fl'l incision
wound YiUrJ1 n~d-.1~ 2 LL~:; 3 i:J 333.78 ± 3.55 LL~:; 379.00± 6.323 1?11d-.1~1~1J Lmt.JuLY1t.Junu '
I I I .:::::l c9. .c:::\1
n'tid-.!Yl 1 'l!'ld-.1 tensile strength 281.30 ± 5.82 (Rita et a/., 2012)
'W!lm1nd'u..:Ji:Jm~Mmnn..:JUYlU1Yl'll'fl'l curcumin 1'Wm~fmnu1~Lbt:J~ t~t.Jmn~d-Jmd-.11ru rl'fl~~1 L'"l'W LL~ :;r1 DJ~d-.!Uii1 'Wn1~~1'W'fl'Wd-J~~M:; l 'W~:;VI~1'l m:;U'"J'Wn1~fm~11J1 (;) bbt:J~ l 'WVI'WYl (;l~'fl'l
q q 'lJ 'U
19
~ ' nt1~m~ni1H~'W albino Wistar strain L~~~ 'fl1t.l 10 L~'fl'W rol1'W"l'W 36 lPl"l L~t.l~~1-..l1J1~LL~~LU~Vl
'IJ 'IJ '
u~L'lru~i..:J'JJ'W1~ 2 cm2 LLU..:J~~LU'W 2 n~~ ~'fl n~~mu~~1~fuJ1Ln~m . .J1~1l911 200 1-JI LL~:;n~~
'VlV1~!l~L~fu curcumin 40 mg/kg m~1l911 200 1-JI LYhn'W LU'W1:;t.J:;L"J~1 12 rJ'W ~~n11Pln'l11~1J~1
curcumin ' ' ~
t!'t.1~1~ru 1:;~1JL'fl'Wl'll~~1'W'fl'W~~~M:;'JJ'fl..:Jn~~V11~fu curcumin LYl~~'W~1nn~1n~~~"l1J~~mh~ Q,l q 'U q q q
~t!'t.1~1~ru ~'f)l9Jnnt1~V1l'i'"l'JJ'fl~LL~~'1J'fl..:Jn~~~1~fu curcumin L;"ln~1'flth~~t!'t.J~1~ru ~ ' ~
tensile strength L~~:f'Wn..:J 129% L~'flL'Ylt.JUnun~~~"lUfl~ (Panchatcharam eta!., 2006)
r.nnm d'Pl n111';Jr.ft.J ~ru~~1J1\'1J'fl..:J~11~ n ~r.nn~'1ll?i'flnt 1n'd:;~'Wn'l'd~1t.I'1J'fl..:J1J1 ~ LL~ ~ 1 'W ' '
~ ~
~l9lrJ'VlV1~'fl..:JU'W trn';J9t.J~U~1~'1l~~1t.J'1l'W~~~'Vlfi~.m~1"l l'i'"l'fl~1Wri'W
.... 3.2.1 U'JlJn
nTj'~n'J11~~'1J'fl..:J asiaticoside ~~nV1l~'"nmJ"lun Centel!a asiastica ~~l?i'flnr'd~1t.J'1J'fl..:J " " punch LL~"l'Vl1 LL~~~"lt.l
~1'd~:::~1t.l asiaticoside 1'WJ1Ln~'fl ~~"l1~L~~~'W 0.05%, 0.1% LL~::: 0.2% m~1ru 20 l~L~'d~l9l'd/
LL~~/~t..:J rJ'Wtl:; 2 ~f..:J LU'WL"l~1 7 rJ'W 'Ci"l'Wn~~WJU~~1.Ji vehicle control ~..:J~~n'lj''VlV1~'fl'l ~u~1 ' '
hydroxyproline LL~:::r-11 tensile
m~1tu~'fl~~1L"'i'WLL~:::m:::~'Wn'l'd~~1'lL~'fl1J~"l1~l.Jl~~n~1n~~~'W 'W'fln"'i1nd' tJ'll~~n'l'd~m~nfi'l ' ' q
~'Vl'if~1'W'fl11~~~~'d:;'1J'fl'l~1'd~rlV1~l~"'i1n'Ci"l'W1U rh'W1U LL~:;'d1n'1J'fl'lU"lUn~"lt.IL'fl'Vl1'W'fltl J1 LL~::: light petroleum ~"lt.J';Jfi diene conjugation formation test L~t.J1.Jim~1'1J~'W'1l'W~ linoleic LL~:::
thiobarbituric acid test (TBA) ~~m'd';J{t.J~1J~1 ~1'd~n~L'fl'Vl1'W'fl~"'i1n'Vln'Ci"l'W'1J'fl'lU"lUn~1~1Hl q
~1'W'fl11~~~M:;1~~~1l'i'"lv11~:::~1t.J'1l'W~~'W 'fl~1'l~t!'t.J~1~blJ'Vl1'l~n~ (p<0.05) (Shukla, 1999)
' ... 3.2.2 'J1'W'VI1'l"'l'i:::L".IJ
nt'd~mn~'Vli'1J'fl..:J~1'W~1..:J"'i'd:;L~1'Wntm'd:::~'Wm'd~1t.J'1J'fl'lLL~~~~"lmr..:J1'W~~~Lu'W LU1~"l1'WL~t.l~~ Wistar rat !Oi:;Qnmdmtl11illU'WLU1~"l1'W~"lt.ln'l'd~~ streptozotocin mmru 70
~tl~nf~Jil'l~nf~ LU'WL"l~1 7 i'W LLU..:J~'W'fl'flnLU'W 2 '1l~ L~'flvl1nt'd~~1..:JU1~LL~~LLuutK~L~'fl'W~~i..:J 'IJ q
J .r:::S I ..:::!il l.l ..:::;::,1 J <V C:
~~ L~'fl~m-11~1 histology Ll~::: wound contraction ~'d'flM1-..lU1~LL~mLU1Jnd~'VlW6"1-..lbL~:::lt.JULL~~ ' ~
L~'fl~ntnr-11 tensile strength "'i1ntT'W9:::LLU'l~'W'fl'flnLU'W'6'l1~n~~ l911~'ll'WV1LL't1 :::';JfinTJ"1ih~1'd Lt?lt.J 'IJ q
20
n~),J~ 1 La.JL~fU~11biP11 n~).J~ 2 Vl1~1'W~1~91:;L~~Lbe.J~ LL~:;n~).J~ 3l~n'W~1'W~1~91:;L~~Lt;J1£m ' ' '
Vl1tnl1P11 n~).J~ 2 Vl1~'"lm'"l~~1'W~1~91:;L~ n~).J~ 3 VJ1m microcurrent LL~:;n~).J~ 4 Vl1L9~~1'W ' ' '
~1~91:;L~i'J).JrlU£J1 microcurrent e.J~n11Vl!Pl~'El~YIU~1LLC-J~'ll'El~~'Wn~).J~ 4 L~1~1:;t.J:; proliferative " ' "
phase 1~L~'1n~1n~).J~'W LL~:;1'ElmLt.Jn~.f'W dermis U!Pll~L~'1n~1n~).J~'W LL~:;tr~il91'W'1'WL"l!~6 fibroblast w~:;w~'El!PlL~'El!Pl~~~1~1~a.i).J1nn~1n~).J~'W LL~!Pl~l~L~'W~1n11L-i!L'"l~~1'W~1~91:;L~i'"l).JrlU
' tl1 microcurrent iJ~VJi~~L~1).Jrl'Wb'Wn11n1:;~'Wn11~1U'll'el~U1!PlLLe.J~LLUULLe.J~LU!Pl (Chithra et a/.,
1998)
3.2.3 Fl'n),Jth
n1JMm~n ~Vlim:;~'Wn1J~1t.J'll'El'l LLC-J~ LIPltJ1~~1J~n ~~'ltJ L'ElVJ1'W'El~'"l1n fiJ1~th L~t.JVJ1U'W LL t.J~~tl.UUr;i'!PlL~'el'W'll'el~~'W Wistar rat ~fl'11).JL~).J~'W 5% LmUULVlt.IUrlUn~).J~VJ1LLe.J~l~l'JUfi1),J
" ' ' ' I d !/ ~ 0 OV
simple ointment base LL~:;n'Cj).JVJVJ1LLC-J~!Pl'"ltl 0 .005% fluticasone propionate L~'ElVl1n11'"l!Pl
m~1ru hydroxyproline fl1 wound contraction LL~:; epithelialization Vln 2 i'W LL~:;r;i'!P)Lif'ElL~'El '
'El'Eln~1~mnin~ru:;VJ1'l histopathology YIU~1 LLC-J~~Vl1 ~'1U~11~n ~91n ~'Wfi11~U19:;~1U L~'J
n11n~).J~'W LL~:::Yiummru collagen fiber, fibroblast LL~:;~~'el!PlL~'el!Pl~~11~~"'W1~~).J1nn11n~).J ' <'I
'El'W(Lodh iet a/., 2006)
3.2.4 LL~tlfi~1tl .. ..::! ~ 0V I !/ V .dil
n1Uln~1 ~Vlfi'll'El~~11~ n !Pl LL~ tJ n~1U t;J'fl n11m:;~'Wn 1:;U'"l'Wn11~1tl'll'El'l LLC-J ~ LLUU t;J !Pl L'U'El'W
L~u1-il~'Wnur.fm LYif!LiJtJ mu 8 iUIP11if ~~~'WVl!Pl~'El~LU'W 3 n~).J ~~1~U11PlLLe.J~~~i~'ll'fl'l~'W l!PlU \.1 .. q \.1 .. 'U
~~n~~~ 1 '"l:;L~fum1VJ1LLC-J~~'1tl methylcellulose ~~n~~~ 2 9:;L~fUn11Vl1LLe.J~~'JtJ~11~rl!Pl ml1U91nlULL~Un~1tJ~fi'11).JL~).J~'W 0.5% LL~:;~'Wn~),J~ 3 '"l:;l~fUn11Vl1LLe.J~~'"ltl~11~rl~~tl1U
" ' 91n bULL~tlm'l1U~ fi'J1).J L~).J~'W 1.5% LL~:::Yi1n11LrlU~'El~~in~ru:;'JJ'fl'lU1 !Pl LLC-J~ LL~:;i ~'ll'W1 !Pli'W~
" LLt.J~'V!n 2 i'WdJ'WL'1~1 16 i'W LL~:;U1).J1A1'W'1rufi1L'U~u~m.J~:;'ll'El'ln11~~1~L~m~~'11~a.J (%
epithelization) fl1L'U~U~'Elu~:;m1~!Pll?i''J'JJ'fl'ILLC-J~ (% wound contraction) LL~:;fi1L'U~U~'Elt.J~:;'JJ'El'l
21
' ' m~YI1tl"ll'tl'lbb~~t~?~m'J:w (% total wound healing) ~'191n~~m~~{t.JY'f!J~1 bb~~YlY111~hm'l1~~rlr;)
LL~tln~1t.JiJm~~~1'1 granulation tissue L~b~'Jbb~:;iJ'l!'I.J1r;)LL~m~n~..:J L~'Jn~1n~:1-J~'J1Jf):I-J LL~:;bL~~~ Yl1~'lt.J~1~~rli?ILL~t.Jn~1t.J 0.5% 9:;iJrh% epithelization, % wound contraction LL~:;% total
wound healing ~'~ n-J1 LLc..J~ 1 \..ln ~ :W~\..lmh'liJutJ~1 ~ C1JY11'1~ii~ (p<0.05) LL~::cl''ln nJj'YJI?l~'flU b ~).JL~:W'Yi1J~1 bb~~~Yl1 ~'Jt.J~1 ~~n (;) bb~t.Jmnm.f..:J~'tl'l ~'l1:1-J b ~:I-J~\..l~1:1-J1Hl'Yi1J m n91n.J'll'tl'l collagen
fiber, fibroblast LL~:;~~'tll?l L~'tl (;) L~'l nrJ 1 bb~ :;;:J m1:1-JYI\..l1 bbkl\..l:I-J1n nrJ1 n ~:1-J ~'l1J fl:I-J ('J ~~i n'rl'bW
P~11h9u, 2554)
4.1 Tetrachlorodecaoxide (TCDO)
Tetrachlorodecaoxide (TCDO) iJ~'tln'l~~h~1 Oxoferin® bU\..l~1~~:;mt.J1'1.Jtl1 bU\..l I I t.t I
activate oxygen ~'I buwK'l Li..:JY11..:J~'JJl1Yi'lloW(;)'tl t~Y~:; d~'tlrPl'Jt.l'l bbmn~:wb~1~Li'l'tlbU'tl"'l:;m:;~'I.J1ihn (;)
m~n~\..l bL~:;vi1~1t.Jbb1J~Yl~t.J (phagocytosis) Y~1'tl~1nLu~mJ~'tl:I-J~\..l 1 vi11i~LL~~~:;'tl1r;)L~'J,}\..l bb'Cl::: i?l'tl ~1\..ln'l~~ (;) b~'tl L'U'Yi1 :;~ rPl'l Li'IY11'1~'lJl1'Yid'bd'tl bbl?ln rPl'J"'l:;L~ metabolite ~'tl oxygen bL~:;
' ' ... chloride iJ~~vi11ilv~:w~'J1:1-J~\..l'll'tl'l oxygen Yl1J11?lbb~~ vi11i~Lnl?lmni..:Jb~'tll?l:I-J1b~t.J'I bL~:;~~1'1
Lrl''tlL~'tl~ bbl?l'l~ 1?11J1b'JDm~~ ;..:JYiJ b ilbn l?ln1~~~1'1 ~'J\..l'tln'll'tl'l Ld''tlb~'tl b~'J,r\..l bb~:;b~\..ll~ b~\..l"Jl 1?1 bb~:; ... ~ ...
~~1'1~'lmr..:J1il~t.J1J hibU\..lLL~~\1\..l LL~:;1ml.) ~..:JU\..l TCDO 9'1iJ~Ylfi~1b~'tl Lb~:;1il Oxygen LLri~'l iJ
~'tlu..:Jb-D1 \..l~th t.1 b1J1YI'J1\..l ~..:J1J11?1 bb~~~n~1t.1-il1 iJ~1 b ~l?l:W191 n m~~u rPl\..l'll'tl..:JYI~'tll?l b~'tll?l~vi11 il 'IJ ' ... ...
b~'tll?l hi~1:1-J1~bl hJ L~t.l'l'tli t.J'J :;~'J\..li?l1'l1 'll'tl'l i1'1n'l t.1 vi11 il~1~'tl'l'YI1nL~ :;'e)'tln~ L 9\..llU L~t.l'l1J1L 'Jill ...
u\..!1 ~r;)U'tlt.J~'~1u~'Jt.J
1J'll?lLLc..l~ LL~::cl''l LU\..!Yl'l'lfh{ !?I'll 'e) 'I L~t.J91n1J1 r;)LL~~ Ld'tl~ruL~t.J~U1~ 'vl1'tlvi1~U1~l~~ (;1~'1 n9:;vi1 'IJ ~
t... i.t I I I
1i~LL~~YI1t.J-il1 LL~:;~I?lb~'tll~~~t.J \..l'tln91ni:J TCDO 9:; b~:I-J~U'Irl1Yi~1Ylb1nvi1mt.J ~'ll?l'tl\..lbLdn LL~~ 'IJ
91n~bbr;)..:J 9:;n~1t.JbU\..l~'ll:I-J~LL~'lrl'tlt.l 1 L~:I-J~U'ILL~:;Ld''tl~Qnvi1~1t.JllJ
n1~~n'l11bmt.J1JbYlt.J1Jn1~'vl1t.J'll'tl'l1J1(;1bb~~1\..l~lhmf'IYI:I-JI?l 60 ~\..! ll?lt.Jb-D TCDO b\..l~tht.i 'IJ 'IJ
29 A\..l Lmt.JUbYlt.JUnum~H PVP-iodine complex b\..l~tht.J 31 ~\..! Ll?1t.Jir;)~~91nm~~l?l'll\..l1(;1'l!'tl..:J 'IJ
... ' Y!'I.JVlLLc..J~Yi1J~1 TCDO iJu~:;~YJfiJl1'YiL\..lm~fm11 complicated wound :W1nn~1'tlti1..:JiJut.J~1~ClJ
... ' \..l'tln"'l1ni:J TCDO U..:J~1:1-J1~mY~i:Jmtl11iliJmn"'l1ClJ'll'tl'l granulation tissue L~'tlti1..:JiJu~:;~YJfiJl1'Yi
mnn-J1m~H PVP-iodine complex Lb~:;1~fum~fu~'tl'lrJ1~1:1-J1~b11-DLU\..l antiseptic L'li\..lb~mnu
PVP-iodine complex (Malik et a/. , 1997)
••
~!te_rf!,_lof:! '!_V~ !)paning:
Turn cap with inner pu1 past the resistance
To be d1spt.>t>sod on rno<J.cal prescription anly
';IU~ 9 Oxoferin® (Tetrachlorodecaoxide) ..
22
m~~m~l~'Vli"ll'il'l Tetrachlorodecaoxide (TCDO) 11.-.l~tht.J oral mucositis grade II- IV
62 11tl ~dlf?1~1n chemotherapy-induced oral mucositis ~'ln~~'Vlf?1'i'l~'l~~1~fu TDCO 1Om I
U'11.-!thm~~'Jn~1.-.li1.-.l'i'l~ 2 ~f'l ~U1.-.l~~t.J~~'1'i'l1 7 i1.-! lf71t.Jif71~'i'l~1n grade of oral pain 'V'lU~1hiiJ ~'11W~lYlnl?i1-.:Jn1.-.l'ilti1-.:JiJ'l!t.J~1r1'b1J'Vl1-.:J~i:l1~h~'El~mt.Ju~Yit~u~~,.,~1'l i1.-.l~ 1. 3, ~~fl~ 7 ~~l?i~~'il
~mtJU~YltJU time to improvement ~~,_,'1.1'ln'i'l·~'Vlf?1'i'l'il'l~~'i'l:::n~~~'1U~~'V'lU~1i1~'11~~~lYlnl?i1-.:Jrl1.-.l q q q
'ilU1-.:Jil'l!t.J~1flbkJ'Vl1'l~~~ (p = 0.02) ~~'i'l:::~~'il~mt.Ju~Yit.Ju percentage of patients with
. . I I I I .:;:;I I
symptomatic Improvement by day 3 ~~'J,I'J1'ln~~'Vlf?1'i'l'il'l~~'i'l~n~~~'1Ufl~'V'lU'11 ~~'11l-mlYlnlY11'l
rl1.-.l'ilU1-.:JiJ'l!t.J~1flbkJ'Vl1'l~i:l~ (p = 0.05) ~1nm~~m~1~~~f71'l1ih~1.-!~1 TCDO m~i:Ju~~'i:mm11.-.l " ~tht.J oral mucositis U1'lnmvh'l!1.-.l (Ma lik eta!, 1997)
4.2 Heparinoid
Heparinoid
glycosaminoglycans mucopolysaccharides
~~1ru ~~U llYl ~b'i'l~'Ji'Jt.J'Ji'il~ bb'll~b'll'i'l;~U1 f?1 b'~U lV1t.JiJM~'V'lAill'Vl1'ltl1 ~'il U~~b'Vl1'illn1~'J,I'i'l'ilV1 b~'ilV1~1 ~ q
1~~':!,.,'1!-.:J'flm~u fl f71Jl1'J~b~'ilP1 rr..:J1~~'J,,r'l vh 1~~'ilm~~'i'l bU1.-.lf71~1-.:Jfl'l bb'i'l~ H'Vl1 b~'il~~ P1~'J,,r'l 'lJ
23
~u-Y1 1 0 Hirudoid® (Heparinoid) ..
n~ 1nm~'fl'fln ~Yli'JJ'fl'ltn le:JUi~"W'fltJ(Y) ~'fl uutT-.:~m~r.Jui?l'J'JJ'fl'l ln~ ~l~'fl~ 1'Wu~v;m.!VI~'fl~
l~'fl~~11~~'JVI'I!'I vl11 iim~1VI~ l1t.J'W'JJ'fl'll~'fl~ 1 'WU1l'"lbl.l~Yl1tn ~~"'W vl11iit~1 n1~U'".l~ 1 'WU1l'"lru~iJ l~'fl~;'llU1U1'1~'1 ~~'fl1n1~U'Jd-J ll~'l 'iJm~U ll~~n1~~'1i?l'J'JJ'fl'I~'JVI'I!'I1'WU1l'"lbl.l~Yl1tn
m~~m11~Yli~l'Wm~umJt~'l skin-flap survival 1'W~rwfYl~~'fl'l (wistar rat) t~mmt.JUlVltJU
m~1~ Hirudoid® 0.5 g. (n= 8) nu topical carrier of Hirudoid® cream without MPS (n= 8) l~t.J ' .dl Q .Q Q/ iJ ' Q.o' Q..:::., (I..'
b'uti'Lm~~YlUJl'".lbl.!VI~'l'JJ'W1~ 10x3 em ~'".lt.J electric clippers ~U'".l1VI~Vjnl9l'".l~'fl~'ll'".ll91VI~'I"'l1n
m~~mn 10 f'Wl~t.JiJ average necrosis area 'JJ'fl'ln~d-J~Yl1~'".lt.J Hirudoid® ~~lU'W 31. 7± 17% '
bb~~ 48 .9 ± 9.6% ~'~lll9ln~1'1n'Wt~ti1'1n,rtl~1fi'C1Jm'~~n~ (p = o.o21)
m~~m11~Yli'JJ'fl'l Heparinoid (HEP) bb~~ troxerutin bmt.JUlVJt.Junun~d-JfPl'JUf)d-J (Normal
saline) 1'Wbb~~ flap necrosis vl1n1~Yl~~t1'11'WVI~ Wistar albino rats n~d-J~~ 10 i?l'J H~~t.J~l'".l~1 " if'IVI:W~ 7 rf'W l~t.Jrf~~~"'l1n Vascular endothelial growth factor (VEGF) levels, blood level of
nitric oxide, mean area of flap necrosis, rates of fibroblast proliferation ~U~1~~i?lu VGEF hi
bbl9ln~1'lrl'Wtlti1'1n'i!t.J~1flrlJYl1'1~n~1'Wlb~~~n~d-J (p=0.30), ~u~~i?lu nitric oxide 1'Wn~d-JfPl'".lUf)d-J
~'ln~1n~d-J~1~fum~fn111tlti1'1noJtJ~1flrlJYl1'l~rl~ (p <0.0001) l~tl~~l?i'U nitric oxide ~~VI~1'1
n~:w~1~fum~fm11~U~1hhlt91nl9i1'1rl'W'fltl1'1ntit.J~1rlrlJYl1'1~n~ 1'W~'".l'W'JJ'fl'l mean area of flap
necrosis ~U~1n~d-J~1~fum~fmnnm~~~~'I'JJ'fl'l area of flap necrosis ~n~1n~d-JA'".lUf):Wt~ti1'1n
'i!t.J~11Plr1JY11'1~n~ (p <0.001) lbt9ihinm1d-Jlbl9lnl9i1'1rl'W~~VI~1'~n~d-J~fum~fm~n (p=0.60) bb~~
rates of fibroblast proliferation 1'Wn~d-JA'JUAd-Jnm~~~~'ld-J1nn~1b~'flblli-t.JubVlt.JUnum1:w~1~fu ' ' '
m~fm11titl1'1iJ'i!t.J~1rlrlJYl1'1~rl~ (p <0.001) ~~n1~~n111bb~~'l1iib~'W~1 troxerutin bb~~ HEP n ...
~Yl~1'Wm~~~ flap necrosis bb~~ improved flap survival (Orner eta!., 2010)
..
.d tJ 'Vl'Vl 3
"ll'£l'l.J L "ll l?l"ll'£1-:J n1'i~~ t1
1. m~~mn~Ylim:;~unT~'VI1tl"1l'fl..:I1J1l?lbbe.J~1uVI~'fll?lYll?l~'fl..:l (in vitro)
1.1 nt~Yll?l~'fl1J~Ylib~:I-J91'W'd'Wb"l!~~ (Cell Proliferation activity)
(Curcuma cf. comosa Roxb.) ~'lt.J1fi WST-1 cell proliferation assay
1.2 m~Yll?l~'fl1J~Ylim:;~um~VI1tl"1l'fl..:I1J1l?lbbe.J~ Ll?lt.JL'ilLL1JlJ91~'fl,m1~VI1t.J'll'fl..:I1J1l?lbbe.J~ (in vitro
"' ..,.., wound healing model) l?l'dt.l'dfi Scratch assay
2. m~~m~1~Ylim:;~um~VI1t.J'll'fl..:I1J1l?lbbe.J~L'W~t1l'JYll?l~'fl..:l (in vivo)
~ ' ~ i,1 0 V .oC::II 4CI/q - I i.f
m~Yll?l~'fl1J ~Ylnm:;[;jun 1 ~VI1t.J'll'fl..:I1J1l?l Lbe..J~'ll'fl-.:1 mmfl ~:I-JYl:I-J~1~~ n l?l'll'fl..:l'd1'WVI:I-J1'l'fl 2.2 SJ 1.1 11 I
n1~[1]~'l"'l:n'Wb oW'flY11..:1"'l~ m t.J'1Jl1 fl ~1~ [1]f'll'fl..:l bU'fl b~'fl (Histology) '
2.3
~1~~rll?lVIt.l1lJ (crude extract) ~'lt.Jr?l'lvh~:;~1t.J ethyl acetate 'll'fl..:lrJ1'WVI:I-J1~'fl (Curcuma
ct. comosa Roxb.) l~"'l1nLflNnt~19t.J~ 1 ~'fl..:! Lflj..:intd19t.J~ 1 ~'fl..:! ntj91bb'Wn"llill?l nfj~rll?l r
bbt.Jn~1m1~Ylfi '
Curcuma cf. comosa Roxb. (Plant identification, extraction, isolation and structural
identification of bioactive compounds from rhizome of a herbal plant, Curcuma cf. comosa
Roxb.) Jl1t.JL~bbe.J'W..:I1'W'19m~mn'W ~'fl n1d~m~1~YliY11..:1~'dJl1'Y'I'll'fl..:i'Vi"ll~~'Wl'Y'Id Curcuma ct.
comosa Roxb. bb~:;~I?J.J'W1bU'W~1f1Jtnfm~1bbe.J~ Ll?lt.J~rll?l"'l1n~'d'Wi'd'll'fl..:i'Vi"ll~:I-J'W1'Y'Id Curcuma ct. '
comosa Roxb. ~'dt.J~fi Fraction percolation ~..:l~~fimd~rll?lL!?lt.J~..:Ib'llU ~'fl i'..:~e.J..:~Vi-n~:I-J'W1'Y'Inb~..:l~ '
lJ!?l'Cl:;b~t.Jl?l b~:1-Jr?f'dvi1~:;~1t.J bb~'d~nl?l'Vi"ll~~'W1'Y'Id~'lmfli'fl..:~ Soxhlet apparatus (Buchi,
Switzerland) LU'Wb'l~1 12 i''dL:I-J-.:1 "'l1ntT'Wm''fl..:l~1d~n~VIt.l1lJ j:;bVIt.lbb~..:l~'dmfli'fl..:~ Rotary
evaporator (Buchi, Switzerland) ~'fln.JVIJlii 50-70 'fl..:l~1b"l!~b~t.l~ "'l'W~1d~rll?lVIt.I11Jbb~..:llJn"'l~..:i1'W ' " '
'll'l~Ldh U'W~nJ1VI'Wn~1d~nl?lVItn1J~Lb~..:i~ilYl diu1'W~LU'W "
..
25
l?l'il'Uvi 1 n1'iflin'lf1t]'Vl ~n'i~ ~'U n1'i~1t:J"ll'il'I1J11?1 LLc.J~ i. 'U ~~'il!Pl'VWI~'il'l (in vitro) "lJ 'il'l'll1'i'lln IPl
tJ1'U~)J11'il
LU'Un'l~~mnlWL'D'Cl~L'Y'j'l:::Liti-:~'Dil~ L929 (mouse subcutaneous connective
tissue) LL'Cl:::L'lf'Cl~m'l:::Liu-:~'D'W~ HeLa cell (human cervical carcinoma) 'l~uni1?1cp.h:::~-:~r1L~'Cl '
th:::Lij'Wth:::~Yl frJl'lyj b 'Wn 1m ~:::~'Wn'ln~U ll?1 bL'Cl::: L~~rol'l'W'J'W'li'Cl'l L'D'Cl~Yl Lll t.J'J~'Cl'lrlUn1~'1i'fl~ LL ']~
U'l~LLeJ'Cl 'i''"l~n'ln'l'i''l-l'ltl'li'Cl'IU'l~LLeJ'ClboW in vitro wound healing model
Autoclave (Kokusan Enshinkimod H-88LL, Japan)
Cell counter (Hand Held Tally Counter, Diamond 24100, Taiwan)
C02 Incubator (Shellab Automatic C02 23237C, USA)
Haemocytometer (Neubaure BOECO, Germany)
Hot air oven (Hearaeus Instrument, T6200, Germany)
Laminar air flow (Holten Mod. HBB 2460, USA)
Micropipette (Tranferpette, Germany)
Microplate reader (Biotek/ELX-808 IU, USA)
Microplate shaker (ELM I shaker ST3, USA)
Microscope (Nikon/Aiphaphot 2, Switzerland),
pH meter (Mettler, Switzerland)
Refrigerated Centrifuge (Labofug 200 S Hettich, Germany)
Reverted microscope (Nikon, ECLIPESE TE 300, Japan)
Ultrasonic bath (521 0, Branson)
Vortex mixer (Thermolyne, USA)
Water bath (Heto-Holten NS BWB22-2, Denmark)
Glassware and Plasticware: 96-well microtiter plates (TPP, Switzerland), 6-well
microtiter plates (TPP, Switzerland), disposable 0.22 IJm membrane filter
(Gelman Laboratory, USA), centrifuge tubes (TPP, Switzerland), cell culture
bottle (TPP, Switzerland)
.... ~1'3'LI"l~
Dimethyl sulfoxide (DMSO) (Sigma, USA),
Doxorubicin (Pfizer, USA)
Dulbecco' Modified Eagle Medium (DMEM) (Gibco, USA),
Ethyl acetate (Merck, Germany),
Fetal Bovine Serum (Gibco, USA),
Penicillin/streptomycin solution (Gibco, USA),
Sodium hydroxide (Merck, Germany),
Sulfuric acid (Merck, Germany),
Trypan Blue Stain (Gibco , USA),
Trypsin EDTA (Gibco, USA),
WST-1 cell proliferation kit assay (Roche, Switzerland)
" .J' L'lf~~L~1~L~ti..:J (Cell cultures)
L'1!'i'l~L'Y'l1:-;L~"tl~l~'"l1n~ru:-;L!l~'1!1"\1~(?1f J..JVI1~'Vlt11~tl!llJ'i'l11'1!fiTW 9TWTW 2 'J!UC?l ~!l q
1. L'J!'i'l~Ltl!lL~!lb~~'d'VItJ~"ll!l~VI'W 'J!UC?l L929 (mouse subcutaneous connective tissue) "
2. L'1!'i'l~J..J:-;L~~1.hnJ..JC?l'i'ln"lJ!l~J..J'W~cl"" 'J!oWC?l Hela cell (human cervical carcinoma) " q
' '
26
.ell .ct. I ,c::l .:;;;:,
Eagle Medium (DMEM) 'VlL(?]J..J 1% penicillin/streptomycin solution LL'i'l:-; 10% FBS lJJ..J'Vl~n.l'VIJJJ..J
37 !l~Pl1L'1!'i'lb~tl~ b'W~Jl1'd:-;~~fl1'1!~1flJ!l'WlC?l!l!lnl'1!!?f5% LL'i'l:-;LU~ti'W'fl1VI11b'VIJ.JVln 1 3 tJ'W
" " ' L 'Y'l1:-;L~ t.J~ L'1!'i'l~~'dtl!l1VI1H~tl~ L'J!'i'l~~J..JlJHU (completed medium) VlU1:-;n!lu ~'d t1
" Dulbecco' Modified Eagle Medium (DMEM), 1% penicillin/streptomycin solution LL'i'l:-; 10%
Fetal Bovine Serum (FBS) UJ..J~~ru'VI~ij 37 °C ~Jl1'd:-;~~n1'1!fl1flJ!l'WlC?l!l!ln1'1!!?15% b'W C02
incubator ~'WL'1!'i'l~L'"l1n.JL~lJl(?l~~'d1J..JVI'W1LLU'W 80-100 % confluent H pipette I?JC?l!l1VI1Hri1!l!ln
b~VIJ..JC?l L~J..J 0.25% Trypsin EDTA llJ..J1n.l 2 ml Yi"~1~ 1 'W1Vl L~J..J completed medium 5 ml 1-D
tlLU(?]C?l C?lLL'i'l:-;Lth~11'i'l:-;'i'l1t.J L~!l HiL'J!~~~ Lm:-;!lti'VI~ C?l!l!ln 1~VI~C?l ~1mf'W rht1~11~:-;~1m'1!~~~~ 1'W " 'II q
conical tube 1111utf'Wl.l.t.Jn~'"ltl Refrigerated centrifuge ~m1J..Jl.~'d 1,500 rpm 15 'W1Vl Vi"~~'d'Wb~
27
l~l.J completed medium 5 ml ~~l.J1ih~1rl\J il11utf\JLLtJn~nr1f'l i'I~'J\JL~l~l.J completed
medium LVI~
n1'i~~~tllJ~1'i~rl ~?UJ'\..11 'Vi'i • l(;]~tJl.J ~~ 'j'~ n ~ ~:W\JL Yi 'j' 1 ~il f)')\l.J l ~ lJ ~\,I 2 l'Vlt"ll!l'l f)') 1'-J L ~ lJ; \,1~1-li'Yl ~ ~!JU ~ ') tJ i?lCJvh
' " ...
~~tnt.J ethyl acetate ll~~ completed medium b~t.JL-ii 1% DMSO -rl'Jt.JL\Jn1'l~~~1tJ 'vl'IU fl'J1l.J
L~lJ~\J~~Yi1m!l'I~:W\JLYi'l~1-ii'Yl~~!luLvhnu 400, 200. 150, 1 oo. 75, 50 LL~~ 25 IJg/ml ' '
a.~ 0 Q, a. cv 'l fi rn1~ '1..1'1..1 n1'i'1~ t1
f' 1 ~ V I 1.'
1.1 n1'i'VIIPI~'£l'Ut]'VIfi~vhJ~1'1..1'1'UL'1!~~ (Cell proliferation activity) "ll'£l'1~1'i~nrn'11'U'VI:W1'J'£l
(Curcuma cf. comosa Roxb.) lPl'1t~1fi WST-1 cell proliferation assay
Cell proliferation assay lU\J~fim'l~m;-1~~"1l!l'l~1'l'Yl~~!lUl'i!lmn91b]dl~Ub(;] ll~~n1'l ' ...
L~lJ91\J'1\J"ll!l'IL'D~;lYi1~l~tJ'l l'YlAUfln1'l'Yl~MlU~'Jt.J~fi WST-1 cell proliferation assay mPi't.J
Vl~n n1'l~ l 'D~;~J:J~~ (;19 ~~1'-J 1 'ltlti!ltJ WST -1 (2-(2-methoxy-4-n itrophenyl)-3-( 4-nitrophenyl)-5-' ...
~" ."1 "" .. "' 'l 0 1""" (2,4-disulfophenyi)-2H-tetrazolium, monosodium salt) bVIlu\J~1'l~lVI~!l'l 'D'l~~~1t.l b\J\J1 ~~ 1 11 11 I I I
LiJ!l1~~1'j'U~'l1u1 \J!l1VI1n~tJ'l L'D~; ll~:::til.J L Yi1:::nul'l!~; lU\Jl'1~1 4 ·11'1 bl.J'l l'D~;V1il~~(;]9:::lu~tJu 11 I I 1.1 11 t..
~1'li?l'l~\J ~!l wsr -1 1uLuw:.J~(;].rlru"YlV1il~LVI~!l'l djm11mVI1'lL~t.J'll'D~;LYi1~L~t.J'lU\JLui~m'l
!?J.~ n~\JLL~'l ~'1 t.1 spectrophotometer Vl1 m'D~ ;ij mn91b]d L~U b(;] l ~l.J91\J'J\J 9~ Ln ~~1 'l~ l Vl~!l'l
lllJ1tl..llJ1n i11L ~n1'l~~n~'WLL~'liJfi1lJ1n Lll'iVI1nL'll~~llJL91ru l~U b(;] VI1!J(;]1t.J~!l19l.J191n A'J1l.J ~ ~
"""" ... '1 fin1'i'VIIPI~'£l'U !PI ':It! WST -1 cell proliferation assay
L9!191'll'D~;lYi1~l~ ... tJ'l~'JtJ completed medium 1~ilA'"l1:Wl;lJ~'W"ll!l'll'D~;whnu 1 X 105
' ... ' !.1 Qoc::!l cv cr"!.l ~ Ct. e- .c:::.l c.l
cell/ml ~'Jt.I'Jn trypan blue assay LL~~\JUl'll~~~'Jt.llA'l!l'l Hemocytometer l(;]lJL'll~~lYi1~l~tJ'I'Yl
uf1Jfl')1l.Jl;lJ;'Wll~')~'lL'W 96 -well plate Vl~l.J~~ 100 IJI Ul.Jl'll~;lYi1~litJ'l~'Qtl..lVI.Qil 37 °C
~111'1~~iln1'1lfl1fu!l\JL~!l!lnL'l!~ 5% L\J C02 incubator LU'WL'J~1 6 iCJt:w'l L~!l1ih'D~;Lm~~~nu
111"ll'U~ Vl~'l91mfu l~lJ~1'l~n~~~'WLYi'lr1'J1l.Jl~:W~'Wl'i1'l1 mlJ1tl..lVI~lJ~:; 100 iJI Lll'i~:;r1'J1l.J l~l.J~\Ji11si'1 3 flf'l Ll~~H Doxorubicin fl'J1lJL~lJ;'W 2 IJg/ml LU'W positive control tim'D~;lL~:;
~1'l'Yl~~!l1J~'Qtl..lVI.QiJ 37 oc lU'Wl'J~1 24 i'JblJ'Il~lJ 5% WST-1 VJnVI~lJ Vl~l.J~:; 50 iJI u:wl'i!l~ 'Qtl..!VI.Qil 37 °C lU'WL'J~1 2 i'Jbl.J'l i~fi1n1'l!?J,~n~'WLl~'l (absorbance) ~'Jt.l microplate reader
28
~~'11~m'1~~u 570 nm t~HJ1-il~'11~m'1~~u 630 nm LUU reference wavelength bb~~NCH'l
bUUA1 Proliferation Index (P.I.) ~~1U'dru1~r.nn~~n1~
Proliferation index (P.I.) = mean absorbance of sample
mean absorbance of control
' ' ~ dl I .<:::ill I dl
b~'il mean absorbance of sample ~'il ~1b'U~HJ"ll'il'1~1n1~~~n'€'lubb~'l"ll'il'1~1~'Vl~~'ilU bb'€'l~
mean absorbance of control ~'il A1b'U~tJ"ll'il'IA1n1~~~n~ubL~'I"ll'tl'ln~~~'dU~~ ~-.:~1~n~1~ ~ ' '
1.2 m'i'Vll?l~'elUCJ'Vl in'i~~u n1'i'VI1tJ"l.l'el-:JU11?1LLe.J~ Ll?lt!"li'm.Ju~1~'eNn1'i'VI1tJ"l.l'el'IU11?1LLe.J~ (in
vitro wound healing model) "l.l'el-:1~1'i~ni?I11U'VI~11''£l (Curcuma cf. comosa Roxb.) ~'Jtl
" ' ' m~1~~ 500 f.JI '€'l'l1u 6-well plate U~b'l1mfL'Y11:;b~tJ'IYl~n.JV1,1JiJ 37 oc Ml1'1~Y1nn1'11
mfu'tlUL~'il'e:!nL'l!!?f5% 1u C02 incubator LUUL'1~1 24 i1t~-.:~ 91nJu1-n pipette tip "l!U1~m~1~~
200 f.JI ~ ~L~U'Y11 ~"ll'd1'1 ~1Un'€'l1'1Vl'€'l~"ll'il'I91UL~"tJ-.:~ L'l1'€'l~LUULL u'1m'1m~~ n1V1u~ L~'e:l Hhii ~ '
'li'e:l'lrJ1'1 (artificial wound) ~1'1LmL'l1'€'l~~Vl'€'l~'il'iln'"l1nm~~~'il'e:ln1u LL~'1U1L'l1'€'l~L'Y11:;L~"tJ-.:~Ju1uti~ '
rlU~1~~rl~rJ1U'Vlm~'il~~'11~b~~~U~1'11 m~1~~ 1,000 f.JI uuVinfl1'Y1b'l1'€'l~b'Y11:;b~"tJ'Ifl1tJ1~n~'il'l
'1'€'l'Vlnf"'u~L'1'€'l1 o, 2. 4, s, LL~:; 16 i1t~-.:~ ~'Vlim:;~um~Vl1tJ"ll'il'ILLe.-J'€'l"ll'il'l~1~~n~~~u1'Y1~ u~:;LiJU91nn1~'€'l~~'l"ll'e:l'l~"u~~'l bUU~rJ1'1~1~~'iltJ~ ~b~'e:l bYltJUnun~~ mu fl~
' '
Animal cages
Metal ear punch diameter 2 mm. (Fisher Scientific, Pittsburgh USA)
Microscope (Nikon/Aiphaphot 2, Switzerland),
Micropipette (Tranferpette, Germany)
Microtome
Scan scope
Vernier caliper
Forceps and scissors
Glassware: Beaker, Slides and cover slips
Ethyl acetate (Merck, Germany)
Diethyl ether (Merck, Germany)
Formalin (Merck, Germany)
Hematoxytin & Eosin (H&E)
Isopropyl alcohol (Carlo, Italy)
Paraffin (Sigma Chemical, USA)
Xylyne (Sigma Chemical, USA)
Zoletil 100 (Virbac, France)
Hirudoid® (Medinova Ltd, Switzerland)
Oxoferin® (Holopack, Germany)
29
Lt?ihn.J~'l~~rlt1l'l-ltl1U~'lti~1~~:::~1\'J ethyl acetate 'JJ!l'l~'l'W'l-1~11!l (Curcuma cf. comosa
'l"'q ll ll Roxb.) b'I-I:W'"''l'l:WL"ll:W"ll'W 0.1% w/v LL'i'l~ 0.2% w/v
. t?i1fu'"'1~V1il~1~~nt1lt-~tl1U~'l\'J~'l~~:::~1\'J ethyl acetate 'JJ!l'l~'l'W'l-1~11!l (Curcuma cf.
comosa Roxb.) 1~~1nlA~'In1~~91'J~ 5 ~!l'ln'l~~I?J.J'W1t?i1fumLl'l1tJ~~~n~'l'W~nt1l~LL~t1l'I~Yli'll!l'l ~'l!~~'Wl'Y'l~ Curcuma cf. comosa Roxb L~!lH1'Wm~fn'jj-1LLe..J~(Formulation development of
bioactive fraction of Curcuma cf. comosa Roxb to promote wound healing) 111tlb~LLe..J'W'l1'W
~9m~tl'lrl'W ~!l m~~n'jj-'l~YliYl'l'l~'J.n'l'Y'l'll!l'l~'l!~~'Wl'Y'l~ Curcuma cf. comosa Roxb. LL~::: ~I?J.J'W'l LU'W~'lfumfn'jj-1 LLe..J~
m~ti'l~1fuA1~iJ~1tJ~:::L~\'Jt1ln'lnl'l1tJ~'Y'l!li'lL'JJU ~'lif
30
"' 1. L!?l~t.J~ Cream base 1,000 g.
Oil phase : ~'i:'l'El~ stearic acid 30 g, glyceryl monosterate 20 g, cetyl alcohol 80
g, cremophor A25 10 g, silicone oil 5 g, isopropyl myristate 30 g [?11~~~~1J uw
water bath '"lt..lnmu~1lihJ~:::~1CU 75°C • 'IJ
Water phase: tu~t.J carbopol ultrez 10 3 g bt..l Propylene glycol 20 g '"lt..lm':::'"llt.J~'l
~~i?l bb~'Jb~h-Jtl1 798.9 ml 91mTt..!'elt..!1~1~'ElCU~1liJU~:::~1ru ?SoC ' ' 'IJ
' ~~~oil phase Lt..! water phase ii11 bb'i:'l:::rm~ihb~~'El '"lt..l'QCU~!JilU~:::J..nru 50°C
' "' "' "' b[?l~ triethanolamine 5 g m~'"l:::b~~ congeal
b~~ germaben-11 8 g bb'i:'l::: a-tocopherol 0.1 g. 1JJI'1'i:'l-.l1l1'llt..l:::
'l iiQ il il
b~~f\'J1~b'l1~'1Jt..l 5%w/w, 1 O%w/w, 15%w/w bb'i:'l::: 20%w/w "lJ'El-.l cream base
1l1t.J~i-.lm~b[?11t.J~~~fur1~,~ ~'fl-.lnm~u~:::bilt..!~1fu~1-.li ~-.lcl' ' "' 0 <V .c:::;:,j .::::.1.:::::.1 (V "'
n1'i'V11Pl?l'fl'UFI':I1~Fl-.lCWJ (stability) "ll'El-.lm~'Ufl~~Yl~~~~~ni?li?l'Jt.J ethyl acetate "lJ'El-.l
rJ1t..l~~~~'fl~'Jt.Jn1n11 heating and cooling cycle li?lt.JU1~1fufl1~~n~~~~ni?l~'Jt.J
ethyl acetate '1J'El-.lrJ1t..l~~~~'Elbrl1Jbt..l~fl'J'Uf)~'QCU~,!Jil~'QCU~,!Jil 45 oc t..llt..l 48 'll~ . ' ~
bb~'l9-.lu11Ubrl1Jbt..!~b~t..!Yl'flCU~1liJ8 oc t..!lt-.!48 'll~. ~i?lbUt-.!1 ~'flU vl1~1 10 ~'flU 'IJ ' 'IJ
m'itl'i::bijuin'l!ttu::'V11~n1Wl1'Vl (appearance) "11'El-.l~1fufl1~~n~~~~ni?l~'Jt.J ethyl
acetate '1J'El-.lrJ1t..l~m~'El~~~t..lm~v11 heating and cooling cycle bmt.J'UbVlt.l1Jrl'U~1fu
fl1~~n~~~~ni?l~'Jt.J ethyl acetate '1J'El-.lrJ1t..l~~~~'El~bnu1~1t..!~ru~.tJil~'El-.l (25°C) li?lt.J
" ~ ' i-.lm[?lin'jj-ru:;bi!'Elfl1~ mnbt.Jnit..l m~[?ln[?l:;n'flt..l ~bb'i:'l:::n~t..l
n1'iU'i::Lijt.IFI':I1~'VIt11Pl (Viscosity) "11'El-.l~1fuf11~~n~~~~ni?l~'Jt.J ethyl acetate "lJ'El-.l
rJ1t..l~~~~'El~~~t..lm~vh heating and cooling cycle bmt.JUbVlt.Junu~1ftm1~~n~~~
~rli?l~'Jt.J ethyl acetate '1J'El-.lrJ1t..l~m~'El~brlU1~1t..!~CU~,!Jil~'fl-.l(25 ° C) li?lt.Jbii
Viscometer (LMS Instruments)
' n1'iU'i::Lijt.IFJ':11~ LUu n'i~P~-~1~ (pH) "ll'El-.l ~1fuf11~V1n~~~~n i?l~'Jt.J ethyl acetate
'11'fl-.lrJ1t..l~~~~'fl~~~t..lm~v11 heating and cooling cycle bmt.J'UbVlt.Junu~1fufl1~~n
31
~~d~nrn~'JtJ ethyl acetate 'll!l--l~1'W\UJ1~!l~Lfiu1~1'W~ru~.!Jil~!l'1(25°C) Lr?1tJL~ pH
paper (Macherey-Nagel, Germany) 'li'J'I pH 3.8- 5.5
m-a1Lfl'i1::lhntihntu~1'i~1~tyl'W~1fur1hJ~i1~1d~nrn~'JtJ ethyl acetate 'll!l--l
~~'W~~~~!l~~~'WnldYil heating and cooling cycle LmtJUlYltJunu ~1fur11~~i1~1d ~nrn~'JtJ ethyl acetate 'JJ!l'I~1'W~~~~!l~Lriu1~1'W~ru~.!Jil~!l'1(25 ° C) LrntJH HPLC
e.Jr1mdtld~Lil'W~1fuf11~ l9i1--l1 !l~L'WLm'lmd1r.ru~ 5 ~!l'~ mdvr[ij_J'WI ~~fumL~1u~r.nn~'J'W
~nrn~ ll~ rn--l~'Vli'JJ!l'l~"ll~d-!'W1'Y'Id Curcuma cf. comas a Roxb l~!l H1'Wmdfm~1lle.Jr1 (Formulation
development of bioactive fraction of Curcuma cf. comosa Roxb to promote wound
healing)
OJ
W\~ mice vr'W( C57BL/6 Arl~l'Y'II"\ !11~ 8 ~tlrn11f U1~1Xntld~J . .nru 20-25 g 9i'W'J'W 36 f?l'J
r.nn~'W~~~1'Vlrnr1!l'lm..l'l"l!1~ ~vn1'Vlm~m.J{:irnr1 ~'Wrlnl~"tJ'~~~!l--l~~1'Vlrnr1!l'I'JJ!l'IAru~Lil~"l!~1~~f " ""
'JJ!l--l~11Xn~~1'Vlrnr1!l--l Llr1~~~J~~r1!lr?1l'Jr11 LU~tJ'W!l1~1nlr1~J~~J.J'Vln 2 i'W LU~tJ'WrJ~rnd!l'I'W!l'W'Vln ' ' '
I ~ ~ I
3 i'W ~n~'J~Y1H1'Wmn~u'~ ~!} !lru\mil 23 ± 2oc A'JIJ.J~'W~J.Jvr'Vlfi 55-60% d~tl~L'Jr11Y1~wr~LL~'~ ' 'IJ
(Ethics committee) (ID #7/2555/research) L1.ln~1J'lfl·m1Ae.J'W'Jn
" n1'i~'i1--l1J1IPILL~'Cl
~rnm~r1U~'W'Vlr?1r11.l'I~'JtJ Zoletil 1 oo 91mT'WLonrnu~L'Jru1u~~'JmL1.lr1n!le:J!l;L~m11r11m~1.l " "
L91~LU\!'ll1.l--l'vn,pf'1~1.l'l~h'I~U~L'Jrunr11'1\! (Center of the cartilag inous part of both ears) L~lU'W l (through-and-through hole) (ltl~ 11) ~'Jtl 2-mm diameter metal ear punch U'WVinili'Y'I
~n'tl'ru~Il1tl'W1.ln'll1.l'IU1r?1Lle.Jr1 Llr1~rJr?1'1J'W1r?1 'll1.l'll~L91~Yl'WYl~'JtJ vernier caliper irn~"l 3 Af'l (hi
~1.l'IUr?1lle.Jr1 llr1~hiilmdHm~1'W9r1~'Y'I ~r1 1.l r?1 n1d'Vlr?1 'i'l 1.l'l ) lL'i'l~tYudj'Wi'W~ o '
32
n1'iV11U11?1 bbe.J~
llU-.1VI'W'Vll91'tl'fl--l'fl'flnbU'W 6 n~J.J n~J.J'tl:: 6 !?i''J 'Vl1~1~'Vll91~'flU~U1l'JbUU11nll~'tl'll'fl-.lbUmf-.l~'fl-.1 'iJ q q 'lJ
' ~ ~ I 'i.l a!J 0 1:1/ ~ ..:::::.la:f Q./ i,l
~1~~nl91VItnUl91'Jt.J ethyl acetate 'll'fl-.l'J1'WVIJ.JI'J'fl Vld'fl l911~Ure1~J.J'VlJ.J~1~~nl91l91'Jt.J ethyl acetate 'll'fl-.1
' '"' "" 'J1'WVIJ.J1'J'fl Vl~'fl positive control
,_, Vi "ll'J1 : 'V\1 (91'Jt.J
negative control
bl91t.JVI~'Vl l91fl'fl,m~J.J t?i1-.11 '"l::Q,n'V\1 ~'Jt.J~1~'Vl l91~'flu lll>l n t?i1-.1 n'W 1?1-:Jtf '
' "" t~'Wn'tlJ.J'YI 1 (n=6) 'lJ q
,_, ,_, '"' 0 ~'ll1-.l'll1t.J : 'YI1l91'Jt.J ethyl acetate
'
"'
2,.1 'V 0,..0 • il ..:::::.1 i.l 'V
0 ~'JJ1-.1'1J'J1 : 'Y11l91'JtJ~1~~nl91VItnU'JJ'fl'l'J1'WVIJ.JI'J'fl'Yire1'J1J.J6'1JJ.J'JJ'W 0.1% w/v
33
VI'Wn'tlJ.J~ 2 (n=6) "' . 0 V1~1~sii1t1 : 'Vl1~'Jtl ethyl acetate
" 0 V1~1~"ll'J1 : 'Vl1~'Jtl~1~~rli?lVIti1U"ll!l~~1'WVIJ.J1~!l~""')1J.Jb~J.J~'W 0.2% w/v
"'
... ... ... 0 ~"ll1~'1l1tl : 'Vl11?l'Jtl ethyl acetate
... ... 0 f . ® 0 Vl"ll1~"ll'J1 : 'Vl11?1'Jtl xo enn "'
VI'Wn'tlJ.J~ 4 (n=6) "' . 0 V1~1~oil1t1 : 'Vl1~'Jtl cream base
"'
' VI'Wn'tiJ.JY\ 5 (n=6) "' .
0 V1~1~sii1t1 : 'Vl1~'Jtl cream base "
' VI'Wn'CiJ.JY\ 6 (n=6) "' .
... ... ... 0 Vl"ll'Wll1tl rm'Vl1i?l'Jtl cream base
"' "' o Vl~1~"ll'J1 : rln'Vl1~'Jtl Hirudoid® cream
"' "
VI'Wn'CiJ.J~ 1-3 'Vl1U11?1bbc.J'i'l~bUVIbbl9J'i'1:::~1~~'Jtl~1~'i'l:::'i'l1tl mJ.J1!?1~ 0.2 ~I ~'J'WVI'Wn'CiJ.J~ 4-6 \.1 'II 'iJ 'lJ ..
' ~
'Vl1U11?1bbc.J'i'lYlbU~bbl9J'i'1:::~1~~'Jti""~J.J mJ.J1W. 10 mg 'Vl1'Y!n'f'W i'W'i'l::: 1 ""f~
n1'itl'i~ Lij'W n1'3~1t~"..l'eNtJ1 !PI u~~
rh t.JJlTvHr n~ru.:::r11 t.J'W!ln"ll !l~U11?1 LLC-J'i'l 1-< t,.t i.l I
~'JJ.Ji~il?1"ll'W11?1b~'We-h~'WtJn'i'l1~"ll!l\lU11?1bbc.J'i'lbl?1tiH vernier caliper 1'1?1~1 3 ""f'l1-wi-wY\ 0, 1, 2, 5,
7, 10, 15, 20, 25, 30 bb'i'l!:~1'W'JW.fl1~!lt.J'i'l:::"ll!l ·-ln1~VI1tl"ll!l~U11?1bLe.J'i'l bl?1t.Jbsii~!?l~
diameter at day 0 - diameter at day n 0 %wound healing= _______ ..:.....::_ _____ ___:....!!._ x 10 diameter at day 0
' .. .:;::,1 ~ Q, t-o
~1~'VlJ.J~'VlHn~::;~-wn1~VI1tl"ll!l\lUc.J'i'l
34
n1'ir;J'i'l"l~UL tl'£lVJ1-:J"l'iW1t~1rn rtfil1~ r;J~"li'£:1-:J L U'£lL~'£l (Histology) •
~~!?l'i'"J'"l:n"uLll'tl1t..vfu~ 0, 2, 5, 10 LL~:; 20 L!?ltJ~1'\.-IU~"ltJ1fiV1~~~1J~"ltJ ether tK!?I'I.-IWW , 'U 'U 'll
~"ltln'i''i'Lm~~1U'll'tl'ILU'\.-I (base of the pinna) L~L't..l 10% buffered formalin pH 7.0 L!?l1tJ~Ld''tlL~'tl .... \1
' ' ... <}' ..:::ij .dl ll d dl ..... ..:::.1
L!?itl Paraffin technique Li"l'tlf'l\mn histology !?I"JtiLfl'i'!l'l~'tl scan scope !?1-:JU lJ !I I I lJ I
'll-'0~ I .::!:J..:;!I dl\1 ..::Y.dl o Washing : b'l.-l't..\1 b'l.-l~e.J1't..lL't..l!lLtl'tl Li"l'tl~1'1 10% buffered formalin '"l1nL't..l'tlLtl'tl
0 Dehydration : ~.:JJ1!l!ln91nL'll~~~"JtJ ethanol L!?itiG~'"l1nfl"l1~L~~~'t..l~1LU~'I ~'tl 25%, 50%, 75%, 100%
0 Clearing : U1 dehydrant !l!ln91mif!lL~'tl L!?itJL-il xylyne dj't..l Clearing agent
0 Infiltration: U1 parafin ~LU't..l embedding agent L~1~Lll'tlL~'tlL~m111~~1tl~!ln1'i'tK!?I 0 Embedding : e!-.:~Lll'tlL~!l~'ll't..l parafin '\.-l~'tl~L'\.-1~'1 (embed) rl!l't..l~9:;U1Lll'tlL~'tl~1tK!?I
L!?lm:;1-ilLLUU (mold) 91nJ't..lLLn:;~~if'tl'tln~1'-l:;L~LU'Win~ru:; parafin
block
0 Sectioning : ~!?l:n"'t..lLll!l~e!.:J'tltiL't..l paraffin block L!?ltJ~fl"J1~'\.-I't..l1 4 micron ~"lmfli'tl.:~ \1
microtome :n"'t..lLif!lL~'tl~L~'"l:;~in~ru:;LU't..l ribbon
0 Adhesion : U1 section ~tKt?\L~~1~t?\~!?11J't..lLLe.iu slide LLfh Lt?\tJU1 section J't..l~1~'tltJ ' ...
L't..l water bath 50 ac Yle.J~~ gelatin 91nt!'t..l-il'tl't..l section ~"JtJ glass
slide LL~"JU1~1~-:JYi ... -.:~1~U't..lLfli'tl--l warmer ...
0 Staining & Mounting : ~'tl~~L~!ll~L~'t..l contrast l't..l tissue ~1n~'t..lL!?itJL-il~
hematoxytin & eosin (H&E} 91nJ't..llhn1'i''\.-ltJ!?I permount LL~"JU!?IvlU ~"JtJ Cover slip Lt?\tJ~.ffu!?l!lUU'tltJ ~~d'
- Deparaffinization : H Xylene 91't..l"J't..l 2 Lbl LU't..ltK"J~:;~1tJ'!.-11'tl~'l wax
medium !l'tln91n:n"'t..lLif!lL~'tl
- Hydration : ~1'1 xylene L~'\.-I~!?ILU91n:n"'t..lLif!lL~!l LL~:;L!?I1tJ~U1~b!if :n"'t..!Lll!lL~'tl~.:J~J1L~'tln1'i'~'tl~~ L!?ltiH ethanol ~ti'I~1~1J91nfl"l1~
'IJ
L~~~'t..l~'ILU~1 ~'f) 100%, 75%, 50%, 25%
- Staining : ~!l~~L!?ItJH~ hematoxytin & eosin (H&E) !,.; J,.t I tl
- Dehydration : ~11~:n't..lLi1'tlLm'tlu't..l~L~!ifunf'll'"l1nu1Lt?\tJL-il ethanol
G~91nm1~L~~~'t..!~11u~-.:~ A'tl 25%, 50%, 75%, 100%
- Clearing : ~11{1:n"'t..lLd'!lL~'tlU't..l~L~!ifunf'll'"l1n dehydrants lt?\tJH
xylene ~'ILU't..l clearing agent L'li't..lL~tJ"JrlUn1'i'L!?I1tJ~:n"'t..lLif'tl Ld'tl'191n
35
xylene ~1l-!1'i'tl~-l"tl'Jn ethanol ~1-iiLu'W(;]'J dehydrant 1~~ ~m.f-lcl'-l
permount
mounting medium ' ~ ~
- Mounting :U1L'fl1 cover glass Vli:J"l!'W11111'1,.,rYn~11'WLi1'ElL~nil'Elt.JU!11
'l'i'U'tl-lU'W1~'WL\f'elL~'el1J'W~L'tl111111lt.J1-ii mounting medium 'l.-lti!11~N1J'W " ' ' dl.c:Y Qo \I qc:; ill vll C' !.1
L'W'elLtl'el VI'Vl1n1'i'tl'ell-l~LM9LL'tl'J'J1-l'Yl1J!11'Jtl cover glass LM""lLL'tl'J
U1LtJmn'tll-11~ mounting medium LL~-l LL~'J scan slide ~'Jtl ' dl dl
Lfl'i''el-ll-l'el scan scope
mnh:::Ln'Wn1'i''l.-l1tl"l!'El-l1J1!11LLt.J'tl Yl""l1HU1""11nf1'J1l-l'l.-l'W1LLki'W'll'El-l polymorphonuclear
(PMN), fibroblast, collagen fiber LL'tl:::'\tl'tl'el!11L~'el!11
LL~!11-lt.J'tln1'i''Yl!11'tl'El-l1'WlULL1J1J mean± standard deviation (S.D.)LL'tl:::~Lf1'i'1:::tfri1'Yl1-l~n~
Ll1lt.J1-ii Student t test rl1 p value < 0.01 : p value < 0.05 n'El~1~'ell-!'tli:Jf1'J1l-!LL[;]nvh'lmh'li:J "
..,:. tJ 'Vl'Vl 4
1.1 t]'VI~L ~~~1'\J'l'WL"l!~~ (Cell Proliferation activity) "ll'fl\l?t1'i?trl~'VIti11J (crude extract) lPl'ltl
~'l'Vh~::~1tl ethyl acetate "ll'fl\ltJ1'1J'VI~1tJ'fl (Curcuma cf. comosa Roxb.)
... . t:-.~-tm1~'Vl ~~!lU~'VlfiLYl~91'W':l'WL'l!~~'ll!l-.l~1~~n ~'VIti1U~':l EJI111Yi1~:::~1EJ ethyl acetate 'JJ!l-.l
. 1 t,l d !.1 t,l I - ciJ
11'W'VI~11!l (Curcuma cf. comosa Roxb.) 'Vlfl11~L'll~'ll'Wm\11 n'W fl!l 400,200, 150, 100, 75,
50 LL'tl::: 25 1-JQ/ml l1i'flL'l!~~LYi1:::Li'EJ\I 2 'l!oW~ A!l L'l!~~di!lL~!lL~~'l'VIU-.l'JJ!l-.l'VI~'lloW~ L929 (mouse
subcutaneous connective tissue) LL~:::L'l!'tl~~:::L;--ll.J1n~~"ln'JJ!l\l~\f11tl 'llU~ Hela cell (human
cervical carcinoma) ~1EJ"1fi WST -1 cell proliferation assay !?1-.l LL~~-.lL'WlU~ 13 u~:::lUVl14
Proliferation of Curcuma extracts againts L929 cells
1.2 *
X Q)
""0 c 0.8 c 0
0.6 :p ro .....
J!! 0.4 ::::; 0 ...
0... 0.2
0 25 50 100 150 200 400 Dox
Cone. (IJg/ml)
~t1Y113 ~'VliL~:J.J91'W'd'WL'l!~~Lcl'!lL~!lL~~'d'VIU\I'll!l\I'VI~'llil~ L929 (mouse subcutaneous
connective tissue) 'll!l-.l~1~~n~~'dEJ ethyl acetate 'JJ!l\1~1'W'VI:J.J11!l (Curcuma cf.
comosa Roxb .) ~fl'd1:J.Jb~:J.J~'Wl1i1-:J1rl'W (* = p< 0.05, LmEJUbVJEJunu control)
37
CJ~ n1j19tJ~'VliL ~l-Jr.i1U'l'\.JLSJl~~'ll!l'l~1'l~rl ~1-lt.nU!~htJ ~'Jvi1~:::~1 tJ ethyl acetate 'll'el'l ~ '
~1\J'I-Il-l1~!l (Curcuma cf. comosa Roxb .) ~!lL'll~~L~1!::L~tJ'I'llil~ L929 (llJY] 13) ~U~1~1j VI I.J.d !.1 i./ Q c!"C::. 0 tr I .:::::.1-0-
~n~'J1\J'I-Il-J1'l!l'Vlfl'l1l-JL'lll-J'lJ\J 50 j.Jg/ml LL~!:: 400 jJg/mll-J~'VlfiL~l-J91\J'l\JL'll~~!lt.n'll-J\JtJ~1fl'kJ
L~!:!LmtJULYltJUrlU~'lWJUfll-l~hJ~~1j~rl~~l-l\J1~j b~tl~fl1 Proliferation index (P.I.) L'YllrlU ' '
1.08 LL~!:: 1.09 1?11l-J~1~U
Proliferation of Curcuma extracts againts Hela cells 1.4 __________________ * _ ___ * ____ _
X 1.2 +------------------1-----. Q)
"'0 c
§ 0.8 :p ~ o.6 ..-----lz~----t.'i~~--
~ 0 0.4 .....
0... 0.2 +--OI;;':~--J.>:t·5f----t:;; ';:1----HlJ'Itl----f±!''lil'l----
0 25 50 100 150
Cone. (1-!g/ml)
200 400 Dox
~tl~ 14 ~'VliL~l-Jr.i1U'l\JL'll~~l-l:::L~'IlJ1nl-J~~n'll!l'll-l'\{lj'tJ 'llil~ Hela cell (human cervical
carcinoma) 'll'fl--1~1j~'Hl!?l~"Jtl ethyl acetate 'll'f:WhW.!fl-l1~'fl (Curcuma cf comosa
Roxb.) ~fl'l1l-JL~l-l~U~1'11nu (* = p< 0.05, LmtJULYltJunu control)
.. ' oC:l~ .c:::t.~ o r - "'vo
CJ~n1j'JfDltJ~'Vln L~l-l91\J'JUL'll~~'ll!l'l~1j~n ~1-lt.nU ~'lt.l [?l'J'Vl1~:::~1tJ ethyl acetate 'll!l'l ~ '
~1\JVIl-J1~!:! (Curcuma cf. comosa Roxb.) ~!:!L'll~~L~1!::L~tl'l'llil~ Hela cell (jlJYJ 14) ~U~1 " ' .. '
lVI i.lo<::li i,.li,.l cl .c:::t,.<:::lt,O frl cl
~1j~n~'l1\J'I-Il-J1'l!l'Vlfl'l1l-JL'lll-J'll\J 200 j.Jg/ml LL~!:: 400 jJg/mll-J~'VlfiL~l-J91\J'l\JL'll~~!lt.n'll-J
-J'tJ~1fln.J L~!:!LmtJULYltJUrlU~'lfl'lUfll-l~1~~~1'l~n ~~~u1~j b~tJ~fi1 Proliferation index (P.I.)
L'YhrlU 1.14 LL~!:: 1.19 [?11l-J~1~U
Doxorubicin (Dox) LU\Jtl1Lfi~U1U~~~~'VliLuu antineoplastic agent '1.11l-l11•ihuu ' ~
positive control L .W!:! fl'lU fll-l LL~ !::[?l'l'l9 ~!luul)mm m j[?l!JU~U!l'l'll!l'l L 'll~~L ~1 :::L~tJ-.1 ~!l~1j
'Vl~~!:!U b~t.l Doxorubicin ~!l'lL~~~~~r.i1U"l\JL'll~~L~1!::L~~tJ'I ~'l~~mj19t.~tl Doxorubicin
..
38 ..,
L~~H'l~ ~?~r.i1'W'l'WL'll~~L Yi1:::L~t.Wl!UI?I Hei...a
Doxorubicin iJrJ'J1~LU'WYh~~!lL'll~~~:::L~~
(in vitro
wound healing model) "ll'fl~?f1'l?Hl~VIti11Jr?i''lti~TVh~~~1tl ethyl acetate "ll'£1\l':hu
Vl~1tJ'tl (Curcuma cf. comosa Roxb.) r?i''lt~1fi Scratch assay
m ~19t.J ~Yl im:::~-wmnn t.J"ll!l~U11?1 LLe.J~ 1-w~~!li?!Yll?i~!l~ L~?Jt.JL-ii LLUUr.i1~!l~ m~~1t.J"ll!l~ 1J11?1LLe.J~ (in vitro wound healing model) ~'Jt.J'ifi Scratch assay ~!l n1~~~1~ artificial wound
'"l1nm~~l?l L~'W'Yi11?1"1J'l1~ ~h'Wn~ 1~~~~"1J!l~'"l1'WL~..,t.J~ L'll~~LU'WLL'W'lt.J1'l L~'t!L ~ Ln ~?J"Ji!l~~1~ LL~'JU~ '
L'll~~L'Yi1:::Lit.J~rl1J~1~Yll?i~!l1J~rl'J1~L~~~'W~1~1 rl'W mn.h:::L~'W~Ylim:::~'Wn1~~1t.J"lJ!l~LLe.J~"lJ!l~ ~1~~r1 ~?~~l.!-w1 Vi ~1~91nm~i~ m ~mJ'~ ~?J~~"ll!l~i-w~~~ Lu-w~~1~ m~J'!lt.J~ ~?~~ L'l~1 ~1~1 now L~!l
.. ~ ~ v ~
e.J~n1J'Yll?l~!l1J ~Ylnm':::l?j'Wn1 ~~1t.J"ll!l~U11?1 LLe.J~"ll!l~~1~~n l?l~t.J11J l?l'l t.J ethyl acetate "ll!l~
' ~1'W~~1~!l (Curcuma cf. comas a Roxb.) Y1 wn~ L ~~~'W~1~1 now ~!l 400, 200, 150, 100, 75,
50 LL~::: 25 fjg/ml ~'t!L'll~~L'Yi1:::L~"m 2 'l!UI?I ~!l L'll~~Llf!lL~!lHlC:hmX~"ll!l~~'W'llUI?I L929 (mouse ..,
subcutaneous connective tissue) LL~:::L'll~~~:::L~~U1m-JI?I~n"ll!l~~'W11tJ 'l!UI?I Hela cell (human .., '
cervical carcinoma) (9)~LL~I?I~L'WtU~ 15 - 30 e.J~n1~'i9t.J'Yi1J~1~1J'~rli?I~1'W~~1~!l~'lt.J ethyl
acetate ~rl'l1~L~~~'W 400 f.JQ/ml iJ~Yli~~~I?IL'Wn1n~~r.i1'W'l'WL'll~~vf~'llUI?I L929 LL~::: Hela
cell1~~1?1n'WL~'l~~I?I~L'J~1 24 -i''l'l~~ '
(A) o hr. (B) 2 hr.
(C) 6 hr. (D) 12 hr.
(E) 24 hr.
~u¥11s ~Vlim:::~'Wn1'i''vl1t.J'll'il'lU1 (?ill~r1lw'vlr1'il (?IVJ (?11'1'il'l ~r.JtJ~fi Scratch assay ln'il hJij
~1'i'~rl(?l~~'Wl'V'l'i' (negative control) lul'llr1~l'V'l1:::l~"tl'l'Jlil(?l L929 ~l'Jr11~1'l1 rl'W I I I I I
(A) 0 ir.JhN (B) 2 ir.Jb~'l (C) 6 ir.Jb~'l (D) 12 ir.Jb~'l (E) 24 ir.Jb~'l
39
40
(A) o hr. (B) 2 hr.
(C) 6 hr. (D) 12 hr.
(E) 24 hr.
' iJ I ~ .:;:,l 'l,.t i./
r?l''Jt.J ethyl acetate 'JJ'E:l~'11'WVI~1'1'E:l (Curcuma cf. comosa Roxb.) 'Vlf1'11~b'JJ~'JJ'W
25 IJQ/mll'Wb'Tifl~b'V'l1::::6~,tJ~"ll'Wr?1 L929 ~b'1fl1~1~1 rl'W I I I L I
(A) 0 '11'1'\:~~ (B) 2 '11'1'\:~~ (C) 6 '11'1'\:~~ (D) 12 '11'1'\:~~ (E) 24 '11'1'\:~~
41
(A) o hr. (B) 2 hr.
(C) 6 hr. (D) 12 hr.
(E) 24 hr.
' i.l ' '11 ~ 2.1 2.1
r?l'dt.J ethyl acetate 'll'fl'l'dTW'\U.Jl'd'fl (Curcuma cf. comosa Roxb.) 'Vlf1'd1~6'll~'ll'W
50 IJQ/ml 1'Wl'll~~6'Y11~l~,..t.J'I'l!Ur?l L929 ~l'd~1l'i1'11 rl'W
(A) 0 i'db~'l (B) 2 i'db~'l (C) 6 i'db~'l (D) 12 i'db~'l ' (E) 24 'Jl'd b~'l
42
(A) o hr. (B) 2 hr.
(C) 6 hr. (D) 12 hr.
(E) 24 hr.
' il I i.l ~ 2,.1 iJ
l9Y.li'J ethyl acetate 'JJ'El~'d1\J'VId-J1'd'El (Curcuma cf. comosa Roxb.) 'VlA'dld-Jb'lld-J'JJ\J
100 IJQ/mlbtJb'll~rlb'IAll:::b~"I'J~'l!oWC?l L929 ~b'd~1t?i1~1 rl\J I I I I t
(A) 0 'lf'dbd-J~ (B) 2 'Jl'dbd-J~ (C) 6 'Jl'dbd-J~ (D) 12 'Jl'dbd-J~ (E) 24 'Jl'dbd-J~
43
(A) o hr. (B) 2 hr.
(C) 6 hr. (D) 12 hr.
(E) 24 hr .
.. ~tl~ 19 ~Vlfim~~'Wn1~VI1 t.J'll'el~U1 ~lli:J'Cll 'WVI'Cl'el ~Vl ~'Cl'el~ ~r;m~fi Scratch assay 'll'el~'6'l1~'6'ln ~
' ~'dt.J ethyl acetate 'll'tl~~1'WVIm~'el (Curcuma cf. comosa Roxb.) Vlfl'"l1~l~~~'W
150 f.JQ/mll'Wl'll'Cl~l'Y'J1~l~"m'll'W~ L929 ~l'd'Cl11?11~1 n'W I I I I I
(A) 0 'll'"ll~~ (B) 2 'll'dl~~ (C) 6 'll'"lld-J~ (D) 12 'll'dl~~ (E) 24 'Jl'dl~~ ..
44
(A) o hr. (B) 2 hr.
(C) 6 hr. (D) 12 hr.
(E) 24 hr.
' ~'HJ ethyl acetate 'll'el..:l'iTWV1:wl~'el (Curcuma cf. comosa Roxb.) Vlfl'J1~b~~~'W
200 f..JQ /mi1'Wb']'6'1~b'Yn~b~"'tWJlU(?) L929 ~b'J'6'11~1'l1 rl'W I I I I I
(A) 0 'll'Jl~..:J (B) 2 'll'Jl~..:J (C) 6 i'Jl~..:J (D) 12 'll'Jl~..:J (E) 24 'll'Jl~..:J
45
(A) o hr. (B) 2 hr.
(C) 6 hr. (D) 12 hr.
(E) 24 hr.
' ~'dt.J ethyl acetate 'll!l'l~TWV1m1'El (Curcuma ct. comosa Roxb.) VlA"ll:hlb~:hl~U
400 IJ Q/mi1Ub'll~£b'Y'11:::b~"t.J'l"lli:ll?l L929 ~b'd~1!?i1'l1 nu I I I I I
(A) 0 i'db:hl'l (B) 2 i"lb:hl'l (C) 6 i'db:hl'l (D) 12 i"lb:hl'l (E) 24 i'db:hl'l
46
(A) o hr. (B) 2 hr.
(C) 6 hr. (D)12hr.
(E) 24 hr .
... ~tl¥1 22 ~'VlfinJ:::~'Wn1J'l.-ntJ'll'il'lU1(?)lle-JflloW~fl'il(?)'Vl(?)fl'il'l~'dt.J~fi Scratch assay 'll'il'l
Doxorubicin l'Wl"ll~~~'Y'Il:::l~"t.J'l'JlU(?) L929 ~l'dfl1l?i1-.11 rl'W
' (E) 24 'Jl'd l:W'l
(A) o hr. (B) 2 hr.
(C) 6 hr. (D) 12 hr.
(E) 24 hr.
~uv1 23 ~'Vlim:::~'L&n15'VI1tJ'lJreJ'I1J1~66~'tl1u·wweJ~'Vl~'tl!h'l~'dtJ~fi Scratch assay 6~reJhJ~~15' ~n~~~Ul'V'J5' (negative control) bW"l!'tl~6'V'J1:::6~"tJ'I'llU~ Hela cell ~6'd'tl1t?i1'11 nu
I I I I
(A) 0 'll'Jl~'l (B) 2 'll'Jl~'l (C) 6 'll'Jl~'l (D) 12 'Jl'dl~'l (E) 24 'll'Jl~'l
47
48
(A) o hr. (B) 2 hr.
(C) 6 hr. (D) 12 hr.
(E) 24 hr.
' i,l I i,/ .:::;::,1 !.f 9,1
W'ltl ethyl aceta te 'JJ'fl'l'd1'WitUJ1'd'fl (Curcuma ct. comosa Roxb.) VlV'I'd1:J-.Jb'JJ:J-.J'JJ'W
25 1JQ/mllwb'llrl~b'Y11::;b~"tl'l'l!UC91 Hela cell ~b'drl1~1'~1 rl'W I I I I I
(A) 0 'll'dl:J-.J'l (B) 2 'll'dl:J-.J'l (C) 6 'll'dl:J-.J'l (D) 12 'll'dl:J-.J'l (E) 24 'll'dl:J-.J'l
49
(A) o hr. (B) 2 hr.
(C) 6 hr. (D) 12 hr.
(E) 24 hr.
' 'V I 'V .:::::1 'V !,1
WltJ ethyl acetate '1J'el~'d1'W'vn..n'd'el (Curcuma cf. comosa Roxb.) 'Ylrl'd1 d-.1 6'1Jd-J'lJ'W
50 ~g/mlb'W6"1l'tl~6'1'11::6~"m'llillY1 Hela cell ~6'd'tl1l'i1~1 rl'W I I I I I
(A) 0 oTI'dld-.1~ (B) 2 oTI'dld-.1~ (C) 6 oTI'dld-.1~ (D) 12 oTI'dld-.1~ (E ) 24 oTI'dld-.1~
. ~
(A) o hr. (B) 2 hr.
(C) 6 hr. (D) 12hr.
(E) 24 hr.
9,_, I i,l c:l iJ 'i.J
CW"Jtl ethyl acetate 'll'il'I'J1t.f'vU.J1'J'il (Curcuma cf. comosa Roxb.) 'Vl~'J1~b'll~'ll'W
100 [Jg/mll'Wb'll'i'l'tlb'vn~b~"tl'l"ll'WC91 Hela cell ~b'J'i'l1~1'11 rl'W I I I I I
(A) 0 'll'Jl~'l (B) 2 'll'Jl~'l (C) 6 'll'Jl~'l (D) 12 'll'Jl~'l (E) 24 'll'Jl~'l
50
51
(A) o hr. (B) 2 hr.
(C) 6 hr. (D) 12 hr.
(E) 24 hr.
' ~'JtJ ethyl acetate 'JJ'fl~'J,1'W'1,.1J-n~'fl (Curcuma cf. comosa Roxb.) VlA'J1l-H~:W~'W
150 ~g/mlb'Wb'l!~;b'Y'I1:;b~"tJ~'Di1f?l Hela cell ~b'J~11?i1~1 rl'W I I 1 I I
(A) o 'll'J'i:w~ (B) 2 'll'J'i:w~ (C) 6 'll'JL:W~ (D) 12 'll'JL:w~ (E) 24 'll'JL:W~ ~·
52
(A) o hr. (B) 2 hr.
(C) 6 hr. (D) 12 hr.
(E) 24 hr.
'1.1 I 'i,l _Q i./ i,l
(?)'Jt.J ethyl acetate 'JJ'il'I'J1'WVIm'J'il (Curcuma cf. comosa Roxb.) 'VlA'J1:Wb'JJ:W'JJ'W
200 f-JQ/miL'Wb'll~~~~1~b~"m'llil(?) Hela cell ~~1~1t?i1-:~1 rl'W I I I I I
(A) 0 ir.Jl:W'I (B) 2 "Jlr.Jl:W'I (C) 6 "ll'Jl:W'I (D) 12 "Jlr.Jl:W'I (E) 24 "Jlr.Jl:W'I
53
(A) o hr. (B) 2 hr.
(C) 6 hr. (D) 12 hr.
(E) 24 hr.
' i.; I 'i.l .:::::1 9,.o 2/
!WHJ ethyl acetate 'lll!'l'd1'W'v1J..J1'dl! (Curcuma cf. comosa Roxb.) VlV'I'dl:Wb'll:W'll'W
400 iJ Q/mll'Wb'lliN'Ci"b'l''n:::b~"t.J'I'lloW1?1 Hela cell ~l'd~N1l'i1'11 rl'W I I I I 1
(A) 0 ir:J l:W'I (B) 2 ir:J l:W'I (C) 6 ir:J l:W'I (D) 12 ir:J l:W'I (E) 24 ir:J l:W'I
. . .
r
54
(A) o hr. (B) 2 hr.
(C) 6 hr. (D) 12 hr.
(E) 24 hr.
I I I I I
(A) 0 'll'Jl:W-:J (B) 2 'll'Jl:W-:J (C) 6 'll'Jl:W-:J (D) 12 'll'Jl:W-:J (E) 24 'll'Jl:W-:J
55
.. oC:t. i.l cv !I' '
m ~'Vl !?l~tlU ~'VI nm'~i?p.m 1 ~V11tJ'lltl'l U1 !?l bbc.l~'lltl'l~1 ~~n !?l"vltnU !?l'dtl ethyl acetate 'lltl'l'dTW
Vl~'l~tl (Curcuma ct. comosa Roxb.) ~~'d1~b~~~1.Ji?l1'l1 rl\J ~tl 0.1% w/v, 0.2% w/v l(?lti~l'l'd
vh~~~ltl ethyl acetate bU\Jl'lr.Jmu~~ (negative control) bb~~H Oxoferin® bU\Jl'l'dbmtlubVltiU
(positive control) .Yh l!?ltl'Vl1~1~'Vl!?l~tlU~U1 !?lbbc.l~'lltl'l"vloii'l'l'll'J'l bb~~'Vl'll'l'd ~'JU ~~~U'l !?lbbe.J~'lltl'l"vl ~ q ~
0, 1, 2, 5, 7, 10, 15, 20, 25, 30 bb~~bb~(?l'lCJ~bU1.Jfh~1ltl~~'lltl'ln1~V11tl'lltl'lU1(?lbbCJ~ (% wound
" heal in g) ~'J~vku\JVi nn1~ ~ n'!1ru:::Jl1tl\JtJn'lltJ'lU1 (?) bbe.J~
CJ ~ n1 ~';jr.f t~lwnJ~ 31 -3 3 bb~ (?)'l!l'l~ ~ n11ru~n1 tJ\J'tl n'll'tl'lU'l (?) bbe.J~~u 1b'J ru 1u"vl'll'tl'l"vl\J ~ ~ ~
'Vl(?)~'tl'll(?)tlbmtJUbVltJUn1~'YI1~1~'YI(?)~tJUrlUl9l''dAr.JU~~ b~'tl~~~\Jn1~~r.ftl (rJ\J~ 0) bb~:::b~tlb'd~1 q
e.h1.J1u 30 rJ\J (rJ\J~ 30)
(A) Day 0 (B) Day 30
~tJ~ 31 ~n'l1n.J~Jl1ti\J'tln'll'tl'lU1(?)bbCJ~~U1b'dbl.JbU~'lJtl'l"vl~'YI(?)~'tl'l 11-Jrf\J~ 0 (A) bb~~rJ\J~ 30 (B)
b~'tl'YilbU\j~'l'l'JJ'd'l~'Jtl~'l~~n(?)~'JtJ ethyl acetate 'JJ'tl'l~1\JV1~1~tl ~'J'l~b~~oii\J 0.1% w/v
bb~~'Vl1bU\j~1'lsii1tJ~'Jtil'l'J'vh~~~~tl ethyl acetate
. . -·
56
(A) Day 0 (B) Day 30
b~oewn1ut~~1~'ll'"l1~'"ltl~1~~n(?1~'"ltl ethyl acetate 'll!l~~1'W'l-nn1!l fl'"l1~b~~~'W 0.2% w/v 'IJ
bb~:;vn1u~~1~~1tl ~'"ltl ~'"lvl1~:;~1tl ethyl acetate 'IJ
(A) Day 0 (B) Day 30
' t:-l~n1~A1'W'"lbWfl1~!lt.J~:;'ll!l~n1~~1tl'll!l~U1f?lbbt:-l~ (%wound healing) b~!l'Vl1U1(?1bbt:-J'Cl~'"ltl
' ~1~~rl(?1~t.J1U~'"ltl ethyl acetate 'll!l~~1'W~:w11!l (Curcuma cf. comosa Roxb.) Vl~'"l1~b~~~'W
0.1% w/v bb~:; 0.2% w/v b(?1t.J~~'"lvl1~:;~1tl ethyl acetate blJ'W~'"l~'"lUf1~ bb~:; Oxoferin® blJ'W~'"l
btntJUbVltJU ~b'"l~11?11~1 nu ~!l iu~ O, 1, 2, 5, 7, 10, 15, 20, 25, 30 bb~(?1~1u~?nn~~ 1
-·
57
IP11'i1.:Ji'11 ~Ylim~~'Wn1J'VI1tl'll!l'I'IJ11?1LL~'tl (% wound healing) L'W~IWJYll?l'tl'fl\l'll!l'l~11~rli?IVIt.I1'1J ' ' i.l I '11 q !.1 iJ ~
l?l'ltl ethyl acetate 'JJ!l'l'l1'WVI~1'l'flYlY'l'l1~ L'll~'ll'W 0.1% w/v LL'tl~ 0.2% w/v L~'fl
LmtJULYltJUnu Oxoferin® L~'flL'l'tl1[;]1'~1 n'W
%wound healing (mean± SD)(n=6)
...... "" q..o I t,.l .<r::li t,.l So'
'l'U'VI ~1oa~ n !11'l1'U ~ ~1'l'El'VI rl'l13J L ".llaJ".ll'U
Oxoferin® Ethyl acetate
0.1%w/v 0.2% w/v
1 13.43 ± 6.94 9.09 ± 0.00 6.15 ± 2.62 10.63±4.15
2 10.45 ± 0.00 3.03 ± 2.62 4.62 ± 2.62 8.04 ± 2.09
5 25.37 ± 6.94 34.85 ± 11.44 10.77 ± 5.25 13.15 ± 4.77
7 35.82 ± 10.50 37.88±13.12 20.00 ± 6.94 16.65 ± 6.98
10 31.34 ± 2.62 28.79 ± 5.25 23.08 ± 2.62 18.15 ± 3.73
15 34.33 ± 2.62 39.39 ± 2.62 33.85 ± 2.62 20.22 ± 1.23
20 38.81 ± 2.62 45.45 ± 4.55 36.92 ± 2.62 23.79 ± 3.60
25 46.27 ± 4.55 56.06 ± 10.50 36.92 ± 2.62 26.12 ± 8.09
30 56.72 ± 9.46 100.00 ± 0.00 36.92 ± 2.62 38.82 ± 32.75
-·
58
L~renl'lt:.J~m~'i9tJ1umn'l~ 1 ~~1'lLU1...lm1rJLL~(;l'lf'l'"l1J.J~J.Jttrufi~~~~1'lrl1~'fltJ~~'ll'fl'ln1~
mtJ'll'fl'lU1(;1LLCJ~ (%wound healing) rlU~:;tJ~L'"J~1r;i1'l1 ~'llU~ 34 Ll't1 :;lU~ 35 t:.l~n1~fi9tJ"tlU~1 ' ~
- 1/ I i,l ..:::::.j !J' l,.t .e:i .c::tt, !.J
~1~~n(;l(;l'"JtJ ethyl acetate 'll'fl'l'"l11...l~J.J1'"l'fl'Ylf'l'"l1J.JL'llJ.J'll1...! 0.2% w/v J.J~'Ylfim~~1...ln1~~1tJ'll'fl'l
U1~LLCJ~~~~~ L~'flLmtJUL~r.JUrlU~1~~rl(;l~'"JtJ ethyl acetate 'll'fl'l~11...l~J.J1~'fl~fl'"J1J.J L~J.J~\..l 0.1%
w/v LL~~ Oxoferin®L(;ltJiJri1 % wound healing Lvhnu 100 A'fl LLt:.J~UV1~il'Yl1urfu~ 30 ~'"J\..l~'Yli
' " " ilc:l !.1!.1 ® 0Q,.' .v.d -"'
~J.J1'"l'fl'Ylf'I'"J1J.JL'llJ.J'll1...! 0.1 % w/v LL~:; Oxoferin ~1J.J~1(;1U Yl'l1...! ~1~~n(;l(;l'"JtJ ethyl acetate 'll'fl'l
~11...l~J.J11'fl i'lf'l'"l1J.JL~J.J~1...! 0.1 % w/v LL~~ 0.2% w/v iJ~'Ylim:;~um~~1tJ'll'fl'lU1(;1LLt:.J~1~~n~1111'"l
fl'"lUf)J.J (ethyl acetate) mh:~iJutJ~1~'1J'Yl1'l~n~ (P < 0.05)
120
100
~ 80 Ll..l
I 0 60 z :::l 0 40 s:
%WOUND HEALING
- Ethyl acetate (negative control) - • - 0.1% Curcuma extract
····A ... 0.2% Curcuma extract Oxoferin {positive control)
.··
... ...: -.a . .. .,..
'#. ~......... . ...... ~·:.· ~-·~~·-·-:.:--·.-
.#"- / ·~·~<!"1·- ;... -.;::t.u-·
20 ,.;.<fl" _..;;,...._;::..· ..... =-·· -------
0~-=..., 0 5 10 15 20 25 30
DAY
'
35
- !.1 I lJ c:l !.1 !I'
~1~~nV1~t11UWJtJ ethyl acetate 'll'fl'l'"l11...l~m'"J'fl'Ylfl'"l1J.JL'lJJ.J'll1...! 0.1% w/v LL~~ 0.2% w/v
LnmWtJUL~tJUrlU Oxoferin® Ln'flL"l~1r;i1'l1 rl\..l
59
120.00
* 100.00
~ 80.00
" _g * "0 60.00 s * :; ~
* ~0.00
20 .00
0.00
*
nim~ -n~~~ nil--~ I 5 10 20 30
Dav
O Ethyl acetate (neganw control) m 0.1% Cun:uma extract 0 0.2°·• Clll'Cuma exn·act ~ Oxofenn (posiuw control)
' ~11~rl~VIt.n1J~"Jt.J ethyl acetate 'll'1l~~TW'\.UJ1~'il'YlfP1':11:1.JL~:I.J~'W 0.1% w/v LL~~ 0.2% w/v
L~'1lLmt.JULYlt.J1JrlU Oxoferin® L~'1lL"J~1l'i1~1 rl'W (* = p< 0.05, Lmt.JULYlt.JUrlU control)
.:f. "' 0 Q.,l .<C:3I ~.c:::tl Q.,l ""
2.2 t)'Vlfin'i~ 1Pp.!n1'i'WUJ"ll'fl~'U1~LLt:J~"ll'fl~ {911'i'U A'i:I.J'Vl:I.J~1'i~n {Pl!Wltl ethyl acetate "ll'fl~
' ... 'l1'WVI:I.J1'l'fl
r ' .<:::Ito i/ 0 IV ~ c:IQ IV i,.t
n11'Yl~~'1lU~'Ylnn1~(;j'Wn11VI1t.J'll'1l~U1~LL~~'ll'1l~m1UfP11:1.J'Yl:I.J~11~n~~':lt.l ethyl acetate ' I 1/ .cl l.l i.l I V dl
'll'1l'I':I1'WVI:I.J1':1'1l (Curcuma cf. comosa Roxb.) 'Ylfl':I1:1.JL'll:I.J'll'W!Wl'l1 n'W fl'El 5% w/w, 10% w/w
b~t.l~ cream base LU'Wl91':1 fPl':IU fPl:I.J LL~~H H irudoid® LU'Wl91':1 Lmt.Ju LYlt.JU vh b~t.I'Yl1~11'Yl~~'1lU~ '
U1~LL~~rJ~91n'll'W1~L~'W~1f"!'WtJnm'l'll'1l'IU1~LL~~1'WrJ'W~ 0, 1, 2, 5, 7, 10, 15, 20, 25, 30 LL'Cl~ " ..
Lb~~'I~~LU'W~i1~'1lt.l~~'ll'1l'ln11VI1t.l'1l'1l'IU1~LL~~ (% wound healing) 1':1:1.Jvl'IU'WYinll1'Y'l~m~ru~
ll1t.I'W'1ln'll'1l'IU1 ~LL~~
~~n11~{t.J1unJ~ 36- 38 LL~~'Ill1'Y'l~m~ru~ll1t.I'W'1ln'll'1l~U1~LL~~~U1L"Jtl.JLUVI'll'1l'IVI'W .. " " 'Yl~~'El'l b~mtJhJULYlt.IUn11'Yl1~11'Yl~~'1l1Jrl1Jl9i'"Jfl':I1Jfl:I.J L~'1l~:I.J~'Wn11~{t.J (rJ'W~ 0) LL~~L~'1lL"J~1
~1uhJ 30 iu (iu~ 30) ~':I'Wll1'Y'l~m~ru:::ll1t.I'W'1ln'll'1l~U1~LL~~1uiu~u1 LL~~'I~~1'Wll1fl~mn
(A) Day 0 (B) Day 30
!.1 !11 6) ll iJ iJ
l'll:W'll'W 5% w/w ll~::;vn b1J~'ll'"Wll1tJr?1r.JtJ cream base 'IJ
(A) Day 0 (B) Day 30
l~1l'Vl11u~~1~'1lr.J1 ~r.JtJ ~1furi~,:w~iJ~1 ~~n r?1~r.JtJ ethyl acetate 'll1l~~1'W~:W1fl'il~ Ar.J1:W
l~:W~'W 10% w/w ll~::;'Vl11u~~h~<ii1tJ~r.JtJ cream base 'IJ
(A) Day 0 (B) Day 30
60
. .
61 I
0 t !l,.l .dij i.l
t:J'tln'l~f11'W'JrurJ1~'eltJ'tl:::'ll'el,m1~'1-11tl'll'el'llJ1~LLt:J'tl (%wound healing) L~'elY11lJ1~LLt:J'tl~'"ltl
~1~ur11~~n~1~~n~~'"lt1 ethyl acetate 'll'el'I~1'W'I-l~1~'el~rl'"l1~L~~~u 5% w/wl 1 O% w/w t~un
cream base LU'W!?i''Jrl'JUfl~ (negative control) LL'tl::: Hirudoid® LU'W!?i''"lLmtiULVltllJ~L'"l'tl11?i1'11 nu
I I
.,. v Q 1 "" rl'el 'J'WYl 01 1, 2, 5, 71 101 151 201 251 30 LL~~\l 'WI?l1n'IY1 2
V l.l I i.l .J !.f il ~ ~1~~n~~'Jtl ethyl acetate 'll'el..:J'J1'W'I-l~1'"l'elYlrl'J1~L'll~'ll'W 5% w/wl 10% w/w L~'el
LmtiULVltllJtllJ Hirudoid® L~'elL'J'tl11?i1'11 tl'W
% wound healing (mean± SD)(n=6)
Hirudoid® Cream base ... .... .... 'VlFI'l1~b"lJ~"lJ'W
5% w/w 10% w/w
1 8.62 ± 2.75 11.86 ± 13.89 3.45 ± 11.55 7.40 ± 2.25
2 14.29 ± 4.76 25.76 ± 2.62 16.67±10.41 9.41 ± 4.92
5 28.57 ± 12.60 30.30 ± 2.62 28 .33 ± 5.77 14.17±6.31
7 33.33 ± 4.76 24 .24 ± 5.25 18.33 ± 12.58 21.62±11.78
10 33.33 ± 0.00 54 .55 ± 0.00 28.33 ± 2.89 25.88 ± 5.43
15 44.44 ± 5.50 33.33 ± 2.62 25.00 ± 5.00 17.41 ± 12.62
20 47.62 ± 0.00 40.91 ± 16.39 28 .33 ± 2.89 21 .39 ± 8.02
25 66.67 ± 17.17 59.09 ± 19.81 26.67 ± 11 .55 31.63 ± 11 .54
30 66.67 ± 4.76 62.12 ± 14.61 18.33 ± 11.55 32.57 ± 12.39
. .
62
L~'fline.J~n1~~rolt.Jb'W~1~1'l~ 2 ~~1'lLU'Wnni~LLM1'lfi'J13J'6{3Jvr'W1fd:::'V1~1'1rl1~'elt.J~:::'ll'el'ln1~
'VI1t.J'll'el'l1.11J;'ILLC-J~ (%wound healing) nu~::;t.J:::L'Jmt;i1..:J 1 LL~J;'I'lb'W~U~ 39 LL~::;~u~ 40 Y11J~1~1f1.1 ' ' .. q c:J~ CV !.1 I t; Q t; !.J Q <Ciro
fi~3JY13J~1~~n J;'IJ;'I'Jt.J ethyl acetate 'll'el'l'J1'W'VI3J1'J'elYlfi'J13JL'll3J'll'W 5% w/w LL~::; 10% w/w 3-J~Yln
m:::~'Wn1~'VI1t.l"ll'el..:J1.11J;'ILLe.J~L~~n~1rPJ'Jfi'J1.1f13J (cream base) 'flti1..:~ih!t.J~1rltl.JY11..:J~ti~ (P < 0.05)
bJ;'It.Jilri1~'elt.J~:::'ll'el..:Jn1~'VI1t.J'll'el'l1.11 J;'ILLC-J~ 1n~ LAt.J..:Jn'W w~:::il~Y1i~n~1 Hirudoid®
%WOUND HEALING
--+--Cream base(Negative control) - • - 5% Curcuma cream
--~-- 1~1. Curcuma cream - .;. Hirudoid(Positive control)
120
100
(.!) 80 z -' s I a 60 z :::J 0 -,. > 40 '?t~
20
• 5 :o 25 3}
DAY
' ' 0 Q..o' Cll Cll.:::l cv 'l/ ' t; q 1.1 !.1
m~1.1fi~3JY13J~1~~nJ;'IJ;'I'Jt.J ethyl acetate 'll'el'l'J1'W'VI3J1'J'elYlfi'J13JL'll3J'll'W 5% w/w LL~::;
10% w/w L~'flLmt.JUbVlt.JUnu Hirudoid® L~'elL'J~1t;i1'l1 n'W
63
120
100
~ so * -;;;
<.) ...<::: "0 60 = ;:> c 3 40 ~ c
20
0
5 10 20 30
Day
0 Cream ba ·c(Negat ivc control) ~ 5% Curcuma cream
~ J 0% Curcuma cream • Hirudoid(Pos itivc control)
r
~tl~ 40 m'1'r'hb'vi'lLL~t?l'l~Ylfim:::~'Wn1~vnt.J'll'fl'llJ1t?lLLt:J'fl (% wound healing) 1'W~I111Ylt?l'fl'fl'l'll'fl'l ' ' 0 CV' q c::,jq CV i,.l I !.I =I !U !/
!111~1Jrln.JYl~~~~~nt?lt?l1t.J ethyl acetate 'lJ'fl'l11'W~~~1'flYlf111~L'lJ~'ll'W 5% w/w LL'fl:::
10% w/w L~'flLmt.JULYlt.llJrllJ Hirudoid® L~'flL1'fl 1 l'i1'l1 rl'W (* = p< 0.05, Lmt.JULYlt.llJ
nu control )
I r I I Q 0 q !.;' ..:::!ij I CV d IV .:::.j
Ylt?l'flll'lYlYl1n1~Ylt?l~lllJ~Ylnm':::l?l'Wn1~~~t.l'lJ'fl'llJ1t?lLLt:J'fl L~'flL1'fl11111'11 n'W flll1'WYl 0, 2, 5, 10 LL'fl:::
20 L~m.h::: LiJ'Wn ~~~1t.l'lJ'fl'llJ1 t?l LLt:J'fl Lt?lt.l~91~ru1 91 n f111~~'W1 LL U'W'llll'l polymorphonuclear
" (PMN), fibroblast, collagen fiber LL'fl:::~'fl'flt?lL~'flt?l !?1\llJ
2.3.1
64
n1'il?l'i'"l"''i'I.JL U'fi'U~'L'Jn.!'U11?1LL~:.J~vi'Vl1~'Jtl?41'i?4rl l?l\UJ1'U ~'H.l ethyl acetate "ll'fl'l
' ... ':1 1 'lJ \UJ1 ':1 'fl
J;' !,1 J;' I I
~:.J~ m J'{;JJ''"l9:Jl'I.JL U'tl'Yl1..:J9 ~mtJ1Jl1 f1 f'\1~ (;lf'll'tl..:J L U'tl L~'tlu1L '"l ruu 1 (?! LLe.J~V11ut.-l'll'tl..:J'VI'W'Yl (?!~ 'tl..:J . " "
L~'tl'Yl1~'ltJ~1J''Ml(?l'VIti1U~'ltJ ethyl acetate "ll'tl\l~1'W'V13-J1~'tl (Curcuma cf. comosa Roxb.) ~ A'l13-JL~3-J~'W{;h'l1 rl'W ~'tl 0.1% w/v, 0.2% w/v L(?ltJ~~'"lYi1~:::~1tl ethyl acetate LU'lJ~'lA'"l'Ufl3-J
(negative control) LL~:::H Oxoferin® LU'W~'"lLmtJ'ULVlti'U (positive control) 1ui'W~ 0, 2, 5, 10
LL~::: 20
I .~
e.J~n1J'm'l9~'WLi'f'tlL'I.Ji'W~ 2 (~u~ 42) L~'tlLmtJULVliJ'UrlU'VI'I~A'"l'Ufl3-J~L'U~hJ1~'Yl1~1J'L~1 L'Wi'W~ 0 (~u~ 41) Y'l'U~1 'VI~Yjnn~3-J~'Yl1~'"ltl (A) ~1J'~rl~'VIti1'U~'"ltl ethyl acetate 'll'tl..:J~1'W'VI3-J1~'tl
' ..:::1 i,l 1J ® I
VlA'l13-JL"ll3-J"ll'W 0.1% w/v, (B) 0.2% w/v, (C) Oxoferin LL~::: (D) ethyl acetate Y'l'U PMN 'VI'W1LL1..J'W
3-J1n~'WL~'tlLmtJuLV1t~unu'VIumuA3-J1uiu~ o LL~:::Y'lu PMN m3-J1ru3-J1n~uL~nu'tltJ'U1L'lru1n~ " . 'U1~LLe.J~ 'i'i'lu91t.J'"ll.J fibroblast LL~::: collagen fiber cl'..:~hi'V'l'U~1~n1n~3-JW3-J1ru:fu9'W~\lLn(;l1~'1l~
'W'tln91 nifcl'..:~ ~911.-J'"lt.JLii ~ L~'tl ~ L 'W'VI~'tl ~ L~'tl ~ L ~3-J~'WL~ nU'tliJ LL~:::cl'\l hi'V'l'U~1~ n1J'~~1\lL 'VI~"ll'tl\lWJ\l fl1Y'l~1 (epithelium)
65
(A) (B)
(C) (D)
' CV i.l I i.l ..::.1 1J \.1
~1j''Vlr?l~'flU (A) ~1j'~nr?lVItJ1UW.ltl ethyl acetate 'll'fl'l'"l1'WVI:w1'"l'fl'Vlfl'"l1:1-JL'li:I-J'li'W
' V !,.1 I !,I ~ lJ !.J
0.1% w/v, (B) ~1j'~nr?1VIti1Ur?l'"ltl ethyl acetate 'll'fl'l'"l1'WVI:I-J1'"l'fl'Vlfl'"l1:1-JL'li:I-J'li'W
0.2% w/v, (C) Oxoferin® LL~:; (D) ethyl acetate
I iJ "' I lJ I I I
91nnJV1 43 e.J~n1j'l?\j''"J91'WLi1'flb'WrJ'WVl 5 'V'IU~1 Li1'flL~'flU1L'"lillU1r?lLLe.J~VlbUVI'll'fl'IVI'WVl'V11~'"ltl " " "
' G.' !;' I i.f ..:;,I i.l !.1 .o::::,.! V
~1j'~nr?1VIt11Ur?l'"ltl ethyl acetate 'll'fl'l'"l1'WVI:I-J1'"l'fl'Vlfl'"l1:1-JL'li:I-J'li'W 0 .1% w/v LL~:;0.2% w/v :1-J~n11ru:;
fl~1tJn'W A'fl 'Y'lU PMN roJ1'W'"l'WVI'W1LLU'W:I-J1ni-.:~~u1L'"lruu1r?lLLe.J~ LL~:;u1L'"lru1n~L~tl'~ LL~:;~m:~-J1ru
~ r?liJ!ltJ~-.:~1'Wu1L'"l ru~1n ~91 nu1 r?lLLe.J~ u~1'Wfl1'V'Ij''"J:I-JLL~'".l9:;~mmruVI'W1 LLU'W:I-J1nn~1 Lif'fl L~!ltln ~ I i.l I ..::J <V .::!1 I 0 I
fl'fl'W"ll1'1:1-J1n L'l!'Wbr?ltl'"lnu fibroblast LL~:; collagen fiber LL~:;VI~'flr?lL~'flr?l 9:;'V'IU'"l191'W'"l'WVI'W1Lb'W'W
. .
66
I t,. I I
L~!lUI?1lJ1 171 LLe.J~ Y'llJ~1LU!l L~'fllJ1L'HUlJ1 1?1LLe.J~~Yl1~'Jt1~1~~n 1?1~tnu~rlt.J ethyl acetate 'll!l-l~TW
m.J1~!l~m1a-JL~a-J~l.J 0.1% w/v ilm~~~1'l1~~n~1~1nH1~P~~m1a-JL~a-J~l.J 0.2% w/v LIPitJilm~~~1'l
.ffu,n.r-lri1Y'l~1ul71u1L'"ln.JU1nLLe.J~lh:;a-J1ru 90% LL~:; 70% 1'11a-J~1!?1'u
(A) (B)
(C) (D)
~1~Yli?1~!llJ (A) ~1~~ni?1~tnlJ~'Jtl ethyl acetate 'll!l-1~1l.J~a-J11'fl~fi'J1a-Jb~a-J~l.J
0 .1% w/v, (B) ~1~~ni?1~tl1lJ~'Jtl ethyl acetate 'll!l'l~1tJ~a-J11'fl~fi'J1a-JL~a-J~tJ
0.2% w/v, (C) Oxoferin® bb~:::: (D) ethyl acetate
. •
67
L~!lLUhJULYltiUflUV1'W~Yl1~'1tl Oxoferin® J-w YIU'JT-i1'W'J'W PMN, fibroblast, collagen "
fiber LL~:;w~!l~ L~!l ~ ~t.IhnruL ~l..lm nn~1V1'1..Wl'1 u fll..l 1 'WrJ'W~ o LLvihJ~ fl'11l..IV1'W1 LL U'WL vh nu'Vl'W~ " ' "
'Vl1~'1tlfl1~fln~V1ti1U~'Jtl ethyl acetate 'll!l~~1'WV1l..l1~!l~fl'11l..IL~l..l~'W 0.1% w/v 1.1.~:; 0.2% w/v
LL~:;~m~fl~1~·n\ .. mtr~n1YI~1th::l..l1n.J 50% LLvlLU'WL~tJ~.ff'WLLm1th::l..l1n.J 1-2 .ff-w ~'J'WV1~~'Vl1~i'JtJ ~'Jfl'JUfll..l ~!l ethyl acetate YIU PMN L~l..lmni'Wn~1V1~L'WrJ'W~ 0 L~ntJ!ltl ~1'W'J'W fibroblast
r.nnnJ~ 44 ~~ n1~[;1~'19;f''Wb\f!l1'WrJ'W~ 10 Ltl!lL~!lU1L'1n.JU1 ~LL~~~1UV1'll!l'l'V1'W~'Vl1 ~'Jtl " 'II "
Go' ll 1 ll J i.l II CJl CV
fl1~fln~V1ti1U~'Jtl ethyl acetate 'll!l~'11'WV1l..l1'1!l'Vlfl'11l..IL'lll..l'll'W 0.1% w/v 1.1.~:; 0.2% w/v l..l~n~ru:;
fl~1tln'W ~!l YIU PMN, fibroblast, collagen fiber LL~::V1~!l~L~!l~~ml..l1n.JV1'W1LLU'W t~m'UYI1:;
u1L'1 ruu1 ~ 1.1.~~ LL~:;~~1'W'1'W~ ~~~ 1 -wu1L'1ru 1n~ L~tl~ LLvltJ~ fl~V1'W1 LL'Il-wn~1V1~ 1 'WrJ'W~ o m~fl~1~ ... ' '
V IV 0 !.1 c:J ll - !.1 I !.I CJl i.l !J
'11'WV1'W~n1YI~1'll!l~V1~'Vl'Vl1~'1tlfl1~fln~~'Jtl ethyl acetate 'll!l~'11'WV1l..l1'1!l'Vlfl'11l..IL'lll..l'll'W 0.1% w/v ...
fl~1~1~1n~L~tJ~nUfl'11l..IL~l..l~'W 0.2% w/v t~tJ~m~fl~1~i-wV1l!~n1YI~1umhmL~~1~ 99% 1.1.~:;
100% ml..l~1~U Ln!lLmtJULVltiUflUV1'W~'Vl1~'1tl Oxoferin® ,_f'W YIU~1~1'W'J'W PMN, fibroblast, 'II
collagen fiber LL~:;Vl~!l~L~!l~ ~mmruV1'W1LLU'Wn~1V1~1'WrJ'W~ o L~niA!ltJ Lt.~:;~~1'W'J'WU1~[;11n~1
V1'W~'Vl1~'1tl~1~~n ~~1'WV1l.J1~!l ~n1~~~1~.r'WV1\X~n1YI'hiJ~tJ1nLb~~!l~1'l~l.JU~nJ 100% fl~1tlflU " "
' ' " v c.~ a 11 o .c:!rl .~C:~t. ~ a 11 , c.1
'J'W'Vl 0 L~n'W!ltl 91'W'J'W fibroblast, collagen fiber LI.~:;Vl~!l~L~!l~LYil..ll..l1n'll'WL~m..\!ltl LL~:;YIU'11l..l
m~~~1~.ff'Wm!-:Jn1YI~1 (epithelium) u~u1~LL~~!l~1~~l..lu~n.r 1 oo% "
' V lt I lJ c:::l i.l ll I
~1~~n~V1ti1U~'Jtl ethyl acetate 'll!l~'11'WV1l..l1'1!l'Vlf1'11l..IL'lll..l'll'W 0 .2%w/v YIU'11 PMN, fibroblast,
collagen fiber LL~:;Vl~!l m~!l~ ~~1'W'J'WV1'W1 LLU'Wl..l1n n~1V1~n~l..l~'W1 t~m.!I~'Y)n ~'11'WrJ'W~ 20 9:;~
m~~~1~V1l!~ri1YI~1u ~u1~mh~~l..lunli1 oo% 1'W'llru:;~ Ltl!lL~'fl'll!l~V1'W~'Vl1 ~'1tJ~1~~n~V1mu ~'JtJ " 'II
' I i.l .o::::!il !.1 iJ ® .c:::,l 0
ethyl acetate 'll!l~'11'WV1l..l1'1!l'Vlf1'11l..IL'lll..l'll'W 0.1% w/v 1.1.~:; Oxoferin l..l91'W'J'W PMN, fibroblast,
collagen fiber LL~:;V1~!lm~!l~V1'W1 LLU'Wl..l1ni-w91n'V1'W1 'WrJ'W~ 0 L~niA'fltJ LI.~:;Yium:;91tJLtJvf'J "
b tl!l b~!l hJ1~~'1l..l ~'1 fl'W 1n ~U 1L'JnJU1 ~ LL~~ vf--1d'V1'W'Vl n n ~l..l~ n1~~~1~V1\X-.1 rl1YI~1U ~U1 ~ LL~~ !l~1~ "' '
. •
(A) (B)
(C) (D)
' ~1J'Vl(YJ~'EJU (A) ~1J~rl(Yl'\.HJ1U~'Jt.J ethyl acetate "ll'E:l~~1'WVI~1~'E:IVlrl'J1~l~~~'W
' 0.1% w/v, (B) ~1J~rl(Y]VItJ1U~'JtJ ethyl acetate "ll'E:l~~1'WVI~1~'E:IVlrl'J1~l~~~'W
0.2% w/v, (C) Oxoferin® ll~:; (D) ethyl acetate
68
. .
·.
69
(A) (B)
(C) (D)
' ~~~'Vl~·MJU (A) ~~~~n~t.H.J'lU~'Jtl ethyl acetate 'll!l-.l~'lU'VIl-J'l~!lVlri'J'll-Jb~l-J~U
' 0.1% w/v, (B) ~~~~rl~'VIti1U~'Jtl ethyl acetate 'll!l-.l~'l'i.J'VIl-J'l~!lVlrl'J'll-Jb~l-J~U
0.2% w/v, (C) Oxoferin® LV~:; (D) ethyl acetate
. ... "lJ ·e:wn u Vll-J 1 'J 'fl
t.f 11 i.l I I
~'tl m ~(;l~'J9~Ub U!l'Vl'l-.:J'"l'tl m tl~ll'l r~ ~~ ~ (;] f'll!l'l L u!l b~!lu 1b 'J ruu 1 ~ bb~ 'tlVJbU'VI'll!l'l'VIU'Vl ~'tl!l'l ' " "
' ' ~ !.1 0 cv q qq cv !.1 ' 11
Ll-J!l'Vl'l~'Jtl(;l'lJUrln.J'Vll-J~'l~~n~~'Jtl ethyl acetate 'lJ!l'l'J'l'i.J'VIl-J'l'J!l (Curcuma cf. comosa Roxb.)
. '
. .
70
~fl'"l1a.JL~a.J~'W!Pi1~l rl'W ~'fl 5% w/w, 10% w/w 'i:~uiJ cream base LU'Wlvl'"lfl'"lUfla..l (negative
control) LL~:;1an Hirudoid® LU'WlYl'"lLUhJuL~t.Ju 1wr'W~ o, 2, 5, 10 LL~:; 20
(A) (B)
/
(C) (D)
' ' ~1~'Vl~~'flU (A) ~1fufl1a..~~iJ~1~~n~~'"ltJ ethyl acetate "ll'fl~~1'W~a.J1~1.l~fl'"l1a.J
' ' 1J !I 0 IV ..:::.1 q..c;:,j CV 1J I 1J ..::I
L"lla.J"ll'W 5% w/w, (B) 1?11~Ufl~a.J'Vla.J~1~~n~~'"lt.J ethyl acetate "ll'fl~'11'W~a.J1'11.l'Vl
1.1 1J • ® fl'11a.JL"lla.J"ll'W 10% w/w, (C) Hirudo1d LL~:; (D) cream base
~~n1~r?l~'19~'WLif'flL'WrJ'W~ 2 (lU~ 46) L~'flLmtJULYltJUrlU~~fi'1Ufla..I~LU~hi1~'Vl1~1~L~l L'WrJ'W~ o (lu~ 41) ~U~1~~V)nn~a..~~'Vl1~'1tJ (A) ~1fufl1a..~~iJ~1~~n~~'1tJ ethyl acetate "ll'fl~~1'W
' 'i-' .-:::::.1 'i-' !i,.<o ® m.J1'11.l'Vlfi'11~L"ll~"ll'W 5% w/w, (B) 10% w/w, (C) Hirudoid LL~:; (D) cream base ~u PMN
. •
•.
71
VI'W1 ml'Wl-!1 n~'WL~'el Lllt.llJ L Vl t.JU rllJ't-I'WfDI'JlJ fOil-! 1 'W'J'W~ 0 bbrl:::~u bU'Wml-!1 n.Jl-!1 n~'Wn~1tJn ~U1L'Jru 'II '
1n~lJ11'lbbe-Jrl ~'J'W91'W'J'W fibroblast LLr~::: collagen fiber cl'-lhi~u~1~n11L~l-lml-l1ru~'WIDl'W~ .. :nn(;l
1~ LL(;]~u collagen fiber u1L'lruthmL~rl Lif'elL~'elbvlt.lvT'lhJ~u~1~91'W'l'WLi11'1L~'ell'lb'WVIrl'ell'lL~'ell'l ' "
L fil-l~'WL~nU'elt.l LLr~:::ci'-3 hi~U~1~n1JI:'l~1-lb t.ni'll'el-3VIU-lfll~~1 (epithelium)
(A) (B)
(C) (D)
4 7 ~rln1 J(;l J'JIDl:f'WL d''el'lJ'el\lL d''el L~'ellJ1L'J n.JlJ1 Vl LL~ rl~blJVI'lJ'el-lVI'Wb 'WrJ'W~ 5 L~'el"r11 ~'Jt.l 'II 'II
l:'l1JY1Vll:'l'ellJ (A) ~1fufDI1l-l~~l:'l1JI:'lrlV1~'JtJ ethyl acetate 'll'el-.l~1t.!VIl-J1~'el~fDI'l1l-l !.J t.J 0 G.' c;::,j Jq CV !,.1 1 t.J .J
L'lll-l'll'W 5% w/w, (B) mmfDIJl-JYll-ll:'l1JI:'lnV1V1'JtJ ethyl acetate 'll'el-.l'J1'WVIl-l1'l'elYl
!.1 !.J ® f0l'l1l-JL'lll-J'll'W 10% w/w, (C) Hirudoid LLr~::: (D) cream base
. .
. '
72
"'l'lnnJ~ 47 ~~n'll(;]i'J"'l~"'WL\f'ilb'l..l'J''W~ 5 'VlU~'l Lif'ilL~'elU1L'JNU'l~LL~~~bU\j'll'il--1'\-H~'V)n "
1 <Ci11 0 .dJ 0 I I
mp.Jl-l"'l'l'W'J'W PMN, fibroblast, collagen fiber LL~:::'ltl~'il~L~'il~"'l'l'W'J'WVI'W1LL'W'Wl-11nn'J'lVI\1fl'JU~l-l
1 'Wi'W~ o t~ tJ'VllJ m:::"'l'lt.Jvf'Jvf--1 L \f'il L~'il t~m'U'Vl'l :::~u1L'JN1J1 ~ LL~ ~"'l:::VI'W'l LL t!'Wn~1 u1L'J ru~'W L~nU'iltJ VI'Wn~3-J~'Vl'l~'Jt.J cream base LL~::: Hirudoid® 'Vl1J~'l~n'liVI'ltJ'll'il--11J'l~LL~~b~~n~1VI'W
" . " n~l-l~'Vl'l ~'Jt.J~1¥u fl13-J~~~~~~n~ ~'Jt.J ethyl acetate 'll'il--1~1'WVI3-J'l~'il t~tJijm~~~~--l.ff'WVIU--1 n'l'Vl~'l
" " mh--1~3-JUinJtJd:::3-J1N 98% LL~::: 100% m3-J~1~1J 'W'iln"'l'lnU~--l~n'ldLLtJn'Ji''W~:::Ln~LLe-l~'il'1ln LVI~'il "
LYitJ--lb~~:::Ln~LL~~VI~~~'tlnLtJLvhtt'W ~'J'WVI'Wn~3-J~'Vl'l~'JtJ ~1¥ufl13-J~ij~~~~n~ ~'JtJ ethyl acetate 'll'il--1~1'WV13-J'll5!l~ f\'J13-J L~3-J~'W
" . 5% w/w ~~L~'VlU~'lijn11'1i'1l3-J LL'll3-J.ff'WVIU--lrl'l'Vl~1 L~tJ L \f'ilL~'1l~'J'W~"'l:::Ll~'W~:::Ln ~ LLe-1~~-.lf\--1 ~~'11~~ U'lm.J'l~LLe-1~ hJijmnmn.ff'W'il'ilnLtJ LLl9iVI'Wn~3-J~'Vl'l~'JtJ~1¥ufl13-J~ij~'l~~n~~'JtJ ethyl acetate
" ' . " . 'll'1l~~'l'WVI:l . .n1'1lVlf\'J13-JL~3-J~'W 10% w/w 'VlU~'lijn11'1i'1l3-JLL'll3-JVIU--lrl'l'Vl~1LL~htJi:::3-J1N 60% LU'1lLU'1l
I I ... 1.1 I I
~'J'WVJ"'l::: LU'W~ :::Ln ~ LLe-J~ ~3-Jij mnLtJ ni'W'il'iln 1tJ L~nu'1ltJ LU'1l LU'il~'J'W~'Wiji m1ru:::fl ~~tJ n'W ~'il 'VlU
PMN, fibroblast, collagen fiber LL~:::VI~'1l~L~'1l~~91'W'J'WVI'W1LLU'Wn~1Un~v~nu'1ltJ t~t.mum:::"'l'lt.J
vf'Jvf~ L \1'11 L~'1l t~u vu'Vl'l:::~u 1L'J ruu1 ~ LLe-J ~"'l:::VI'W'l LL U'Wn~'lu1L'J ru~'WL~ nil 'flu
"'l'lndU~ 48 LL~:::nJ~ 49 e-J~n'li(;]i'J9~"'WL\f'1lb'Wrf'W~ 10 LL~:::i'W~ 20 'VlU~1VI'Wn~3-J~'Vl'l~'JtJ \J \J \J q
" cream base LL~::: Hirudoid® ijn11VI'lt.J'll'il--1U'l~LL~~t~~n~'l'Vlnn~3-J L~t.J~n'l~~~~--li'W'l-n!--ln'l'Vl~'l
' ' 'ilt.h~~3-JUinJ 100% ~'J'W~:::Ln ~LL~~VI~ ~~'iln LtJLL~'J L\f'il L~'ilU1L'JNU1 ~LL~~ijinl1ru:::1n~L~tJ--lrlU
" ' Ltfm~mJn~ 'VlU PMN, fibroblast. collagen fiber LL~:::VI~'il~L~!l~91'W'J'WVI'W1LLU'Wn~'lun~L~nil'ilt.J 1~tmun 1:::"'11 m~''Jvf--1 L tl'il L~'fl
I I t,l I
~'J'WVI\1n~3-JVl'Vl'l~'JtJ ~1¥ufl13-JV]ij~~~~n ~~'Jt.J ethyl acetate 'll'il--l~'l'W'ltll-l'l~'il 'VlU~'l LU'il LU'il
~'J'W~9::: LU'W~:::Ln ~ LLe-~ ~~--1 f\--1 ~ ~'ilti~tJ~n m ~ LL~~ t~u~:::Ln ~ LLe-~ ~'ll'1l--1VI'W~'Vl1 ~ 'Jt.J~~ ¥u '"'13-J~ij~~~ " "
" . . ~ ~ .t;::llt. I ..:::!11 .::::J Q..<l i.f IV .odj
100% L'W'1lLtl'1l1JdL'JNU'l~LL~~~'J'W'iltm~nl1ru:::f\~1t.Jn'W f\'il'Vl1J PMN, fibroblast. collagen fiber
LLt1:::91'W'J'W'ltl~'il~ L~'il~VI'W'l LLkl'W Ln'1lLmtJ1JLVltJUrl1JVI'Wn~3-Jf\'J1Jf\3-JL'W1''W~ 0 ( IU~ 41). n~3-J~'Vl1 'U q q \J q .
i,.l I .:::1 ,... ® mtJ cream base LL~:::n~3-J'Vl'Vl'l~'Jt.J Hirudoid
'
73
(A) (B)
(C) (D)
' ' !J !J 0 CV ..:::1 q.Q V i,l I !.1 ..::::1
L'll~'ll'W 5% w/w, (B) l?n'l'1Jfl'l'~Vl~~1'l'~nrMl'dtl ethyl acetate 'll!l-l'd1'W"v1~1'd!lVl
'1; !.1 • • ® fl'd1~L'll~'ll'W 10% w/w, (C) H1rudo1d LL'l'l::: (D) cream base
74
..
(A) (B)
(C) (D)
' ' 0 CV Q Cli..::::.l IV tl I iJ ..::::.1
'(l'lj'YWI'tl!llJ (A) (;l'lj'lJ~j':l.JYl:W'tl'lj''(ln"HWJtl ethyl acetate 'JJ!l'I'JTW'VI:l.J'l'J!lYlWJ'l:W
' ' L~:w~u 5% w/w, (B) ~1fu~1:wY1i:J'tl1j''tlrll?l~'ltl ethyl acetate 'JJ!l-.1~1\J'VI:W'llJ!lYl
"' "' . ® ~'l'l:WL'JJ:W'JJU 10% w/w, (C) H1rudoid w~:; (D) cream base
. .
.·
. '
...:. 'UVIVI 5
'El .ihJ 'i1 tJ ~ fl n1 '31 r.t tJ
~1u:){u d~ULI"lNm~:){u~ 2 ~'fl~fl1tJUiLL~U~1U:){u ~'fl~ n11~n'li1~'Vlivn~:J!'lll1~"11'fl~ ~"ll~1u"ffu~1VIJ..i Curcuma cf. comosa Roxb. LL~~"tfoo.JU1LU'W~11'umfm~nLLeJ~ (Study of
Biological Activities of a New Specie Herb, Curcuma cf. comosa Roxb., and Development
for the Use as Wound Healing Promotion) ~~lh~n'fltJ~'ltJLI"l~~m~:){ml'fltJ 5 ~'fl~ ~'fl ~ . ~
n1~~n~ LLtJn~1~'lJ1~'Vlfl LL~~ ~n'tt1LI"lN~'f1~LI"llJ"ll'fl~~1~VllJ~'Vlfl'Vl1~:J!'lll1~
r.nmVI~1"11'fl~~"ll~aJUL~~ Curcuma cf. comosa Roxb . • (Extraction, isolation and structural identification of bioactive
compounds from rhizome of a herbal plant, Curcuma cf. comosa
Roxb.)
m~~mi1~'Vlim~~um~VI1tl"ll'fl~LL~~"ll'fl~~"ll~~u1~~ Curcuma cf. comosa
Roxb.
(A study on wound healing effect of a herbal plant, Curcuma cf.
comosa Roxb.)
m~~n'li1~'Vli~1U"~~:n~"ll'fl~~"ll~~u1~~ Curcuma cf. comosa Roxb.
(A study on antimicrobial activity of a herbal plant, Curcuma cf.
comosa Roxb.)
m ~~n'li1~ 'Vli~hutl5mu1'tl'fln;n L~'li'u LL~:;~hun1~'rfn L~utl'l ~"ll'fl~Yl"ll ~aJUL~~ Curcuma cf. comosa Roxb . •
(A study on antioxidative, anti-inflammatory and analgesic activities of
a herbal plant, Curcuma cf. comosa , Roxb.)
n1~Yi'(;J.J'W1~11'uu11'n'li1LLe.J~~1n~1~~n~~iJ~'Vli'Vl1~:n'lll1~"11'fl~Yl"ll~~u1~~ Curcuma cf. comosa Roxb .
(Formulation development of bioactive fraction of Curcuma cf. comosa
Roxb to promote wound healing )
. .
. •
76
Lti'el\191nLflNn11'l4'u~ 1L~vhmn~rlP'ILl-l~1"11'el\1~1U'IUI1~'el~lrltJI1l'lvl1~~~1tJ"ll'ilP1~1\11
~'el Ll-lfi1U'el~ LreJfi~reJ~OULIWVl LL~~Le:~m"llu 1\huu~11~nP1 LL~~'i:m-1n11'l4'u~ 3t~Pim~1~V1i~1u q .... I J,l t.l IV g :: Gilt. I - .oill!lr, oQ .d .:'U
'"t~"l!Vi"ll'el-.1~11~ n P'l'l1Ul-ll-J1'l'el P'I'JtJ[;l'JV11 ~ ~~ 1 tJVl\1 3 "l!UP'I ViU'J1~11~ n P1 L'elfi~'el~"l! L[;]Vll-J ~Vl fi [;111..1
,~~Vi~~~P'I ~1l-!11ClUUtT-.1L:f'elLLUflViL1u1~ 4 "llilP'I ~'el Bacillus cereus, Campylobacter jejuni, .. ... "
Shigella sonnei, LL~~ Streptococcus pyogenes LL~~il~VlflUUU\IL:.HreJ11 Aspergillus niger 1~ "" 1 .. I
(cUIIJ'l~ LL~~fln.l~. 2549) LL~~-11Ufl{tJUU\IL~L~'eln~11~rlP'In\IU1~Vlfl"ll'el\l~'lU~rlP'IL'elfl~'el~~L[;]V1Lrl'el ~ ' .
Pimt1~Vli~1u,~:n'V'l~n~'Ju (mfl-ln1Cl.J9U LL~~r~ru~. 2550) Lfl:Nn11'l4'u~ 4 1~Pin'l!l'1~Vli~1'W tlfimm'el'el non LP'I"li'W LL~ ~~1'Wn1 1tln L~U U'l P'l"ll'el\1~11~ « P1~1'Wl-ll-J 1~'el ~'J tJ i'lvl1~!:~ 1 tJ"llilP'I ~1-11 ~ 'el .. ...
Ll-lfi1'W'el~ L'elfl~'el~OUL[;]Vl LL~~Le:lnL"ll'W LP'ItJVlP'I~'elU~VJfiuuu..:~n11LnP'I lipid peroxidation Y'IU~1~11~rlP'I ... ' ...
"' ..:::;1 ... ..... Cll .qq 0 A. ff'
P'I'JU ethyl acetate l-lfl'l1l-!~1l-!11CltlUtJ-.1n11LnP'I lipid peroxidation P'!Vl"'P'I ('W1Vl'V'ltl LL~!:fln.l~. .. ...
2548) ~'JU~Vlfl~1'Wn11U'JP'Ivi-1L'Wl-l~'elP'IV1P'I~'el-1 (in vitro) LL~~'lul[;]tJVJP'I~'el-.1 (in vivo) Y'IU~1Y'IU~1 ... .. ..
~11~rl P1~1'Wl-ll-!1~'elL ui'lvl1~~~1Uvi-1 ~ 13-J"llilP'Iil ~Vlfl ~1'Wn11U'l p) 1n ~ L~tl-.1 nu ibuprofen LL~ ~il ~Vl fi ... ...
~1'Wn11tlnL~ULP'ItJil~VlfiUUU-1L'el'WL"ll~ COX-I LL~~ COX-II 'W'eln91nil Teerawatanasuk LL~~fln.l~ .. .... I 11V i.r IV 0 t.l I t.l .d A.i.l W
(2003) tJ..:JY'IU'J1 ~1r~nP'IP'I'JtJI?l'JV11~~~1tJ ethyl acetate 91nLl-l'l1"11'el-l'J1'Wl-ll-!1'J'ell-l~Vlfim'Wl-l~LN
LP"~Uil~Vliumf'lmn~l-!~1'W'l'W (anti-proliferative effect) "1l'el-1L"ll~;l-l~L1'lLVi'U"ll'el-ll-l~ (murine
hepatocellular carcinoma) "llilP'I CB0140C12 LL~~il~VJiuutY..:~mrlin~n (anti-invasion) ~Vli~1'W "
n11'el'V'ltJY'l (anti-migration) "11'el\IL"ll~;l-l~L1\IL'Wl-l~'elP'IV1P'I~'el..:J (in vitro) LL~~tJUU'ln1nm~~P'I"ll'el-1 ... ..
L"ll~;l-l~L1'l~'el extracellular matrix proteins 'W'eln91nilu..:~il~Vlfi anti-angiogenic activity LP'Itl
uutf..:~ hepatic sinusoidal endothelial (HSE) cells L'Wn11~~1'l tube-like structures Ld'elLiu..:~L"l!~; L'W matrigel-coated substrate 91nc.J~m'l'1~u"....tJ'lLflNn11'1~u~1'll ih..:J~'W ~..:JLU'WLl-l"li:-1~
... .. ~UU~11'W L ~LflN n 11'fl{uuL~'el n~11~n P1 ~'l tJ LVJ'lvl1 ~~~1tl ~'ltl L'elfl~'el~sn LI?1Vll-l1vl1 n 11Vl P'l~'elU ~Vl fi
' ... m~U'J'Wn11l-l1tl"ll'el'l LLC.J~ (wound healing) Luum~u'Jun11V11\1~11'lV1tJ1 'li..:~ Ln P'I~ULU'W
' ' ' 0 Go' I .dj .V ..:;;;!! .c:rl t.l ..::::Y .c:rl CV
~1P'IUI?l'elL'W'el'lnU 4 1~U~ fl'el1~tl~V11: 1~tl~l-l13-JL~'elP'I (hematostasis) 1~tl~Vl 2: 1~tl~n11'elnL~U
(inflammatory phase) 1~tl~~ 3: 1!:tl~n11LLU\IL"l!~;LL~~"li'ell-lLL"l!l-JU11ilLLt.J~ LL~~1~tl~~ 4: 1~tl~n11 ... ' ..
tlfu'i:r~N~~1'lLU'elLU'el (Remodeling phase) (Beanes et a/., 2003) n11~n'l!l'1~Vlfim~~'Wn11\-11tl
t;' I I I
LLU'lL"l!~;LL~~"li'el~LL"l!~U11ilLLt:.J~ L!ilm~u~il L"l!~; fibroblasts "'l~Lfl~'elUVl~1tJ\IU1L'ln.!U11ilLLt.J~ LVireJ 9.1 I I I tJ
L~1~1~tl~n11LLU..:JL"l!~; ilm1L"'l1Cl.J"....'el..:J LU'elLU'el LVireJ"li'el~ LL"l!~1J1t11LLC-J~ m~U'J'Wn11~1'11 vhn t1l~'WLU 1~u:::cl'1~LLfi n11~~1'lW~'elt11L~'elt111mJ (angiogenesis) n11~~1..:JLif'elL~reJ'lmJ (granulation tissue
.. . •
77
formation) ~~1.h~n~u~'ltl fibrin, fibronectin, collagens, proteoglycans, glycoproteins ~L~~
glycosaminoglycans (GAGs) L"li'W hyaluronan LU'W~'W i:ln'l~~:;~~FI~~~'lL'"l'W (collagen ... I
deposition) ~1'l..:ILil~L~~'4~'lL'VI~ (epithelization) LL~::n11'VI~tol'l'1l~~LLt:J~ (wound contraction)
(Bale and Jones, 2006) "' .. ~ .dA
.. :nu'fl{u\hh:: Li:i'W~'Vlfln1::~'Wn'l~'V11 ti'11~~U'l ~ LLt:J~ L 'Wn1::U'l'Wn11'VI'lt1'11~~ LLt:J~ 1::ti::'Vl 3 Fl~
1::u::m 1LLU~ L "11 ~£fLL~::"Ji~~ LL"l!~U1 ~ LLt:J~'11~~~'l1~rl ~'VI tl'lU ~'l tl L ~fi~~:;~ Lln'Vl'11~~~1'W'VI~'l~~ L~tln'l 1
'Vl~~~U~'Vl'ifn1::~'Wn11'VI'lt1'11~~U1~LLt:J~L'W'VI~~~'Vl~~~~ ~~tj~'"l1nFI'l'l~~'l~11riL'Wn11L~~~'l'W'l'W L"l!~t'fLVn::Lfu~ (Cell Proliferation activity) ~'lu'lfi WST-1 cell proliferation assay LL~~~t:J~LU'W
proliferation index (P.I.) 'VI'ln~'l1'Vl~~~ui:lCJ'VliL~~~'l'W'l'WL"l!~t'f 1'11 P.l. Fl'l1i:ll'i'l~'lnn~1 1 LL~::L-il o . . , 1.1 o~::~t.CII ~
LLUU'"l1~~~n11'VI'lti'11~~U1~LLt:J~ (m v1tro wound healing model) ~'ltl'lfi Scratch assay Fl~ m1
... I
Ln~"Ji~~~1~ LL~'lU~L"l!~t'fLrn::L~tl~rlU~'l1'Vl~~~U~FI'l1~L~~~'W!?h~1 rl'W LL~'l~~Lnlnn'l1~~~'l'11~'l il:!4'U .JJ J I Cil .J I CV ~ .c::i tV I .c:i .: !,/
V'I'W'Vl"l!~ LU'W'Vl'l'l..:Jin'l~1~ti'11~'Vl L'l~11Pl'l'l1 nu L~~L'VltiU nun~~ Fl'lU '"1~ 'VI'ln~11'Vl ~~~U~CJ'Vlnm::l?lu
n1 1'VI'ltl"ll~'l U'l~ LLt:J~ Fl'l ~i:J Fl'l'l ~ ~'l~11CIL 'Wn11L~~~'l'W'l'WL "11~~1 ~:jj ~rl'WL;'l~~~
u~ n'"l1 nd ..:J'lu'fl{uL~'Vl ~~~u 'J'Vl im~~'Wn11'V11 U"ll~..:JU'l ~ LLt:J~ 1u~ l?l'J'Vl ~~~'~ ~11~iJ 'J'Vl i 1.1 I I 'U I
m::~um1'VI'lU"ll~'~ LLt:J~ '"l~Li..:Jre1 1?11'ln11~'f'l..:J Lil~ L~~ 1'VI~~1":d~~LL"ll~umLLt:J~~ Ln~~u ~'~~1~1~r~ ' "
1.h:: Li:i'WL~'"l'l n"ll'W'l ~lL '"l'l ~~ L~ n~..:J Lri~ L'l ~'l &.huhJ LL~~tf~~'l~11mh::Li:i'W~'l U~1 n11":d~~ LL"l! ~
U'l~LLt:J~Ju YhL~Ln~LLt:J~LU'W (scar) ~1nil~mLfiL'VI'W~'lU mnh~Li:i'Wn11'V11U"ll~..:JU'l~LLt:J~tj~'"l'ln .. 1~u~~"ll~'ln11'VI'l U"ll~..:JU'l ~ LLt:J~ (% wound healing) 'VI'l n~'l1'Vl ~~~uiJ ~'Vlfim::~'Wn11'VI'l U"ll~'l
!II t,l t,l I
L vh nu 1 0 0% LL~ ~'l~'l LLt:J~U ~~U'Vl ~'l'Wn1~1?11'l'"l:Ji'WL U~'Vl'l'l'"l~ n'l u"lrn Fll'll'l~ ~nf"ll~'l L ilm~~ '
(Histology) L ~ ~ 1.h:; L ij 'W n 11 'VI1 U "11 ~ 'l1J 1 ~ LL t:J ~ L ~ U ~ ~ L n In '"l 'l n F1 'l 'l ~'VI 'W 'l LL U 'W "11 ~..:!
polymorphonuclear (PMN}, fibroblast, collagen fiber LL~~'VI~~~L~~~~~'f'l..:JL'VI~ ~'l~'l1'Vl~~~ui:l .. oQ. 1.1 ai 0 I
~'Vlfin1::1?l'Wn11'VI1tl"ll~'l1J'l~LLt:J~ Fl'l1~'"l'l'W'l'W~1nn'l'l !.t I I tr
'W~n'"l'lnU tJ..:JiJ'flfi'Vl ~~~1J~'W1 Lfi~1h::Li:i'W~'Vlfln1~~'Wn11'VI1 U"ll~'l1J'l ~ LLt:J~"ll~'l~'l~'Vl ~~~1J
1u~l?ltf'Vl~~~'~ L"liu m1~1u~u~~:a~1:::1u1:::'VI~1..:Jm::u'lum1'VI'lU"ll~..:JLLt:J~. mn~~m~1n.m~:::m1 '"
I ...
~..:! LF111:::'\.f Fl~~~'l L'"l'WLL~:: hydroxyproline, n'l~LYi~n11'VI~f'l'11~'l"11~1J LLt:J~ (wound contraction)
'"l'lnfll1L'U~u'f~u~::m1'VI~I?i''l"ll~'lLLt:J~ (%wound contraction) t~u~11~1J1~LLt:J~LLUUn1~~'VI~..:JLL~:::
78
l,,j vh 1 ~~13J1~r:J~~1-:J L~'fl1i~'J l'l-1,j11nl"l~3JlJ1~LL~~ U1Li'J~l.l r.nnrh L'1l~U1'flu~::"ll'fl-:Jm 1~11-:~ L~'fl
1.!~'lb'!-1a.l (% epithelization)
~11'Yi !"ln.!n11fn'l!t1 LL(:.I~"ll'fl~~1l.IV13J1~'fl~"l!1'Jthl.IU1 L Vl~1~ ~ (rhizome) 3J1 t.l1l.ILUl.ILLc.il.l 1 " I
lJ1~1 LL~'J~l.l~'ellJlJ1~LL(:.I~~~vhn~,1n"ll'fl-:JiJI"l3J~~~n Vi'l1'lLUl.IL'l~1 3-5 tfl.l L~'flLU~'fl'fln~ 'V'IlJ~'flU
LL(:.Ifl L~'fl3Jnl.IL~~ L 'l-1~'fl L YlU-:J~'flU LL~-:1 LLfl::L,jilm~~ ~ L=f'fl 'fl1,'flfilJ1 U 1111 n!J n11n.11~,1 n (:.1 fln11ilr.l uif
~'YilJ~1 ~1l.IVI2-11~'flil~'Ylim::~l.lm1V11U"ll'fl~lJ1~LL(:.Ifl 'l:~uil~'YliL~3J~1l.I'Jl.IL"l!~; ~13J11mi~i~11 " 1 I
n11~11-:J L U'fl LU'flL Vl,j3J1"1i'fl3J LL"l!2-lil ~lJ1 ~ LL(:.I~ m::~l.ln1 n Vi2-1~1l.I'Jl.l"ll'fl'l"'J'fl-:J polymorphonuclear •
(PMN}, fibroblast, collagen fiber LL~::VI~'fl~L~'fl~~~~1'lLVI,j l.l'flnr.nnifu-:~il~'Yli~l.l1 ~~uu~~l.l n11VI1 U"'J'fl~lJ1 ~ LL(:.I~,1n11U-:11l.ln11ilrolU"'J'fl'l Ll"lN n11il4'u Ll.ILL(:.Il.l'l1l.IL~m nl.l mVi ~'Yli ~1'1..1"\~~'Yi
.. " (ru~11?iJ LLfl::l"ln.!::, 2549; n1'fl'ln1(]J,U LL~::r1ru::, 2550) ~'Ylfi~1l.l11!jmm'fl'flnanL~"Jil.l (lhVi'YiU LL~:: .. r1ru::, 2548; fi1f'lln~ 'l-:J1fl1l.l LL~::r1n.~::, 2550)
(:.l~n11il4'u~'YilJ~1~11~n~~'lm'flfifl'fl::anL~'Yl"ll'fl'l~1l.IVI2-11~'fl (Curcuma cf. comosa Roxb.) .. "
iJn'Ylfin1::~l.ln11VI1 U"'J'fl'llJ1 ~ LL(:.I fl ~'fl ~ rl~'fl'l nu 11U'l1l.ln11ilr.l U"ll'fl'l~"l!~3Jl.IL 'Yi1"lli1l.I"Jil.l Curcuma .I • • ' " ..
longa ~'l'fl~Ll.I~1:::Qfl Curcuma 'J'lPl Zingiberaceae L"lil.lnl.l L~U"lli1l.I"Jil.lii~'Ylfim::~l.ln11'1-11U"ll'fl~ "' col
lJ1~LL(:.IflLl.li~tf'Yl~~'el~ vi~Ll.l excision LLfl:: incision wound models 2-1 % excision wound LL~:: " .. '
tensile strength ~n~1n~2-lrl'llJ!"l3J (Rita et a/., 2012) l.l'fln,1ni1 curcumin U'lil~'YlfiLYi2-lm3J1n.! ' " oiClf c::i Cil Q..ll' IV r;S I ~ ~ .o:::S c;;[lj ~
rl'flflfl1L,l.ILLfl::~L'fll.IL'fl 2J'el~11n11VI~~'J"'J'eNLL(:.IflL1'Jn'J1 2-1 tensile strength L'Yi2J"lll.l LL~::2J!"ltl.l~3JlJ~
Ll.ln11~1l.l'fl'4~fl:SM:: Ll.I1:::VI~1'l m::u'Jl.lm 1fmnu1 ~ LL(:.Ifl ~n~1n ~2-1 rJ'JlJ !"l2-l'flth-:~ iluu ~ 1 Fi' (]d
(Panchatcharam et a/., 2006)
'l1l.lilr.lu01-ii Oxoferin® Vli'fl Tetrachlorodecaoxide (TCDO) LUl.IIK'JLmuuLYiuunu~11
~n ~VIU'llJ ~'J u L'flfifl'fl :;an L~'Yl"ll'fl~~1l.IVI2J1 ~'f) L ~'fl~,1 n LUl.l(:.l~ L'l.rlru'¥1111:: L11'Yl ~11~ ::~1 u~ii~1wt..h u
Ll.!Vl'fl'lL'lfl1~ Oxoferin® il~'Ylim::~l.l phagocyte 1ih11fl1ULL1Jf'1YJ~U LL~::LL~nl?i'JL~ oxygen vh1~ II t.f I 1.1
.-, I .ro!il .-.::i II c!l .:oil ~ .OC::.IV .cl I IV 1,.1
Ln~n11LN L~'fl~2-11 LflU'l LL~::~11'l Ll.l'el LU'fllJ1L'J CWLL(:.Ifl ,1n (:.lfln11'J,Ul.I'YilJ'l1 ~11~n ~VIU'llJ~'JU
L'flfi~'fl ::an L~'Yl"ll'fl'l~1l.IVI3J1~'flil ~'Ylim::: ~l.ln 11VI1 U"ll'fl'llJ1 ~ LL(:.I~ ~n~1 Oxoferi n ® 'l:~uil r-h1'flu~:::"ll 'fl~ n11VI1U"ll'fl-:JlJ1~LL(:.I~ (% wound healing) 2-11nn~1 L~UL'U'Yi1:::~11~n~~l"l'J13JL;]3J;]l.l 0.2% w/v
~12-111r:JVhL~LL(:.I~il~~il'Y1L~Ll.l1'l.l~ 30 (% wound healing = 100) LLLJlLUl.l~'l.hi~Ln~~~in'l!tru::
m:til ~"ll'el'l1J1 ~LL(:.I~ Ld'fl'Y11 ~'JU~11~n~ , ::'Yiu~::Ln ~LL(:.I~ LUl.lfl'fll.ILL-ii~ Ll.l"llru:::~ Oxoferi n ® iJ rh1'flu
~::"ll'fl~n11V11U"ll'fl'l LL(:.I~U'flUn~1 ~'fl 36.92 ± 2.62 1wfl.l~ 30 LLLJl~'JVIU'llJ1L'lrulJ1 ~LL(:.I~,:;~uu 1,j
LUl.ILL(:.I~~l.l L~'fl'l,1n oxoferin ® ~12-11:tm~3JVIU'ln1Yi11~tJnvh~1u LL~::vh 1~Ln~m:t~11'l~'Jl.l'fln " ' dl dl
"ll'fl-:1 Ll.l'flLU'fl (Malik et a/., 1997)
79
iK'lLmUULVJUU~L~hm1-ilL'Wfi11VI!1\~'ElU~VIin1~~'Wn1~'1-11U'li'El'lU1!1\LL&Jt'l'11'fl'l(;hfufl1~~iJ ~'l'W&J~~'11'El'l~11~nl1l~hm'Elfit'l'El~onLI1lVI'li'El'l~1'W'VI~1~'El ~'El 1"11~ Hirudoid® 'VI1'El Heparinoid ~'ILU'W 'El"*'ri'u('li'El'l Heparin LU'W~1nh~tJ1VI glycosaminoglycans 'VI1'El mucopolysaccharides •Jhu1~
~ I
fOI'l1~,~0D'WtlUL'llt'li "Ji'lUL'llt'lt'!1~L"'l"1t1JL~ULI11 LLt'l~"Ji'lU"Ji'El~LL'li~L"llt'liVJU1!1\L~U "'l1fi&Jt'ln11flr.fU'YiU~1
LP111ufl1~~il~11~tl!11'11'El'l~1'W'VI~1~'Elil~VIim~~um1'VI1U'11'El'lU1!11LL&Jt'l~n~11"11~ Hirudoid® L!11UL'W I I
.... cl 0 .., ..::::.1 .Gi!IC!il .... I !.1 Q I !.1
'l'WVI 30 11l1~Url~~VI~~11~fi!11'11'El'l'l1'W'VI~1'l'El 5% w/w LLt'l~ 1 O% w/w ~f0111'ElUt'l~'11'El'ln1~'VI1U'11'El'l
u1~LL&Jt'l Lvhnu 66.67 ± 4.76 LLt'l~ 62.12 ± 14.61 11l1~~1~u 1u'llru~~ Hirudoid®ilrh'f'Elut'l~'li'El'l n11'VI1U'11'El'IU1!11LL&Jt'IL~U'l 18.33 ± 11.55 U'ElUfi~1iK'lfl'lUf'l~~'El cream base (% wound healing
I ~ .,.
= 32.57 ± 12.39) ~'ln11rl!1\L~'ElniK'lLmUULVJUUU 'El1"'lhiL'VI~1~~~ hi~1~11mmUULVJUU~VIfi
n~~~'Wn11'VI1U'li'El'IU1~LL&Jt'IL~ L'Yi11~nt'!Lnn1J''fl'fln~VIjf'11'El'l Hirudoid® fl'El uutT-:Jm14"UiK'l'11'El'ILn~111 I ...
L~'El~L'WU1L'ltu'VIt'l'El111L~'El~~1L~e:i'l'VIU'l vh1~n11L'VIt'IL1Uu'11'El'IL~'El!11L'WU"iL'ltuVJVI1U1~~u vh1~
'El1n11U'l ~ 1uu1L'ltu~iJ L~'El~;'ILU1U1'lt'l'l t'l ~'El1n11U'l~ LL~-3 'flm~u LLt'l~n11~-:JlJi'l'11'El'le:i'l'VIU'IL'W I II II I
u1mlJVJVJ1m ~-:Juu 'VI1nilm1~mnilr.fu1ul"lf'l'VIU1 r~'ln~'ElniK'lLmuuLViuuViL'V1~1~~~ ilnt'!Lnm1 .,. ...
'El'Elfi~VIfim~~um~'V11U'11'El'IU1~LL&Jt'l L"liu m1ih Oxoferin® ~1lJi-:JLU'WLPl1i'UI"l1~~'lU cream base
~'I LUUlJi'l rl'lU f'l~L 'Wn11VI ~t'l'El-3 ll 1.1 II I I
t.Jt'l n11111~'l"'l:nuL U'ElVI1'l"'lt'l muiln1 r~ P\1~ 11lf il~'El~'l Ln 11l~1 LU'El LU'ElU 1-L'l ruu1 ~ LL&.~t'!Vi1ut~'11'El'l'VI'W ' 'IJ ..
I I dl II .... !I .::::1t. Q. 1 ,_, 0 .... ..::::.1 .r:::li..:::iil ...,. !I
Vl~t'l'El'l L~'ElVI1 ~'lU~11~n ~'VIU1U ~'lU L'Elfit'l'El~'liLI11VI'11'El'l'l1'W'VI~1'l'El LLt'l~l1l11U I"IJ'~VI~~11~n ~11\'l U
L'Elflt'ltl~"RLI11VI'11'El'l~1'W'VI~1~'El "'l~'Yi'l.JW~1tuLii~L~'El~'111'l'liU~ polymorphonuclear (PMN) ~'ILU'W
L"ll'tl'tl1'Wn1~U'l'Wn11'flnL~U'VI'W1LLU'W~1nn~1n~~rl'lUrl~ LLt'l~n~~~L~f'l.JlJi'lLmUULVJUU Oxoferin® ' ' '
LU'W~11~tl~'VIU1U u'lhiL~LLUfiL~LU'W~11U1~VIi~i1L'tl'Yi1~~11~1rlt1J r.i-:Ji1~11~'W1 (impurities)~
'El1"'l1umun"t~Lnm1m~~um1'V11U'11'El'IU1~Lu.Jt'l 'i:~um~~um~u'lum1'flm~u 9 .. n111~Y-~u PMN ' '
I ...
... "" '1 -~ ... ® ® ... Vl'lJ1L'ltu'l.J1!1\LL&Jt'l b'Wu1~1tu~1n 'Wtlfi"'l1fi'W Oxoferin LLt'l~ Hirudoid (Livaoglu et at., 2010) ~
.. ...
~VIflL'Wfi11UUU'ln1~'l.J'l'Wn11'flm~u (Angelika et at., 2000; Tiziana et at., 2012) 9-rvh1~'Y'I'l.J 1.1 I I !.f ..... !I Q. Q. 1 f.l 0 cv .c::il .Jq
PMN 'WtlUfi'l1n~~QfiV11!1\'lU~1~~n~'VIU1U!1\'lUL'ElVIt'ltl~'liLI1lVI'11'El'l'l1'W'VI~1'l'El LL'tl~I1111Uf011~VI~~11 ... ...
~n ~ ~'lmtJVit'l'El~iR Ll1lVI'lltJ'l~1u'VI~1~'El ~-:Juu m~~r.fmh~un~ hJ1utJu1 r~ (;] m~iln1nLumru~11
~n ~'VImu L~'El~n ~ LLun 1 ~ Luu~11u1~VIi Luum1ri1r.f~~'.lu1h~n'Elu~u1 ~hi1"li~11~1 R't1J~tl1"'l Luu
U"'l9U1Un'.l'Wfi1~U'J'Wn11'VI1U'11tl'l'l.J1~LLt.Jt'l
. ..
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n1~Plm~1~'Vlin~~~un1~VI1tl"ll'El.:ltJ1~LLl!J~"ll'El.:!Yl'li~"!'WL~~~1UVI'-l1~'El (Curcuma ct.
comosa Roxb.) m~Q~ Curcuma 'Nfll Zingiberaceae L~'El~OO-IU1LU'IJ[;'ht'umt'mnLLl!J~ vhL~tl ~ ~
'Vl~~'El1J~'Vlfin~~~Un1~VI1tl"ll'El-:!1J1~LLl!J~"ll'El-:!~1~~rl~VIti11J~'ltiL'Elfl~'El~~L[;]'V1"1J'El-:!~1UVI'-l1~'El vl.:ILU
VI~'El~'Vl~~'El-:1 (in vitro) LL~~L'Wi[;]tJ'Vl~~'El-:1 (in vivo) LL~~'Vl~~'El1J~'Vlim~~Un1~VI1tl"ll'El-:!1J1~LLl!J~LU i [;]tJVI ~~'El.:l"ll'El.:lf1i1t'U fl1'-l~ ~~'l'Wl!J~l.J"ll'El-.3~1 ~~ rl ~ ~'l tl L'Elfl~'El~~ L[;]'Vl"ll'El-.3~1'WVI'-l1~'El
c ..:::.cv 1 ..... II ..:::.. .Q I ll c:l II !.1
l!J~ n1 ~'l~ t1~1J'l1 ~ 1~~ n ~VIti11J ~'ltiL'Elfi~'El::;'l! L[;]'Vl"ll'El.:!'l1'WVI'-J1'l 'El'Vl fl'l1'-l L "11'-J"ll'W 400 IJQ/ml
~~'Vlin~~~un1~VI1tl"ll'il-:!1J1~ LLl!J~ LUVI~'El~'Vl~~'il-:1 (in vitro) L~ti~~'VliL ~l.l~1U'l'WL'l!~6 (proliferation activity) i.:~L'l!~6L~1:::Liti..:JLif'ElL~'ilL~Cl'lVIU.:I"ll'il..:JV11,J'llil~ L929 LL~~L'li~6'-J~L~~1.hn
l-l~~n"ll'il~l-l'41=1'U'llil~ Hela cell 'Elth~~Ut1~1rlqj'V11-:!~ti~ (p < 0.05) Lri'ilLmtltJLVltltJrltJ!K'lmtJ'"}l-l
~hi~~1~~n~~l.JUL~~ LL~~~n'Vlim~~Un1~V11tl"ll'il-:!1J1~LLl!J~~~~~ Lri'EIL~LL1J1J~1~'il~n1~VI1tl"ll'il~ .. .I -. ,..
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U'iln~1nU L~'il'Vl~~'il1J~1~~rl~VIti11J~'ltiL'ilfl~'il~;nL[;]'Vl"lJ'il'l~1'WVI'-l1~'ilVlfl'l1'-JL~l.J~U 0.1% w/v LL~~
0.2% w/v ~1J~1 ~1~~n~~fl'l1'-JL~l-l~U 0.2% w/v ~~'Vlim:::~Un1~VI1tl"ll'il~1J1~LLl!J~L'Wi[;]tJ'Vl~~'il~ (in vivo) ~~~~ L~tl~~'iltl~~"ll'il'ln1~VI1tl"ll'il'ltJ1~LLt:J~~n~1~1~~n~~fl'l1'-lL~l.l~u 0.1% w/v IK'l • fl'ltJ'"ll-l LL~~ Oxoferin® 'Elti1-:!~Ut1~1rlq)VI1'l~ti~ (p < 0.05) LL~~~1'-l1~fi'Vi1L~tJ1~LLl!J~U~~\l'V1L~
1wru~ 3o
L~'ilU1~1~~rl~VIt111J~'ltiL'ilfl~'E!::;;nL[;]'Vl"ll'il~tJ1'WVI'-l1~'ill.J1~00-IU1Llj'Wf1i1t'Ufl1'-l~~~'l'Wl!J~l.J
"ll'il-:!~1~~n~~fl'l1'-JL~l.J~U 5% w/w LL~~ 10% w/w L~'il'Vl~~'il1J~'Vlim~~Un1~VI1tl"ll'il~1J1~LLl!J~LU l[;]tJ'Vl~~'il-:1 (in vivo) t:J~n1~~{tlwu'l1 ~17ufl1'-l~~~1~~n~~'lm'Elfi~'il~~L[;]'Vl"ll'il~tJ1UVI'-l1~'il~
fl'l1'-JL~l.J~U 5% w/w LL~~ 10% w/w ~~'iltl~:::"ll'il~n1~VI1tl"ll'il~1J1~LLl!J~Lnlh~tl~rl'W ~~~n~11K'l
fl'ltJ'"}l-l LL~~ Hirudoid® 'Elri1-:!~Ut1~1rlq)VI1'l~ti~ (p < 0.05)
Cii"'d' .Q tf" J.J ell J .Q
l!J~n1~[;]~'l~'l!UL'W'il'V11'l"i~n1tl'lll1 M11~ [;]~"lJ'il'l LU'El Ltl'iltJn'lCW1J1 ~ LLl!J~ L ~'El~~tl1 fi~ll1~ . ~
I-' I !I I I df ,dl I df df .Q q 1.1 V SJ' .Q Cl 1 !.1
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LL~:;mnn1!1'1~mu1~"1l'WI?l'W. ~2-.I~PlN'VI 3. n~-:JL'VI~l..I'V11'WPl~ : '1~'VI'tl-:J; 185.
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Ruangphisan M, Wijitsoontornkul Hand Rungseevijitprapa W. and Prasitpuriprecha C. 2010.
Development of Probiotics Entrapped in Microencapsulation. The Second Annual
Northeast Pharmacy Research Conference 2010. 108.
Sirima Suwannakut, Chutinun Prasitpuriprecha, Kandanai Jansong, Chantima Phiantham
and Paramee Pookhaow. 2010. Characterization and Identification of Potentially
Pro biotic Lactobacillus Strains. The 1st current drug development international
conference, Phuket, 6-8 May 2010: 162-166.
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nntn~CU1ftnu ~a.J'l~fif LL~:::"lf~U'UVJ U'i::i'VI~1l~....rn"lf1. 2555. nVJim:::~'Wn11i~Lf1111:::,XLa1~~ "'I ~ • ~ ul q
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Annual Northeast Pharmacy Research Conference 2012 (Proceeding).vl'ib 125-134.
Damkleang, E., Rungseevijitprapa, W. and Prasitpuriprecha, C. 2012. Preparation and
characterization of ovalbumin loaded chitosan nanoparticles for intranasal vaccine
delivery. The Forth Annual Northeast Pharmacy Research Conference 2012. 296.
Aranya Manosroi, Puxvadee Chaikul, Pisit Chainonthee, Laksana Charoenchai, Chutinun
Prasitpuriprecha, Wandee Rungseevijitprapa, Worapaka Manosroi and Jiradej
Manosroi. 2012. Effects of the plant crude extracts and fractions loaded in niosomes
on melanogenesis in B16F1 0 melanoma cells. Natural Products Research and
Development Center (NPRDC), Science and Technology Research Institute (STRI),
Chiangmai University, Chiangmai, 28-30 Nov. 2012 .
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The ih Northeast Pharmacy Research Conference of 2015, KhonKaen University,
KhonKaen, 7-8 March,2015 (proceeding), in lsan Journal of Pharmaceutical
Sciences val 11 (supplement), p 54-60.
Suparat Chanluang, Pommavong Salikabkeo, Jainuch Kanchanapoo, Rawiwun
Kaewamatawong and Chutinun Prasitpuriprecha. 2016. Anti-oxidative Effects and
Toxicity of Bauhinia penicillilobaEthanolic Leaf Extracts. Chiangmai J. Sci.; 43 :1-12.
Pattra Chanchai, Sirima SuvarnakutaJantama, Chutinun Prasitpuriprecha, Sunthorn
Kanchanatawee and Kaemwich Jantama, 2016. Effects of the Food Manufacturing
Chain on the Viability and Functionality of Bifidobacterium animalis through
Simulated Gastrointestinal Conditions.PLOS ONE,
DOI:10.1371/journal.pone.0157958 June 22 2016, p.1-17.
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Teerawatanasuk N., Nakamura ES, Wangmaneerat A, Saiki I. Anti-invasive and anti
angiogenic activities of a Curcuma sp. J Traditional Medicine 2004.
Teerawatansuk, N ErbB protein tyrosine kinases: A novel target for cancer treatment.
Srinagarind Medical Journal2003 (In press)
93
Teerawatansuk, N. Role of COX-2 inhibitors in cancer prophylaxis and treatment. Thai Jamal
of Pharmacology 24(2), 2002.
Teerawatansuk, N. and Sripanidkulchai, B. Anti-angiogenesis: New approach in cancer
treatment. Srinagarind Medical Journal2001 ;16:1-10.
Teerawatanasuk N, Skalnik DG, Carr LG. CCAAT displacement protein (CDP) negatively
regulates transcription of the human tryptophan hydroxylase gene. Journal of
Neurochemistry 1999;72: 29-39.
Teerawatanasuk N, Carr LG. CBF/NF-Y activates transcription of the human tryptophan
hydroxylase gene through an inverted CCAAT box. Molecular Brain Research
1998;55: 61-70.
Teerawatanasuk N, Reed GE, Eichholtz S, Carr LG. The mouse tryptophan hydroxylase
(m THP) promoter interacts with members of the steroid/thyroid hormone nuclear
receptor superfamily. Alcoholism 1997;21 (197): 75A.
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94
1. Vajragupta 0, Toasaksiri S, Boonyarat C, Wongkrajang Y, Peungvicha P, Watanabe
H, Boonchoong P. chroman amide and nicotinyl amide derivatives: inhibition of lipid
peroxidation and protection against head trauma. Free Radic Res 2000, 32, 145-
155.
2. Vajragupta 0, Boonchoong P, Wongkrajang Y. Comparative Quantitative Structure-
Activity Study of Radical Scavengers. Biorg Med Chem 2000, 8, 2617-2628.
3. Vajragupta 0, Monthakantirat 0, Boonchoong P, WatanabeH and Peungvicha P .
Neuroprotection of Hydroxylnicotinyl Amide 18 against Lipid Peroxidation and
Memory Impairment. Mahidol J Ph arm Sci 2002, 29 ( 1-2), 33-43.
. ,.
95
4. Vajragupta 0, Boonchoong P. New leads of HIV-1 integrase inhibitors. Mahidol
University Journal of Pharmaceutical Sciences. 2002, 29, 33-43. Mahidol J Pharm
Sci 2002, 29 (3-4), 1-10.
5. Vajragupta 0, Boonchoonga P, Sumanonta Y, Watanabeb H, Wongkrajanga Y and
Kammasuda N. Manganese-Based Complexes of Radical Scavengers as
Neuroprotective Agents. Biorg Med Chern 2003, 11, 2329-2337.
6. Vajragupta 0, Boonchoong P, Watanabe H, Tohda M, Kummasud N and Sumanont
Y. Manganese Complexes of Curcumin and Its Derivatives: Evaluation for the
Radical Scavenging Ability and Neuroprotective Activity. Free Rad Res 2003, 37
(supple 1 ), 49.
7. Boonchoong P, Vajragupta 0 and Berliner LJ. Manganese complexes of Curcumin
Analogues: Evaluation of Hydroxyl Radical Scavenging Ability, Superoxide
Dismutase Activity and Stability towards Hydrolysis. Free Rad Res 2003, 37 (supple
1 ), 51.
8. Vajragupta 0, Boonchoong P, Watanabe H, Tohda M, Kummasud N and Sumanont
Y. Manganese complexes of curcumin and its derivatives: evaluation for the radical
scavenging ability and neuroprotective activity. Free Rad Bioi Med 2003, 35 (12),
1632-1644.
9. Vajragupta 0, Boonchoong P, and Berliner LJ. Manganese complexes of Curcumin
Analogues: Evaluation of Hydroxyl Radical Scavenging Ability, Superoxide
Dismutase Activity and Stability towards Hydrolysis. Free Rad Res 2004, in press.
Presentations
1. Oral presentation. Classical and three-dimensional quantitative structure-activity
relationship studies of antiradical agents. The third annual national symposium on
computational science and engineering. Faculty of Science, Chulalongkorn
University, Thailand. March 24-26, 1999.
2. Poster presentation, RGJ Conference 2003. Pattaya, Thailand. April 25-27, 2003.
3. Gave special lecture on "Manganese complexes of curcumin analogs: evaluation of
their radical scavenging ability and neuroprotective actions" and "Active binding
96
modes of curcumin on HIV-1 protease and integrase" at Curie Institute, Paris,
France. June 23, 2003.
4. Poster presentation. Free Radicals and Oxidative Stress: Chemistry, Biochemistry
and Pathophysiological Implications - Meeting of the Society for Free Radical
Research, European Section, loannina, Greece. ,June 26-29, 2003.
1. Opa Vajragupta, Preecha Boonchoong, Arthur J. Olson and Garrett M. Morris.
Active Site Binding Modes of Curcumin in HIV-1 Protease and lntegrase. Submitted
2. David Siegel, Daniel Gustafson, Donna L. Dehn, Jin Y. Har, Preecha Boonchoong,
Lawrence J. Berliner and David Ross. NAD(P)H:quinineoxidoreductase 1 (NQ01 ):
Role as a superoxide scavenger. Submitted
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4) Trichinosis: epidemiology in Thailand.World J Gastroenterol. 2006 Oct
28;12(40):6440-5.
5) Human cholagiocarcinoma and its pathologic anatomy in ubonratchathani.
11'i'~1'i'~r.i't1 2-.l"ll.il "<L <Uuu~ 5 'VIib <L<LI:li-<L&-""' ~1Lilum'i''i':::Wl1..:JL~'Elu2-.lm1r12-.l ... .. Cl..:J Yl'lPI'"ln1ti'W I!::J&'&'o
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8) The ultrastructure of the brain of adult liver fluke,
OpisthorchisviverriniPublicated in International Journal of Parasitology
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lllflllill f Jl 'I'IW:!Pf" 'IIIII l"'SIP""'
J !I; 1 ..., ~ 0 ..., Q .J"'J qnBn1~~un11~1V~B~U1~uua "~~1n~aa~~u~~11U~1~n~~1uu~~~u~ •
~11~fl~tJ1tJLBiiau~tUL~n11U~ C. cf. comosa Roxb.
CU I
un~~VB
Curcuma ct. comosa Roxb.
fl'D''!I11UJ1\Wt1~', Lfl~Hi'll1ll fl1.,)h 1, i'91'"1 L;munnm2
, m'D'11JtU"i.,)1, 1'ilin1 flil1iiil'"1
, 'D'fi\l'wn lh::iYtftiim'D'11 "' • , .. ,. :&I 1 •
1. lni7fff1nfiift'1u, VU1flJJ 2558; 11 (nuuwmH) : 54-60
umh: Curcuma cf comosa Roxb ~~a·"i1Wlnn1a· LilutnJul-vnlf"'f11U1"U.lumf'IEhnuua::ti~::L 'Ylflnli'Vl"'f1ll1L~.5'1Htl . . ,. t ...... • ... .cflld-.. .. 1 .-,J4 J ., , _,. ..... ,::( dA1
111 'lf1mt11J1tlU~a -11'U.1'i tJ'U.111 f1 ~ u1::H-1f'IL Vl£lfln1~1tl'Yllin1::~'U.n1~~1tJ"JJ £l-11J1tiU~a ~ 'U.H911'Yltla£J-1"JJ £1-1f1111Jf'I111'Yl11H1'U.~ H11"JJ£1-1 .... ... A A AA o A A... - • - .... ..l..t ._. .... A A
H1~Hnt~mEJL£Jlla£J::"lfL91'Yl1l£1, C. cf comosa Roxb 1tin1~fl1l\U,n11 .. 'Hlll: Vltii'U.1fl1~1Jf'l~li'Yl11H1~Wntlmm£Jlla£J::"lfL91'Yl1JEI-1 C.
cf. comosa Roxb fl1111Lilli'U. 5%, 10%, 15% ua:: 20% w/w 'Ylt1H£l1Jfl1111fl-1tl1"JJ£J,thi'ufl~llfi1-1 ") fi1tJi~ heating-cooling
cycle 1h::Lijum111fl, fl1'i1 ni mfm::m, m um'Vl m111Lil umfl-fi1, m111~ Uti ua::~1m111 tU.Hl~thfi'tylfflm, ~al uthi'uti'1 tJ
High Pressure Liquid Chromatography fltl La an thi'ufl~lllf Sjm11lf'l,1fl1~ 111'Yl t1 H£11J q 'Ylim::iu.m1~1 U"JJ £1,11J1tl u~ a h • tfflf'Yltlfl£1,1 1t~tJ li'~'U.Hltl~Wrf C57BU6 £11U 8 tf'tlm\{ L'i1:: \u~i,H£J-1"Ji'1,1\iLi)"U.11J'U.1tiLa'U.~hfiU.UOa1,1 2 ih'I~Lllfl1~1tl metal
'U , , ,. • ,.
ear punch m~11m111tU. 10 mg lf1u~u~L1tU.U1t~ur.~a 'Yln1u1ua:: 1 flf, LiJU.L1fl1 30 1u 1fltJml'l"li'1,oU1tJi1tJtl1fl11Jflllfl£1 11 , 'II ,
cream base m~"Ji'1,"ll11i1tJihi'uf'l~lllfSjH1~Hnfl (n=6/mi11) uuiinmw\u~ ua::lt11JU.1flLH'U.~ht'I'U.Una1,"JJ£1,1J1tiU~a LLHtl,1~fl ,. , ,. , il
.. .:'d', J .. d'J, ... d L \.lo~£JtJa::•t1fi,1011~1U"JJ£1,1J1flLLf.la ua::f!11v'JI'U.L'U.£1"JJ£1HlJ~ L'Vi£1~WU11iHI11'Vi"JJ£1,1L'U.£1LU£1 ~'U.1'U.'Yl 0, 2, 10, 20 f4afl1"i
Flmt1='iiu: th~ufl~lllfSjH11Wnt~m111L"Ii'11-llu 5% ua:: 1 Oo/o w/w Sjm111fl,1fl1~ni1vh~ufl~lllfSja-11Hnt~m111L"Ii'11-llu. 15% LLa::
20% w/w i,amtm::m,m umw ua::m111~ ut~ a1U.fl1111 Lllumt~-fi1,ua::tl~111 ma-1~~1fi ru lffl,L ~ ~a'Yl mi1~ul~Sjm111 .. . • "' A 4 A ..., a ttlta ..., ""' " J' ,... 'I &... ~
U910fi1,10'U. 'i-1LS£10YI111Jfl~11'YI11H1~HOtlf'I1111L"JJ11"JJ'U. 5% LLa:: 10% w/w 111'YltiH£11Jtl'Yllln1::Vl'U.n1~~1tJ"JJ£1,1lJ1tiU~a ~ U.Hf11'Yltliil£1,1
l'llJi1thi'ufl'111ilijH11'«nflfl<111~'1i111iu 5% ua:: 10% wlw ii~aua::"lle..,m~l'I1U"lJthl1J1flUI'H'IlnaLfi u~nu. tie ~ewa:: 66.67 LLa::
62.12lu1u.lf 30 fl111thiu ~,ilni1tl1f'I1Uf111£JU1.1SjutJ~1fi'rum,wi19i (p<0.05) ~an11f111'iiudla"JJa-11u~VllJi1 Lilmfla . .. . "" d " • ..., a da ""'" .., ., d ,;. A' ~ 4 .a.
1J1L 1tll.lJ1flU~a1JEJ~~\t 'YI'YI1t11 tlfl1~1Jf'I111'Yl11H11H0tlf'I1111L"JJ11"JJ'U. 5% Uiil:: 1 Oo/o w/w 11011LW11"JJU."JJ8,1L11t1LS£1ti"JJ11'JI'\.Iofl PMN,
fib bl A tl A ... o ... .. d d ... ., A A d
1 ro ast, collagen fiber LLa::~aatnaat~ ~1 uan1"i'l\lll: fl111Jfl111'Yl11H1~Hntlfl'ltJL£11iaa::•JHfl'YI"llil,1 c. cf. comosa Roxb n
fl1111L"Ii'li"Ji'"U. 5% ua:: 1 Oo/o wlw Sj ct'Ylin~::t1u.m~~1 tJ"JJ a,u1t1 u~ a l"U.it~1'Ylflaa, Sjt1ntJm'Vlhm~ll1 lt1~1111u.1 LilutJ11 ~ ~1 u • £l'U.1fl91
1 ... ~ ... - -fltll::lllft'lff118fl1 11'ft11fltnav~uan'llli1'14 2.. - f" ... ... ...
1nV1ftVILVtnVfl18fi11La:n11R1li11t\Linl 11~11YIV1ilV01Jan'llli1'14 . . * Q I .... ~
ilflfiOt;!'llYI'Illi: [email protected]
·-
The 7'" Annual Nollheast Pharmacy Re:sean;h Conference of 2015
"Challenges and Opportunities of the Pharmacy Profession in Post-M~Iennium DevelOpment Agenda"
Faculty of f'llarmlla!ullcl Sciences, Khon Kaen University, Thailand Jll!~
102 lllil! '!I lil'illldlll Ill 1 ttwrrwrwm r
Abstract
In vivo Wound Healing Effects of Formulated Creams Containing Ethyl Acetate Extract of Curcuma cf. comosa Roxb
Pitchaya Samanmit', Settiyanee Duangjai', Rattana Leksomboon2, Preecha Boonchoong'. Warisda Sila-on', Chutinun Prasitpurlprecha'•
/JPS, March 2015; 11(Supplement): 54-60
Introduction: Curcuma cf. comosa Roxb or 'Wan-ma-wor" is a traditional herb that commonly used in Northeastern
part of Thailand and Cambodia for the treatment of wound. The objective of this study was to investigate for in vivo wound
healing effects of formulated cream containing ethyl acetate extract of C. cf. comosa Roxb. Materials and Method: Extract
was formulated to creams with the concentration of 5%, 10%, 15% and 20% wlw. Those formulas were tested for stability
by heating-cooling cycle method. They were evaluated for appearance, pH, viscosity and amount of active substances
remaining in formulas by High Pressure Liquid Chromatography. The formulas having good stability were chosen to study
for in vivo wound healing effects. Both ears of 8 weeks old of C57BU6 mice were punched to make a hole with 2-
millimeters of diameter by metal ear punch. Creams were applied to wounds once a day for 30 days. Left ear was applied
with the control of cream base. Right ear was applied with formulated cream containing ethyl acetate extract of C. cf.
comosa Roxb (n=6/group). Wound healing effects were determined by taking photographs and measuring the diameter of
the wounds that be calculated into o/o of wound healing. The pathological effects of histology of these ears were also
examined at day 0, 2, 10, 20. Results: The formulated creams containing 5% and 10% w/w of curcuma extract had the
appearance and viscosity better than that of 15% and 20% wlw. But the results of pH and amount of active substances
remaining in formulas were not different. Then 5% and 10% wlw of curcuma extract creams were chosen to study for in
vivo wound healing effects. Results showed that 5% and 10% wlw of curcuma extract creams had indifferent of %wound
healing about 66.67% and 62.12% at the day of 30, respectively. But both formulas had significantly wound healing effects
better than the control (p<0.05). The histological effects expressed that tissues around the wound area of treatment groups
were found PMNs, fibroblasts, collagen fibers and vessels.· Conclusion: The formulated creams containing ethyl acetate
extract of C. cf. comosa Roxb with the concentration of 5% and 10% wlw had in vivo wound healing effects that might be
developed to a potential drug in the future.
Keywords: wound healing effects, formulated creams, ethyl acetate extract, Curcuma cf. comosa Roxb.
• Corresponding author: [email protected]
:=1 • .¥ d ..
Curcuma cf. comosa Roxb Lu'l.l11'\.U'I'I.H11D.:J 1.:11'1
Zingiberaceae ~~D'YiD.:~riui1 "i1U~11TlD" ~~D "i1u~111 U1" am~tU:Il1 [JUDn~f11111fla1 [JtlS.:JnlJi11.li'n 11iH'ln {C. .. comosa Roxb) utim1L;[J.:J~1tllJ'JJD.:Jil1fla1DlYlti'uflmh.:~
v1ni1ui'mJflenm.h.:~uau 2 ti1LL~\4.:~ {fl11vLDnan'!Ym
tl1Un11iLfl11:l\ DNA 'JJD.:J 18s ribosome 1flu Professor
Katsuko Komatsu, Toyama Medical and Pharmaceutical
University, Japan) (Teerawatanasuk, 2003) ~.liiu~.:~H~D .. ... l11.:J~'Hln1:tf'l1tffl111 C. cf .comosa Roxb (cf = look like)
rit~um,-V.a~u.Yhnnu~ulfl~~ .. , '1!'11U1uhi-:~~1fiDlJ a11'151i1Ulla:lh:ll1f'ln11W'Il'1 , ,
~EJ3Jl\1 C. of. oomo~o Roxb a.nH1m:t11J1fiU~H~ LfiEJH1U
l~S1Hfl (rhizome) li]uLL~UlJ1.:J ') ~U1DlJlJ1fiU~a~Lnm1n
-55-
'Jla.:~nfllliflan LLa1~uti1mhtlm11J Yi'ArLu'l.lna1 3-5 1'1ol J tJ . J ~ .. - .. .. L11DL fiDDnfl ';I::Wl.l111tl[JLL~l'lL'Il'D1JnULflfl L~atlLW[J.:J')tl[J .a., .. j' d
Ufi.:JUa: L111Jn11flfiL'tffl (Prasitpuriprecha, 2004) 'lf.:JnU.:J1U
n11iiU'JJD.:J Panchatcharam (2006) Wl.li1W'Il'fl1:na • • ... tr;" .J' J ~
Curcumm Yl11~ l.l1fl LLf:.l a~1 u L 11'JJU~1 n t}l11in11n1:flu , collagen u a:: a fl reactive oxygen species f:.l1 i [J l flij n 11
.r .. J .c ... .. 'Ylfltffll.lLl.lil.:JfiULW£1~1 t}'il'lin1:f1Un11~1[J'Jl £l.:Jll1fl LLf:.l a 1 U , ~aDfi'YlflaD.:J'Jlil.:JH11ffnfi~U11.l'JJD.:J c. cf. comosa Roxb
Wl.li1 ff11ffnfl~q'ilfm:tium1~1U'Jla.:Jl.l1fiLL~a 1MH11 , .... ... A A dJ' .,
ffn flfl1 [J LD'Iil'l D:'lfL fl 'illl t} 'il1in11n1:fl Un11~ 1 [J'Jl D .:1 ll1 fl LL~ a
~ni1H11ant~ti1mh. L1l~aut~anan~a LLa:LnnL'If'l.l fi.:~Ju «-Ji [J~.:J tm 1 ,.,~t.IJ 'lol1 fi111.lfl~1J~ ij H11Hn flfl1 [J LDb D:::ftLfi'Yl .. .. .c ... 'JlD.:J C. cf. comosa Roxb LLa:f'lnH1t}'il1im::~Un11~1U'JJD.:J
ll1fiU~a1uifi1'YlflaD.:J'JJD.:Jti111.lfl~1ltl.:Jn~11
The 7" Annual Northeast Pharmacy Research Conference of2015
"Challenges and Opportunities of the Pharmacy Profession in Post-Millennium Development Agenda"
FacuRy of Pharmaceutical Sciences, Khon Kaen Univeil;ip Thailand
llllillRR' II' ___ EE_il&-Jli&IIII!III!IIIWII~~&di! If J F ~ii!::i!!l!illlllll.lllliii'Pa!llllllliiiJ!!flll!l11jR1PiXWilll'ilil!!lllilli..,lill•ll!li!llilll'lli!IIIIRIIWIIIIillllllll"!lli!lll!l!illll&lfi!U!'<! .. IIIBIIIIII
d A4.Q....,
"i::ltnJ1J1fi1'il !.1
211"i21rlfliUJ'"lrn '
tn"iifrlflfl1tltl1'~ha::a'ltJLEJiaa:::aLfiYI"'IB~ c. cf.
comosa Roxb \ti~1nmuntHhml'IS1 ti1v1i percolation
tn"i'Rrltlitltiii o/o yield = 10.24% wlw .. .., • .., 4
fl'l"iih1fl1"iUR'UI .. ,J... t . 'd Lfl"i£lll cream base Ylll H1 'I.UHUI"JJ a~ s eanc act ,
glyceryl monosterate, cetyl alcohol, cremophor A25,
silicone oil, isopropyl myristate, carbopol ullrez, propylene
glycol, triethanolamine, germaben II ua:: a- tocopherol
Ua::Lf11£Jllti1i'Ufl1ll~ ij R1"i'Rrlfl fl'liU L Yl"i lt~ [I~ Hllfl'1"i'Rrltl • nu cream base Ll'lih.fill1tll:tl11~L'li~'liu"JJEI~tn"ilUltlLfi1nu 5%, 1 Oo/o, 15% ua:: 20% w/w
n1"infl21uUR11~R~~~"JJu~~1i'u - • - ... .ld - ....
Ylflfl'CIUtl11lltl~f11"JJEI~fi11Utl"iliYI~fl'11Rnflfi1U Ao A ,J ... ... I
Lvlav::'lfLfln"JJa~ C. cf. comosa Roxb Yltl11liL"JJli"JJUf11~ ') A I Ad •
ua:: cream base Lllv~1Un"i"i~11 Heatmg and cooling
cycle "1 Tt~mnut'hi'utl~llluimutl~EHU l'lnD (Heraeus, ,. , , . Germany)~ 45 "C U1U 48 i1 t~~ LLa1~~'1hLtiLJiulufiLUU ~atU.l'lnD 8 "C U1U 48 i1t~~ titu11u 1 "iEIU ,,,a; 10 • • 1au LmtJlJLfi UlJnUthi'Ufl~~~ Lnu~atU.l'lniiia~ (25"C) . .. lw~::u::na1Lthnu 1.h::Liium1~mi1"11E1~thi'utl~~ i~d
... ... - .l" d - amtm::tn"n1tJnTW H~Lnflam;m::LUvtl"ill m1uun
~ 111/J ~ 'lfuua::t~nfl::nau " ua::nau
- A11~'Hitfl 1fli1U Viscometer (LMS Instruments)
- A11~Lih~n"ifl-fi1" (pH) 1flfi'1U pH paper 'i!1~ pH
3.8-5.5 (Macherey- Nagel, Germany)
- tl~~1fl'it1"ithA'!J 1Lfln::il'l1tfi~1tu."JJEI~tn1Rnfl
i1maiaa::tHLfl'rl"JJa~ c. cf. comosa Roxb luthi'utl~~~1U 4
Lfl~tH High Pressure Liquid Chromatography (Waters,
t . 11 ,J USA) tltl01l'IUflL u marker n 1-6 ~1n Chromatogram
"JJa"R1"ifl'fltlf111liL'Jili'JiU 0.5 mg/ml ~tl11lltl11tl~U 254 .J
nm ("itln 1) •
• 14 1 ""' ... A A JlJn LLfl'tH Chromatogram "JJa"R1~Hnflfi1ULalaa::IJfLfll1
C ,J "' "' "IICI~ . cf. comosa Roxb l1f111liL"JJ~"JJU 0.5 mg/ml
-56-
LLRfl ~~a 1 w~ll o/o Recovery ~ ~ d] u fa t1 a ::"II a .J .. t1~111 tU.«1"iR1tl ru~ wu 111 Ul'l a~';l1 n Ltl~u 3J L11uthi'1Jfi13J
Lii vunm.fill1 cu.;1,.ihtl' '!! L~3J tiu ~1«1 uth i'ufl1ll f11u 1 cu.
-;nnafl"i .. o/o Recovery"' lA111W.R1~ihA!Y~I'l!Jl'lol.JLti1t111LUUthi'!.lfl~11 x 100
tfi111tuR11ihAIYL,11tiu{\Muth~ufl~11
d 11 ~ -~ • u ua:: % Found 'lf~L u~eua::"JJEI~1J"ill1tli.H1"ifl1tlty
~wuluvhi'ufl~ll~~hum~"1 heating and cooling cycle
Lll~umfiuunmhi'ufl~ll~ Li)u~ a ml'lnijia~ (25"C) 1 u . ..
%Found: m111tuR11i1filJI1Ufl'i11~U1U heating and cooling cycle X 100
m111tul11i1f1ryl"fl'l11~Lnu~~w."~iii'o.J (25"C)
J "" I) .... 4' f)nfin"i::fJ~n1"i'H1U"JJ iNU1 fl ue.~a L ~it fl1n fla EJ~ .... .. itfl1nflaD"
- .- ....... ., r l'IU mice R1tJWUI C578L/6 ci1U 8 «ufl1l'l "1Ja';l1n .. . .
f3UUHflf'rlfla a~LL ..;~'lf1fi Lifu~~i e~~tiDUim"ii'frfn fl a a~ ""' ~ A - d
"IIEI~tl tu.::L11R'lffi1Hfl"i lll'I11Y1 m auaua~1'lfi1U RllTl::m~ • ,:¥ ...... .. ..J A
UW~f11llll1fl"il1U"JJEI~I'IUU fl'tl1Ylfl ~ El~ YIEI tU.l'lllll 23±2"C .., ,. , 'V
m1ll.:aui:JJ~l1'1f 55-60% 1 iEI1l'I11Lijflif1 L~'im..lua:: ti,i'1 • f1 a a fl n a 1 t t1 u "l fi' i' u au ~fi 1 il i i' t1 f n ., a a H 1 n
'
.. .l -~ua"i1'lfi1U La"JJl1n1"ia~llfl 7/2555/research)
(Clark, 1998)
RaUl'\ U~1 u U1 Ufl Zoletil (Virbac, France) n1 fl11ll • A 1 ~ .. 1 U ..
H::El1flU~L1nL Ul'lfl1tiLLEI~nEltJEla L'il1:: U~l'l~REI~"JJ1~ .. .. u 'i L 1 tU. n a 1 ~ l'll i L 11 u ~ ti 1 EJ metal ear punch (Fisher .. " ScientifiC, USA) "'IU1flLiu~hfluuna1~ 2 ij~aL~fl~ "l1itlfl .. LL~a ua::hilit11ti1u'ila:awflaaflm"inflaEI~ mtl~lllfill1tU. • 10 mg ~1ul'lu'inmu1fiLL~i'l 1ua:: 1 flf~ nn1u L1luna1 .. . 30 1u lfltll11l'l'li1~'1i1Uti1Utl1tl1Utlll ~El cream base 'rl1l'l .. . ..
" "' e ...., A tJ~ ._, t
"111~"1111fi1Uf11~Umlln~H1"i'Rnfl (n=6/n~ll)
m~'li1~"1111ti1u • - .d dtt:t f11"ilJtl"illl1liR1~ fl1 U cream base
Rnfl
;
The 7"' Annual Northeast Pharmacy Research Conference of 2015
"Challenges and Opportunities of the Phannacy Profe55ion in Po&t-Millennium Development Agenda"
Faculty of Pharrnaceullcal ScienCeS, Khan Kaen University, Thailand
lllliiiBIBIII ________ I&IililFI!IIlli' i!ll"illlMIIIIII,..iiiiJIII l!llllllilllililii l!lil\illiiiT771liiiiiZIIII!iiFiilli!Uli~~-· il T 'Ttl 7Z~i 1U 51 d .Zll lllli
m·nh::lil\\n1"i'H1fJ11ih11J1tlLl~il - .d: .... J, ... u-unnm~an~m::n1tJOUan'lla~lJ1tlLL~an Lll'l'l ua::1f1 •
'll'U1flLiouf-i1fiOUUna1~'lltl~ll1fiLL~ail1 EJ vernier caliper 1fl ~ ~ .
•1h 3 A~" 1wr-u~ o. 5, 10. 20. 30 ti1ou1mfhi"aEJa:'lla-1
n1"i'I'I~£J'lla~lJ1fiLL~a (o/o wound healing) 'i1n~tl,-
o/o wound healing = diameter at day0 - diameter at day. x 100
diameter at day0
L~D diameter at day0 flil LltUU1fiU!i'na1~'1lil~lfltlLLUi1LUiU~ 0
diameter at day. f\a Liuu,;u£fmn~'llii~U1tlLLUi11uiuil n . .. ..r
fl1"itl"n\Jih~LlUlf11-t Histology (Deodhar, 1997) .:- ..r 1 A A ..r A ,
tl"i1'i'lf\U 'UEJ'liEJ~ ll'l'l I 'I-'4Eltl'I"HJ11ift'n1'1-'4'll EJ~I 'Utl LtHl L 'U ~ J • '"., ..r A "' 1'Ul'l 0, 2, 10 ua:: 20 1tlEJitl"iEJ:UI'UEJUJEJtl1EJ Paraffin
. ,. ~ .¥ A .., A techmque f'IOl:nan~~meUJEJfi1EJLfl"itl~ Scan ~ope CS
(Aperio, USA) ~'i1"it\1.1n1"i'1'11El'liEJ~ll1tlU~a'i1nn1"ii~:U~'U 'll a" Polymorphonuclear leucocytes (PMN), Fibroblast,
Collagen fiber ua::'l'laat'llaaflll~nm1nall1tlu~a ua::faEJ .. , • .,. 0 .. d ... ~ .r
a::'!la~m"ia""i1,'1L'I'I11'lla"'l'l""m'l-'4"i1 'll~'i:::Uu"i111tli.:U1n'llou
tl111"i::£J::m~1 (Midwood, 2004) A f' AA
n1"i11A"i1::'Hfl1\ltHlil
LLft'fl-1~aLiloufi1 mean± so ua::11fl"i1::ifl11:U
II t1 n fi 1 "l11 ~a- ii 9i il1 EJ Independent t-test 11 a:: Mann
Whitney test EJ a11~1J"i::ilu fl1111 11 t1 n til ~l11 ~ a-ii 9i mh ~n ~ • a.. d
OUEJft'lfltyl'l p value < 0.05 (*)
~am'l;in
1. m1aJA-til1'21il-tih~uA"i&~~iia1"it'lnflti1mnfiiiEJ::~ Ltlfl'2lil-t C. ct. comosa Roxb
1.1 an~lll::fl1..1fl1Sn1Yi a.. ~ ~ d
an~m::m~m EJ111'1-'4l'l ~m111 L '1'1 a1'1l a~ 1 -uam11 m"i
LLEJn.ffou flT"itlfltl::naou a ua::n~OU'lla~th~llfl111~ija'1"ift'nfl .... A A J " .... mma1iaa::'lfLtll'l'lla~ C. cf. comosa Roxb l'lfl1111L'Il11'UOU
5%, 10%, 15% ua:: 20% w/w L~mh-unwrh Heating and -~ .d .... ..:: ... .ca .I 111 ,l - ....
cooling cycle LlJ"itllJL 'YI VlJn1J••1"illfl"i11'YILnlJl'latll.'l'ln11'1'1a" • • 1
J • ... , J .. 0 .,. .. .~ .. 'U"i::EJ::L1ft1'Y1Ll11n"U Ua'tl~~a L14tl1"i1~'YI 1 'IS~tl1"i1Jfl~11l111
ft'1"ia"lltltl1EJR11"Ul'l-'4"i~fl1111L"li11"liou 15% ua:: 20% w/w 'l.li Q .... , ~ " 11fl1111flr3tl1'YI1~mEJn1'1-'4 ~1Jn1"iLLEJn'lfouLLa::tlnfl::no-u1'1'1 1 ~ ouLllou'itl'll11n"i::~1tJ1 "' dafl1ll tbouiLLa::n~ou'lla~th~u • fl~11 ~:: L ~11~1491111 \B111 mfl1111 L "Ji 11"Ji'U'll a~ a'1"i«nfl a'll14 L 'l-'4"i •
1.2 A113J'H\itt
fl1111'1'1 i'lfl'll a~th ~u fl111 ~ ii a-1 "i«nfl il1 mala o:::nLfll'l J ....
'Ua~ C. cf. comosa Roxb l'lfl1111L'Il11'll'U 5%, 10%, 15% A
ua:: 20% w/w L11af.i1"Un1"ilh Heating and cooling cycle
LmEJm ii EJunuti1i'ufl~1lyf uiu~ am 'l'ln iii a~ 1" "i::EJ:: 11 a1yf . .. L Yhn-u 'l-'4ll'h 'l.liLLt~nfi,_,n-uar.h~ii\tmhfl'rum""'Hi 1fiEJ ., fl1111'1'1 Ufl'll a~th~1Jfl~11 ij LL OU11 U111'1fl a ,'I ~~ aL~ 11 f\1111
L"lill"liou'lla~ft'l,.«nfl«~"'~,. quy{ 3)
l:- m ~----~~~ ---
' ....... .. -.... '
r-----~·~-~-~·-..... I ,_ . ---·------~"'-·-- --
1-1-------t-
:~ I ··.11
• L,__ __ _.,_ ____ ___L.,_ ___ _.__ __ _
I a • • -IIIINl
.• .~ 3 .. • ~ .. .... - -;'jiJfl fl11:U~\4fl'IJ!J~tn~1Jfl~:U'YI:Uft'1~Rnflfl1!JL!l1ililil::'lfltl'YI'IJ!l~
C f. R b .l ~ ~ ' J .I •
• c. comosa ox 'Yifl"l1:UL11:U11\Lt11~ ") L:UEIH1lLn1'nn Heating and
cooling cycle LmuuLiiutJn1lthi'tJA1:u~u'itJ~Ot\L'I'II1iiia~ (25.C) . .
tl1"i1..1ii 1 .!';! ... ~ .,. ... J .. LJJ"iElllll'IEJllan~m::m~mEJn1~'Ua~ti1')1Jfl"i:Ul111ft'1"ift'nflil1EJ ethyl acetate 'UEJ~ C. cf. comosa Roxb A • .l A -.-~A
111mnlll'l~tli.'1'11J:U 25"C (A) nllfl1"illfl"i:ULliCHhoum"i'Yh Heating and cooling cycle (B)
tnll'llll thfufl;aJYILnUl'lfliii'II-Qii 25•c (A)
11111rlfl i lllthi'u
f1111JI'IIi1 n1111VO ni'll fl11all'llil1
'll!l~f11aJ i'll .r
'llii~L\lD
(o/owlw) fl~IJ
0 '1111 t'llli!!'IIA l'lliiD'IIA l'llii!l'IIA l'llii!l'IIA
5 + ,r,,nalia'll + 1'1\U!l'IIA L'I\U!l'IIA 1'1\U!l'll A l'lliia'IIA
10 + Jl1tna + l'lliia'll A l'llii!l'IIA I'IIUD'IIA l'lliJO\lA
15 H ,r,,naL'Iia~ + l'lliiB'IIA 11vni'll ui'IILiJ'II>f1'11111~nli'ao ihillil''llni1 A t'lliia'IIA .:... ,¥ _,
m:•1Dl'l11'11!lll1lJ
20 +++ Jl1111ill'lilJ + l'lliiu'll A II!Jni'll lii'IIIU'14,f1'1111'h'll1'14 ihillil',mi1 A l'lliJi1'14 A .!. ,¥ ..
lJ1nn,:>1!Jl'I11Uilf1,U
'H&I A • ~ J ..r 1liL'Hfl L11EJ'i1"1'U + LLa'fl~"i::fi1Jl1111n"ll"U
-57-
The 7"' Annual Northeast Pharmac;y Research Conference ol2015
"Challenges and Opporlun~ies of the Pharmacy Profession in Post-Millennium Development A9enda"
Faculy of Ptoannaceutical Sciences, Khon Kaen University, Thailand
J II iilaMTIMRBillllliilllllllllliiMIIIII'II1lln~lfll!llltvll n · rrmrmx Di!iii ili!IW',.H1D6Rn
iJ I
1.3 A,1a.n ~mA- f11.J (pH)
fl11l.l Lilun~fl-fi1-3'1l u-3th1'1J fl~l.l~ ij R1Utlfltl1 u - - ~ .., ... LuJaa:'lflfiYI'll£1~ C. cf. comosa Roxb Ylfl11l.ll'IJ~'11U 5%,
10%, 15% U.ft:: 20% w/w Lrl El ~h u n1 ~~ 1 Heating and
cooling cycle L1flD1Jlll!JlJtllJf111'lJmlJ~L01J~£1tU.~Ilijie-3 . .. wu-h1liu.flntt1-3rlU
100.00
50.00
0.00
% ReOO\Iefy
~---~·--~.----~--~~·--,-~ ':. - .1:: J:: .. ·.:r: .11 .. :;· . ~·j Marker 1 Market" 2 Marker 3 Marker 4 Marker 5 Marker 6
•s% curcuma cream !110% curcuma cream
1115% curcm~a cream •20% curcuma cream
Jtl~ 4 o/o Recovery 111l~tn111Cli.IM'I'mi'fl~Vi111l1Vl'IS~n1'1' Ul~vuthi"ut~;u,h~ 'l L~llLYiuunumu1mW1'1'i1t1cy L~uti'"~M'"thi"ufl;u
%Found
140.00
90.00
40.00
-10.00
• 5% curcuma cream D 10% cu-cuma cream
a 15% curcuma cream •20% curcuma cream
Jll~ s o/o Found 11a~m111tl411a~~n,.anfl~Viul'"'h-rumlJ~ 1-11'"
n1'1'lh heating and cooling cycle LlfiumYiuunuth-rufl;ll~ .. J. -~ t .~
LnlJ:ri~CUl'I.IJliM~ (25'C) '"'S:U:L'la1Lmn'"
"'U.l 4 ' cu • " a .la ,.. ,: Yl YilJ11 7o Recovery "!Jfl;Jfl1~1Jfl'UJYI1Jft1'51fnfl
fl1111L"li1liuti1-3 'J ~fh lnaLfiv~nu ii1u·n.Jif s YilJi1 % .. Found "!la~th11Jfl111ifijff17fftlflfl11 ~Li11'2i'Uti1-3 ') ijth
lniLAU-3L"liunu
d ._, d d e .... ,::& dd .., ... A A
11fl1111fl.1fl1tl flu fl111Jfl71JY!11 R1'Hmflm tlLClli«B:"li'UlYI
"!JCJ-3 C. cf. comosa Roxb ~fl1111Li11iu 5 % LLfl: 10% w/w
J "' 1 ..... 111'Yifl Rfl1JC)Yllin~: ~UnT')~ 1 U'll !l-3lJ1 fl LUHI 'Ufffl1 Y1 fl fl a -3
fi!lLl.l
~58~
J .. 1 ..... 2. l]l11ifl~::fJ~flTi'H1V'Vil"lJ'lfiLUH~ \oU'ftt,nflau.J
" .l A 1 'i1n~m:tm:rnuuBn'lla-31J1fiU.~H~Yl1J'H1tU. 1J~11a-'~U .. ..
d ... e .... .. jp~ .... " A A
YJnnmtnVfl17lJtl111'Yil.lft11A"Mlf\1ULaliae:'II'LtiY!"lla-3 c. cf. .. .l "' .. J. ..
comosa Roxb Yltl111Jl"JUJ'IJU 5% Ufl: 10% w/w LlJOLl1tJ1J .... I ~ .. • ...
n1J cream base Vi1J'l1 "11U1f\"!JEJ-31J1flll~Hmnn'Y11f\1Vfl111J .. .d da - A « A a ..,
tl1l.I'YI11R11Rnfl 1:11"llU1fiLflnM L11al'YitJ1Jn1J cream base
.l :'1 ... .l.l tl.l Y!LuUfl1tl11Jfl11 l~uYI 6 LUI: 1 'Y1 7} , 'V ..
.. '~ . (A) DayO (B) Day 30
~tl~ 6 1J1flU.~-Ja~lu11a~y,'" Ldam1Ul'l'lh~1111fl1tlthiufl;uilu u .. ..
81'Sft'nfl811'"1l'l'Sfl1111LilliU 5% w/w LLa:m luYiiWrltl • • ~ t~J. ~.J mv cream base '"1wn 0 (A) u.a:1wn 30 (B)
(A) Day 0 (B) Day 30
_,J. 7 .. t J. 1 ~ ~ ~ ...... Jun 1J1flU.~-Jan 1J11£l~l'l'" Lllam 1Jl'l111~1111fl1Vthmfl'l'11'1111 . . W1'1'anflft'll'"tVi'l'fl11lJL'lilJ'li'" 10% wtw u.a:n1hJY1'1i1~-ilu . . ~ 1 ... J. .....
fl1V cream base '"1'":rl 0 (A) u.a::1'":n 30 (B)
100
* Ol 80
-~ * 'iii * Cll 60 ..
Bl .r: u c 40 * *· :::> 0
nil ?: ~ 20 0
0
10 20 30
Day 0 c .. om t>ase(No9a!M conuct) r.a 5'!1. Curcum • cream g 10'!1. Cur<urna aeam
'I'll~ 8 'Yo w d h ,. .. ~ • ~ .. J... ~ ~ u • oun ea mg 1lll-1YI~n:n1fl1Vt11'1'\Jfl'l'lJ'I1111M'I'II'nflfl1V
.C. A ,l ~ _.. La1iaa::'IILfll'l1la-1 C. cf. comosa Roxb :rlfi1111L1111'll'" So/o,
10% wtw u.a:: cream base LU'"L'la1 30 1'" (' = p < o.os LYiuunuti'1muf11J)
•
The 7"' Annual Northeast Pharmacy Research Conference of 2015
"ChaDenges and Opportunities of the Phannacy Profession in Post-Millennium Development Agenda'
Faculy of Phannaceutlcal Sciences. Khon Kaen University, Thailand
111n~1.l~ 8 fhi'aua::'1Ja~n1~"'1U'11CJ.:JlJ1fiU~H~ (o/o • ..:.¥ J...:.Kt J. wound healing) 1::LVUJ11Uf112J~::U::L1S1'YILW11'1JU fl[J..,~'YI
mti1 u thi'ufl'iJJ~ihrmn1fl 11::iifi1i'e u a::11 e.:Jm~..,, U'11£l.:J
• J. .. .J.i'J .. lJ1flU~Sll1nn11"'1,t'YI'YI1fl1U cream base 'YIL Ufl1fl1ll~11
mh~ihtmtuity'YI1.:Jwii~ (p<O.OS) LLfiL~mlfivuL fi vui'eva:: ~ .., • ..., a da
11 e ~m ~"' 1 U'IJ e ~u1 flu~ a'lJ e~"' u n n1 fl1 u m~u fi'UJ n11 1f1~ • tillflti3.1UlW~fl11llL'Jill'JiU 5o/o Wlw JllJ 10% wlw WlJi1iJfh , 1naLAU-'!JlU
.. J ... .J: - d d~:t - d ... ... 'DEl-'! LlJ"''YI tln'YI1fl1 Ut~1~1Jfl~ll'Yillif1~1fnfi'Yif'l11lll'1JJJ'1JU 5o/o, .. .
, .. .l • 10% wlw ua:: cream base LU1U'YI o. 2, 10, 20 Wll11
1.l'ill1nUiifiLaE!fl'lJ11'1fUfl polymorphonuclear leucocyte
(PMN). fibroblast, collagen fiber ua::..,fHJfiLaElflll'imulna
mt~u'-ls iithlnaLAU-'!nu ufill1nni1i1fi11Jf'lll (cream , base) R1ui'eua::11El.:Jn1~Hi'1-1 1 "'~11a.:Jwlt~rhwi'1ilf\1hi
LLflnfi1.:Jnu tflu1u1u~ 10 11::iim~Hi'1-1wlt.:Jn1wi'1ifuhui • <' .I J.
£l[J1.:JHll1,J~nL (l1J'YI 9-10)
"" .1"! .. .. ..¥ "" .. Lll£1 LIJ~EJlJL 'YIUlJWU11iH1)1W'lJ£l-'l L U£ll[J £llJ~L 1 t\L1J1f1Uj:j a
'11£1-'~ 1u"' "'u ~ n nY11ti1 uthi'uf'l'i11~ ii «1~«nflti1 u La ia e::.ii ,. .. ,. J. .. "
Lfi'YI"lJ£1-'1 C. cf. comosa Roxb 'Yifl'l111L'1J11'1JU 5%, 10% w/w
~~::U::L1a1fi1.:J') W1Ji1 1.l'ill1t\LLiiflla£lfi'IJ11'1fUfl
polymorphonuclear leucocyte (PMN), fibroblast, collagen
fiber ua::..,i.'lElfiLa£lfllJ'iL1tU1nalJ1fiLLj:jfl iith 1naLAV.:JnU uti
111 n n ·h i1 fl11J flll (cream base) "1 u fa £J a ::"lJ a., n1~ , Hf1.:Jh1i'1Je-'l"u"rhwi1iifi1l~utln~1-'~rlU tMlu1u~ 10
";J::iim~Hfl.:J"' U~n1Wf1ifu1"' 1i £1 ti1-'1HlllJ~nt ,.
-59-
(A) (B)
(C) (D)
J. .¥.J. 1 1"J ,-t.Jn 10 Jl1~LU!JlUOU1"l1tulJ1flLU:Jii1Jfl~ 1J1'1 (A) U1U'n 0 {B) n1 u •
..... .. • .... ... J,. "" f11U cream base (C) l'l1fl1Uf11,.1Jfi111'Yillln,.ft'nfl 5% wlw (D) l'l1
il1uthi'ufl';ll~ii~n,-ft'nfl 10% wlw 1u1u~ 20
oihJ-nuua::ilf·nJ~an1"i;iu '
.... e .... d dd - .,_ A A
n1~Wtll U1 f11~1Jfi~11Ylll H1~Hn flfl1 U L£11ii.'l £l::'l1~f1Yl'11£l.:J
C. cf comosa Roxb liiif'l111lf'l-'lfl1'1JEI-'Itl1i'1Ji1 fie-11-i
tl'ill1 t\L «1 ~Hrl fl ~ fl11ll L 'Ji ll'Ji'U 5% ua:: 1 Oo/o wlw ttw Sj .... .ld d. d .¥ #:1 ~~~ I
i.'lnl:it\L::'Y11-'In1U1)1ViYlfl f'l£1 llfi1111L"'i.'l11J£l-'ILUE!fl1ll Lllll1n
l~iiuun-iu l~t~nt~::nau utiiiiua::n~u'1JEJ.:JH1nnfl «llULW~"lffiL'iU lhUfl11ll..,Ufl fl11llLUUn~fl~1~ ua:: , 1.1'1111 t\La'1~i1fi' ty 1 uthi'lJL~Ltl~vuuua.:JluL~£1~1UnTnh heating and cooling cycle
.... ,Jl,.., .I .f " ~1U1';J[JU 'lfl1JU1JlJ'1J£l-'ln1~YlfiH£llJ(]'Y11im::~Un1~"'1[J
1 .. .. ..¥ "" J. '1J£l-'11J1f1Uj:ji.'l UHtl1'rlfla£l.:J ';11 nn1~Hll1ULLj:j iil'11£l~L U£1 LU£l'YI
u'inmlu"' -i-1 Clark (1998) wui1ii'li'efini1m~~hliLiifl • .. .. .. "" "" 1 iJ .. lJ1fiLL~i.'llJ~L1t\L~1..,U-l£lU LU£l-'!';11n 1J"''i::H1ll1~tl fla'U'rl ,. "' o A :'1 A ..¥ "'... o .,/. -~ <t .r LllLnfi~Lj:ji.'lLuU LLi.'l::flfiL'IfCl Lf1V1nn11 LllEIL1J~[JlJL'YIUlJ(]'r11i
m::fi um~..,, U'll e~ll1fl LLj:J a 1 uH't~fnfl a £1.:1'11 £J., ihi'ufl'ill~ ii , .... .... A A J.
«1~«nflfi1VLEI'liiil£l::'l1Lfi'YI1J£l-'! C. ct. comosa Roxb 'Yif'l11ll
L'li'll"llu 5% wlw nu 10% wlw wu111m~'Lfiu.:~nu utitini1 d .r .. ..
cream base 'lf.:J(]Yltin~::~um~"'1V1Ja.:JlJ1f'IU~a1Ja.:Jif1~Hnfl
tl1UL£Jn-aa::~Lf1Yl"IJ£J.:J C. cf. comosa Roxb a1';1LUU~all1
';11nnalnn1~n~::fiu collagen LLa::afl reactive oxygen • species "lJa.:JVi'lffl~::na Curcumin (Panchatcharam, 2006) .. uam1nU!T-1ilqnlfi1U";Ja<ifw ~.:JYh 1 imfl Uj:Ji.'l..,, vlti'L~1iu , (Hwang, 2000)
;
The 7"' Annual Northeast Pharmacy Research Conference of 2015
"Challenges and Opportun~ies of the Pharmacy Profession in Post-Millennium Development Agenda"
Faculty of Pharmaceutlcal Sciences, Khon Kaen UnlversitlD~aOand
J r 1 wrr 'P '7-Ff ·r'3"11111!111iilllllkliiiF 11IM!li HIM liillli=· i!f!'§F::WJ Jail
.r~,f u~.., a·nJltii1 fhi'tJfl111 ~ ~ a1~anflti1 u La ia a ~.:a ' . .1. ... ... lw
LflYI'llil~ C. cf. comosa Roxb Ylfi1111L'llli'IIU 5% w LLa::
10% wlw iifl'nutnri lum11l1 LlJ~IJU1Llluu1 hui1uaU1flfl
References
Clark L, Clark R, Heber-katz E. 1998. A new murine
model for mammalian wound repair and
regeneration. Clinical immunology and
immunopathology; 88(1), 35-45.
Deodhar AK, Rana RE. Surgical physiology of wound
healing: a review. J Postgrad Med. 1997;43(2):52-
6.
Hwang, JK, Shim JS, Pyun YR. Antibacterial activity of
xanthorrhizol from Curcuma xanthon'hiza againt
oral pathogens. Fitoterapia, 2000; 71 : 321- 23.
Midwood KS, Williams l V, Schwarzbauer JE. Tissue
repair and the dynamics of the extracellular
matrix. lnt J Biochem Cell Bioi. 2004;36(6):1031-7.
Panchatcharam M, Miriyala S, Gayathri VS, Suguna L.
2006. Curcumin improves wound healing by
modulating collagen and decreasing reactive
oxygen species. Mol Cell Biochem; 290(1-2):87-
96.
Prasitpuriprecha C, Suttipan N, Werawattachai N,
lntaranongpai. 2004. The usage of medicinal
plants in Ubonratchathani, UBU J.; 6(2): 117-138.
Teetawatanasuk N, Nakamura ES, Wangmaneerat A,
Komatsu K. Saiki I. Anti-invasive and anti
angiogenic activities of C. sp. Extracts.
J. Trad.Med, 2003; 20: 1-8.
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