V i e t n a m J o u m a l of Physiology 16(1), 4/2012 ISSN: 1 8 5 9 - 2 3 7 6
7. MikI T, Mitamura K, Ross MA, et al.
(2007) Identification of stem cell mariner- positive cells by immunofluorescence in temi human amnion. J Reprod Immunol.
75(2):91-96.
8. Niknejad H, Peirovi H, Joijani M, et al.
(2008) Properties of the amniotic membrane for potential use in tissue engineering. Eur Cell Mater. 15.88-99.
D. Parolini O, Soncini M, Evangelista M, et dl. (2009) Amniotic membrane and amniotic fluid-derived cells: potential tools for regenerative medicine? Regen Med.
4(2):275-291.
lO.Toda A, Okabe M, Yoshida T, et al.
(2007) The potential of amniotic membrane/amnion-derived cells for regeneration of various tissues. J Phamnacol Sci. 105(3):215-228.
SUMMARY
isolation, culture and characterisation of stem cells from human amniotic membrane
Do Minh Trung^, Pham Van T r a n \ Nguyen Bao T r a n \ Tran Hai A n h \ Toshiko Y o s h i d a ^ Toshio Nikaido^
^Military Medical University, Vietnam,
University of Toyama, Japan Received 12 March 2012; revised 25 March 2012;
accepted 10 April 2012 Stem cells are defined by their capacity of self-renewal and generate functional differentiated cells that replenish lost cells thnuughout an organism's lifetime. In recent years human amniotic membrane has been recognised as a possible resen/e source of stem cells that may be useful for clincal application in regenerative medicine. The aim of this study was to establish a new cell source that can be used for research and treatment; therefore, this study made isdatlon, prolifemtion cultures, and kientification of expression for the mariners of the stem cells isolated from human amniotic membrane. The results showed that the isolated cells in culture medium had a typical shape, and they expressed positive markers for stem cells and amniotb epithelial cells as Oct-4; SSEA4, Sox2, Vimentin, CK18. Those findings indicated that stem cells from human amniotic membrane were isc^ated and klentified.
Key words: epithelial stem cell, amniotic membrane, human amniotic membrane stem cell
TAC DyNG D6I vai VI K H U A N TAI CHd T 6 N THU-ONG B 6 N G
C O A K E M H E R B A V E R ALam Thj Dan C h i \ Trdn Ngpc Tudn^, 'Wg/J B d n g Quoc gia
^Hgc vign Quan y Nhgn bdi 17/1/2012; su-a bai 18/3/2012; chap nhgn dang 10/4/2012
Nghien cwu dwac tiin hdnh tren 50 bgnh nhdn bdng ndng dieu tri ndi trO tgi Khoa chu-a Bdng Ngwdi l&n ti> thdng 4/2011 den thdng 11/2011 M^c tidu cua cdng trinh ndy Id ddnh gid tdc dicing uc chi vi khuin tgi bi mgt vet thwang bong cua kem Herbavera. Kit qua cho thay kem Herbavera cd tdc dijng uc chi si/phdt trien cua vi khuan tgi cho vet thwang bdng ndng.
Tu' khda: kem Herbavera, vi khuan tgi vet thwang bong
Vietnam Joumal of Physiology 16(1), 4/2012 ISSN: 1 8 5 9 - 2 3 7 6
I . D A T V A N D e
D i l u trj tai cho vet bong nham d y phdng vd chlng nhiem khuan. tao dieu kign tot cho qud tflnh tdi tgo vd b i l u md hod [4]. Mpt so thuoc khdng khuan thdng thudng a l u tri tgi cho v l t thuang, vet bong do si> dyng rpng rdi vd trong mdt thdi gian ddi nhu cdc thudc Betadin 10%, Siiver-sulfadiazin 1%, sulfamlon... tren lam sdng cho thay cd hi#n tuyng khdng thulc. M$t khde. cdc thuoc ndy h l u h i t deu phai nhdp k h i u nen gid thdnh cao. Khi n j a hogc d i p v l t thuang, v l t bdng cdn gdy xdt hoac gdy d$c cho mpt so ca quan nen khdng the dung trdn didn r|ng trong thdi gian ddi [1], [2], [3]. Kem Herbavera Id san phIm duyc bdo c h l ti> cdy Cd Ido, Id Ld hpi vd cdy Bo cdng anh. Cdc duyc lidu ndy dd duyc chu-ng minh ed tdc dyng tot tren vet bdng, cd tdc dyng khdng khuan, tieu viem va Idm tdng qud trinh bieu md hda v l t thuang bong 19]. Chung tdi tien hdnh nghidn cu^i tdc dyng cua kem Herbavera trdn v l t thuang bdng vdi myc tieu ddnh gid tdc dyng <rc c h l vi khuan tgi b l mdt vet thuang bdng cua kem Herbavera.
2. D 6 | TU-i^NG VA PHU'ONG P H A P N G H l i N Cl>U
2.1. D l i t u y n g nghidn cO'u
Nghidn cu-u trdn 50 bgnh nhdn bdng iSlu trj ndi tnj tai Khoa Bdng Ngudi Idn - Vign bdng Quoc gia tiF thdng 4/2011 den thang 11/2011, trong dd cd 37 bgnh nhdn bdng ndng vd 13 bgnh nhdn bdng sdu.
- TiSu chuan chpn bdnh nhdn: khdng cd dien bien ngng nhu s i c bdng. soc nhiem khuIn
huyet, khong cd bgnh mgn tfnh vd bdnh tmyen nhiim kdm theo, t y nguygn thsm gia vdo nghien cu'u.
2.2. C h i t ligu nghidn ciKi
- Thuoc so sdnh: kem silver sulfadiazin 1% (SSD 1 %) do hdng R^tako&erett (An Dp) sdn xuat.
- Thudc nghidn cuu: kem Herbavera do Tnjng tdm Nghidn cu-u ung dyng Sinh-Y-Duyc - Hpc vien Qudn y san xuat.
2.3. P h u o n g phap nghien cO'u Nghien cuu tdc dyng dipt vi khuan cua kem Herbavera tren v l t bdng, cd d l i chO-ng, so sdnh tru-dc vd sau d i l u trj, so sdnh vdi thuoc doi chu'ng tOng cap trdn cung mpt bgnh nhdn.
- Xdt nghigm vi khuIn tai v l t thuang bong:
cdc xdt nghigm vi khuan d u y c thyc hidn t^i khoa Cgn Idm sdng - \^dn Bdng Quoc gia Bgnh pham d u y c lay d nhdm bgnh nhdn bdng ndng d hai vung nghidn cOu, vdo hai thdi diem trudc vd sau nghidn cO'u 7 - 1 0 ngdy. Chi tieu theo ddi gom:
+ Khd nang O'c che sy phdt t n i n vi khuSn trong 24 gid in vitro: theo phuang phdp dim vi khuan sau khi u mang nghidn cu'u 24 gid.
+ Djnh loai vd djnh luyng vi khuan theo phuang phdp cua Ivanov N.A. vd Danilova E,G. (1984). Lay bgnh phIm vd x u IJ- bgnti pham tren cung mpt vj tri bdng d ca hai lan xdt nghigm, trudc vd sau nghien cuu 7 ngdy.
+ Ldm khang sinh do: thyc hign tren hg thing VITEK 32.
3. K £ T Q U A NGHIEN CCPU Cdc k i t qua nghidn cO'u duyc trinh bdy trdn cdc bang 1-'-5.
Bang 1. Hoat tinh khdng khuan cua kem Herbavera vdi mdt so chung vi khuIn tgi be mgt vet thuang bdng.
LoSi vi kiiuan S. aureus P. aeruginosa E. coli
Di/crng kinti vong vo Ittiuan Kem Herbavera (n = 10)
16,5 ±1,6 14,2 ±0,8 24,3 ± 1,7
SSD 1% (n = 10)
23,5 ±2,4
20,6 ±1.4
15,1 ±1.1
V i e t n a m J o u m a l of Physiology 16(1), 4/2012 ISSN: 1 8 5 9 - 2 3 7 6
K i t qua trdn bang 1 cho t h i y d l i vdi E. va P. aeniginosa dudng kinh vdng vd khuan c(^i kem Herbavera c6 du-dng kinh vdng vd cua kem Herbavera kem han so vol thuoc khuan han h i n so vdi SSD1%, Vdi S. aureus chO'ng.
Bang 2. S y phdn bo cdc lodi vi khuan tgi v l t thuang bdng ndng.
Lodl vi khuSn
P. aeruginosa S. aureus S. epidermidis Entefobacter E. coli Kh6ng mgc Tong cong
S6 Ian cay khuSn Vtjng A (n =
L a n i 4 2 1 1 0 12 20
Lan 2 9 4 1 3 1 2 20
30) Lin
3
5 2 3 1 0 9 20
VCing B (n = L a n i
3 3 0 1 1 12 20
Lan 2 8 4 2 3 0 3 20
30) Lan 3
4 3 2 1 0 10 20
T6ng cong (%)
33 (45,8) 18(25) 9 (12,5) 10(13,9) 2 (2,8) 48 (41) 120(100)
Bang 2 cho t h i y hai lodi vi khuan chilm ty Ig tuan dieu tri, so lan x u ^ hign vi khuIn P.
cao nhat Id P. aenjginose vd S. aureus Sau 2 aeruginosa vd S. aureus giam ro vdi p < 0,05.
Bang 3. S l luyng vi khuan P. aeruginosa vd S. aureus tgi be mgt v l t thuang bdng ndng.
Lodi vi ktiuin
P. aeruginosa S. aureus P
So luvng trung binh vi khuan (x10'/cm^) Vung A
L a n i 6e.4±165.2 163,3±202,2
>0,05 Lan 2 8676±207.2 550.3±892,3
>0,05 Lan 3 124,4±36,7 161,6148,1
>0,05
VUngB L a n i
99,4±19,1 284,(1±139,3
>0,05 Lan 2 762,1±170,2 579,7±284,1
>0,05 Lan 3 162,1±170,2 191,3±363,7
>0,05
Cdc SO lidu trdn bang 3 Cho thay SO luyng thay doi tuang duang nhau sau 10 ngdy hai iogi vi khuIn P. aeruginosa vd S. aureus nghidn cuu d ca hai viing vdi p > 0,05.
Bang 4. Phdn bo cdc lodi vi khuan tai v l t thuang bong sdu.
Lodi vi khuIn
P. aenjginosa S. aureus Enterobacter Khflng moc Tong cOng
S6 l^n cay khuan Vung A ( n =
L a n i 6 3 1 0 10
Lan 2 6 3 1 0 10
60) Lan 3
5 3 0 2 10
Vijng B (n = L a n i
6 4 0 0 10
Lan 2 6 4 0 0 10
50) Lan 3
6 5 0 0 10
Tong cong (%)
34 (68,6) 22 (37,9) 2 (3,4) 2 (3,3) 60(100)
Vietnam J o u m a l of Physiology 16(1), 4/2012 ISSN: 1 8 5 9 - 2 3 7 6
Ket qua tren bang 4 cho thay hai lodi vi chiem ty Ig cao d nhdm bgnh nhdn bong khuan Id P! aerruginose vd S. aureu van sdu.
Bang 5. Thay d l i ve s l luyng vi khuIn tgi be mgt v l t thuang bdng sdu.
Loai vi khuIn
P. aeruginosa
S. aureus Enterobacter P
So lu-ong trung binh vi khuin (xio^/cm^) Viing A
L i n l
1979.8±830,6 1839,0±858,2 3576,0±0<0,05 Lan 2 1227,1±157,1 1626,5±239,2 637.0±0
<0,05 Lan 3 112,3±234,1 272,8±41,6
0
>0,05
Viing B L a n i 2297,8±938,6 2477,2±760,2
0
<0,05 Lan 2 461,0±173,4 1659,6±62,4
0
>0,06 Lan 3 93,6±48,5 95,5122,3
0
<0,05
TO- bang 5 t h i y rd s l luyng vi khuan giam dang ke sau khi cat hogi t ^ d ca hai vCing nghien cOu vdi p < 0,001, Qua tO'ng lan c l y khuan vCing doi chdng d u y c dieu tri bang SSD 1 % so luyng vi khuan ca hai lodi P.
aeruginosa vd S. aureus giam rd han so vdi vCing duyc d i p kem Herbavera.
4. BAN LUAN
Theo Le The Tmng [4] quan t h i vi khuan tgi cae vet thuang bdng phy thupc vac tO-ng thdi ky t i l n triln cua ton thuang bdng, tinh chat hogi ti> bdng, dign tich bdng sau, vj tri cua v l t bdng, phuang phdp dieu trj toan thdn vd tgi c h l . Da ed nhieu cdng trinh nghidn cOu v l quan the vi khuan tgi v l t thuang bong d cdc thdi diem. Tuy s6 lidu nghidn cu'u khde nhau nhung deu ed chung k i t ludn Id cdc vi khuan thudng ggp tren v l t bong gom S.
aureus, P. aenjginosa, Enterobacter vd mpt s l true khuan Gram duang cd bdo tO-. Vdo hai thdp nidn cuoi cua t h i ky 20, P.
aeruginosa, S. aureus. Enlerococcus Id nhung nguyen nhdn chu y l u gay nhiem khuIn vet thuang, vet bdng vd nhiem khuan bgnh vign [1]. [2], [3]. Trong nghien cu'u nay d nhdm bdnh nhan bdng nong vdi 72 lan cay khuan duang tinh d ea hai vung. D i l u ndy cho thay P. aeniginosa Id vi khuan du-ng hdng d I u , chiem 45,8%, S. aureus chilm 25%, t i l p
theo Id Enterobacter, c h i l m 13,9% vd S, epidermidis c h i l m 12.5%, c u l l ei!ing Id E. coli chiem 2,8%. So lan cdc vi khuan xuat hidn a hai viJng nghidn cuu tgi cdc thdi d i l m tucng duang nhau.
O nhdm bgnh nhdn bdng sdu hai lodi vi khuan P. aeruginosa vd S. aureus chiem u-u the lan luyt Id 58,6% vd 37,9%, tuang duang d ca hai viing nghidn cO-u. 6" nhdm ndy ty 1^
khdng mpc vi khuan chilm ty Id nhd (3,3%).
Teplit Z.C Id ngudi de xuat can xac ^nh so luang vi khuan de ddnh gid mOc dp nhiem khuan bdng d ngudi bj bdng vd d dpng vgt d u y c gdy bdng. Tdc gia cho ring so luyng vi khuan cd trdn mpt dan vi d i f n tich ca the Id y l u t l quylt djnh mu'c dd nhiem khuan, cho phep tidn luyng nhiem khuan tai cho. So luyng vi khuan d vet bong phan dnh sy cdn bang giua s y xdm nhgp vi khuan va ca che bao vg cua vgt chu (theo 14]).
So luyng vi khuan phat triln tgi be mat ton thuang bdng phan anh muc dd nhiem khuan.
Khi so luyng vi khuan > lOVcm^ b l mgt vet bdng hoac 1 gram md to chOc thi nguy ca nhiem khuan lan trdn se xay ra. Khi so luang ndy > lOVcm^ thi nguy ea nhiim khuan huyet, nhilm khuan toan thdn rIt cao vd da ghdp khdng song d u y c (7], [8]. So luyng vi khuIn tgi cho v l t bdng Id mpt trong nhung chT tidu
Vietnam J o u m a l of Physiology 16(1), 4/2012
ISSN: 1859-2376
xdc djnh mO'c dp nhiem khuIn, giup cho thay thulc cd bidn phap d i l u trj thich hyp. Day cung Id tidu chuIn ddnh gid tdc dyng cua mpt logi thulc ho$c mdt phuang phdp d i l u tq tgi chS v l t thuang bdng.
C nhdm bgnh nhdn bdng ndng, xdt nghigm vi khuIn (VK) cho t h i y s l luyng vi khuan trung binh/cm^ tdng cao d ngdy thO 5+10 sau b6ng. Sau 7 ngdy d u y c ffliu trj b i n g kem Herbavera sd luyng vi khuIn P. aeruginosa giamtu- 867,5 ± 207,2 x 10^ VK/1cm^xu6ng cdn 124,4 ± 86,7 vd S. aumus g i i m tO- 550,3
± 892,3 xuing cdn 161,6 ± 4 8 , 1 . K i t qud nayf tuang duang so vdi d nhdm chi>ng d u y c dilu trj bang SSD 1 % .
Trdn v l t thuang bdng sdu t h u l c cOng cd tdc dyng Idm gidm s l luyng cua P.
aemginosa vd S. aureus sau khi d i p kem Herbavera tu' 5 - 6 ngdy, nhung rd rdt han khi v l t thuang duyc cat hoai tu'. Tgi vOng doi chOng SSD 1 % Idm giam s l luyng hai lodi vi khuan P. aeruginosa vd S. aureus rd han so vdi vCing d i p kem Herisavera vdi p < 0,01.
Dd cd mdt so cdng trinh nghidn cu-u ve thuoc d i p tgi chS v l t bdng so sdnh vdl SSD 1 % nhu nghidn cO-u cua Nguyen Ngpc Tuan 15] so sdnh vdi Maduxin d i l u trj v l t bdng do vdl tdi ndng, cua Do Luang Tuan [8] so sdnh dieu trj v l t thuang bdng bang AT-04. Cdc tac gia d I u cho t h i y so luyng vi khuln/cm' dign tich bong tdng cao nhat vdo ngdy thu- 7 vd giam sau 14 ngay dieu tri. Dk logi bd trigt d l sy cd mdt cua vi k h u I n tgi c h l vet thuang bdng sdu phuang phdp eat hogi tu* dd td ro uu t h i noi bdt [5], [6].
Nhu v&y, kem Herbavera c6 tdc dyng khdng khuIn khi d u y c dieu tri v l t thuang bong nong d0 li, III, hidu qua tuang duang so vdi nhdm chung d i l u trj b i n g SSD 1 % nhung tren vet thuang bong sdu tdc dyng ndy kdm han so vdi SSD 1 % .
5. K £ T LUAN
0 v l t thuang bdng ndng so luyng vi khuan trung binh/cm^ tang cao d ngdy thO 5+10 sau bdng; sau 7 ngdy d u y c d i l u Iq bang kem Herbavera so luyng vi khuan P.
aeruginosa giam tO 867,5 ± 207,2 x 10^ vi khuan/lem^ xuong cdn 124,4 ± 86,7 vd S.
aureus giam tO* 550,3 ± 892,3 xuong cdn 161,6 ± 48,1. Tren vet thuang bdng sdu kem Herbavera cung cd tac dung Idm giam so luyng cOa P. aeruginosa vd S. aureus sau khi dap tO" 5 - 6 ngay, nhung rd rdt han khi v l t thuang duyc c i t hogi t u .
T A I L i e u THAM K H A O
1. Ld Thu Hdng, Nguyen Dinh Bang (1994) MOe dp khdng khdng sinh cua cdc chung vi khuan phdn lap tO- bdnh nhan bdng tgi Vign Bdng Q u l e gia. Trong: Mgt so cdng trinh nghidn cu'u v l dp nhgy cam cua vi khuan vdi thuoc khdng sinh Vign Thdng tin y hpc Taing uang - Ha Npi, tr.14-29.
2. N g u y i n Thai San, Nguydn Dinh Bang (1996) Can nguyen vi khuan gay nhiem khuIn vet bong vd muc dp khdng khang sinh cua Chung. Thdng tin bdng, so 1, tr. 34-36.
S.Nguyen T h i n g , Ndng ich Dong (2000) Nhiem trung v l t thuang bong. Tap chi thdng tin Y Duyc, Hd Npi, tr. 83-88.
4. Le T h i Trung (2003) Bdng - nhung kiln thO-c chuydn ngdnh. NXB Y hpc, Hd Ndi.
5. N g u y i n Ngpc Tuan (1998) Nghidn cu-u dac diem Idm sdng va tdc dyng dieu tri tgi cho v l t bdng do vdi tdi ndng cua thulc m d Maduxin. Ludn an T i l n sy Y hpc. Ha Npi, tr.
122-123.
6. D 5 L u a n g Tudn (1998) Ddnh gia tac dyng d i l u trj tgi cho vet thuang bdng bang AT-04 cd so sdnh vdi m d Silver sulfadiazin 1%.
Lugn vdn Thae sy khoa hoc Y-Duyc, Hd Npi.
7. Carsin H, et al. (1995) Infections nosocomiales in burns patient. Infections
Vietnam Joumal of Physiology 16(1), 4/2Q12 ISSN: 1 8 5 9 - 2 3 7 6
nospcpmiales and preventions. Care med, p: 313-327.
8. Heggers JP, Robson MC (1996) Infection control in bums patients. Clin plastic surg.
13:39-47.
9. Visuthikosol V, Chovirchuen B ( ) Effect of aloe vera gel to healing of burn wound a clinical and histologic study. J Med Assoc Thai. 78(8) 403-409.
SUMMARY
The effect o f Herbavera cream on bacteria at burn w o u n d s
Lam Thi Dan C h i \ Tran Ngoc Tuan^
^National Institute of Bum
'Vietnam Military Medical University Received 17 January 2012; revised 18 March 2012;
accepted 10 April 2012 A study was conducted on 50 patients with shallow bum wound at the Department of Adult Bums from April 2011 to November 2009. The aim of the study was to evaluate the inhibiting effect of Herbavera to bacteria at bum wound surface. The results showett that Herbavera cream can inhibit the proliferation of bacteria at the superficial burn wound.
Key words: Herbavera cream, bacteria at bum wounds
N G H I E N c C f U T A C O g N G D I & U TR! TAt C H 6 C O A K E M H E R B A V E R A T R t N V ^ T T H U a N G B O N G N O N G
Lam Thj Dan C h i \ Trdn Ngpc Tuln^
V / ^ n Bdng Quoc gia 'Hgc vi^n Quin y Nhgn bdi 17/1/2012; su'a bdi 18/3/2012; chap nhgn dang 10/4/2012
Nghien cO'u di/pc tien hdnh trdn 37 bgnh nhdn dieu trj ngi trO tgi Vign Bdng Quoc gia tO' thdng 4/2011 den thdng 11/2011, tuoitw 15 din 60. Muc tieu cua cdng trinh ndy Id danh gid tac dijng kich thich tdi tai bieu mo hda vd lam lien vet thwang bdng ndng cua kem Herbavera. Kit qud cho thay kem Herbavera cd tac di^ng kich thich tdi tgo md vd idm liin vet thwang. Sng thai cd tdc dijng i>c che vi khuan tgi vit thwang bdng ndng twang duang voi nhdm chwng dum dieu trj b^ng silver sulfadiazin 1%.
T(f khoa: vit thwang bdng ndng, kem Herbavera
I.DATVAND6
Vigc dieu trj tai cho v l t bdng cd mpt vj tri rat quan trpng trong viec d y phdng va chong nhilm khuan, tgo d i l u kign tot cho qua trinh tai tgo va b i l u md hod [1], [3]. Ke thOa vd phdt t r i l n cac che pham d u y c bdo c h l tu' nguin d u y c lidu trong nude nham tao ra san p h I m thuoc dap tgi cho dieu trj v l t thuang bdng cd higu qua vdi gid thdnh thap, Hpc vign Quan y dd nghien cuu bdc che kem Herbavera tu- cdy Cd ido, Id Ld hpi vd cdy Bo
cdng anh. Cdc d u y c lieu ndy da duyc chi>ng minh cd tdc dyng tot trdn v l t bong, c6 tdc dyng khdng khuan, tieu viem vd Idm tdng qua trinh b i l u md hda v l t thuyng bdng qua n h i l u cdng trinh nghien cu-u trong vd ngodi nude [2], [4], [8], [7], [11], Chung tdi tien hdnh nghien cuu tdc dung cua kem Herbavera tren vet thuang bdng vdi myc tieu ddnh gid tdc dyng kich thich tdi tao b i l u m6 hda, lien v l t thuang bdng ndng eua kem Herbavera.
