Nghiin ciyu - Ky thujit
Nghien cii-u hieu qua ghep te bao goc trung mo tach tir mang day ron ngvol tren v§t thuong bong nhiet thyc nghiem
Dinh V f i n H&n, Phan M i n h H o ^ n ^ , L i r i m g Q u a n g A n h Hoc vi^n Qudn v
Gh6p t6 bio di tJu-g'c dung kh^ phd biAn trong di6u tr^ cdc v4t thiro-ng bding (3$c bi$t cdc t6n thifang cli§n tich rOng ho$c cdc v6t thuwrig mgn tinh' . Nhiing nghiSn CLTU V ^ t6 bdo g6c di chCrng minh, hdng ngdy ca t h ^ con ngu-d-i duiqc dfii mdi thay th4 nhiing l6 bdo gid ch6t Id do ngu6n cdc \h bdo g6c bi$t hod thdnh nhOng t6 bdo chLPC ndng. Khi bj thtrang, cdc \k bdo g6c se di CU' ti> tuy xuwng, ILF cdc mfl khdc nhau vd ngay ca bin cgnh ton t h u o n g d^n dk bi$t hod thdnh cdc ih bdo chu'c ndng Idm li^n vdt thuwng '^^', Tuy nhien, viec gh6p tk bdo g6c d ^ di^u trj v6t IhiKyng bj hgn che bb'\ vkn de s6 lu'p'ng t^
bdo n6u Id t^ bdo goc triFdrng thdnh hodc bdi van dh y du-c neu Id \.k bdo g6c phdi thai.
Gan day, cdc nhd khoa hQc tai OH Qu6c gia Singapore da phdt hi$n ra cong ngh0 tdch, nhdn rpng hai logi te bdo g6c ttf mdng ddy r6n Id l ^ bdo goc tnjng mo va te bdo g6c bieu m6 vd chLcng mmh chung co kha ndng bi^t hod thdnh nhieu logi te bdo chiJc ndng, dac bi^t Id cdc te bao cua da'^'. Nguon t^ bdo goc tii' mdng ddy ron ddp Lfng du-p-c ca hai yeu cau la s6 luyng (k bao va khong vi pham y diFc nhu'ng li^u chung cd kha nang lam Idnh v4t thu'ong tren Idm sdng khong thi can phai ti^p tyc du'oc nghifin cdu, Vi vdy, Chung toi tren hdnh nghien c d u ndy nhdm danh gia tac dyng ghep te bdo goc trung mo mdng day ron ngut^i {TBGTMMDRN) tr^n qud trinh li^n vet thuo-ng bong nhiet thyc nghidm, dong thdi ddnh gia anh hu'dng cua cay ghdp trong vi^c gay nhiem khuan vet thuong vd tinh smh mien djch cua te bao ghep,
D o i t u - a n g v a p h u - o - n g p h a p n g h i e n CCPU
Ooi t u v n g nghien CLPU
- TBGTMMDRN thudc dong te bdo P53 du-oc phan lap ti> mang day ron cua tre so sinh va mdi
trud'ng chuydn dyng nudi c^y TBGTMMDRN do Ngdn hdng T 6 bdo g6c MekoStem cung cdp.
- Thd di/c 15 con, cdn n^ng tnjng binh 2,0 ± 0,2 kg do Ban Cung ekp ddng v$t thi nghidm HQC vi$n Qudn y cung cdp. Thd d i r ^ c nudi ddm bdo theo tidu ehudn t h ^ c nghi$m.
- Cdc hda chdt dung cho nudi cdy td bdo vd hda chdt d$t tidu ehudn nudi cdy md vd tieu ehudn phdn tich do cdc hang Invitrogen, Merck, Sigma cung cdp.
P h u v n g phdp n g h i d n c i n i
TBGTMMDRN du'ac cdy v d i m$t dg 5x10* t4 bdo/cm^ trong mdr tnj'dng DMEM-F12 len mang Tegaderm vd nudi trong tu dm d didu kien ZVC, 5% CO2. Djnh ky thay mdi tardng nudi cay cdch nhau 2 ngdy cho den khi cdc td bdo mpc kin trdn bh m$t gid d d (vdo ngdy thi> 4 - 6 sau cay) thi thu hogch tdm td bdo dd cay ghdp.
Thd d u v c gdy bdng tren lung vdi dd sdu todn t)0 da theo phuo'ng phdp cua Halovec vd Poaado, Ghdp tdm td bdo l&n vdt bdng: tdm te bao da ehudn bj s^n dutye d^t len vdt lidng vung A vdo ngdy thu- 5 sau khi gdy bong. Sau khi dat, dung mdt Idp gge vaselin dd dp vd c6 dinh tdm te bao I6n vet thuxyng bdng. €)6i vdi cdc vet bdng vCing B chi phu mdng Tegademi khdng cd te bdo goc, sau dd ddp ggc cd tdm thudc Sivirin (sulphadiazin bgc) cue An Dd. Sau dd Idy mSu lam cac xet nghi6m:
- Xet nghidm vi khudn vung vet thuong nghidn ci>u v d doi chi/ng du'i?c tien hanh tgi 3 thdi didm khdc nhau: tru'dc luc ghep te bao, ngdy thLP 5 sau khi ghdp te bao (D5), ngay thi>
l O s a u g h d p t d b a o ( D I O ) .
- Xdt nghi$m md hpe: Sinh thiet md vet b6ng bang dung cy sinh thidt vao cdc thdi diem tnfdc ghep td bdo, D5, D10 Quan sat hinh thai mfi hoc va so lu'p'ng te bar viem, nguy&n bdo spi, t l
TAP CHi DUgtC HOC-08/2012 (SO 436 NAM 52)
• Nghien clfu - Ky thugt
tdm td bdo gdc, sau dd bdo qudn d -80°C vd tim khdng thd thd khdng TBGTMMDRN bdng phuo'ng phdp ELISA. Dpc kdt qud d Ijudc sdng 450 nm trdn hd thdng DTX 880 cua Badtman Coulter, bao sQ-i trdn mOt don vj didn tieh (DVDT) dudi
kinh hidn vi quang hpe.
- Huydt thanh thd duvc Idy tgi cac thdi didm DO (ngay sau ghdptd bdo), D15, D30vd D60 sau ghdp
Ket qua va b ^ n l u $ n Dien bidn Idm sdng thd nghidn ciki Diin biin t§ti ch6 vung t6n thwong
Bang 1: Dien bdn hinh inh lim sing t^i chd v4t thuvng bdng thue nghidm cd ghip t4 bio goc (viing A) vi viing khdng ghip tS bio gSc (viing B)
Thdi diem Viing A VOng B
5 ngdy ddu Vet thu'O'ng vi&m nd, xung huydt 66. Nen vdt Vdt thu'ong viSm nd, xung huydt d6. Ndn sau gdy bdng thu'ong ho^i tO trSng rdi rdc. B6' m6p vdt vdt thirang dd, ddi ch5 hoai tu' trdng. Bd
thiFcng khdng vi§m nd m6p vdt thu-ang kh6ng vi6m nd.
gid mac bdm d l bdc, Ndn vdt thirong dm, gi^ mac mSu vdng nhgl b^m ndn vdt thux>ng, tidt d|Ch vifa, ddi chd hoai ti> tr^ng 86' m6p vdt thyang khdng vi&m n^
djch il, V^y tidt d l Trud'c gh6p Ndn vdt thuwng dm
y\&m nd giSm, cdn hoai tCr trdng Bd m6p vdt thuo'ng khdng viSm nd.
Ngay thCp 5 sau gh6p
Vdt thu'ong sgch khd, khdng tidt djch Vdy tidt Vdt thu'ong sach.
kho mdu vdng sgm, ego sgch ldp vdy tidt b6n bdc, mdu vSng nhgt, ego sach Idp vliy ti^t difdi rudm mau Ndn vdt thicang mau hong b§n du-di rudm miu. Nen vet thu-cng mau nhat. Bd m6p vet thu'O'ng khdng vi§m nd hdng. Bd m6p vdt thu'ong khdng viem ne.
Ngay thd 10 Di^n tfch vdt thu'ong thu nhd, nen vet thu'ang Di^n tich vet thucng co nhd Igi nhung Idn sau gh6p sgch, mdu hdng, mo hat dd, bieu md hda tu' hen viing A. Men vet thuung am, sgch, nen bd mep vet thu'ong, mgch mau tang sinh vet thu'ong mdu hdng nhat, djch tidt vifa.
manh. Bd mep vet thu'ong khdng viem ne. Md hgt dd, bidu md hda tif bd mdp vet thuang cham... Bd mep vlt thuong khdng viem nd.
Ngay thd 15 Ndn vdt thuong phdng, ndng, khong co dum. Ndn vet thuong phang, rudm mau, s ^ co sau ghdp Bidu md hda tCf bd mdp vet thuong tdt. Vdt diim. Dien tich vet thuang dudi 5cm^ nen
thuong co bin da khdi. vet thuong mSu hdng nhgt Bang 2: Dien tich biSu mo hoa trung binh (em^) vi ty li % liSn vet thwa/ng theo thai gian
Thai gian Vung A(n = 15)
cnr
Vung B(n = 15)
D10 14,83 ±2,06
Nhan xet: Qua bang 1 va 2 nhan thay toe dp bieu m d hda cua vung A nhanh hon toe do bieu mo hda cua vung B. Tuy nhien s u thay ddi tdc dp bieu md hda giua viing A va vung B trong 5 ngay dau sau ghep chua c6 y nghTa thdng ke ddi vdi ca dien tich \§n ty )# vdi p>0,05. Nhung neu so sanh s u thay ddi tdc dp bieu md hda gida viing A va vung B trong khoang thdi gian tai ngay t h u 10 sau ghep te bao gdc thi s y khdc nhau cd y nghla thdng ke vdi ddi vdi ca dien tfch l i n ty Id (p<0,05) Dieu nay chdng td cang ve sau thi tdc dp bieu md hda vung A cang nhanh han vung B. Didn bien lam sang vung bdng du'p'c ghep te bdo gdc dien ra thuan Ip-i
eho qud trinh lien vet thuong rd ret han so v d i viing chdng.
Bang 3: Ty le v4t thuxmg khoi hoan toin giira vung A va viing B trong tirng giai doan
Th&i gian Viing A (n = 15)
Viing B (n = IS)
Di^&i 15 ngay sau bong Tir 16-20 ngay
sau bong Trgn 20 ng^y
sau bong Sd tho 0 11 4
7y(#
%
0 73,3 26,7
Sd thd 0 g 6
Tyl^ "
%
0 > 0,05 60 > 0,05 40 > 0,05
TAP CM\ nTr0.C_Hr»r'_n8y>ni-» / c o Aie. MAIV
Nghidn ci>u - Ky thugt
Bdng 5: S6 v4t bdng eOa hai vung bi nhi§m vdi cic chOng vl khuin
Th6i gian
L i m Lin 2 L i n 3 TAng
Vimg A(n- 15) Sdmiu
(*>
1 5 3 9
Tylf
%
6,67 33,33 20,00 60
VOng a SdmSu
(*)
3 5 2 10
fn • 15) Tylt
%
20 33,33 13,33 66,67
Chiing vl khuin E.coli E. cloeceae
S. aureus P. mlrebilis K. pneumoniae Aci. baumennii SN. boydl
VOng A 3 3 1 1 1
VungB 4 3
2 1 Nh$n xit: Trong gial do^n 16-20 ngdy ty' 1$ vdt
thuvng bdng vung A t j l§ vdt thu'ong eOa vOng A khdi hodn todn (73,3%) cao han so vdi vung B (60%), tuy chLca ed y nghTa t h i n g kd (p > 0,05).
Xdt n g h i d m v i khudn t^l vdt t h i r a n g vCing n g h i d n ci>u vi v d n g d d i chi>ng
Bdng 4: Tin suit xu4t hiin vi khu4n qua cic thdi di4m nghiin cO-u
Nh$n xit: Sd vdt bdng vCsig A vd vCing B tij nhidm khudn Idn \UK^ Id 9 vd 10 mSu. Nhu' v|y s6 vdt bdng bj nhidm khudn cua vung A vd vung B tuang d u v n g nhau. Didu ndy chimg td viOc ghdp td bdo toing md khdng dnh hudng ddn tinh t r ^ nhidm tnjng eua vdt bdng. Chu ydu dc vdt bdng bj n h i S m £ c o / / v d f 0/030636(13^19 vdt bdng).
Kdt qua n g h i d n ci>u m d hpe tren tidu bdn s i n h thidt
Bdng 6: Hinh thii md giira 2 viing A vi vung B tru>&c vi sau ghip t4 bio gSc trung md Thdidiim
Tmdc ghdp tdbdo
Ngdy thu 5 sau ghep Ngay thCr 10 sau ghdp
VungA VungB Bd m|t tdn ttiucng khdng cd hogi td, dupe Bd m^t tdn thuang khdng cd hoai td. dupe phO
phu mdt Idp to huydt mdng, MS phia dudi mdt Idp ta huydt mdng. Md phia dudi cd it djch cd It djch phii. xen ke it td bdo vidm chu ydu phu, xen kd it td bdo vidm, chu ydu Id bach cdu Id bach cdu da nhdn bung tinh. rdi rdc cdc da nhdn trung tfnh. rdi rdc cdc dai thi/c bdo, dai thi,K; bdo, nguydn bdo SQ'I. nguydn bdo sgn.
Md Tien ket mdng cd it td bdo vidm. Md tidn kdt ddy. nhidu td bdo vidm, nguydn bdo nhidu nguydn bdo spi vd tdn mach, s^i vd tdn m^ch, cdn phii vidm nhp khdng cd djch phiJ vidm.
Md lidn ket It td bdo vidm, nguydn bdo Md hdn kdt cd ntiidu td bdo vidm nhimg it hon sp-i. te bdo spi, tdn mach. Cd 8/15 vdt tardc, khdng cd phu vidm nhidu nguydn bdo thuong dd duoc che phu bdi Idp bidu bl sp-i, nhidu tdn mgch. Cd 5/15 vdt thuang da cd mdng nhung chua du 4 Idp Ldp td bdo bidu bi che phu, cd su phdn chia cda td bdo bidu bi lidn kdt ch$t che vdi chdn bi, Idp mdm.
td bdo mdm tdng phdn chia.
Bang 7: Thay doi s6 IwQ-ng t4 bio viim vet bdng
_ S6 lu^fmg t4 b^o vi§m/DVDT(122400prn^) ~^^ ~^
Vung A Vung B
(270ti^ubin) (270 ti6u bin) ^
12.7 ±3,9 13,6 ± 4 ^ _ > 0,05 14,6 ±8.2' " 16,3 ± 9,7 " ' > 0.05 4,7 ±3,4 _?-6±6^3 <0,05
>0,05 " >b,b5
< 0,05 < 0,05
< 0,05 < 0.05 r h d i diSm
Trudc ghdp (1) _ D5 sau_gh6p (2) DIP sau ghdp (3)
P1.2
Nh$n xit: Qua bang 6 vd 7 nhgn thdy tru'dc khi ghep te bao gdc trung md, sd lu'ong te bdo viem a hai vung nghien CLFU tu'ang du'ong nhau.
Ngay t h d 5 sau ghep te bao gdc trung md, sd
lup'ng td bao viem cua cd 2 vung deu tang nhe so vdi trud'c luc ghep te bdo gdc, vung B tang han so v d i vung A nhu'ng khdng cd y nghTa thdng kd. Ngay t h d 10 sau ghdp td bdo goc
TAP CHI DirOC HOC - 08/2012 fSO i^t. NAM 52)^
• Nghien cifu - Ky thugt
trung mo, sd lup-ng td bdo vidm ed hai vung ddu giam mgnh. Trong dd vung A ed sd lup'ng td bdo vidm it han so vdi vCing B ed >' nghTa thdng kd vdi p<0,05. Cd thd thdy vide ghdp td bdo gdc
Bdng 8: Sw thay dSi s6 IwQng t4 bio spi, nguyin bio soi d hai vOng A vi vung B trwde vi sau ghip t4 bio gdc trung md
tmng md Idm eho didn bidn md hpe tgi vdt bdng thu$n Ip-i han cho qud trinh lidn vdt thuang so vdi nhdm chdng.
Th64 dl4m T r u ^ c g h ^ p f l ) D5 sau gh6p (2) D10saugh6p(3)
Pl,2 P t j P i j
S6 Itrtjmg NB VimgAjim'iiSu bin)
41,2±11,7 51,2±11,4 54,7 ±12,8
>0,05
<0,05
>0,05
S, TBS/BVDT (122400 pm') VimgWlm mu bin)
45,6 ±5,2 49,2 ±12,1 50,6 ±11,2
>0,05
<0,05
>0,05
P
>0,05
>0,05
> 0,05
Nhin xet- Sd lup'ng td bdo sp'l vd nguydn bdo sp'l d cd 2 viing A vd B khdng khdc nhau d ede thdi didm trud-e ghdp te bdo gdc vd sau ghdp 5, 10 ngdy vdi p>0,05. 0 thdi didm sau
Ket q u a mien djch hoc
ghdp 5 ngdy sd lup'ng nguydn bdo sp'l vd td bdo sp'l khdng thay ddi so vdi trudc khi ghep, nhung d thdi didm sau ghdp 10 ngdy thi s y thay ddi cd y nghla thdng ke vdi p<0,05
02 H o 15
SB'S
B^^BlPtgSSwJS^^SsS^^^Sml
az D i 3 D30 D'ld - • — i;5O0 - • - 1/100 1/50 1 1/10
bao nguyen ven d ca 3 thdi diem D15, D30 vd D60. Tuy nhien chi thdi diem D15 la cao han ed y nghTa thdng ke vdi p<0,05.
Hinh 1 : Hoat tinh khang thi tho khang TBGTMMDRN nguyin ven va sieu nghien Nhan xet: C/ thdi diem DO ngay sau khi ghep
te bao, khdng cd mau huyet thanh thd nao cd hoat tfnh khang the khang td bao. Vdi khang the khang td bao nguyen ven nhan thay d thdi diem D15 da xuat hidn khang the khdng te bao d u a n g tfnh vdi dp pha loang huyet thanh 1/100, hoat tfnh khang the d cac thdi diem D30, D60 giam xudng chi cdn phan ung du'ong tfnh d cdc mdu huydt thanh pha loang 1/50 ldn, d thdi diem D15 da xuat hidn khang the khang te bao d u a n g tfnh vdi dp pha loang huyet thanh 1/100.
Vdi khang the khang te bao sieu nghien nhan thay hoat tfnh khdng the a cac thdi diem D30, D60 giam xudng chi cdn phan dng du'ong tfnh d cac mdu huyet thanh pha loang 1/50 lan. Ket qua cho thdy hoat tfnh khang the phan dng vdi khang nguyen sidu nghien manh han so vdi td
filtiil
Bieu do 1: So sanh kha ning phat hien khing the tho khing ti bao goe voi khing nguyen la ti t}ao sieu nghiin va te bao nguyen ven
T^P CHI Tiitac H o r - (ts/2ni7. i^sh <«« MAM ^-J^
Nghien CIFU - Ky thujit
Kdt lu$n
TrSn cdc vdt bdng thi^e nghi$m d thd, td bdo gde trung md cOa dfly rdn ngu-di dd cd nhO-ng bidu hi$n Idm tdng qud trinh lidn vdt thu-ang so v d i nhdm ehdng. Tdc dyng cCia td bdo gdc trung md trong vide hgn chd nhidm khudn t^l vdt bdng chu'a thd hi$n rd rdng. Trong khi dd vide ghdp td bdo gdc trung md Idm cho didn bidn md hpe t^i vdt bdng thudn Icri han cho qud trinh lidn vdt thuwng. Cdc TBGTMMDRN khi d u v c ghdp dj lodi vdo vdt bdng trdn thd ed kich thich thd sinh khdng thd khdng TBGTMMDRN. Hogt tinh khdng thd khdng mgnh vd gidm nhanh sau khi ngung cdy ghdp td bdo.
Summary
The effect of transplantation of human umbilical cord lining mesenchymal stem cells (hCLMSC) on healing expenmental bum wounds was investigated and their immunogenicity was evaluated in 15 healthy rabbits with experimentally induced deep thermal bums on the back, two burn wound areas each. Of the two bum wound areas, one was covered with hCLMSC fixed on Tegaderm membrane on the 5"" day after bum induction.
Wound healing rate, infection state and
histological examination were regularly determined at Interval of 5 days. The hCLMSC improved bum wound healing through the effectiveness on celt regeneration, but were ineffective in limiting infection. In rabbits, antibodies to hCLMSC were produced; the activity of the serum specific antibodies was found highest on the Is"" day of transplantation, then lower and tower on the 3Ct and 6(f day
Tdi li$u tham khdo 1. Dinh Vdn Hdn, Luang Quang Anh, Nguydn VSn Hud (2009), "Nudi cdy keratinocyt trong md trudng khdng huydt thanh vd thu nghidm ghdp keratinocyt ti/
thdn nudi cdy didu tr) vdt thuang bdng", Tpp chl Y hpe thi,ie h6nh, (652+653), tr. 30-34
2 Nguydn Thj Thu Hd (2004). "Td bdo gdc vd Cmg dgng trong y sinh hpe" Tap chl Nghi6n cim y duqc /ipc32{6),tr. 13-26.
3 Binvanlou A H el al (2003), 'Stem cells, setting standards for human embhonic stem cells', Science. 300. pp. 913-916-
4 Mareschi K (2001), "Isolation of human mesenchymal stem cells* bone marrow versus umbilical cord blood", Heamatotogica 86 (10), pp 1099-100.
5. Toan Thang Phan, Ivor Lim (2008). 'Isolation of stem celts-Progenitor ceils from amniotic memt)rane of umbilical cord". UK Patent GB 2432166.
Nghien cuu su dung methyl jasmonat lam chat kich thich sinh tong hop paclitaxel
trong nuoi cay sinh khoi te bao thong do {Taxus wallichiana Zucc.)
Dat van de
Thdng do (Taxus wallichiana Zucc ) phan bd d cao nguyen Da Lat, Lam Dong vdi sd lup'ng ca the nho. Day la du'p'c lidu ghi trong sdch dd can du'p'c luu y bao vp'^' Tii' thdng dd chidt xuat