中华生物医学工程杂志
中華生物醫學工程雜誌
중화생물의학공정잡지
CHINESE JOURNAL OF BIOMEDICAL ENGINEERING
2009年
1期
14-19
,共6页
万鹏霞%王智崇%马萍%高楠
萬鵬霞%王智崇%馬萍%高楠
만붕하%왕지숭%마평%고남
羊膜%接触镜%干细胞移植%角膜缘干细胞缺损%上皮,角膜
羊膜%接觸鏡%榦細胞移植%角膜緣榦細胞缺損%上皮,角膜
양막%접촉경%간세포이식%각막연간세포결손%상피,각막
Amnion%Contact lenses%Stem cells transplantation%Limbal stem celldeficiency%Epithelium,corneal
目的 利用眼表生物膜固定装置(BMFD)与羊膜(AM)制成接触镜,输送人表皮干细胞至角膜缘干细胞缺损(LSCD)的兔眼表,并评价其重建角膜上皮的效果.方法 制作去角膜上皮及角膜缘干细胞的雌性LSCD兔模型,分为3组:羊膜加人表皮干细胞移植组(n=20),将男性人表皮干细胞悬液注射到BMFD-AM接触镜(简称羊膜接触镜)与眼表之间;羊膜遮盖组(n=20),戴羊膜接触镜;对照组(n=20),单纯药物治疗.裂隙灯显微镜结合角膜荧光素钠染色观察并评价角膜修复情况.随访至角膜完全上皮化后,取角膜组织行病理学检查和免疫组织化学鉴定,PCR检测人Y-STR基因鉴定细胞来源.结果 羊膜加人表皮干细胞移植组角膜上皮修复快,平均(5.60±0.46)d达到完全上皮化,另外两组角膜上皮修复迟缓.羊膜遮盖组(9.25±0.51)d、对照组(12.45±0.65)d才完成角膜上皮化(P均<0.05).组织病理学示羊膜加人表皮干细胞移植组角膜上皮细胞形态接近正常,K3/K12(+)、Mucin 5AC(-)、K4(-).并可检测到人Y-STR基因.组织病理学示另外两组角膜上皮结膜化,K4(+)、Mucin 5AC(+)、K3/K12(-).结论 羊膜接触镜联合人表皮干细胞悬液注射能够重建LSCD兔角膜上皮.
目的 利用眼錶生物膜固定裝置(BMFD)與羊膜(AM)製成接觸鏡,輸送人錶皮榦細胞至角膜緣榦細胞缺損(LSCD)的兔眼錶,併評價其重建角膜上皮的效果.方法 製作去角膜上皮及角膜緣榦細胞的雌性LSCD兔模型,分為3組:羊膜加人錶皮榦細胞移植組(n=20),將男性人錶皮榦細胞懸液註射到BMFD-AM接觸鏡(簡稱羊膜接觸鏡)與眼錶之間;羊膜遮蓋組(n=20),戴羊膜接觸鏡;對照組(n=20),單純藥物治療.裂隙燈顯微鏡結閤角膜熒光素鈉染色觀察併評價角膜脩複情況.隨訪至角膜完全上皮化後,取角膜組織行病理學檢查和免疫組織化學鑒定,PCR檢測人Y-STR基因鑒定細胞來源.結果 羊膜加人錶皮榦細胞移植組角膜上皮脩複快,平均(5.60±0.46)d達到完全上皮化,另外兩組角膜上皮脩複遲緩.羊膜遮蓋組(9.25±0.51)d、對照組(12.45±0.65)d纔完成角膜上皮化(P均<0.05).組織病理學示羊膜加人錶皮榦細胞移植組角膜上皮細胞形態接近正常,K3/K12(+)、Mucin 5AC(-)、K4(-).併可檢測到人Y-STR基因.組織病理學示另外兩組角膜上皮結膜化,K4(+)、Mucin 5AC(+)、K3/K12(-).結論 羊膜接觸鏡聯閤人錶皮榦細胞懸液註射能夠重建LSCD兔角膜上皮.
목적 이용안표생물막고정장치(BMFD)여양막(AM)제성접촉경,수송인표피간세포지각막연간세포결손(LSCD)적토안표,병평개기중건각막상피적효과.방법 제작거각막상피급각막연간세포적자성LSCD토모형,분위3조:양막가인표피간세포이식조(n=20),장남성인표피간세포현액주사도BMFD-AM접촉경(간칭양막접촉경)여안표지간;양막차개조(n=20),대양막접촉경;대조조(n=20),단순약물치료.렬극등현미경결합각막형광소납염색관찰병평개각막수복정황.수방지각막완전상피화후,취각막조직행병이학검사화면역조직화학감정,PCR검측인Y-STR기인감정세포래원.결과 양막가인표피간세포이식조각막상피수복쾌,평균(5.60±0.46)d체도완전상피화,령외량조각막상피수복지완.양막차개조(9.25±0.51)d、대조조(12.45±0.65)d재완성각막상피화(P균<0.05).조직병이학시양막가인표피간세포이식조각막상피세포형태접근정상,K3/K12(+)、Mucin 5AC(-)、K4(-).병가검측도인Y-STR기인.조직병이학시령외량조각막상피결막화,K4(+)、Mucin 5AC(+)、K3/K12(-).결론 양막접촉경연합인표피간세포현액주사능구중건LSCD토각막상피.
Objective To introduce contact lens made up of bio-membrane fixing device (BMFD) and amniotic membrane (AM) as a cell delivery system on ocular surface and evaluate its efficiency in transplanting human epidermal stem cells (Epi-SCs) to the limbal stem cell deficiency (LSCD) rabbits' ocular surface for corneal epithelium reconstruction. Methods The female LSCD rabbit models with the limbal stem cells and corneal epithelium removed were established, and were divided into 3 groups. The concentrated suspension of male human Epi-SCs was injected into the space between the LSCD rabbits' ocular surface and the BMFD/AM contact lens (AM contact lens for short) in AM + Epi-SCs group (n=20), wearing the AM contact lens was performed in AM patching group (n=20), and only drug treatment was performed in control group (n=20). All the rabbits were followed up with slit-lamp examination and corneal sodium fluorescent staining, and the corneal restoration was evaluated. Pathological and immunohistochemical stained tissue sections were examined till completely re-epithelization. Human Y-STR gene was detected to trace the implanted Epi-SCs by PCR. Results The complete re-epithelization time of cornea in AM + Epi-SCs group was (5.60±0.46) d, which was much shorter than that of the other two groups [AM patching group: (9.25±0.51) d; control group: (12.45 ± 0.65) d, all P<0.05]. The reconstructed epithelium in AM + Epi-SCs group showed K3/K12 (+), Mucin 5AC (-), K4 (-) detected by fluorescent immunohistochemistry, which indicated a normal phenotype of corneal epithelium. And human Y-STR gene was found too. By contrast, the corneal epithelium showed conjunctiva phenotype in control group with K4 (+), Mucin 5AC(+), K3/K12(-). Conclusion AM contact lens can serve as a cell delivery system and help corneal epithelium reconstruction in LSCD rabbits by combined with the human Epi-SCs suspension injection.
