羊膜接触镜输送人表皮干细胞重建兔角膜上皮

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.

Reconstruction of rabbits' corneal epithelium with human epidermal stem cells delivery by amniotic membrane contact lens

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.