壳聚糖复合共混膜培养兔角膜缘上皮及其移植

Cultivating limbal corneal epithelial cells on chitosan complexes and transplanting in rabbit

目的 探讨壳聚糖、胶原、透明质酸钠复合共混膜作为兔角膜缘上皮细胞体外培养载体及其自体移植的可行性。方法 活体取兔左眼角膜缘浅层小块 ,置此共混膜上 ,常规培养 8天。将兔右眼角膜浅层基质分离成囊袋 ,用 3mm环钻去除角膜中央浅表层 ,将含培养角膜缘上皮细胞的共混膜植入自体角膜囊袋内 (6只兔 ) ,对照组则植入不含细胞的共混膜 (6只兔 )。术后观察 1月。结果 角膜缘小块在共混膜上培养生长良好 ,移植手术后 40h ,实验组 5 /6角膜植片荧光素染色阴性 ;对照组术后 4天角膜植片荧光素染色仍为部分阳性。实验组组织学检查显示材料表层上皮完整 ,AEl/AE3免疫组化染色阳性 ,材料部分降解。结论 壳聚糖胶原透明质酸钠共混膜可作为培养角膜缘上皮细胞及其移植的载体。

Objective To investigate the feasibility of chitosan-collagen-sodium hyaluronate complexes as substrates for cultivating limbal corneal epithelial cells and transplanting them onto rabbit eyes.Methods From the limbal region of the left eyes of six rabbits,small biopsy specimens of corneal epithelial cells were taken and cultured for 8 days on the blended biomaterials of chitosan-collagen-sodium hyaluronate. A circular blended biomaterial with diameter of 6.5 mm was inserted into corneal pocket with a 3 mm central window. Six eyes received grafts of blended biomaterials containing autologous cultured epithelial cells.Six eyes received grafts of blended biomaterials without epithelial cells and served as controls. Transplanted eyes were examined daily for up to 30 days. Epithelialization of cornea,histological examination and immunohistochemistry for AE1/AE3 were performed.Results The cultivated corneal epithelial cells on the blended biomaterials of chitosan-collagen-sodium hyaluronate grew well.5/6 of rabbit eyes that were grafted with the cultivated epithelial cells on the blended biomaterials were all epithelialized 40 h after surgery. Corneas that were grafted with blended biomaterials alone were epithelialized 5-6 days after surgery.Histology revealed that experimental eyes achieved epithelialization.Epithelial cells were positively immunostaied with AE1/AE3 monoclonal antibody. The biomaterials degraded.Conclusion The blended biomaterials of chitosan-collagen-sodium hyaluronate may be as suitable substrate for cultivating limbal corneal epithelial cells and transplantation.