角膜体外重构异种生物载体材料植入性实验研究  

Study on animal intracorneal implantation model with xenogeneic swine corneal stroma

作　　者：吴静[1] 郝念[1] 李姝燕[1] 徐锦堂[1] 王彦平[1] 

机构地区：[1]暨南大学医学院眼科研究室,广东广州510632

出　　处：《中国病理生理杂志》2005年第6期1193-1197,共5页Chinese Journal of Pathophysiology

基　　金：广东省科技计划基金资助项目(No.2KM05203S);国家自然科学基金资助项目(No.39870855)

摘　　要：目的:分析测定异种(猪)角膜基质组织免疫原性,观察其角膜层间植入后与受体角膜愈合情况,以评价该材料生物相容性,并在此载体上体外重构角膜内皮组织,探讨其作为角膜重建载体材料可行性。方法:(1)将新鲜、脱水两种猪角膜基质植片分别植入F344大鼠角膜基质层间,术后12、90d对外周血进行CD25和CD4/CD8双色免疫荧光标记,流式细胞仪测定分析。(2)猪角膜基质植入新西兰白兔角膜层间,定期临床观察植片愈合情况,并取受体兔角膜进行组织学观察。(3)将猫角膜内皮细胞接种于保留后弹力层的脱水猪角膜基质上,加入培养液培养7d,组织学观察体外重构的角膜内皮组织形态结构。结果:(1)测得新鲜组、脱水组大鼠外周血T淋巴细胞CD4+CD25+、CD8+CD25+双阳性表达率及CD4+/CD8+比值与同基因移植组、阴性对照组比较无显著差异(P>0.05)。(2)兔眼临床观察:全部植片存活,12只术眼未见有角膜水肿混浊、角膜新生血管、排斥反应发生,新鲜植片在2个月左右已透明,脱水植片在6个月后透明。兔角膜组织学观察:新鲜植片4个月时与兔角膜基质相融愈合,脱水植片经角膜细胞再分布、胶原纤维改建重塑于8个月后与兔角膜基质相融愈合,两组植片愈合过程中未见有淋巴细胞浸润及新生血管生成。(3)体外重构的内皮组织形态结构与正常内皮层相似。结论:异种(猪)角膜基质免疫原性低,具有良好的生物相容性,是目前较为理想的角膜体外重构载体材料。AIM: To study the immunogenicity and biocompatibility of xenogeneic swine corneal stroma as biological carrier for cornea reconstruction and to reconstruct corneal endothelial tissue with this carrier in vitro. METHODS: (1) The lymphocytes from the peripheral blood of F_(344) rats were immunologically labeled by anti-rat CD25-FITC and anti-rat CD4/CD8-PE, then determined by flow cytometry (FCM) at 12^(th), 90^(th) day after intracorneal implantation with fresh and dehydrated swine corneal stroma. (2) The fresh and dehydrated grafts made of swine corneal stroma were implanted intralamellarly in corneas of New Zealand rabbits. Clinical examinations were performed monthly and histological examinations were made at 14^(th), 30^(th), 60^(th), 120^(th) and 240^(th) day. (3) The cat corneal endothelial cells were seeded on the Descemet's membrane of the dehydrated swine corneal stroma, then cultured in the medium with epidermal growth factor and laminin for 7 days. The morphology of reconstructed tissue was tested by microscope. RESULTS: (1) Compared to isograft group and negative control, the expression CD4^+CD25^+, CD8^+CD25^+ and CD4^+/CD8^+ of xenograft rat group implantation with swine corneal stroma did not appear significantly different in statistics (P>0.05). (2) In the total 12 rabbits, all the cornea grafts survived without rejection reaction, corneal haze or corneal neovascularization. The fresh grafts got transparent after 2 months, and the dehydrated grafts got transparent after 6 months. Histological study demonstrated both fresh and dehydrated stroma grafts had fused with rabbits'corneal stroma very well without lymphocytes infiltrating. (3) As shown in histological observations, the reconstructed cat corneal endothelial tissue was similar to the nature tissue. Cultured cat endothelial cells connected tightly to each other and attached to the Descemet's membrane closely. CONCLUSION: Swine corneal stroma has low immunogenicity and satisfying biocompatibility,it is an ideal biological carrier for cornea rec

关 键 词：角膜基质 生物相容性材料 猪 猫 

分 类 号：R363[医药卫生—病理学]