异种猪–兔角膜基质透镜植入可行性及安全性评价

The Safety and Feasibility of Xenogeneic Intrastromal Lenticule Inlay in a Pig-to-Rabbit

目的:本研究通过探究异种猪兔角膜基质微透镜作为植入材料的可行性,为角膜微透镜的个性化定制提供了实验基础。方法:取20只猪角膜基质微透镜植入新西兰白兔角膜基质囊袋中,均选右眼为实验组,一组10只眼联合角膜交联(CXL组),另一组10只眼为非CXL组,每个时间段(一周,一个月,3个月,6个月和1年)每组取两只实验兔进行研究,另选3只新西兰白兔作为空白对照组。裂隙灯检查兔眼角膜的透明度并照相,病理组织切片分析HE染色、免疫组化及超微电镜结果,观察术后1年猪兔异种角膜基质微透镜植入后的排斥反应和愈合反应,探究异种微透镜作为基质植入材料的可行性。结果:异种猪–兔角膜基质透镜植入CXL组和非CXL组术后1年内不同时间点分别观察结果显示:除一例非CXL组兔眼角膜混浊外,其余19例裂隙灯检查兔眼角膜透明,未见新生血管;眼前节照相可见透镜居中透明;HE染色植入的微透镜与周围基质组织之间的边界明显,角膜全层未见明显炎性细胞浸润和新生血管形成;免疫组化CXL组和非CXL组均未观察到I型胶原异常高表达。术后1年CXL组微透镜中I型胶原蛋白的表达较植床明显减少,也较非CXL组微透镜中I型胶原蛋白表达少;电镜结果两组植入的微透镜内的胶原纤维排列与对照组相似,胶原纤维在分界处呈梭形且不规则。结论:异种猪兔角膜微透镜植入可行,猪兔角膜组织相容性好,未见排斥反应。

Purpose: To evaluate the safety and feasibility of xenogeneic intrastromal lenticule inlay in a pig- to-rabbit with different interventions. Method: Twenty New Zealand white rabbits were divided evenly into experimental group and control group (10 rabbits each group). Corneal intrastromal pockets were made by femtosecond laser, and corneal stromal lenticules extracted from porcine eyes were implanted into rabbit eyes. Corneal collagen crosslinking (CXL group) and non-CXL treated (Non-CXL group) were all performed in right. Two rabbits from each group were euthanized at 1 week, 1 month, 3 months, 6 months and 1 year after surgery, respectively. Another 3 New Zealand white rabbits were selected as the control group. The structural changes of corneas after lenticule implantation were observed by hematoxylin-eosin staining and transmission electron microscopy. Corneal healing and rejection reactions were observed using immunohistochemistry and immunofluorescence staining. Results: The results showed that except for one case of xenogeneic porcine-rabbit corneal stromal lenticule was epinephelos in non CXL group, the cornea of the other 19 cases were transparent without cell infiltration by slit lamp examination. Corneal collagen was arranged regularly, the boundary between implanted lenticules and surrounding stroma was clear, and no inflammatory cell infiltration was detected under light microscopy. The collagen fibers at the junction were fusiform while fibers inside the lenticules are arranged regularly. Collagen I inside the implanted lenticules was arranged regularly, and there was no obviously high expression of collagen in both CXL and non-CXL groups. Conclusion: Pig-to-rabbit intrastromal lenticule xenotransplantation is safe and feasible. Implanted lenticules are compatible with surrounding tissue, with stable wound healing and no rejection reaction.