角膜基质脱细胞处理与相容性实验研究

Experimental study on preparation of acellular corneal stroma and its compatibility

目的寻找有效适宜的脱细胞方法对猪角膜进行处理,制备角膜生物支架材料。方法利用胰酶和胶原酶消化结合液氮冻融的方法对猪角膜基质进行处理,60Co消毒,冻干保存。经组织学观察、细胞培养附和与动物移植检验其生物特性。结果获得的脱细胞角膜基质,显微结构观察显示板层结构完整,胶原纤维间组织结构疏松,未见细胞残留,整个基质成网状结构,在上皮面保存有完整连续的基底膜。细胞培养附和实验观察到种子细胞可以在基质上良好生长。动物移植实验未见明显的炎症反应与排斥。结论经该法脱细胞处理后得到的角膜基质无细胞毒性,组织相容性好,可以成为适宜的组织工程支架材料。

Objective To investigate an effective and compatible way to obtain acellular cornea, which should be a suitable scaffold for construction of tissue engineering cornea. Methods Fresh cornea of swine were treated by taking enzyme digestion combined with frozen in liquid nitrogen and then thawed. Collected after the process, swine cornea became acellular stroma. Treated cornea were sterilized with 60 Co and preserved in plastic bags after a freeze-drying process. Histologic examination, seed cell compatibility and transplantation to the rabbit were performed to assess biocompatibility. Results Histologic diagnostic results showed no cell residues. Collagen fibers were arranged in a loose fashion, and lamina structure resembled that of the normal. It had complete continual epithelium basement membrane and seed cells could be cultured well on it. No rejection was observed after transplantation onto the surface of the rabbit＇s eye for months. Gonclusion Acellular corneal stroma prepared by this method has a good biocompatibility and atoxity for cells. Results of the experiment indicate that this method can be a suitable tissue engineering scaffold for cornea.