摘要
目的:观察以脱细胞角膜基质作为支架材料、以骨髓间充质干细胞作为种子细胞复合构建组织工程化人工活性角膜的可行性。方法:实验于2004-09/2005-06在南昌大学江西医学院烧伤研究所完成。①取兔新鲜角膜,浸入1.25g/L胰蛋白酶和0.1g/L乙二胺四乙酸溶液中作用24h,然后浸入1%曲拉通X-100溶液中作用120h,制备成脱细胞角膜基质,进行常规组织学苏木精-伊红染色观察。②采用密度梯度离心结合贴壁法分离纯化人骨髓间充质干细胞,用含10μg/L表皮生长因子的Dulbecco最低必需培养基进行诱导培养。③将经诱导培养的骨髓间充质干细胞种植于脱细胞角膜基质上,复合构建组织工程化人工活性角膜,观察细胞与材料的黏附、细胞增殖情况。结果:①脱细胞角膜基质主要由胶原纤维组成,未见细胞成分残留。②经表皮生长因子诱导培养的人骨髓间充质干细胞体外增殖良好,24h后贴壁细胞增多,7～10d细胞多为梭形,呈有规则的集束辐射状排列;14～21d左右细胞基本融合。③将人骨髓间充质干细胞种植于脱细胞角膜基质上,细胞铺展和增殖良好。结论:以骨髓间充质干细胞作为种子细胞复合脱细胞角膜基质支架材料可成功构建组织工程化人工活性角膜,有望成为一种修复角膜缺损的新技术方法。
AIM： To investigate the feasibility of construction of tissue engineered actively artificial cornea with acellular cornea matrix as scaffold and bone marrow-derived mesenchymal stem cells （MSCs） as seed cell.METHODS： The experiment was conducted in the Bum Institute of the First Affiliated Hospital of Jiangxi Medical College, Nanchang University from September 2004 to June 2005. （1）The fresh corneas from rabbits were put into the solution including 1.25 g/L trypsin and 0.1 g/L ethylene diamine tetraacetic acid （EDTA） for 24 hours. After that, those corneas were immerged into the solution with 1% Triton X-100 for 120 hours to prepare the acellular cornea matrix and were observed with routine histological haematoxylin-eosin staining. （2）The human MSCs were isolated and purified by combining density gradient centrifugation with adherent method, and were induced and cultured with 10 μg/L epidermal growth factor in Dulbecco＇s modified Eagle＇s medium. （3）The induced and cultured MSCs were implanted into the acellulas matrix to construct the tissue engineered cornea. The biological attachment and growth of cultured MSCs were observed. RESULTS： （1）The acellular cornea matrix was mainly ,composed of collagen fibers without cell pieces. （2）The human MSCs induced and cultured by epidermal growth factor grew well in vitro. The adherent cells increased after 24 hours, turned to be fusiform shape with order clusterradial arrangement between the 7^th to 10^th days, and the cells fused well between the 14^th and 21^th days. （3）The human MSCs were implanted into the acelllular cornea matrix, where the cells spread and proliferated well. CONCLUSION： The tissue engineered cornea can be reconstructed successfully by combining MSCs with acellulas cornea matrix, which may be a new technological method for the repair of corneal defect.
出处
《中国临床康复》
CAS
CSCD
北大核心
2006年第25期54-56,共3页
Chinese Journal of Clinical Rehabilitation
