同种异体气管软骨脱细胞基质构建组织工程化气管的初步研究  

Experimental study on construction of tissue-engineered trachea with allogenic acellular matrix of tracheal cartilage

作　　者：李尚滨[1] 于庆平[2] 张文浩[3] 霍然[1] 

机构地区：[1]济南山东大学附属省立医院烧伤整形美容外科,250021 [2]山东省菏泽市立医院烧伤整形外科,274000 [3]山东省济宁市第一人民医院烧伤整形科,272000

出　　处：《中华损伤与修复杂志（电子版）》2009年第6期11-14,共4页Chinese Journal of Injury Repair and Wound Healing(Electronic Edition)

基　　金：山东省自然基金资助项目(Y2004C14)

摘　　要：目的初步探寻同种异体气管软骨脱细胞基质与向软骨诱导的骨髓间充质干细胞(MSCs)共同构建组织工程化气管的可行性。方法分离并诱导宿主犬MSCs向软骨细胞分化,通过内蛋白多糖含量测量及Ⅱ型胶原免疫组化染色观察初步诱导效果。使用曲拉通化学消化法对供体犬气管软骨进行脱细胞处理,根据消化时间分组,观察气管软骨脱细胞基质制备效果。将诱导后MSCs与同种异体气管软骨脱细胞基质分别进行体外与体内复合培养,组织学及电镜下观察诱导细胞的生长情况,以进行构建组织工程化气管可行性初步分析。结果宿主犬MSCs软骨方向诱导后诱导组细胞培养液内蛋白多糖含量为(42.48±2.32)μg/ml,较未诱导组(19.62±1.30)μg/ml明显增高(p<0.01),诱导组细胞Ⅱ型胶原免疫组化染色阳性。HE染色及扫描电镜均可见120小时组供体犬气管组织黏膜细胞及软骨细胞完全消失,软骨基质结构完整。体外联合培养7天内可见诱导后MSCs在脱细胞软骨基质表面生长,14天时细胞逐渐凋亡;体内联合培养可见诱导细胞在软骨基质浅层生长,但基质内部未见细胞成分生长,并无新生软骨形成。结论使用曲拉通化学消化法可成功制备结构完整的同种异体气管软骨脱细胞基质,但作为构建组织工程化气管的支架材料,其空隙率和贯通性仍需进一步改进。Objective To explore the application value of the allogenic tracheal acellular matrix in the construction of tissue-engineered trachea.Methods The allogenic trachea rings were processed by improved-Courtman method to remove all cells in the tissue.The chondrogenic induced MSCs taken from host canines were cultured with allogenic tracheal acellular matrix in vitro and in vivo t o observe the adherence and proliferation.Results All cells in the allogenic tracheas were removed completely by improved-Courtman method without damage to the structure of the tracheal acellular matrix.The induced MSCs survived and proliferated on the surface of matrix in a short period.But thecells failed to grow into the internal part of the matrix and form new cartilage tissue.Conclusions The improved-Courtman method can remove all cells of the trachea samples completely.But the features of void ratio and perforating ability need to be further improved before the allogenic tracheal acellular matrix is used for the scaffold material of tissue-engineeredtrachea.

关 键 词：犬 脱细胞基质 气管 软骨 组织工程 

分 类 号：R318[医药卫生—生物医学工程]