骨髓基质细胞膜片复合聚乳乙醇酸支撑体体外构建管状软骨  

Constructing tissue engineered trachea-like cartilage graft in vitro by using bone marrow stromal cells sheet and PLGA internal support： experimental study in bioreacter

作　　者：张浚睿 陈富林[2] 吴炜 魏建华 封兴华 毛天球 

机构地区：[1]第四军医大学口腔医院颌面外科,西安710032 [2]陕西西安西北大学生命科学院

出　　处：《中华整形外科杂志》2009年第2期124-128,共5页Chinese Journal of Plastic Surgery

基　　金：基金项目：国家“973”课题项目（G1999054308）;军队临床高新技术重大项目-骨与软骨组织工程技术

摘　　要：目的研究利用骨髓基质细胞膜片复合聚乳乙醇酸（ploy of lactic-co-glycolic acid，PLGA）支撑体，在生物反应器条件下体外构建管状软骨的可行性。方法分离兔骨髓基质细胞，高密度连续培养，转化生长因子-1诱导构建成干细胞膜片，制作圆柱状PLGA支撑体，将细胞膜片均匀缠绕在表面。静置孵育14d，使细胞膜片与PLGA相互贴附后，进入生物反应器动态培养8周后，取出标本。从大体形态、组织学结构、蛋白多糖含量以及生物力学性能等方面评价形成软骨的理化特性。结果通过此策略构建的软骨外观与天然软骨组织非常相似，保持着良好的管状外形，颜色呈乳白色，有光泽，质地均匀，弹性好，具有中等偏软的硬度。组织学结果显示总体结构呈现软骨样结构，HE染色可见软骨样细胞分布于细胞陷窝之中，周围是均匀的细胞外基质，番红-O染色可见细胞外基质着色为鲜红色，提示蛋白多糖含量丰富，有大量软骨基质物产生。结论细胞膜片复合支撑体策略能形成管状形态的软骨组织，为气管软骨的再造提供了新的方法，有可能解决气管缺损的临床难题。Objective To explore the feasibility of constructing tissue engineered trachea-like cartilage graft in vitro by using bone marrow stromal cells（BMSCs） sheet and PLGA internal support. Methods Rabbit BMSCs were expanded and induced by transforming growth factor-1 to improve chondrocyte phenotype of BMSCs. BMSCs sheets were obtained by continuous culture and wrapped the PGLA scaffold in the shape of cylinder. The constructs were incubated in spinner flask for 8 weeks and cartilage formation was investigated by gross inspection, histology, glyeosaminoglyean and mechanical strength content. Resdts After in vitro culture, cartilage like tissue in cylindrical shape had been regenerated successfully. Stiff, shiny, pearly opalescence tissues were observed. Histological analysis showed engineered trachea cartilage consisted of evenly spaced lacunae embedded in matrix, cells stationed in the lacunae could he noticed clearly. Safranin-O staining on the sections showed homogenous and positive red staining, which demonstrated that the engineered tissue was rich in proteoglycans. Conclusions Based on the cell sheet and internal support strategy, trachea-like cartilage in cylindrical shape could be successfully fabricated which provided a highly effective cartilage graft substitute and could be useful in many situations of trachea-cartilage loss encountered in clinical practice.

关 键 词：组织工程 生物反应器 细胞膜片 气管 软骨 

分 类 号：R318.0[医药卫生—生物医学工程]