组织工程化软骨组织形成的最佳细胞浓度和最佳形成时间的实验研究

THE EXPERIMENTAL STUDY ON OPTIMAL CELL DENSITY AND FORMATION TIME OF TISSUE ENGINEERED AUTOLOGOUS CARTILAGE

目的研究探讨自体组织工程化软骨在动物自体内形成的最佳细胞浓度和最佳形成时间。方法取贵州香猪部分耳廓软骨组织体外消化分离软骨细胞，与可注射性生物材料Ｐｌｕｒｏｎｉｃ混合形成复合物，浓度为１０、２０、３０、４０、５０、６０、７０×１０６／ｍｌ，将细胞－生物材料复合物接种于猪自体腹外侧壁皮下。第６周取材，分别称重，并作ＨＥ、ＳａｆｒａｎｉｎＯ染色，评价软骨组织形成的最适细胞浓度。参考上述实验结果，制成最适软骨细胞－生物材料复合物，接种到猪自体腹外侧壁皮下，分别在接种后第１、３、６、９和１５周取材，大体观察，并作ＨＥ、ＳａｆｒａｎｉｎＯ染色，确定软骨组织形成的最佳时间。结果在动物自体内形成的软骨组织，与正常软骨组织有相似的组织学特性，形成软骨组织最适细胞浓度为５０×１０６／ｍｌ，最佳形成时间是第６周。结论成功地利用组织工程技术在具有免疫功能的自体动物体内形成了软骨组织，并确定了软骨组织形成的最佳细胞接种浓度和接种的软骨组织形成最佳时限。

Objective This paper aims to investigate the suitable cell density and the best formation time of tissue engineered autologous cartilage and to provide theoretical basis and parameters for clinical application.Methods The chondrocytes isolated from mini swines' ears were mixed with injectable biocompatible matrix (Pluronic),and the density of cell suspensions were 10,20,30,40,50,60,70×10 6/ml.The chondrocyte polymer constructs were subcutaneously injected into the abdomen of autologous swine. The specimens were observed grossly and histologically after 6 weeks, and investigated the suitable cell density. Then the chondrocyte polymer constructs with suitable cell density were transplanted into the abdomen of autologous swine and evaluated grossly and histologically in 1, 3, 6, 9, 15 weeks after transplantation to investigate the best formation time of tissue engineered cartilage.Results The experiments demonstrated that the tissue engineered autologous cartilage was similar to the natural cartilage on animals with normal immune system in histological characteristics. The optimal chondrocyte density is 50× 10 6/ml , and the proper harvest time is the sixth week.Conclusion With tissue engineering skills, we have identified the optimal chondrocyte density and the proper harvest time.