真皮多能干细胞复合聚乳酸支架修复关节软骨全层缺损的实验研究.DOCVIP

临床医学论文-真皮多能干细胞复合聚乳酸支架修复关节软骨全层缺损的实验研究 ????????????作者：夏亚一，鲁茂森，袁凌伟，邢帅，汪静，王翠芳，汉华，沈海丽? 【摘要】? ［目的］ 探讨真皮多能干细胞在软骨缺损修复中的可行性，为扩大软骨缺损修复中种子细胞的选择提供实验依据。［方法］ 以新生青紫蓝兔为研究对象，机械法直接分离得到真皮组织，采用酶消化法获取细胞，利用干细胞贴壁粘附生长的特性获取高克隆细胞群，并进行传代培养。使细胞浓度达到1×106/ml与聚乳酸支架材料共培养一周后植入兔膝关节软骨全层缺损处，用3～5个月龄青紫蓝兔30只，60个关节，随机分为真皮多能干细胞/聚乳酸支架材料、聚乳酸支架材料、空白对照三组，每组20个关节，于术后12周空气栓塞处死实验兔，对修复组织进行取材，行HE、甲苯胺蓝染色，根据软骨缺损修复情况进行大体、组织学评分，并进行统计学分析，比较各组的评分差异是否具有统计学意义。［结果］大体及组织学观察显示，真皮干细胞/支架材料实验组修复组织表面光滑、呈透明样,与周围软骨及软骨下骨整合良好，而聚乳酸支架材料组有少许透明样软骨,主要以纤维软骨修复,空白对照组则表面有缺损,以纤维软骨修复。大体及组织学观察后进行统计学评分分析，显示A组修复效果最优、优于B、C组（P0.05），B组优于C组（P0.05）［结论］真皮多能干细胞修复关节软骨缺损效果明显，恢复正常关节软骨结构，可作为组织工程化软骨种子细胞之一。 【关键词】? 真皮多能干细胞; 聚乳酸; 关节软骨； 全层缺损; 软骨修复 ??? Abstract: ［Objective］To investigate the feasibility of the dermal pluripotent stem cells in the repairing of cartilage defects. It was aimed to provide experimental basis in view of cartilage defects of expanding the repair of cartilage defects seed cells selection. ［Method］Neonatal cyanotic blue rabbits were used. Dermal tissue directly isolated through mechanical method and? cells? through enzymatic digestion were obtained. The growth characteristics of adherent adhesion of stem cells were used to obtain high cloned cells, which were subcultured. The cell concentration of 1×106/ml was cultured with polylactic acid scaffold for one weeks, and the obtained result was implanted into full-thickness articular cartilage defects of rabbits. Thirty cyanotic blue rabbit (60 joints )for about 3～5 months were randomly divided into three groups:dermal pluripotent stem cell /? scaffold polylactic acid in group A,polylactic acid scaffold in group B, controls in group C. Twenty joints for each group. The rabbits were killed by air embolization at 12 weeks, restoration organization was extracted and stained by HE and toluidine blue .According to the repairing result of cartilage defect, gross and histologic scoring was made, and? analyzed by statistics. The comparison of score difference between each groups was performed statist