壳聚糖水凝胶复合脂肪间充质干细胞修复兔关节软骨缺损

摘要

Objective To fabricate a novel tissue-engineered cartilage with adipose-derived stem cells (ADSCs) seeded on the chitosan hydrogel scaffold to repair articular cartilage defect.Methods Adipose tissue and costal cartilage were harvested from New Zealand rabbits,and ADSCs in passage one and chondrocytes were obtained after the samples were digested and cultured in vitro.ADSCs were digested,suspended,seeded onto the sterile chitosan gel,and cultured in vitro for 1 week to fabricate the tissue-engineered cartilage.The defects were respectively filled with the tissue-engineered cartilage (composite group),chondrocyte suspension (cell group),chitosan gel (material group) and nothing at all (control group).At postoperative 12 weeks,cartilage repair was evaluated using the gross examination,histological staining,immunohistochemical staining and international cartilage repair society (ICRS) histological score.Results Effect of cartilage repair in composite group was significantly better compared to other groups.The regenerated tissue in composite group seemed tightly bound in normal tissue,with similar structure and extracellular matrix secretion.ICRS histological score in composite group was (13.89 ± 0.14) points,which differed significantly from (7.06 ± 0.19) points in control group,(7.14 ± 0.22) points in cell group and (7.46 ± 0.26) points in material group (P ＜0.01).Conclusion The tissue-engineered cartilage with ADSCs seeded onto the chitosan hydrogel is effective for repair of articular cartilage defect.%目的 探讨利用脂肪间充质干细胞(ADSCs)复合壳聚糖水凝胶支架构建的组织工程软骨修复兔关节软骨缺损的效果. 方法 分别取新西兰大白兔皮下脂肪和肋软骨,消化后体外扩增培养分别得到P1代ADSCs和软骨细胞.将ADSCs消化后制成细胞悬液,并种植于灭菌后的壳聚糖水凝胶上,体外培养1周构建组织工程软骨,将构建的组织工程软骨植入到兔的关节软骨缺损处.实验分为复合组(缺损处植入ADSCs复合新型壳聚糖水凝胶支架)、细胞组(缺损处注射一定浓度的软骨细胞悬液)、材料组(缺损处植入单纯的壳聚糖水凝胶)和对照组(缺损处未做任何处理),各组分别于术后12周取样,通过大体观察、HE染色、番红-O及Ⅱ型胶原免疫组化染色等方法观察缺损关节软骨的修复情况,并用国际关节软骨修复协会(ICRS)制订的评分法进行组织学评分. 结果 复合组缺损软骨的修复效果明显优于其他3组,新生组织与正常组织结合紧密,且结构和细胞外基质的分泌情况类似于正常组织.对照组、细胞组、材料组和复合组ICRS评分分别为(7.06±0.19)分、(7.14±0.22)分、(7.46±0.26)分和(13.89±0.14)分,复合组与其他各组差异有统计学意义(P＜0.01). 结论 ADSCs复合壳聚糖凝胶支架构建的组织工程软骨对缺损关节软骨具有良好的修复作用,且修复是一种结构性的修复.