温敏性壳聚糖水凝胶支架联合骨髓间充质干细胞治疗兔膝关节软骨缺损  

Thermosensitive chitosan hydrogel scaffold combined with bone marrow mesenchymal stem cells for treatment of knee cartilage defects in rabbits

作　　者：李明静 李凡[1] 徐剑 向江 郭井泉 LI Mingjing;Li Fan;Xu Jian;Xiang Jiang;Guo Jingquan(Department of Pediatric Orthopedics,Wuhan Fourth Hospital,Puai Hospital,Tongji Medical College Affiliated to Huazhong University of Science and Technology,Wuhan 430033,China)

机构地区：[1]武汉市第四医院华中科技大学同济医学院附属普爱医院小儿骨科,武汉430033

出　　处：《中华手外科杂志》2021年第4期283-289,共7页Chinese Journal of Hand Surgery

基　　金：湖北省自然科学基金(2017CFB385);武汉市临床医学研究项目(WX17Q20)。

摘　　要：目的制备壳聚糖/Ⅱ型胶原/聚乳酸复合水凝胶支架,以支架为骨髓间充质干细胞载体,评价修复兔膝关节软骨缺损的可行性。方法将壳聚糖、Ⅱ型胶原、聚乳酸联合β-甘油磷酸钠盐制备复合水凝胶,检测凝胶形成时间、弹性模量及细胞相容性;制作新西兰兔关节软骨缺损模型,分为四组。空白组:正常软骨,未作任何处理;模型组:旷置骨软骨缺损作为对照;凝胶组:单纯以水凝胶填充软骨缺损;凝胶+干细胞组:以骨髓间充质干细胞及水凝胶复合物填充软骨缺损。分别于术后8周取材,根据大体观察及组织染色,比较各组膝关节软骨缺损修复效果。结果壳聚糖/Ⅱ型胶原/聚乳酸复合溶液在37℃下形成凝胶,成胶时间(7.50±0.41)min,弹性模量(6.96±0.43)kPa;骨髓间充质干细胞在凝胶中具有较好的增殖能力,且软骨分化相关基因的表达明显增高;动物实验大体观察及组织染色结果表明凝胶+干细胞组软骨缺损修复效果明显优于凝胶组及模型组,其新生软骨与正常软骨边界模糊,其内可见软骨细胞,Ⅱ型胶原蛋白含量与正常软骨类似。结论壳聚糖/Ⅱ型胶原/聚乳酸复合水凝胶支架具有良好的温敏性及细胞相容性,联合骨髓间充质干细胞可有效修复兔膝关节软骨缺损。Objective To prepare chitosan/typeⅡcollagen/polylactic acid composite hydrogel scaffold used as a carrier for bone marrow mesenchymal stem cells and to evaluate the feasibility of repairing cartilage defects of rabbit knee joints by bone marrow mesenchymal stem cells.Methods The chitosan hydrogel was synthesized from chitosan,typeⅡcollagen and polylactic acid combined withβglycerophosphate.The gel formation time,elastic modulus and cytocompatibility were detected.The articular cartilage defect model of New Zealand rabbit was made into four groups.Blank group:normal cartilage without any treatment;model group:bone cartilage defects as control;gel group:pure gel filled cartilage defects;gel+stem cell group:bone marrow mesenchymal stem cells and hydrogel complex filled cartilage defects.At 8 weeks after operation,the repair outcomes of knee joint cartilage defects in each group were compared according to the gross observation and tissue staining.Results The chitosan/type Ⅱ collagen/polylactic acid composite solution formed gel at 37℃,gel time(7.50±0.41)minutes,elastic modulus(6.96±0.43)kPa.The bone marrow mesenchymal stem cells had good proliferation ability in gel,and the expression of cartilage differentiation related genes increased significantly.The animal gross observation and tissue staining showed that the effect of gel+stem cell group on cartilage defect repair was significantly better than that of gel group and the model group.The boundary between the new cartilage and the normal cartilage was fuzzy,and chondrocytes were found in the cartilage.The content of typeⅡcollagen was similar to that of normal cartilage.Conclusion The chitosan/type Ⅱ collagen/polylactic acid composite hydrogel scaffold has good thermosensitivity and cytocompatibility,and combined with bone marrow mesenchymal stem cells can effectively repair rabbit knee cartilage defects.

关 键 词：间充质干细胞 膝关节 软骨缺损 壳聚糖水凝胶 温敏性 

分 类 号：R318.08[医药卫生—生物医学工程]