丝素蛋白-壳聚糖支架复合骨髓间充质干细胞体内构建组织工程化软骨的生物相容性

文题释义： 生物相容性：是指生命体组织对非活性材料产生的一种性能，一般是指材料与宿主之间的相容性，包括组织相容性和血液相容性。 检测相容性的方法：是将支架材料与种子细胞在体外共培养，检测支架毒性、细胞活性、细胞增殖及细胞与支架的黏附情况等指标，该方法具有客观性强、可重复性强、影响因素相对简单及敏感性高等特点。 背景：课题组前期的研究中发现，丝素蛋白-壳聚糖支架材料复合诱导后骨髓间充质干细胞在兔体内能修复缺损的软骨组织，但对于该组织工程化软骨组织的生物相容性还未进一步研究。 目的：研究丝素蛋白-壳聚糖支架材料复合骨髓间充质干细胞在体内构建组织工程化软骨的生物相容性。 方法：使用丝素蛋白-壳聚糖按1∶1比例混合制备三维支架材料，提取兔骨髓间充质干细胞，将诱导后的骨髓间充质干细胞与丝素蛋白-壳聚糖支架构建修复体，再将修复体移植到兔关节软骨缺损模型中修复软骨组织。实验分为3组，实验组植入诱导后骨髓间充质干细胞+丝素蛋白-壳聚糖支架，对照组植入丝素蛋白-壳聚糖支架干预，空白组未植入修复体。 结果与结论：①实验成功制备丝素蛋白-壳聚糖三维支架材料及提取骨髓间充质干细胞，并构建软骨缺损的修复体，将修复体植入兔体内能成功修复缺损的软骨组织；②建模后2，4，8，12周，3组血常规、降钙素原、血沉、C-反应蛋白结果提示无明显的全身感染征象，3组血常规及肝肾功能各时间段比较差异无显著性意义(P > 0.05)；③一般观察、苏木精-伊红染色及扫描电镜观察：建模后12周，相比其他两组，实验组软骨缺损已修复，支架材料已吸收，修复组织周围未见炎性细胞，修复组织已正常组织整合良好；④结果证实，丝素蛋白-壳聚糖支架复合骨髓间充质干细胞在体内构建的组织工程化软骨具有良好的生物相容性。 ORCID: 0000-0002-8139-1175(佘荣峰) 中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Biocompatibility of tissue engineered cartilage constructed in vivo by silk fibroin-chitosan scaffold carrying bone marrow mesenchymal stem cells

BACKGROUND:Our previous studies have found that silk fibroin-chitosan scaffold carrying bone marrow mesenchymal stem cells can repair cartilage defect in rabbits,but further exploration on the biocompatibility of tissue engineered cartilage is yet to be done.OBJECTIVE:To explore the biocompatibility of tissue engineered cartilage that is constructed in vitro by silk fibroin-chitosan scaffold with bone marrow mesenchymal stem cells.METHODS:Three-dimensional silk fibroin-chitosan scaffolds were prepared in a ratio of 1:1.Rabbit bone marrow mesenchymal stem cells were extracted,induced and seeded onto the silk fibroin-chitosan scaffold to construct the cell-scaffold composite.The composite was then implanted into a rabbit joint defect model for cartilage repair.There were three groups in the present study:experiment group with implantation of induced bone marrow mesenchymal stem cells+silk fibroin-chitosan scaffold into the cartilage defect model,control group with implantation of silk fibroin-chitosan scaffold into the cartilage defect model,and blank group without implantation.RESULTS AND CONCLUSION:The three-dimensional silk fibroin-chitosan scaffolds were successfully prepared and combined with bone marrow mesenchymal stem cells(BMSCs)to construct the tissue engineered cartilage for repair cartilage defects in rabbits.Blood routine parameters,procalcitonin levels,erythrocyte sedimentation rates and C-reactive protein levels detected at 2,4,8,and 12 weeks post-implantation indicated no obvious signs of systemic infection,and there was no damage to liver and kidney functions in the three groups.There were also no significant differences between the three groups in terms of blood routines and liver and kidney functions(P>0.05).As shown by gross observation,hematoxylin-eosin staining and scanning electron microscope,in the experimental group,cartilage defects were repaired,with scaffold degradation,no presence of inflammatory cells,and good integration with surrounding tissues.Therefore,tissue engineered cartilage constructed in vitro by silk fibroin-chitosan scaffolds carrying bone marrow mesenchymal stem cells has good biocompatibility,which provides an experimental basis for tissue engineering approaches to cartilage repair.