新型聚己内酯/磷酸钙支架的制备及生物相容性研究

摘要

To investigate the preparation and biocompatibility of porous polycaprolactone/calcium phosphate cement ( PCL/CPC) scaffolds in uses for bone marrow stromal cells (BMSCs) ,the scaffolds were fabricated by solution blending method,and water soluble salt crystals were used as pore-forming agent. The dynamic mechanical test (DMA) was conducted using Q800 dynamic mechanical analyzer,with the porosity measured by drainage method. BMSCs were implanted and cultured with PCL/CPC scaffolds in vitro,PCL and CPC scaffolds were taken as controls. The attachment,proliferation and differentiation of BMSCs were detected through morphological characters,growth curve,the activity of alkaline phosphatase (ALP) and osteocalcin ( OC) and entopic bone formation in nude mice. Results showed that,storage modulus of the composite scaffold reached the maximum when the ratio of PCL to CPC was 7 :3,the pore size of the material was 250 ~ 350μm,and the porosity of the scaffolds reached 70% ~ 80%. BMSCs attached to and grew better on the surface of porous PCL/CPC and CPC scaffolds than PCL scaffolds ( P < 0. 05) ,and BMSCs proliferate,and function more actively on porous PCL/CPC and CPC scaffolds than on PCL scaffolds ( P < 0. 05 ). More bone form in nude mice of PCL/CPC 13.78% ± 1.60% and CPC scaffolds 15.29% ± 1.20% than PCL scaffolds 7.56% ± 2.20% (P < 0. 05). In summary,PCL/CPC scaffolds have good biocompatibility for BMSCs to fabricate tissue-engineered bone in vitro.%探讨新型聚己内酯( PCL)/磷酸钙(CPC)复合材料支架的制备方法及对骨髓基质细胞(BMSCs)的生物相容性.采用溶液共混法,利用可溶盐晶体做造孔剂,制备PCL/CPC复合材料支架,以单纯PCL和CPC支架为对照组,Q800型动态力学分析仪进行动态力学性能试验(DMA),采用排水法测量孔隙率;灭菌后通过与犬BMSCs体外共同培养后细胞形态、生长曲线、碱性磷酸酶(ALP)染色和半定量及骨钙素(OC)半定量等方法检测细胞在支架材料上的黏附、增殖及成骨分化情况,动物体内异位成骨检测其成骨情况.结果显示,复合材料的储能模量在PCL/CPC比例为7:3时达到最大,制得的材料孔径为250～350μm,多孔支架的孔隙率为70％～80％;BMSCs在新型PCL/CPC组、CPC组支架表面分布均匀,生长增殖明显较PCL组活跃(P＜0.05);PCL/CPC组、CPC组BMSCs成骨行为与PCL组之间有显著差异(P＜0.05).动物体内异位成骨检测提示,4周时PCL/CPC组为13.78％±1.60％、CPC组BMSCs为15.29％±1.20％,成骨显著强于PCL组BMSCs的7.56％±2.20％(P＜0.05),表明PCL和CPC的复合明显改善了两种材料的缺陷,获得的PCL/CPC支架具有良好的生物相容性,可与BMSCs共同构建具有成骨能力的三维立体组织工程化骨.