纳米晶重组类人胶原基/聚乳酸复合支架材料的表征及细胞相容性研究

Characterization and biocompatible evaluation of nHA/RHLC/PLA composite scaffold

目的制备纳米羟基磷灰石/重组类人胶原基/聚乳酸复合支架材料(nano-hydroxyapatite/recombinant human-like collagen/polylactic acid,nHA/RHLC/PLA),观察材料的形貌特征,探讨材料对骨髓基质干细胞(BMSCs)增殖、黏附及分化等生物学行为的影响。方法制备nHA/RHLC/PLA复合支架材料,应用X光衍射分析(XRD)、红外光谱分析(FTIR)、ZWICK Z005测试机对样品的化学成分、机械性能测试和压缩强度进行测试,通过扫描电镜检查等方法观察材料的表征;将犬骨髓基质细胞(BMSCs)接种在支架材料上培养,检测材料-细胞的黏附情况及材料对细胞生长增殖的影响。结果 nHA/RHLC/PLA复合支架材料压缩强度均大于1MPa,达到了天然松质骨的最低强度。扫描电镜结果显示:支架材料呈三维多孔结构,孔为不规则多边形,孔的走向多样,纵向和横向孔隙互为交通,孔径在几十微米到300微米不等,孔隙率为75%～83%。nHA/RHLC/PLA复合支架材料表面BMSCs的黏附、生长良好;而BMSCs的增殖能力与对照组相比,差异无显著性意义(P>0.05)。结论 nHA/RHLC/PLA复合支架材料符合组织工程骨支架的力学要求,具有良好的微观结构,无细胞毒性,细胞与支架生物相容性良好。利用重组类人胶原代替动物源性胶原制备纳米晶骨修复材料,规避了动物胶原交叉感染的风险,有望成为一种理想的骨组织工程支架材料。

Objective To explore the characteristics and biocompatibility of nano-hydroxyapatite/ recombinant human-like collagen/polylactic acid（nHA/RHLC/PLA） composite scaffold.Methods The morphologic features of nHA/RHLC/PLA scaffold were observed under scanning electron microscopy.The porosity of the scaffold was determined via isopropyl alcohol displacement method.The canine BMSCs were separated and cultured in vitro.The third passage of cells was seeded into the scaffold for analyzing the adhesion and proliferation ability of BMSCs on scaffold.Results Scan electron microscopy showed that,the surface of scaffold was porous.BMSCs infiltrated into the pores of scaffold,adhered,spread,and proliferated on the surface of scaffold pore within one week.However,we did not detected the significant difference of adhesion and proliferation of BMSCs between nHA/RHLC/PLA scaffold culture and normal culture（P0.05）.Conclusions The nHA/RHLC/PLA composite scaffold has favourable microstructure,which is biocompatible to BMSCs.