多孔型活性CPC复合骨髓基质细胞构建组织工程骨的实验研究

Experimental study of constructing tissue engineering bone with Active cpc and BMSCs

目的:观察多孔型活性磷酸钙骨水泥用作骨组织工程支架材料的可行性。材料与方法:抽取成年毕格犬的骨髓,贴壁法获得骨髓基质细胞,经成骨诱导培养液体外培养、扩增、诱导后观察细胞增殖情况。将培养的第3代细胞接种于多孔型活性磷酸钙骨水泥,进行超微结构观察,并将多孔型活性磷酸钙骨水泥/骨髓基质细胞复合物植入毕格犬背部皮下,2,4周后进行组织学检测。结果:碱性磷酸酶染色呈阳性;VonKossa染色可见钙结节形成;骨钙素免疫细胞化学染色呈阳性;超微结构观察可见细胞生长附着于材料网孔内表面;组织学检测提示4周时复合物内有新骨形成。讨论:活性多孔CPC/BMSCs骨修复材料孔径250μm,孔隙率为70%,易于BMSCs的渗入发育成骨。结论:多孔型活性磷酸钙骨水泥/骨髓基质细胞复合物显示良好的成骨活性,多孔型活性磷酸钙骨水泥可以用于骨组织工程支架材料。

Purpose:To study the probability of Active pcpc that be used as a scaffold for bone tissue engineering.Material and Methods:After drawing out the bone marrow from the Beagle Dog,the bone marrow stroma cells were separated by adherent methods and induced with the conditional medium,the phenotype of the BMSCs was evaluated by immunocytochemistry.The induced passage 3 cells and active cpc were mixed to form a complex.After the SEM scanning,the complex was implanted into the subcutaneous parts of the beagle dog,histological observation were taken after 2 and 4 weeks.Reslts:ALP staining is positive,Von Kossa staining proofed the formation of mineralization nodules,immunocytochemical proofed the express of collagen I,Osteocalsin;Scanning electron micrograph showed the cells adhered to the inner surface of the Active pcpc;HE staining showed the new bone formation and vascularization 4 weeks later in group of complex.Dicussion:Activity CPC aperture is 200-500μm,small aperture is 1-5μm,porosity 70%.Active cpc is easy for osteoblast activity of BMSCs.Conclusion:Complex(Active cpc/BMSCs)showed new bone formation and vascularization in Beagle dog,the Active cpc can be be used as a scaffold for bone tissue engineering.