物理联合化学或化学方法处理去抗原异种松质骨支架与骨髓间充质干细胞的细胞相容性

摘要

背景:研究证明去抗原异种松质骨支架具有良好的理化性能和三维立体多孔结构,但应用于临床还需要考虑其安全性和生物相容性.目的:比较物理联合化学及化学方法处理去抗原异种松质骨支架与羊骨髓间充质干细胞的细胞相容性.方法:用梯度密度离心法分离培养羊骨髓间充质干细胞,四甲基偶氮唑盐比色法检测物理联合化学及化学方法处理去抗原异种松质骨支架材料对骨髓间充质干细胞增殖的影响.取第3代骨髓间充质干细胞,接种在两种支架上共同培养,倒置相差显微镜和扫描电镜观察细胞在两种支架材料上的形态、黏附、生长和增殖情况.结果与结论:物理联合化学组细胞毒性为0或1级,细胞能在材料上良好地黏附、增殖、生长,细胞活性未受到支架材料的影响.化学组细胞毒性为3级,细胞在材料上生长受到抑制,支架孔隙内无细胞黏附.提示经过物理联合化学处理的去抗原异种松质骨支架材料与羊骨髓间充质干细胞具有良好的生物相容性;单纯经过化学处理的支架材料生物相容性较差,不符合生物材料安全性标准.%BACKGROUND: Antigen-extracted xenogeneic cancellous bone has been shown to be characterized by well physicochemical performance and three-dimensional stereo porous structure, however, the biocompatibility and safely still need to be further studied in clinical application.OBJECTIVE: To study the cytocompatibility of bone marrow mesenchymal stem cells (BMSCs) and antigen-extracted xenogeneic cancellous bone scaffold treated by chemical or physicochemical method.METHODS: The gradient density centrifugation was used to separate and cultivate sheep BMSCs, the influence of antigen-extracted xenogeneic cancellous bone scaffold materials treated by chemical or physicochemical method on the proliferation of the BMSCs were measured by four thiazole salt colorimetric experiment (MTT method). The third generation of BMSCs were taken and seeded on two scaffolds, the morphology, adhesion, growth and proliferation of the cells on the two scaffold materials were observed by inverted phase contrast microscope and scanning electron microscope.RESULTS AND CONCLUSION: The cytotoxicity was 0 or 1 level in the physics joint chemical group, the cells exhibit well adhesion, proliferation and growth, cell activity was not affected by the material. The cytotoxicity was 3 level in chemical group, the cells were restrained, no cell adhesion in scaffold pores. The antigen-extracted xenogeneic cancellous bone materials treated with physical combined with chemical methods have good cytocompatibility to BMSCs. However, the scaffold materials treated by chemical methods have poor biocompatibility and it is not qualified for the safety standards of biological materials.