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目的探讨以煅烧羊椎骨松质双相陶瓷粉末/聚乙烯醇(polyvinyl alcohol vinylalcohol polymer,PVA)生物高分子聚合物凝胶为原材料的新型三维打印组织工程骨支架成型方法,及支架的生物相容性。方法取健康、清洁级新疆阿勒泰大尾羊骨髓液培养羊骨髓间充质干细胞;取新鲜市售羊椎骨,去皮质骨,脱蛋白质、脱脂、煅烧后松质双相陶瓷精细研磨,钴60放射灭菌,与体积分数15%PVA溶剂体积比1∶2混合,制备成型材料。将建立好的STL格式文件导入固高GXYZ303010-XYLE三维打印系统,装配快速成型材料,采用熔融堆积成型法完成三维打印组织工程骨支架的成型。将羊骨髓间充质干细胞与灭菌三维打印组织工程骨支架在体外复合培养,扫描电镜观察羊骨髓间充质干细胞在三维打印组织工程骨支架中的生长情况;制备支架材料浸提液,采用MTT法检测三维打印组织工程骨支架对羊骨髓间充质干细胞增殖的影响,采用皮内刺激试验评价三维打印组织工程骨支架材料的生物相容性。结果三维打印法建立的组织工程骨支架具有良好的通透性,孔隙率为68.3%,孔隙间连通率为84.0%,具备一定的力学性能;三维打印组织工程骨支架浸提液对羊骨髓间充质干细胞的细胞毒性为0~1级,无明显细胞毒性;羊骨髓间充质干细胞可紧密附着于支架材料表面,黏附生长、增殖,并有突起向支架内部空隙伸展;皮内刺激试验结果表明三维打印组织工程骨支架对新疆阿勒泰大尾羊无刺激性。结论利用羊椎骨松质煅烧双相陶瓷粉末/PVA生物高分子聚合物成型的三维打印组织工程骨支架无刺激性、无明显细胞毒性,具有良好的生物学功能,有望应用于骨组织工程学。
OBJECTIVE: To investigate a new method of forming a three-dimensional tissue engineered scaffold using calcined sheep bone spunbonded biphasic ceramic powder / polyvinyl alcohol (PVA) biopolymer gel as raw material, and to evaluate the biocompatibility of scaffold Sex. Methods Sheep bone marrow mesenchymal stem cells were cultured in the bone marrow of Altay, Great-Tail sheep in Xinjiang with a clean and healthy grade. The commercially available sheep vertebra, cortical bone, deproteinized, degreased, Sterilized, and the volume fraction of 15% PVA solvent volume ratio of 1: 2 mixed to prepare a molding material. The established STL format file is imported into the GIGABYTE GXYZ303010-XYLE 3D printing system, and the rapid prototyping material is assembled. The forming of the three-dimensional printed tissue engineering bone scaffold is completed by the fusion deposition modeling method. Sheep bone marrow mesenchymal stem cells and sterilized three-dimensional printing tissue engineering scaffold compound in vitro culture, scanning electron microscopy sheep sheep bone marrow mesenchymal stem cells in three-dimensional printing tissue engineering bone scaffold growth; preparation of scaffold material extract, using MTT assay was used to detect the effect of three-dimensional printed tissue engineered scaffolds on the proliferation of sheep bone marrow mesenchymal stem cells. The intradermal stimulation test was used to evaluate the biocompatibility of three-dimensional printed tissue engineered scaffolds. Results The tissue engineering scaffolds established by three-dimensional printing method had a good permeability, with a porosity of 68.3% and a viable rate of 84.0%. The three-dimensional printed tissue engineering scaffolds had good permeability to goat bone marrow The cytotoxicity of MSCs was from 0 to 1 grade, with no obvious cytotoxicity. Sheep bone marrow mesenchymal stem cells could adhere tightly to the surface of scaffold material, adhere to the growth and proliferation, and the protrusions extend to the internal space of the scaffold. The intradermal stimulation test results That the three-dimensional printing tissue engineering scaffold on Xinjiang Aletai Tail Sheep non-irritating. Conclusion The three-dimensional printed tissue engineered scaffold made of sheep vertebrate cancellous calcined biphasic ceramic powder / PVA biopolymer has no irritation and no obvious cytotoxicity. It has good biological function and is expected to be applied to bone tissue engineering.