【摘要】 背景:淫羊藿苷可促进骨髓间充质干细胞的增殖与成骨分化,具有良好的促成骨性能。脱钙骨基质具备良好的骨传导与骨诱导能力,可以单独或联合其他材料广泛用于骨修复中。目的:观察3D打印淫羊藿苷/脱钙骨基质材料的缓释性能、细胞相容性与体内成骨性能。方法:利用3D打印技术制备淫羊藿苷/脱钙骨基质材料与脱钙骨基质材料,检测淫羊藿苷/脱钙骨基质材料的体外缓释性能。将骨髓间充质干细胞分别接种于两种材料表面,以单独培养的细胞为对照,于设定的时间点进行活/死染色、MTT、碱性磷酸酶活性与骨钙素含量检测。取新西兰大白兔30只,建立股骨髁骨缺损模型后分3组处理:对照组不植入任何材料,一组植入脱钙骨基质与同种异体兔骨髓间充质干细胞复合物,另一组植入淫羊藿苷/脱钙骨基质材料与同种异体兔骨髓间充质干细胞复合物,术后4,12周进行Micro-CT、组织学与力学性能检测。结果与结论:①3D打印淫羊藿苷/脱钙骨基质材料具有缓释性能,在28d时淫羊藿苷释放量达总量的(54.9±7.9)%;②活/死染色显示,接种1 d后两组材料表面的细胞数量较少,随着培养时间的延长,细胞数量明显增多,至7 d时细胞形态良好,并且淫羊藿苷/脱钙骨基质材料表面的细胞更加均匀、数量更多;③MTT检测显示,淫羊藿苷/脱钙骨基质组培养7,10,14 d的细胞增殖快于其余两组(P <0.05);④淫羊藿苷/脱钙骨基质组培养7,10,14 d的碱性磷酸酶活性与骨钙素含量均高于其余两组(P <0.05);⑤术后12周Micro-CT显示,植入材料两组均可见大量的骨小梁,其中淫羊藿苷/脱钙骨基质组骨小梁数量更多、骨小梁厚度增加、骨小梁分离度更小;⑥术后12周组织学显示,植入材料两组可见大量骨组织形成,其中淫羊藿苷/脱钙骨基质组新生骨量最多;⑦淫羊藿苷/脱钙骨基质组抗压强度高于其余两组(P <0.05);⑧结果表明,3D打印淫羊藿苷/脱钙骨基质具有良好的缓释性能、细胞相容性与骨诱导性能。更多还原

【Abstract】 BACKGROUND: Icariin can promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells, and has good osteogenic properties. Decalcified bone matrix material exhibits good bone conduction and bone induction. It can be used in bone repair alone or in combination with other materials. OBJECTIVE: To investigate the sustained-release performance, cytocompatibility and osteogenesis of 3 D-printed icariin/decalcified bone matrix material. METHODS: Icariin/decalcified bone matrix materials and decalcified bone matrix materials were prepared by 3 D printing technology. The in vitro sustained-release performance of the icariin/decalcified bone matrix material was detected. Bone marrow mesenchymal stem cells were inoculated on the surfaces of the two materials respectively. The cells cultured separately were used as the control. At the designated time points, the live/dead staining and MTT assay were performed, and alkaline phosphatase activity and osteocalcin content were determined. Thirty New Zealand rabbit models of femoral condyle defect were prepared and then divided into three groups. In the control group, no implant was used. In the simple decalcified bone matrix group, decalcified bone matrix and allogeneic rabbit bone marrow mesenchymal stem cells composite were implanted. In the icariin/decalcified bone matrix group, icariin/decalcified bone matrix and allogeneic rabbit bone marrow mesenchymal stem cells were implanted. At 4 and 12 weeks after surgery, micro-CT and histological and mechanical properties evaluation were performed. RESULTS AND CONCLUSION:(1) 3 D-printed icariin/decalcified bone matrix material exhibited sustained-release performance. At 28 days, the release of icariin was(54.9±7.9)% of total release amount.(2) The live/dead staining revealed that the number of cells on the material surface in simple decalcified bone matrix and icariin/decalcified bone matrix groups were decreased after 1 day of inoculation and significantly increased with the prolongation of culture time. At 7 days, cells grew well, and the cells on the surface of icariin/decalcified bone matrix material were more evenly distributed and increased in number.(3) MTT test showed that cells in the icariin/decalcified bone matrix group proliferated faster after 7, 10, and 14 days of culture than those in the other two groups(P < 0.05).(4) The alkaline phosphatase activity and osteocalcin content in the icariin/decalcified bone matrix group were higher than those in the other two groups after 7, 10, and 14 days of culture(P < 0.05).(5) At 12 weeks after surgery, micro-CT revealed that a large amount of bone trabeculae were observed in both simple decalcified bone matrix and icariin/decalcified bone matrix groups. The number of bone trabeculae was higher, bone trabeculae were thicker, and defect cavity was smaller in the icariin/decalcified bone matrix group than those in the simple decalcified bone matrix group.(6) At 12 weeks after surgery, histological results revealed that a large number of bone tissue was formed in both simple decalcified bone matrix and icariin/decalcified bone matrix groups, but the amount of newly formed bone was higher in the latter group.(7) The compressive strength of bone defects in the icariin/decalcified bone matrix group was higher than that in the other two groups(P < 0.05).(8) These results suggest that 3 D-printed icariin/decalcified bone matrix material exhibits good sustained-release performance, cytocompatibility, and osteogenesis.更多还原