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选区激光熔化 Ti6Al4VELI 多孔支架的孔径和孔隙率对细胞增殖、成骨和骨长入的影响。

Influence of the pore size and porosity of selective laser melted Ti6Al4V ELI porous scaffold on cell proliferation, osteogenesis and bone ingrowth.

机构信息

Department of Orthopedics, The Third Affiliated Hospital of Southern Medical University, No.183, Zhongshan West Avenue, Tianhe District, Guangzhou, 510630, Guangdong, PR China; Academy of Orthopedics, Guangdong Province, PR China; Orthopedic Hospital of Guangdong Province, PR China.

National Engineering Laboratory for Modern Materials Surface Engineering Technology, The Key Lab of Guangdong for Modern Surface Engineering Technology, Guangdong Institute of New Materials, Guangzhou, 510651, PR China; ICB UMR 6303, CNRS, Univ. Bourgogne Franche-Comté, UTBM, F-90010, Belfort, France.

出版信息

Mater Sci Eng C Mater Biol Appl. 2020 Jan;106:110289. doi: 10.1016/j.msec.2019.110289. Epub 2019 Oct 7.

Abstract

This paper systematically investigates the biomedical performance of selective laser melted (SLM) porous Ti6Al4V ELI scaffolds for bone implantation through in vitro and in vivo experiments. Scaffolds with pore sizes of 500 μm, 600 μm and 700 μm and porosities of 60% and 70% were manufactured in order to explore the optimum pore size and porosity. Rat bone marrow mesenchymal stem cells (rBMMSCs) were used in the in vitro experiments. Cell Counting Kit-8, live/dead staining and scanning electron microscope were used to assess the cytotoxicity of the porous scaffolds. DNA content quantification was performed to investigate cell proliferation on the porous scaffolds. The osteogenic differentiation of cells was measured by alkaline phosphatase (ALP) activity and osteogenic gene expressions, including bone morphogenetic protein-2 (BMP-2), collagen type 1α1 (COL-1), osteocalcin (OCN), osteopontin (OPN) and runt-related transcription factor-2 (RUNX-2). The Sprague-Dawley (SD) rat models with distal femoral condyles defect were used in the in vivo experiments. Micro-CT analysis and histological analysis were performed after implantation surgery to reveal the bone ingrowth into the porous scaffolds. All in vitro data were analyzed by one-way ANOVA followed by Tukey post hoc tests, in vivo data were analyzed using Kruskall-Wallis ANOVA and Conover-Inman post-hoc test. Based on the in vitro and in vivo experiments, it is found that the porous scaffolds manufactured by SLM did not induce a cytotoxic effect. Among all the porous scaffolds, the scaffold with a pore size of 500 μm and porosity of 60% showed the best cell proliferation and osteogenic differentiation (in vitro experiments) and bone ingrowth (in vivo experiments).

摘要

本文通过体外和体内实验系统研究了选择性激光熔化(SLM)多孔 Ti6Al4V ELI 支架的生物医学性能,用于骨植入。制造了孔径为 500μm、600μm 和 700μm 以及孔隙率为 60%和 70%的支架,以探索最佳孔径和孔隙率。在体外实验中使用了大鼠骨髓间充质干细胞(rBMMSCs)。使用细胞计数试剂盒-8、活/死染色和扫描电子显微镜评估多孔支架的细胞毒性。通过 DNA 含量定量来研究细胞在多孔支架上的增殖。通过碱性磷酸酶(ALP)活性和成骨基因表达,包括骨形态发生蛋白-2(BMP-2)、胶原类型 1α1(COL-1)、骨钙素(OCN)、骨桥蛋白(OPN)和 runt 相关转录因子-2(RUNX-2)来测量细胞的成骨分化。在体内实验中,使用 Sprague-Dawley(SD)大鼠股骨远端髁缺损模型。植入手术后进行 micro-CT 分析和组织学分析,以揭示多孔支架中的骨向内生长。所有体外数据均采用单因素方差分析(ANOVA),随后进行 Tukey 事后检验,体内数据采用 Kruskall-Wallis ANOVA 和 Conover-Inman 事后检验。基于体外和体内实验,发现 SLM 制造的多孔支架没有引起细胞毒性。在所有多孔支架中,孔径为 500μm、孔隙率为 60%的支架在细胞增殖和成骨分化(体外实验)和骨向内生长(体内实验)方面表现最佳。

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