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孔隙几何形状对选择性激光熔化 Ti6Al4V 骨支架上接种的人骨膜来源细胞体外生物学行为的影响。

The effect of pore geometry on the in vitro biological behavior of human periosteum-derived cells seeded on selective laser-melted Ti6Al4V bone scaffolds.

机构信息

Department of Mechanical Engineering, Division of Production Engineering, Machine Design and Automation, Katholieke Universiteit Leuven, Belgium.

出版信息

Acta Biomater. 2012 Jul;8(7):2824-34. doi: 10.1016/j.actbio.2012.04.001. Epub 2012 Apr 7.

Abstract

The specific aim of this study was to gain insight into the influence of scaffold pore size, pore shape and permeability on the in vitro proliferation and differentiation of three-dimensional (3-D) human periosteum-derived cell (hPDC) cultures. Selective laser melting (SLM) was used to produce six distinct designed geometries of Ti6Al4V scaffolds in three different pore shapes (triangular, hexagonal and rectangular) and two different pore sizes (500 μm and 1000 μm). All scaffolds were characterized by means of two-dimensional optical microscopy, 3-D microfocus X-ray computed tomography (micro-CT) image analysis, mechanical compression testing and computational fluid dynamical analysis. The results showed that SLM was capable of producing Ti6Al4V scaffolds with a broad range of morphological and mechanical properties. The in vitro study showed that scaffolds with a lower permeability gave rise to a significantly higher number of cells attached to the scaffolds after seeding. Qualitative analysis by means of live/dead staining and scanning electron micrography showed a circular cell growth pattern which was independent of the pore size and shape. This resulted in pore occlusion which was found to be the highest on scaffolds with 500 μm hexagonal pores. Interestingly, pore size but not pore shape was found to significantly influence the growth of hPDC on the scaffolds, whereas the differentiation of hPDC was dependent on both pore shape and pore size. The results showed that, for SLM-produced Ti6Al4V scaffolds with specific morphological and mechanical properties, a functional graded scaffold will contribute to enhanced cell seeding and at the same time can maintain nutrient transport throughout the whole scaffold during in vitro culturing by avoiding pore occlusion.

摘要

本研究的具体目的是深入了解支架孔径、孔形和渗透性对三维(3-D)人骨膜源性细胞(hPDC)培养物体外增殖和分化的影响。选择性激光熔化(SLM)用于生产六种不同设计的 Ti6Al4V 支架,具有三种不同的孔形(三角形、六边形和矩形)和两种不同的孔径(500 μm 和 1000 μm)。所有支架均通过二维光学显微镜、三维微焦点 X 射线计算机断层扫描(micro-CT)图像分析、机械压缩测试和计算流体动力学分析进行了表征。结果表明,SLM 能够生产具有广泛形态和机械性能的 Ti6Al4V 支架。体外研究表明,渗透性较低的支架在接种后附着在支架上的细胞数量明显增加。通过活/死染色和扫描电子显微镜的定性分析显示,细胞呈圆形生长模式,与孔径和形状无关。这导致在 500 μm 六边形孔的支架上发现的孔闭塞最高。有趣的是,发现孔径而不是孔形显著影响 hPDC 在支架上的生长,而 hPDC 的分化则取决于孔形和孔径。结果表明,对于具有特定形态和机械性能的 SLM 生产的 Ti6Al4V 支架,功能梯度支架将有助于增强细胞接种,同时通过避免孔闭塞在体外培养过程中维持整个支架的营养物质运输。

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