三维多孔支架孔径调节人间充质干细胞成骨分化的研究
首发时间:2013-12-13
摘要:本研究为探究多孔支架孔径对干细胞成骨分化行为的影响,首先通过制备不同粒径的明胶微球,以此为致孔剂来制备孔径大小可调的聚己内酯三维多孔支架;进而以人间充质干细胞为研究对象,考察其培养14d后的成骨分化效果。基于扫描电镜表征发现,聚己内酯多孔支架能够很好的复制致孔剂的尺寸和形态,且按孔径大小分为四类,分别具有100-200 μm,200-300 μm,300-450 μm和450-600 μm的孔径。在成骨诱导培养后,通过扫描电镜、茜素红和ALP染色、钙含量和ALP酶活性检测、基因表达分析,发现孔径为200-300μm的支架最有利于人间充质干细胞的成骨分化,孔径越大,成骨分化效果越差。该结果为骨组织工程支架材料的构建提供了基础数据。
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Effects of Pore Size of Three Dimensional Porous Scaffolds on Osteogenic Differentiation of Human Mesenchymal Stem Cells
Abstract:The objective of this paper was to study the effects of pore size of three dimensional porous scaffolds on the osteogenic differentiation of mesenchymal stem cells. In order to fabricate porous polycaprolactone scaffolds, gelatin microspheres with different diameters were first prepared, which were used as porogen. Based on scanning electron microscopy (SEM), scaffolds were able to replicate the size and morphology of the porogen, resulting in four types of scaffolds with different pore size: 100-200 μm,200-300 μm,300-450 μm and 450-600 μm. After 14 d osteogenic induction, based on SEM, Alizarin red S staining, ALP staining, calcium quantification, ALP activity assay, and gene expression analysis, it was found that human mesenchymal stem cells demonstrated highest osteogenic differentiation in scaffolds with 200-300 μm. These data definitely provide guidance for fabricationg bone tissue engineering scaffolds.
Keywords: tissue engineering porous scaffolds pore size mesenchymal stem cells osteogenic differentiation
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