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首页> 外文期刊>Acta biomaterialia >Ingrowth of human mesenchymal stem cells into porous silk particle reinforced silk composite scaffolds: An in vitro study.
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Ingrowth of human mesenchymal stem cells into porous silk particle reinforced silk composite scaffolds: An in vitro study.

机译：人间充质干细胞向多孔丝颗粒增强的丝复合支架中的生长：一项体外研究。

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Silk fibroin protein is biodegradable and biocompatible, exhibiting excellent mechanical properties for various biomedical applications. However, porous three-dimensional (3-D) silk fibroin scaffolds, or silk sponges, usually fall short in matching the initial mechanical requirements for bone tissue engineering. In the present study, silk sponge matrices were reinforced with silk microparticles to generate protein-protein composite scaffolds with desirable mechanical properties for in vitro osteogenic tissue formation. It was found that increasing the silk microparticle loading led to a substantial increase in the scaffold compressive modulus from 0.3 MPa (non-reinforced) to 1.9 MPa for 1:2 (matrix:particle) reinforcement loading by dry mass. Biochemical, gene expression, and histological assays were employed to study the possible effects of increasing composite scaffold stiffness, due to microparticle reinforcement, on in vitro osteogenic differentiation of human mesenchymal stem cells (hMSCs). Increasing silk microparticle loading increased the osteogenic capability of hMSCs in the presence of bone morphogenic protein-2 (BMP-2) and other osteogenic factors in static culture for up to 6 weeks. The calcium adsorption increased dramatically with increasing loading, as observed from biochemical assays, histological staining, and microcomputer tomography (muCT) analysis. Specifically, calcium content in the scaffolds increased by 0.57, 0.71, and 1.27 mg (per mug of DNA) from 3 to 6 weeks for matrix to particle dry mass loading ratios of 1:0, 1:1, and 1:2, respectively. In addition, muCT imaging revealed that at 6 weeks, bone volume fraction increased from 0.78% for non-reinforced to 7.1% and 6.7% for 1:1 and 1:2 loading, respectively. Our results support the hypothesis that scaffold stiffness may strongly influence the 3-D in vitro differentiation capabilities of hMSCs, providing a means to improve osteogenic outcomes.
机译：丝素蛋白蛋白具有生物可降解性和生物相容性，对各种生物医学应用均显示出优异的机械性能。但是，多孔三维（3-D）丝素蛋白支架或丝海绵通常无法满足骨骼组织工程的初始机械要求。在本研究中，用丝微粒增强丝海绵基质以产生具有所需机械性质的蛋白-蛋白复合支架，以用于体外成骨组织形成。已经发现，对于以干重计的1：2（基质：颗粒）的增强载荷，增加丝微粒的载荷导致支架的压缩模量从0.3 MPa（非增强）显着增加到1.9 MPa。使用生化，基因表达和组织学分析方法研究由于微粒增强而增加复合支架刚度对人间充质干细胞（hMSCs）体外成骨分化的可能影响。在静态培养中，在骨形态发生蛋白2（BMP-2）和其他成骨因子存在下长达6周，增加的丝微粒载量可增强hMSC的成骨能力。从生化分析，组织学染色和微型计算机断层扫描（muCT）分析中观察到，钙吸附量随装载量的增加而急剧增加。具体而言，对于基质与颗粒的干质量装载比分别为1：0、1：1和1：2，支架中的钙含量在3至6周内分别增加了0.57、0.71和1.27 mg（每杯DNA）。 。此外，muCT成像显示，在第6周，骨体积分数从非增强骨的0.78％分别增加到1：1和1：2负荷的7.1％和6.7％。我们的结果支持以下假设，即支架刚度可能会强烈影响hMSC的3-D体外分化能力，从而提供改善成骨结果的手段。

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