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用I型胶原蛋白或羟基磷灰石修饰的可吸收支架:对人间充质干细胞的体外研究

Resorbable scaffolds modified with collagen type I or hydroxyapatite: in vitro studies on human mesenchymal stem cells.

作者信息

Rumian Lucja, Wojak Iwona, Scharnweber Dieter, Pamuła Elżbieta

机构信息

AGH University of Science and Technology, Faculty of Materials Science and Ceramics, Department of Biomaterials, Kraków, Poland.

出版信息

Acta Bioeng Biomech. 2013;15(1):61-7.

Abstract

Poly(L-lactide-co-glycolide) (PLGA) scaffolds of pore size within the range of 250-320 μm were produced by solvent casting/ porogen leaching method. Afterwards, they were modified through adsorption of collagen type I and incubation in simulated body fluid (SBF) to allow deposition of hydroxyapatite (HAp). The wettability of the scaffolds was measured by sessile drop test. Scanning electron microscopy (SEM) evaluation and energy dispersive X-ray analysis (EDX) were also performed. SEM evaluation and EDX analysis depicted the presence of HAp deposits and a collagen layer on the pore walls on the surface and in the bulk of the scaffolds. Wettability and water droplets penetration time within the scaffolds decreased considerably after applying modifications. Human mesenchymal stem cells (hMSC) were cultured on the scaffolds for 28 days and cell morphology, proliferation and differentiation as well as calcium deposition were evaluated. Lactate dehydrogenase (LDH) activity results revealed that cells cultured on tissue culture polystyrene (TCPS) exhibited high proliferation capacity. Cell growth on the scaffolds was slower in comparison to TCPS and did not depend on modification applied. On the other hand, osteogenic differentiation of hMSC as confirmed by alkaline phosphatase (ALP) activity and mineralization results was enhanced on the scaffolds modified with hydroxyapatite and collagen.

摘要

采用溶剂浇铸/致孔剂沥滤法制备了孔径在250 - 320μm范围内的聚(L - 丙交酯 - 乙交酯)(PLGA)支架。之后,通过吸附I型胶原并在模拟体液(SBF)中孵育对其进行改性,以使羟基磷灰石(HAp)沉积。通过静滴法测量支架的润湿性。还进行了扫描电子显微镜(SEM)评估和能量色散X射线分析(EDX)。SEM评估和EDX分析表明,在支架表面和内部的孔壁上存在HAp沉积物和胶原层。改性后,支架的润湿性和水滴在支架内的渗透时间显著降低。将人间充质干细胞(hMSC)在支架上培养28天,并评估细胞形态、增殖、分化以及钙沉积情况。乳酸脱氢酶(LDH)活性结果显示,在组织培养聚苯乙烯(TCPS)上培养的细胞具有较高的增殖能力。与TCPS相比,支架上的细胞生长较慢,且不依赖于所应用的改性。另一方面,经碱性磷酸酶(ALP)活性和矿化结果证实,hMSC在经羟基磷灰石和胶原改性的支架上的成骨分化得到增强。

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