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通过逆转录聚合酶链反应(RT-PCR)分析评估成骨细胞对多孔生物活性玻璃(45S5)基质的反应。

Evaluation of osteoblast response to porous bioactive glass (45S5) substrates by RT-PCR analysis.

作者信息

Effah Kaufmann E A, Ducheyne P, Shapiro I M

机构信息

Department of Bioengineering, School of Dental Medicine, Center for Bioactive Materials and Tissue Engineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.

出版信息

Tissue Eng. 2000 Feb;6(1):19-28. doi: 10.1089/107632700320856.

Abstract

Previous studies have shown that neonatal rat calvaria osteoblasts elaborate substantial amounts of extracellular material with bone-like characteristics when cultured on porous bioactive glass substrates in vitro. However, the osteoblastic response to this material has not been fully characterized. The objective of this study was to characterize osteoblast response to porous bioactive glass substrates following the expression of the classical markers for osteoblast differentiation. In this study we synthesized porous bioactive glass substrates, seeded them with osteoblast-like cells (ROS 17/2.8) and followed the temporal expression of alkaline phosphatase (AP) activity, as well as the expression of mRNA for collagen type I (Coll-1), osteonectin (OSN), osteopontin (OPN), osteocalcin (OCN), and bone sialoprotein (BSP). The data confirm that porous bioactive glass substrates are capable of supporting the in vitro growth and maturation of osteoblast-like cells. At a porosity of 42% and an average pore size of 80 microm, the substrates promote the expression and maintenance of the osteoblastic phenotype. The results additionally suggest that there is both a solution-mediated and a surface-controlled effect on cell activity.

摘要

先前的研究表明,新生大鼠颅骨成骨细胞在体外多孔生物活性玻璃基质上培养时,会合成大量具有骨样特征的细胞外物质。然而,成骨细胞对这种材料的反应尚未得到充分表征。本研究的目的是在成骨细胞分化的经典标志物表达后,表征成骨细胞对多孔生物活性玻璃基质的反应。在本研究中,我们合成了多孔生物活性玻璃基质,接种成骨样细胞(ROS 17/2.8),并跟踪碱性磷酸酶(AP)活性的时间表达,以及I型胶原(Coll-1)、骨连接蛋白(OSN)、骨桥蛋白(OPN)、骨钙素(OCN)和骨唾液蛋白(BSP)的mRNA表达。数据证实,多孔生物活性玻璃基质能够支持成骨样细胞的体外生长和成熟。在孔隙率为42%且平均孔径为80微米时,这些基质促进成骨细胞表型的表达和维持。结果还表明,对细胞活性存在溶液介导和表面控制的效应。

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