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MC3T3-E1成骨样细胞在涂覆碳/碳复合材料的胶原/纳米羟基磷灰石支架上的体外矿化

In vitro mineralization of MC3T3-E1 osteoblast-like cells on collagen/nano-hydroxyapatite scaffolds coated carbon/carbon composites.

作者信息

Cao Sheng, Li Hejun, Li Kezhi, Lu Jinhua, Zhang Leilei

机构信息

State Key Laboratory of Solidification Processing, School of Materials Science and Engineering, Northwestern Polytechnical University, Xi'an, Xi'an, Shaanxi, China.

出版信息

J Biomed Mater Res A. 2016 Feb;104(2):533-43. doi: 10.1002/jbm.a.35593. Epub 2015 Nov 3.

Abstract

Collagen/nano-hydroxyapatite (collagen/nHA) scaffolds were successfully prepared on carbon/carbon composites as bioactive films using the layer-by-layer coating method. Surface characterizations of collagen/nHA scaffolds were detected by scanning electron microscope (SEM), X-ray diffraction (XRD), and Fourier transform infrared (FTIR) spectroscopy. Compressive strengths of the scaffolds were evaluated by a universal test machine. In vitro biological performances were determined using scaffolds seeded with MC3T3-E1 osteoblasts-like cells and cultured in mineralization medium for up to 21 days. In addition, cellular morphologies and several related gene expressions of MC3T3-E1 cells in the scaffolds were also evaluated. Chemical and morphological analysis showed that the scaffolds had uniform pore sizes and unified phase composition. Mechanical testing indicated that the collagen/nHA scaffolds had the highest compressive strength in 50% of strain condition when the proportion of collagen and nano-hydroxyapatite was 1:3. Cellular morphology observations and cytology tests indicated that MC3T3-E1 cells were adhered on these scaffolds and proliferated. SEM photographs and gene expressions showed that mineralized MC3T3-E1 cells and newly formed extra cellular matrix (ECM) filled up the pores of the scaffolds after the 3-week mineralization inducement. Nano-sized apatite particles were secreted from MC3T3-E1 cells and combined with the reconstructed ECM. Collectively, collagen/nHA scaffolds provided C/C composites with a biomimetic surface for cell adhesion, proliferation and mineralized extra cellular matrices formation.

摘要

采用层层涂覆法,在碳/碳复合材料上成功制备了胶原/纳米羟基磷灰石(collagen/nHA)支架作为生物活性薄膜。通过扫描电子显微镜(SEM)、X射线衍射(XRD)和傅里叶变换红外(FTIR)光谱对胶原/nHA支架进行表面表征。用万能试验机评估支架的抗压强度。使用接种了MC3T3-E1成骨样细胞的支架并在矿化培养基中培养长达21天来测定体外生物学性能。此外,还评估了支架中MC3T3-E1细胞的细胞形态和几种相关基因表达。化学和形态分析表明,支架具有均匀的孔径和统一的相组成。力学测试表明,当胶原与纳米羟基磷灰石的比例为1:3时,胶原/nHA支架在50%应变条件下具有最高的抗压强度。细胞形态观察和细胞学测试表明,MC3T3-E1细胞附着在这些支架上并增殖。扫描电子显微镜照片和基因表达表明,经过3周的矿化诱导后,矿化的MC3T3-E1细胞和新形成的细胞外基质(ECM)填充了支架的孔隙。纳米尺寸的磷灰石颗粒从MC3T3-E1细胞中分泌出来并与重建的ECM结合。总体而言,胶原/nHA支架为C/C复合材料提供了一个仿生表面,用于细胞粘附、增殖和矿化细胞外基质的形成。

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