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用于引导骨再生的三种壳聚糖/聚阳离子复合膜的表面表征及细胞相容性

Surface characterization and cytocompatibility of three chitosan/polycation composite membranes for guided bone regeneration.

作者信息

Zheng Zhenhuan, Wei Yujun, Wang Gan, Gong Yandao, Zhang Xiufang

机构信息

Department of Biological Sciences and Biotechnology, State Key Laboratory of Biomembrane and Membrane Biotechnology, Tsinghua University, Beijing 100084, People's Republic of China.

出版信息

J Biomater Appl. 2009 Sep;24(3):209-29. doi: 10.1177/0885328208095825. Epub 2008 Nov 5.

Abstract

Guided bone regeneration is a promising surgical procedure for reconstructing bone defects. In this study, three chitosan/polycation composite membranes for guided bone regeneration are produced by blending chitosan with poly-L-lysine, polyethyleneimine, and poly-L-ornithine. For all composite membranes, the surface characteristics including surface topography, chemistry, and wettability are examined by atomic force microscopy, X-ray photoelectron spectroscopy, and contact angle assay. Their cytocompatibility is also evaluated with MC3T3-E1 osteoblast-like cells at cell, protein, and gene levels through cell biology assays, western blot, and RT-PCR analysis. On chitosan/poly-L-lysine composite membrane, MC3T3-E1 cells present well-developed cytoskeletal organization and significantly higher adhesion, proliferation, and differentiation than those on chitosan and the other two composite membranes. Furthermore, MC3T3-E1 cells on chitosan/poly-L-lysine membrane exhibit increased phosphorylation levels of focal adhesion kinase and extracellular signal-regulated kinase 1/2, and achieve an enhanced mRNA expression of fibronectin, Runx 2, RhoA, integrin alpha 5, and integrin beta1. From our results, we conclude that chitosan/ poly-L-lysine composite membrane possesses improved cytocompatibility with osteoblasts when compared to chitosan and holds potential for guided bone regeneration in the near future.

摘要

引导性骨再生是一种用于修复骨缺损的很有前景的外科手术方法。在本研究中,通过将壳聚糖与聚-L-赖氨酸、聚乙烯亚胺和聚-L-鸟氨酸混合,制备了三种用于引导性骨再生的壳聚糖/聚阳离子复合膜。对于所有复合膜,通过原子力显微镜、X射线光电子能谱和接触角测定来检测其表面特性,包括表面形貌、化学性质和润湿性。还通过细胞生物学试验、蛋白质印迹和逆转录-聚合酶链反应分析,在细胞、蛋白质和基因水平上用MC3T3-E1成骨样细胞评估它们的细胞相容性。在壳聚糖/聚-L-赖氨酸复合膜上,MC3T3-E1细胞呈现出发育良好的细胞骨架结构,并且与壳聚糖膜和其他两种复合膜相比,其黏附、增殖和分化能力显著更高。此外,壳聚糖/聚-L-赖氨酸膜上的MC3T3-E1细胞显示出黏着斑激酶和细胞外信号调节激酶1/2的磷酸化水平增加,并且实现了纤连蛋白、Runx 2、RhoA、整合素α5和整合素β1的mRNA表达增强。从我们的结果来看,我们得出结论,与壳聚糖相比,壳聚糖/聚-L-赖氨酸复合膜与成骨细胞具有更好的细胞相容性,并且在不久的将来具有引导性骨再生的潜力。

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