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用于骨组织工程的光交联壳聚糖-丙交酯水凝胶的体外评价

In vitro evaluation of photo-crosslinkable chitosan-lactide hydrogels for bone tissue engineering.

作者信息

Kim Sungwoo, Kang Yunqing, Mercado-Pagán Ángel E, Maloney William J, Yang Yunzhi

机构信息

Department of Orthopedic Surgery, Stanford University, Stanford, California.

出版信息

J Biomed Mater Res B Appl Biomater. 2014 Oct;102(7):1393-406. doi: 10.1002/jbm.b.33118. Epub 2014 Feb 6.

Abstract

Here we report the development and characterization of novel photo-crosslinkable chitosan-lactide (Ch-LA) hydrogels for bone tissue engineering. We synthesized the hydrogels based on Ch, LA, and methacrylic anhydride (MA), and examined their chemical structures, degradation rates, compressive moduli, and protein release kinetics. We also evaluated the cytotoxicity of the hydrogels and delivery efficacy of bone morphogenetic protein-2 (BMP-2) on osteoblast differentiation and mineralization using W-20-17 preosteoblast mouse bone marrow stromal cells and C2C12 mouse myoblast cells. NMR and FTIR revealed that the hydrogels were formed via amidation and esterification between Ch and LA, and methacrylation for photo-crosslinkable networks. Addition of a hydrophobic LA moiety to a hydrophilic Ch chain increased swellability, softness, and degradation rate of the photo-crosslinkable Ch-LA hydrogels. Changes in Ch/LA ratio and UV exposure time significantly affected compressive modulus and protein release kinetics. The photo-crosslinkable Ch-LA hydrogels were not cytotoxic regardless of the composition and UV crosslinking time. Higher alkaline phosphatase activities of both W-20-17 and C2C12 cells were observed in the less-crosslinked hydrogels at day 5. Mineralization was enhanced by sustained BMP-2 release from the hydrogels, but was cell type dependent. This photo-crosslinkable Ch-LA hydrogel is a promising carrier for growth factors.

摘要

在此,我们报告了用于骨组织工程的新型光交联壳聚糖-丙交酯(Ch-LA)水凝胶的研发与特性。我们基于壳聚糖(Ch)、丙交酯(LA)和甲基丙烯酸酐(MA)合成了水凝胶,并检测了它们的化学结构、降解速率、压缩模量和蛋白质释放动力学。我们还使用W-20-17前成骨细胞小鼠骨髓基质细胞和C2C12小鼠成肌细胞评估了水凝胶的细胞毒性以及骨形态发生蛋白-2(BMP-2)对成骨细胞分化和矿化的递送效果。核磁共振(NMR)和傅里叶变换红外光谱(FTIR)显示,水凝胶是通过Ch与LA之间的酰胺化和酯化以及用于光交联网络的甲基丙烯酰化形成的。在亲水性Ch链上添加疏水性LA部分可提高光交联Ch-LA水凝胶的溶胀性、柔软度和降解速率。Ch/LA比例和紫外线照射时间的变化显著影响压缩模量和蛋白质释放动力学。无论组成和紫外线交联时间如何,光交联Ch-LA水凝胶均无细胞毒性。在第5天,在交联程度较低的水凝胶中观察到W-20-17和C2C12细胞的碱性磷酸酶活性均较高。水凝胶持续释放BMP-2可增强矿化,但这取决于细胞类型。这种光交联Ch-LA水凝胶是一种很有前景的生长因子载体。

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