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用于增强原代人半月板细胞增殖的丝素-聚乙烯醇混合纳米纤维支架

Silk-PVA Hybrid Nanofibrous Scaffolds for Enhanced Primary Human Meniscal Cell Proliferation.

作者信息

Pillai Mamatha M, Gopinathan J, Indumathi B, Manjoosha Y R, Santosh Sahanand K, Dinakar Rai B K, Selvakumar R, Bhattacharyya Amitava

机构信息

Tissue Engineering Laboratory, PSG Institute of Advanced Studies, Coimbatore, 641004, India.

Advanced Textile and Polymer Research Laboratory, PSG Institute of Advanced Studies, Coimbatore, 641004, India.

出版信息

J Membr Biol. 2016 Dec;249(6):813-822. doi: 10.1007/s00232-016-9932-z. Epub 2016 Oct 11.

Abstract

In this study, silk fibroin nanofibrous scaffolds were developed to investigate the attachment and proliferation of primary human meniscal cells. Silk fibroin (SF)-polyvinyl alcohol (PVA) blended electrospun nanofibrous scaffolds with different blend ratios (2:1, 3:1, and 4:1) were prepared. Morphology of the scaffolds was characterized using atomic force microscopy (AFM). The hybrid nanofibrous mats were crosslinked using 25 % (v/v) glutaraldehyde vapor. In degradation study, the crosslinked nanofiber showed slow degradation of 20 % on weight after 35 days of incubation in simulated body fluid (SBF). The scaffolds were characterized with suitable techniques for its functional groups, porosity, and swelling ratio. Among the nanofibers, 3:1 SF:PVA blend showed uniform morphology and fiber diameter. The blended scaffolds had fluid uptake and swelling ratio of 80 % and 458 ± 21 %, respectively. Primary meniscal cells isolated from surgical debris after meniscectomy were subcultured and seeded onto these hybrid nanofibrous scaffolds. Meniscal cell attachment studies confirmed that 3:1 SF:PVA nanofibrous scaffolds supported better cell attachment and growth. The DNA and collagen content increased significantly with 3:1 SF:PVA. These results clearly indicate that a blend of SF:PVA at 3:1 ratio is suitable for meniscus cell proliferation when compared to pure SF-PVA nanofibers.

摘要

在本研究中,开发了丝素蛋白纳米纤维支架以研究原代人半月板细胞的附着和增殖情况。制备了具有不同混合比例(2:1、3:1和4:1)的丝素蛋白(SF)-聚乙烯醇(PVA)共混电纺纳米纤维支架。使用原子力显微镜(AFM)对支架的形态进行了表征。混合纳米纤维垫用25%(v/v)戊二醛蒸汽交联。在降解研究中,交联纳米纤维在模拟体液(SBF)中孵育35天后重量缓慢降解20%。用合适的技术对支架的官能团、孔隙率和溶胀率进行了表征。在纳米纤维中,3:1的SF:PVA共混物显示出均匀的形态和纤维直径。共混支架的吸液率和溶胀率分别为80%和458±21%。从半月板切除术后的手术碎片中分离出的原代半月板细胞进行传代培养,并接种到这些混合纳米纤维支架上。半月板细胞附着研究证实,3:1的SF:PVA纳米纤维支架更有利于细胞附着和生长。3:1的SF:PVA使DNA和胶原蛋白含量显著增加。这些结果清楚地表明,与纯SF-PVA纳米纤维相比,3:1比例的SF:PVA混合物适合半月板细胞增殖。

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