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纤维环椎间盘细胞在聚-D,L-乳酸/45S5生物活性玻璃复合膜上的体外附着、分化及基质产生的研究

In vitro studies of annulus fibrosus disc cell attachment, differentiation and matrix production on PDLLA/45S5 Bioglass composite films.

作者信息

Wilda Helen, Gough Julie E

机构信息

Materials Science Centre, School of Materials, The University of Manchester, Manchester M1 7HS, UK.

出版信息

Biomaterials. 2006 Oct;27(30):5220-9. doi: 10.1016/j.biomaterials.2006.06.008. Epub 2006 Jun 30.

Abstract

The aim of this study was to investigate the potential of using PDLLA/45S5 (PDLLA--poly(D,L-lactide)) Bioglass composite films for the culture of annulus fibrosus (AF) cells in vitro with a view to a tissue engineering application. PDLLA films incorporated with different percentages (0, 5 and 30 (wt%)) of Bioglass particles were prepared by solvent casting and characterized by scanning electron microscopy (SEM), water contact angle and white-light interferometry. Bovine AF cell morphology and attachment were analysed using SEM. Cytoskeletal organization was determined by actin labelling with FITC-phalloidin using fluorescence microscopy. The amount of sulphated glycosaminoglycan (sGAG) and collagen produced by AF cells were quantified using the 1,9-dimethylmethylene blue (DMMB) and Sircol assays after 4 weeks in culture. Composite films of PDLLA filled with Bioglass are an appropriate substrate for annulus cells and these films promote the production of an extracellular matrix (ECM) containing abundant sGAGs and collagen. These findings provide a basis for the understanding of the production of ECM molecules by cells cultured on 2D PDLLA/45S5 Bioglass composite films. The results will provide new insights into the design and development of composites containing Bioglass and resorbable polymers as scaffolds for intervertebral disc tissue repair.

摘要

本研究的目的是探讨使用聚(D,L-丙交酯)/45S5生物活性玻璃(PDLLA-聚(D,L-丙交酯))复合膜在体外培养纤维环(AF)细胞用于组织工程应用的潜力。通过溶剂浇铸制备了含有不同百分比(0、5和30(wt%))生物活性玻璃颗粒的PDLLA膜,并通过扫描电子显微镜(SEM)、水接触角和白光干涉测量法对其进行了表征。使用SEM分析牛AF细胞的形态和附着情况。通过用异硫氰酸荧光素(FITC)-鬼笔环肽进行肌动蛋白标记,利用荧光显微镜确定细胞骨架的组织情况。在培养4周后,使用1,9-二甲基亚甲基蓝(DMMB)和Sircol检测法定量AF细胞产生的硫酸化糖胺聚糖(sGAG)和胶原蛋白的量。填充有生物活性玻璃的PDLLA复合膜是环细胞的合适基质,并且这些膜促进含有丰富sGAG和胶原蛋白的细胞外基质(ECM)的产生。这些发现为理解在二维PDLLA/45S5生物活性玻璃复合膜上培养的细胞产生ECM分子提供了基础。这些结果将为含有生物活性玻璃和可吸收聚合物作为椎间盘组织修复支架的复合材料的设计和开发提供新的见解。

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