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

摘要

The aim of this study was to investigate the potential of using PDLLA/45S5 (PDLLA-poly(D,L-lactide)) Bioglass (R) 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 (R) 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 (TM) assays after 4 weeks in culture. Composite films of PDLLA filled with Bioglass (R) 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 (R) lot composite films. The results will provide new insights into the design and development of composites containing Bioglass (R) and resorbable polymers as scaffolds for intervertebral disc tissue repair. (c) 2006 Elsevier Ltd. All rights reserved.