Bioactive glasses particles direct fibroblast towards better wound healing

摘要

Introduction: Lately, some studies have shown that bioactive glass take an active role in stimulating angiogenesis, indicating its potential application in soft tissue repair, like skin regeneration. Some early pioneering efforts have found inspiring results for the bioactive glass on skin repair in animal models like pigs, rats, dogs. However, the specific regulation of BG on skin cells are still unclear and demand more investigation. Herein monodisperse spherical BG particles and normal human foreskin fibroblasts were cocultured to systematically study the wound healing related cellular response of fibroblasts to BG particles. Materials and Methods: The acid-alkali co-catalyzed methodology were used to fabricate 90S bioactive glass particles. The obtained BG particles was characterized using SEM, LPSA and ICP-AES. Fibroblasts were seeded on 6-well plates allowed to attach for 24h before BG particles containing medium was added. The regulation of BG particles on fibroblasts were evaluated by CCK-8, immunofluorescent microscopy, flow cytometry, time-lapse video microscopy, qRT-PCR and western blotting. Results and Discussion: Time-lapse imaging revealed a promoted fibroblasts motility stimulated by BG particles. Results on the expression of extracellular matrix (ECM) related gene illustrated BG particles reduced the synthesis capacity for critical ECM molecules including type Icollagen, type Ⅱ collagen and fibronectin, while increased the expression of tenascin-C. More importantly, the fibroblasl-to-myofibroblast differentiation was greatly inhibited for the fibroblasts cocultured with BG particles. Further analysis on the intracellular signaling pathways demonstrated that BG particles down-regulated myofibroblastic differentiation via TGF-β1-Smad signaling evidenced by decreased level of TGF-β receptor I and its downstream effector Smad2. We also found that even in the TGF-β1 abundant environment, the myofibroblastic differentiation were still suppressed by BG particles. Conclusion and Significance: In this study we showed how BG particles regulates the cellular responses of fibroblasts related to wound healing. It is clear that BG particles promoted fibroblast migration, modulated the synthesis of new ECM molecules and inhibited the myofibroblastic differentiation. TGF-beta 1 -smad2 signaling may be involved in the regulation of fibroblast-to-myofibroblast differentiation. Our study not only provided a comprehensive knowledge of the specific regulation of BG particles on fibroblasts, but also offer a potential therapies aiming for a better wound healing with less scar.