Abstract:
During the physiological healing of skin wounds, fibroblasts recruited from the uninjured adjacent dermis and deeper subcutaneous fascia layers are transiently activated into myofibroblasts to first secrete and then contract collagen-rich extracellular matrix into a mechanically resistant scar. Scar tissue restores skin integrity after damage but comes at the expense of poor esthetics and loss of tissue function. Stiff scar matrix also mechanically activates various precursor cells into myofibroblasts in a positive feedback loop. Persistent myofibroblast activation results in pathologic accumulation of fibrous collagen and hypertrophic scarring, called fibrosis. Consequently, the mechanisms of fibroblast-to-myofibroblast activation and persistence are studied to develop antifibrotic and prohealing treatments. Mechanistic understanding often starts in a plastic cell culture dish. This can be problematic because contact of fibroblasts with tissue culture plastic or glass surfaces invariably generates myofibroblast phenotypes in standard culture. We describe a straight-forward method to produce soft cell culture surfaces for fibroblast isolation and continued culture and highlight key advantages and limitations of the approach. Adding a layer of elastic silicone polymer tunable to the softness of normal skin and the stiffness of pathologic scars allows to control mechanical fibroblast activation while preserving the simplicity of conventional 2-dimensional cell culture.
摘要:
在皮肤伤口的生理愈合过程中,从未受伤的邻近真皮和较深的皮下筋膜层募集的成纤维细胞被瞬时激活为肌成纤维细胞,首先分泌然后收缩富含胶原蛋白的细胞外基质,形成机械抗性瘢痕。瘢痕组织在损伤后恢复皮肤完整性,但以美学差和组织功能丧失为代价。僵硬的疤痕基质还在正反馈回路中将各种前体细胞机械激活为肌成纤维细胞。持续的肌成纤维细胞活化导致纤维胶原的病理性积累和肥厚性瘢痕形成,叫做纤维化。因此,我们研究了成纤维细胞到肌成纤维细胞的激活和持续的机制,以开发抗纤维化和促愈合治疗。机械理解通常始于塑料细胞培养皿。这可能是有问题的,因为成纤维细胞与组织培养塑料或玻璃表面的接触总是在标准培养物中产生成肌纤维细胞表型。我们描述了一种直接的方法来产生用于成纤维细胞分离和持续培养的软细胞培养表面,并强调了该方法的关键优势和局限性。添加可调节正常皮肤的柔软度和病理性疤痕的硬度的弹性硅酮聚合物层允许控制机械成纤维细胞活化,同时保持常规二维细胞培养的简单性。