Ramklowan D S H, Snyman C, van de Vyver M, Niesler C U
Discipline of Biochemistry, School of Life Sciences, University of KwaZulu Natal, Scottsville, South Africa.
Division of Clinical Pharmacology, Department of Medicine, Faculty of Medicine and Health Sciences, Stellenbosch University, South Africa.
Biochimie. 2023 Apr;207:96-101. doi: 10.1016/j.biochi.2022.10.017. Epub 2022 Nov 4.
Fibroblasts function to secrete and modify components of the extracellular matrix. During wound healing, fibroblasts migrate to the site of injury and differentiate into contractile myofibroblasts; this differentiation is characterised by an increased contractile capacity. Fully differentiated myofibroblasts can be distinguished from fibroblasts via the higher expression of α-smooth muscle actin as well as a denser cytoskeleton. Impaired wound healing has been characterised by a lack of myofibroblasts; as a result, tissue does not fully regain its strength and function. Under pathological conditions, this may be associated with the effect that a pro-inflammatory microenvironment has on fibroblast and skeletal muscle progenitor cell migration and differentiation. Given their distinct roles in tissue maintenance and repair, the communication between fibroblasts versus myofibroblasts with other cellular mediators of repair is likely to influence cell behaviour and the outcome of wound repair. An in vitro test model is required to investigate this intercellular influence, but the establishment of such a model is hampered by the difficulty in retaining the dedifferentiated fibroblastic phenotype under regular serum-containing cell culture conditions. We present a model that supports the establishment and retention in culture of fibroblast and myofibroblast phenotypes for use in a simple, inexpensive, yet relevant in vitro 2D assay. This model is then applied in a co-culture setting to determine whether the presence of myoblasts affects the ability of fibroblasts versus myofibroblasts to close an in vitro wound. Our results emphasize the importance of considering the impact of paracrine communication between all cells during wound healing.
成纤维细胞的功能是分泌和修饰细胞外基质的成分。在伤口愈合过程中，成纤维细胞迁移到损伤部位并分化为收缩性肌成纤维细胞；这种分化的特征是收缩能力增强。完全分化的肌成纤维细胞可通过α-平滑肌肌动蛋白的高表达以及更密集的细胞骨架与成纤维细胞区分开来。伤口愈合受损的特征是缺乏肌成纤维细胞；因此，组织无法完全恢复其强度和功能。在病理条件下，这可能与促炎微环境对成纤维细胞和骨骼肌祖细胞迁移及分化的影响有关。鉴于它们在组织维持和修复中的不同作用，成纤维细胞与肌成纤维细胞之间与其他修复细胞介质的通讯可能会影响细胞行为和伤口修复的结果。需要一个体外测试模型来研究这种细胞间的影响，但在常规含血清细胞培养条件下难以维持去分化的成纤维细胞表型，这阻碍了此类模型的建立。我们提出了一个模型，该模型支持在培养中建立和保留成纤维细胞和肌成纤维细胞表型，用于简单、廉价但相关的体外二维分析。然后将该模型应用于共培养环境中，以确定成肌细胞的存在是否会影响成纤维细胞与肌成纤维细胞闭合体外伤口的能力。我们的结果强调了在伤口愈合过程中考虑所有细胞之间旁分泌通讯影响的重要性。
