Li Zhuqing, Bratlie Kaitlin M
Department of Materials Science & Engineering, Iowa State University, Ames, IA 50011, USA.
Department of Materials Science & Engineering, Iowa State University, Ames, IA 50011, USA; Department of Chemical & Biological Engineering, Iowa State University, Ames, IA 50011, USA.
Mater Sci Eng C Mater Biol Appl. 2021 Mar;122:111915. doi: 10.1016/j.msec.2021.111915. Epub 2021 Jan 29.
In tissue regeneration, the goal is to regenerate tissue similar to what was damaged or missing while preventing fibrotic scarring, which may lead to decreased mechanical strength and dissimilar tissue characteristics compared to native tissue. We believe collagen orientation plays a critical role in wound contraction and scarring and that it is modulated by myofibroblasts. We used macrophage conditioned medium to simulate complex events that can influence the fibroblast phenotype during the wound healing process. In addition to examining the effect of macrophage phenotype on fibroblasts, we inhibited focal adhesion kinase (FAK), Rho-associated protein kinase (ROCK), and myosin II for fibroblasts cultured on both tissue culture plastic and methacrylated gellan gum to understand how different pathways and materials influence fibroblast responses. Collagen orientation, α-SMA expression, focal adhesion area, and cell migration were altered by inhibition of FAK, ROCK, or myosin II and macrophage phenotype, along with the substrate. An increase in either focal adhesion area or α-smooth muscle actin (α-SMA) expression correlated with an aligned collagen orientation. Gellan gum hydrogels upregulated α-SMA expression in ROCK inhibited conditioned media and downregulated the FAK area in FAK and ROCK inhibited conditioned media. Myosin II had no impact on the α-SMA expression on the substrate compared to coverslip except for M2 conditioned medium. Gellan gum hydrogel significantly increased cell migration under FAK and Myosin II mediated conditioned media and unconditioned media. Collectively, our study examined how macrophage phenotype influences fibroblast response, which would be beneficial in controlling scar tissue formation.
在组织再生中，目标是再生出与受损或缺失组织相似的组织，同时防止纤维化瘢痕形成，因为纤维化瘢痕可能导致机械强度降低以及与天然组织相比组织特性不同。我们认为胶原蛋白的取向在伤口收缩和瘢痕形成中起关键作用，并且它受肌成纤维细胞调节。我们使用巨噬细胞条件培养基来模拟在伤口愈合过程中可能影响成纤维细胞表型的复杂事件。除了研究巨噬细胞表型对成纤维细胞的影响外，我们还抑制了在组织培养塑料和甲基丙烯酸化结冷胶上培养的成纤维细胞的粘着斑激酶（FAK）、Rho相关蛋白激酶（ROCK）和肌球蛋白II，以了解不同途径和材料如何影响成纤维细胞反应。FAK、ROCK或肌球蛋白II的抑制以及巨噬细胞表型和底物改变了胶原蛋白取向、α-平滑肌肌动蛋白（α-SMA）表达、粘着斑面积和细胞迁移。粘着斑面积或α-平滑肌肌动蛋白（α-SMA）表达的增加与排列的胶原蛋白取向相关。结冷胶水凝胶在ROCK抑制的条件培养基中上调α-SMA表达，在FAK和ROCK抑制的条件培养基中下调FAK面积。与盖玻片相比，除了M2条件培养基外，肌球蛋白II对底物上的α-SMA表达没有影响。结冷胶水凝胶在FAK和肌球蛋白II介导的条件培养基和非条件培养基下显著增加细胞迁移。总的来说，我们的研究探讨了巨噬细胞表型如何影响成纤维细胞反应，这将有助于控制瘢痕组织形成。
