School of Biomedical Engineering, Western University, London, ON N6A 5C1, Canada.
Department of Physics and Astronomy, Faculty of Science, Western University, London, ON N6A 3K7, Canada.
ACS Appl Mater Interfaces. 2023 Apr 26;15(16):19817-19832. doi: 10.1021/acsami.2c22381. Epub 2023 Apr 17.
The substratum topography of both natural and synthetic materials is a prominent regulator of cell behaviors including adhesion, migration, matrix fibrillogenesis, and cell phenotype. Connective tissue fibroblasts are known to respond to repeating groove topographical modifications by aligning and exhibiting directed migration, a phenomenon termed contact guidance. Although both reside in collagen rich connective tissues, dermal and gingival fibroblasts are known to exhibit differences in phenotype during wound healing, with gingival tissue showing a fetal-like scarless response. Differences in adhesion formation and maturation are known to underlie both a scarring phenotype and cell response to topographical features. Utilizing repeating groove substrates with periodicities of 600, 900, and 1200 nm (depth, 100 nm), we investigated the roles of integrins αvβ3 and β1 associated adhesions on contact guidance of human gingival (HGFs) and dermal fibroblasts (HDFs). HGFs showed a higher degree of orientation with the groove long axis than HDFs, with alignment of both vinculin and tensin-1 evident on 600 and 900 nm periodicities in both cell types. Orientation with grooves of any periodicity in HGFs and HDFs did not alter the adhesion number or area compared to smooth control surfaces. Growth of both cell types on all periodicities reduced fibronectin fibrillogenesis compared to control surfaces. Independent inhibition of integrin αvβ3 and β1 in both cell types induced changes in spreading up to 6 h and reduced alignment with the groove long axis. At 24 h post-seeding with blocking antibodies, HGFs recovered orientation, but in HDFs, blocking of β1, but not αvβ3 integrins, inhibited alignment. Blocking of β1 and αvβ3 in HDFs, but not HGFs, inhibited tensin-1-associated fibrillar adhesion formation. Furthermore, inhibition of β1 integrins in HDFs, but not HGFs, resulted in recruitment of tensin-1 to αvβ3 focal adhesions, preventing HDFs from aligning with the groove long axis. Our work demonstrates that tensin-1 localization with specific integrins in adhesion sites is an important determinant of contact guidance. This work emphasizes further the need for tissue-specific biomaterials, when integration into host tissues is required.
天然和合成材料的亚表层形貌是调节细胞行为的重要因素，包括细胞黏附、迁移、基质纤维生成和细胞表型。众所周知，结缔组织成纤维细胞会对重复的沟槽形貌变化做出反应，通过定向排列和定向迁移来进行响应，这种现象被称为接触引导。尽管真皮和牙龈成纤维细胞都位于富含胶原蛋白的结缔组织中，但在伤口愈合过程中，它们的表型已知存在差异，牙龈组织表现出类似于无瘢痕的反应。黏附形成和成熟的差异是导致瘢痕表型和细胞对形貌特征反应的基础。我们利用具有 600、900 和 1200nm 周期性（深度为 100nm）的重复沟槽基底，研究了整合素 αvβ3 和 β1 相关黏附在人牙龈（HGFs）和真皮成纤维细胞（HDFs）接触引导中的作用。与 HDFs 相比，HGFs 与沟槽长轴的定向程度更高，两种细胞类型在 600nm 和 900nm 周期性时，vinculin 和 tensin-1 的排列都很明显。与光滑对照表面相比，HGFs 和 HDFs 中任何周期性的沟槽定向都不会改变黏附数量或面积。与对照表面相比，两种细胞类型在所有周期性表面上的生长都会减少纤维连接蛋白纤维生成。在两种细胞类型中，独立抑制整合素 αvβ3 和 β1，在 6 小时内会引起细胞扩展的变化，并减少与沟槽长轴的对齐。在接种阻断抗体 24 小时后，HGFs 恢复了定向，但在 HDFs 中，阻断β1 整合素，但不阻断 αvβ3 整合素，会抑制定向。在 HDFs 中，阻断β1 和αvβ3 整合素，但不在 HGFs 中，会抑制与 tensin-1 相关的纤维状黏附形成。此外，在 HDFs 中抑制β1 整合素，但不在 HGFs 中，会导致 tensin-1 募集到αvβ3 焦点黏附，从而阻止 HDFs 与沟槽长轴对齐。我们的工作表明，在黏附部位，特定整合素与 tensin-1 的定位是接触引导的重要决定因素。这项工作进一步强调了在需要整合到宿主组织中时，需要使用组织特异性生物材料。
