Centre de recherche du CHU de Québec-Université Laval, axe médecine régénératrice, Québec, Québec, Canada; Centre LOEX de l'Université Laval, Québec, Québec, Canada; Département, d'ophtalmologie et ORL-CCF, Université Laval, Québec, Québec, Canada.
Centre de recherche du CHU de Québec-Université Laval, axe médecine régénératrice, Québec, Québec, Canada; Centre LOEX de l'Université Laval, Québec, Québec, Canada.
Exp Eye Res. 2021 May;206:108532. doi: 10.1016/j.exer.2021.108532. Epub 2021 Mar 6.
Mechanicals forces are known to influence cell behavior. In vivo, the corneal endothelium is under the influence of various mechanical forces, such as intraocular pressure (IOP) and fluid flow. In this study, we used a corneal bioreactor to understand the effect of these hydrodynamic forces on the transcription of intercellular junctions associated genes in the corneal endothelium. Native and tissue-engineered (TE) corneal endothelium were cultured in a corneal bioreactor for 7 days with 16 mmHg IOP and 5 μl/ml of medium flow. RNA was harvested, and gene expression was quantified. Cells that were used to reconstruct the TE corneal endothelia were also seeded on plastic to characterize their morphology by calculating their circularity index. For native endothelia, hydrodynamic forces increased gene expression of GJA1 (connexin 43), CDH2 (N-cadherin), TJP1 (ZO-1), ITGAV (integrin subunit αv), ITGB5 (integrin subunit β5) and CTNND1 (p120-ctn) by 1.68 ± 0.40, 1.10 ± 0.27, 3.80 ± 0.56, 1.82 ± 0.33, 1.32 ± 0.21 and 3.04 ± 0.63, respectively. For TE corneal endothelium, this fold change was 1.72 ± 0.31, 1.58 ± 0.41, 6.18 ± 1.03, 1.80 ± 0.71, 1.77 ± 0.55, 2.42 ± 0.71. Furthermore, gene transcription fold changes (hydrodynamic/control) increased linearly with TE corneal endothelium cells population morphology with r = 0.83 for TJP1 (ZO-1) and r = 0.58 for CTNND1 (p120-ctn). In fact, the more elongated the cells populations were, the greater hydrodynamic conditions increased the transcription of TJP1 (ZO-1) and CTNND1 (p120-ctn). These results suggest that hydrodynamic forces contribute to the maintenance of tight and adherens junctions of native corneal endothelial cells, as well as to the formation of tight and adherens junctions of corneal endothelial cells that are in the process of forming a functional endothelial barrier.
机械力被认为会影响细胞行为。在体内，角膜内皮细胞受到各种机械力的影响，如眼压（IOP）和流体流动。在这项研究中，我们使用角膜生物反应器来了解这些流体动力对角膜内皮细胞中细胞间连接相关基因转录的影响。天然和组织工程（TE）角膜内皮细胞在角膜生物反应器中培养 7 天，眼压为 16mmHg，培养基流量为 5μl/ml。提取 RNA，定量基因表达。用于重建 TE 角膜内皮的细胞也接种在塑料上，通过计算其圆形指数来表征其形态。对于天然内皮细胞，流体动力使 GJA1（连接蛋白 43）、CDH2（N-钙粘蛋白）、TJP1（ZO-1）、ITGAV（整合素亚基αv）、ITGB5（整合素亚基β5）和 CTNND1（p120-ctn）的基因表达分别增加了 1.68±0.40、1.10±0.27、3.80±0.56、1.82±0.33、1.32±0.21 和 3.04±0.63。对于 TE 角膜内皮细胞，这种变化倍数分别为 1.72±0.31、1.58±0.41、6.18±1.03、1.80±0.71、1.77±0.55、2.42±0.71。此外，基因转录倍数变化（流体动力/对照）与 TE 角膜内皮细胞群体形态呈线性关系，TJP1（ZO-1）的 r=0.83，CTNND1（p120-ctn）的 r=0.58。事实上，细胞群体越细长，流体条件对 TJP1（ZO-1）和 CTNND1（p120-ctn）的转录增加就越大。这些结果表明，流体动力有助于维持天然角膜内皮细胞的紧密连接和黏附连接，并有助于正在形成功能性内皮屏障的角膜内皮细胞的紧密连接和黏附连接的形成。
