Institute for Science and Technology in Medicine, School of Medicine, Keele University, Stoke-on-Trent, United Kingdom.
Mid Cheshire Hospitals NHS Trust, Stoke-on-Trent, United Kingdom.
Invest Ophthalmol Vis Sci. 2019 Feb 1;60(2):605-614. doi: 10.1167/iovs.18-25335.
Surgical techniques such as trabeculectomy aim to treat glaucoma by making an incision into the scleral tissue, to create an alternative drainage pathway for aqueous to flow into the sub-Tenon's/subconjunctival space. However, tissue fibrosis and wound healing occurring after the procedures can reduce the success rate. This study aims to investigate the synergistic effects of aqueous humor in combination with shear stress on the fibrosis response occurring in Tenon's capsule and conjunctival tissue (TCCT) after glaucoma surgery.
Two-dimensional (2D) and 3D in vitro TCCT models were constructed by seeding porcine Tenon's capsule + conjunctival fibroblasts in collagen gel. These were used to investigate key growth factors (singular and natural form) with shear stress, which are believed to influence tissue fibrosis after glaucoma surgery. In addition to cell proliferation assessments, a nondestructive assay to quantify neocollagen synthesis in TCCT models, in response to these factors, has been applied up to 14 days.
TCCT fibroblast proliferation increased significantly with doses of TGF-β, TNF-α, and VEGF, in comparison with the control. Furthermore, fibroblasts exposed to 50% aqueous humor had significantly increased proliferation and actin expression. Shear stress-induced mechanotransduction was also found to promote metabolic activity across experimental conditions. Neocollagen labeling cross validated the fibrosis process.
Shear stress appeared to enhance the influence of key growth factors and further promoted fibrotic response within the model. These findings offer a useful insight for further study into the wound-healing response triggered by aqueous fluid outflow after glaucoma surgery.
外科手术技术，如小梁切除术，通过在巩膜组织上做切口，为房水提供另一条进入腱膜下/结膜下间隙的引流途径，从而治疗青光眼。然而，手术后发生的组织纤维化和伤口愈合可能会降低手术成功率。本研究旨在探讨房水联合切应力对青光眼手术后Tenon 囊和结膜组织（TCCT）纤维化反应的协同作用。
通过在胶原凝胶中接种猪 Tenon 囊+结膜成纤维细胞，构建二维（2D）和三维（3D）体外 TCCT 模型。这些模型用于研究在青光眼手术后可能影响组织纤维化的关键生长因子（单一和天然形式）和切应力。除了细胞增殖评估外，还应用了一种非破坏性测定法来定量 TCCT 模型对这些因子的新胶原蛋白合成，时间长达 14 天。
与对照组相比，TGF-β、TNF-α 和 VEGF 的剂量使 TCCT 成纤维细胞的增殖显著增加。此外，暴露于 50%房水中的成纤维细胞增殖和肌动蛋白表达显著增加。还发现切应力诱导的力学转导促进了实验条件下的代谢活性。新胶原标记物验证了纤维化过程。
切应力似乎增强了关键生长因子的影响，并进一步促进了模型中的纤维化反应。这些发现为进一步研究青光眼手术后房水流出引发的伤口愈合反应提供了有用的见解。
