Eugene and Marilyn Glick Eye Institute, Department of Ophthalmology, Indiana University School of Medicine, USA.
Eugene and Marilyn Glick Eye Institute, Department of Ophthalmology, Indiana University School of Medicine, USA; Department of Pharmacology and Toxicology, Indiana University School of Medicine, USA.
Exp Eye Res. 2022 Jan;214:108891. doi: 10.1016/j.exer.2021.108891. Epub 2021 Dec 9.
The human anterior segment perfusion culture model is a valuable tool for studying the trabecular meshwork (TM) and aqueous humor outflow in glaucoma. The traditional model relies on whole eye globes resulting in high cost and limited availability. Here, we developed a glue-based method which enabled us to use human corneal rims for perfusion culture experiments. Human corneal rim perfusion culture plates were 3D printed. Human corneal rims containing intact TM were attached and sealed to the plate using low viscosity and high viscosity glues, respectively. The human corneal rims were perfused using the constant flow mode, and the pressure changes were recorded using a computerized system. Outflow facility, TM stiffness, and TM morphology were evaluated. When perfused at rates from 1.2 to 3.6 μl/min, the outflow facility was 0.359 ± 0.216 μl/min/mmHg among 10 human corneal rims. The stiffness of the TM in naïve human corneal rim was similar to that of perfusion cultured human corneal rim. Also, the stiffness of TM of corneal rims perfused with dexamethasone was significantly higher than the control. Human corneal rims with glue contamination in the TM could be differentiated by high baseline intraocular pressure as well as high TM stiffness. Histology studies showed that the TM tissues perfused with plain medium appeared normal. We believed that our glued-based method is a useful tool and low-cost alternative to the traditional anterior segment perfusion culture model.
人眼前节灌流培养模型是研究青光眼小梁网(TM)和房水流出的有价值的工具。传统模型依赖于整个眼球,导致成本高且可用性有限。在这里,我们开发了一种基于胶的方法,使我们能够使用人眼角膜缘进行灌流培养实验。人眼角膜缘灌流培养板采用 3D 打印技术制成。含有完整 TM 的人眼角膜缘分别使用低粘度和高粘度胶附着并密封到板上。使用恒流模式对人眼角膜缘进行灌流,并用计算机系统记录压力变化。评估流出效率、TM 硬度和 TM 形态。当以 1.2 至 3.6 μl/min 的速率灌流时,10 个人眼角膜缘的流出效率为 0.359 ± 0.216 μl/min/mmHg。未处理的人眼角膜缘的 TM 硬度与人眼角膜缘灌流培养的 TM 硬度相似。此外,用地塞米松灌流的角膜缘 TM 的硬度明显高于对照组。TM 中有胶污染的人眼角膜缘可以通过高基础眼压和高 TM 硬度来区分。组织学研究表明,用普通培养基灌流的 TM 组织外观正常。我们相信,我们的基于胶的方法是传统眼前节灌流培养模型的有用工具和低成本替代方法。