Department of Medical Biotechnology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Department of Electronics, School of Electrical and Computer Engineering, Tehran University, Tehran, Iran.
Brain Behav. 2020 Oct;10(10):e01799. doi: 10.1002/brb3.1799. Epub 2020 Aug 16.
We developed a glaucoma-on-a-chip model to evaluate the viability of retinal ganglion cells (RGCs) against high pressure and the potential effect of neuroprotection.
A three-layered chip consisting of interconnecting microchannels and culture wells was designed and fabricated from poly-methyl methacrylate sheets. The bottom surface of the wells was modified by air plasma and coated with different membranes to provide a suitable extracellular microenvironment. RGCs were purified from postnatal Wistar rats by magnetic assisted cell sorting up to 70% and characterized by flow cytometry and immunocytochemistry. The cultured RGCs were exposed to normal (15 mmHg) or elevated pressure (33 mmHg) for 6, 12, 24, 36, and 48 hr, with and without adding brain-derived neurotrophic factor (BDNF) or a novel BDNF mimetic (RNYK).
Multiple inlet ports allow culture media and gas into the wells under elevated hydrostatic pressure. PDL/laminin formed the best supporting membrane. RGC survival rates were 85%, 78%, 70%, 67%, and 61% under normal pressure versus 40%, 22%, 18%, 12%, and 10% under high pressure at 6, 12, 24, 36, and 48 hr, respectively. BDNF and RNYK separately reduced RGC death rates about twofold under both normal and elevated pressures.
This model recapitulated the effects of elevated pressure over relatively short time periods and demonstrated the neuroprotective effects of BDNF and RNYK.
我们开发了一种青光眼芯片模型，以评估高眼压对视网膜神经节细胞（RGCs）活力的影响，以及神经保护的潜在作用。
我们设计并制作了一个由聚甲基丙烯酸甲酯片组成的三层芯片，其中包括相互连接的微通道和培养孔。孔的底面通过空气等离子体进行修饰，并涂覆不同的膜，以提供合适的细胞外微环境。通过磁辅助细胞分选，从新生 Wistar 大鼠中纯化出高达 70%的 RGCs，并通过流式细胞术和免疫细胞化学进行鉴定。培养的 RGCs 在正常（15mmHg）或升高的压力（33mmHg）下暴露 6、12、24、36 和 48 小时，同时加入脑源性神经营养因子（BDNF）或新型 BDNF 模拟物（RNYK）或不加。
多个入口允许在升高的静水压力下将培养基和气体引入孔中。PDL/层粘连蛋白形成了最佳的支撑膜。在正常压力下，RGC 存活率分别为 85%、78%、70%、67%和 61%，而在 6、12、24、36 和 48 小时时，高压下的存活率分别为 40%、22%、18%、12%和 10%。BDNF 和 RNYK 分别在正常和升高的压力下使 RGC 死亡率降低约两倍。
该模型在相对较短的时间内再现了升高压力的影响，并证明了 BDNF 和 RNYK 的神经保护作用。
