Abstract
We developed a glaucoma-on-a-chip (GOC) model to evaluate the viability of retinal ganglion cells (RGCs) against high pressure and neuroprotective treatments. A three-layered chip consisting of interconnecting microchannels and culture wells was fabricated based on simulation of physical parameters. The bottom surface of the wells was mechanically modified by air plasma and coated with different membranes to model an extracellular microenvironment. SH-SY5Y used as model cells to determine the best supporting membrane which was revealed to be PDL/laminin. RGCs were isolated from postnatal Wistar rats and purified by magnetic assisted cell sorting up to 70%. The cultured RGCs were exposed to normal (15 mmHg) or elevated pressure (33 mmHg) for 6, 12, 24, 36 and 48 hours, with and without adding brain-derived-neurotrophic factor (BDNF) or a novel BDNF mimetic (named RNYK). 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 hours, respectively (P<0.0001). BDNF and RNYK reduced the rate of RGC death under both normal and elevated pressures, two-fold approximately (p<0.01-0.0001).
Footnotes
Summary statement: Glaucoma-on-a-chip offers the advantages of allowing controlled experimental conditions, preliminary targeting of a specific cell type or pathway involved in glaucoma and investigating putative neuroprotective agents prior to assessment in animal models.