Purchase this article with an account.
Jonathan Pieter Vande Geest, Jr-Jiun Liou; Effect of genipin on the metabolic activity of porcine lamina cribrosa cells. Invest. Ophthalmol. Vis. Sci. 2021;62(8):1649.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
To investigate the effect of scleral stiffness on the susceptibility to glaucoma, Kimball et al. treated mouse eyes with glyceraldehyde alone via subconjunctival injections and found no effect on retinal structure or function. However, in mice with experimental glaucoma, glyceraldehyde significantly increases intraocular pressure and axon loss at 6 weeks. It therefore may be that de-stiffening of the sclera can rescue or halt glaucomatous damage. The effect that stiffening or de-stiffening agents have on the health of lamina cribrosa cells has not been fully quantified in the literature. In this preliminary study, we evaluated the effects of genipin on optic nerve head cells, specifically, lamina cribrosa cells.
Porcine lamina cribrosa (LC) cells were isolated from three fresh pig eyes purchased commercially. Cell characterization was performed using anti-alpha smooth muscle actin (αSMA) and anti-glial fibrillary acidic protein (GFAP) antibodies from Abcam. Cells at passage 3 were seeded onto a 24-well plate and cultured in DMEM medium with 10% Fetal Bovine Serum, 2.5% HEPES, 1% Amphotericin B and 1% Penicillin-Streptomycin. Genipin was reconstituted in medium at a final concentration of 10 mM. MTS assays were performed on day 3, day 7, and day 10 to quantify the metabolic activity of these cells.
Porcine LC cells are characterized as αSMA positive and GFAP negative (Figure 1A). Genipin at 10 mM significantly reduces the metabolic activity of LC cells in monolayer cultures on day 3, day 7, and day 10 (Figure 1B). The metabolic activity of genipin group is no different than the negative control (Figure 1B).
Genipin at 10 mM reduces the metabolic activity of LC cells. Future investigations are needed to examine the effects of genipin at a lower concentration (1 mM or 5 mM) in order to conclude the cytotoxic effects of genipin. We are currently working on eye culture with genipin treatment and examine the stiffening effects via whole globe inflation tests.
This is a 2021 ARVO Annual Meeting abstract.
This PDF is available to Subscribers Only