Abstract
Purpose :
To evaluate the protective effect of plasma rich in growth factors (PRGF-Endoret) over blue-light treated retinal pigment epithelium cells cytotoxicity
Methods :
Eye drops were obtained using the Endoret (PRGF) Ophthalmology kit (BTI Biotechnology Institute, S.L., Vitoria, Spain). Briefly, after informed consent was obtained, blood from healthy donors was collected into 9 mL tubes. Blood was centrifuged at 580g for 8 minutes, whole plasma column was drawn off avoiding the buffy coat, incubated at 37 oC for one hour, and the released supernatants were collected and filtered. Retinal pigment epithelium (ARPE-19) cells were exposed to blue light (470 nm) for 24 hours. Then, cells were incubated with PRGF or with routine culture medium (control) for another 24 and 48 hours maintaining the exposure to blue light. The cytoprotective effect of PRGF on blue light exposed ARPE-19 cells was evaluated by measuring the cell viability, the ROS production, the mitochondrial health status (JC-10) and the expression of some proteins related to the cellular stress like heme-oxygenase 1 (HO-1), catalase and superoxide dismutase (SOD-1)
Results :
The cell viability increased significantly at 24 and 48 hours after PRGF treatment comparing to the control group. The synthesis of ROS was significantly diminished in cells treated with PRGF but very elevated in the case of control. Also, after 24 hours of PRGF treatment, the JC-10 expression that reflects mitochondrial membrane potential was decreased. HO-1 levels measured by immunofluorescence and western blot was significantly reduced after PRGF treatment at both times of treatment. Furthermore, a significant increase of catalase expression was observed after PRGF treatment at 24 and 48 h regarding to the control group. However, no significant changes were observed in SOD-1 expression between both groups
Conclusions :
The present results suggest that PRGF treatment exerts cytotoxic protective effects on blue light-oxidative stress in an in vitro model of pigment epithelial cells
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.