Abstract
Purpose :
Proliferative Vitreoretinopathy (PVR) is common cause of surgical failure in up to 10% of patients who have undergone vitrectomy surgery. We have previously shown that our hydrogel-based vitreous tamponade (termed vitreogel) has dual functions: (i) to act as an endo-tamponade agent for retinal detachment surgery, and (ii) biodegradable scaffold for vitreous reformation. In this study, we explore the anti-PVR effects of vitreogel using an in-vitro retinal pigment epithelial (RPE) model.
Methods :
Using a human retinal pigment epithelial line, ARPE-19 (ATCCÒ CRL-2302™), we induced formation of PVR-like contractile masses by co-treatment of TNF-alpha (TNF-α) and TGF-beta 1 (TGF-β1). ARPE-19 cells were cultured in the presence or absence of vitreogel during co-treatment. Morphological changes were observed using phase contrast microscopy. Cell viability test for biocompatibility was monitored using Cell Counting Kit-8 (CCK-8) assay. Gene and protein expression of RPE-specific and mesenchymal markers were studied using quantitative reverse transcription PCR (RT-qPCR) and immunofluorescence (IF) respectively.
Results :
TNF-α and TGF-β1 (TNT) co-treatment synergistically activates epithelial-to-mesenchymal (EMT) in ARPE-19 to form 3D masses by 72 hours. 3D mass formation is inhibited in groups exposed to vitreogel alone. In vitreogel treated cells, there was a reduction in RPE-specific markers with no corresponding gain in mesenchymal markers. In addition, a scratch wound assay performed showed suppression of APRE-19 migration in groups exposed to vitreogel.
Conclusions :
Vitreogel prevents EMT transition and cell migration in ARPE-19 cells treated with TNT. This suggests that vitreogel may lower the risk of PVR. Future studies will be focussed on assessing vitreogel’s ability to prevent PVR formation in an in-vivo rabbit model.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.