Antiproliferative and apoptotic effect of anti-VEGF DRUGS (aflibercept, ranibizumab, brolucizumab) on human retinal pigment epithelium (ARPE-19) and human umbilical vein endothelial cells (HUVEC)

Age-Related Macular Degeneration (AMD) is a progressive disease characterized by visual loss, mainly affecting the posterior pole and increasing in incidence with age. The interactions between retinal pigment epithelium (RPE), choroid, and photoreceptors, as well as various mechanisms such as oxidative damage, immune system activation, and inflammation, are involved in the pathogenesis of AMD. With the understanding of the importance of angiogenic pathways and vascular endothelial growth factor (VEGF) in the pathogenesis of AMD, anti-VEGF agents have become the first-line treatment. This study aims to compare the antiproliferative and oxidative stress-induced apoptotic effects of Aflibercept, Ranibizumab, and Brolucizumab in ARPE-19 and HUVEC cultures. For this purpose, the drug-dose selection was made using the MTT test. The apoptotic and necrotic effects were evaluated with Flow Cytometry and DAPI staining method. For all three drugs, the VEGF-A gene expressions and protein levels were evaluated after 24-hour incubation. Total oxidant and antioxidant capacity (TOC and TAC) were determined by the colorimetric test, and Nrf2 & NF-kB levels were measured by ELISA. In our study, anti-VEGF drugs increased apoptosis and decreased cell proliferation in both HUVEC and ARPE-19 cell cultures. TOC increased, and TAC decreased, consistent with decreased Nrf2 and increased NF-kB levels. All three drugs decreased VEGF-A gene and protein expression. Even though anti-VEGF drugs provide anatomical improvement, they may cause increased RPE atrophy, inflammation, and oxidative stress. For this reason, they should be used at minimum effective doses and the lowest possible frequency. The discovery of new treatment agents or combination therapies with reduced side effects is crucial. Keywords: Aflibercept, Ranibizumab, Brolucizumab, Age-Related Macular Degeneration, oxidative stress