The effects of anti-vascular endothelial growth factor agents on human retinal pigment epithelial cells under high glucose conditions.

Published on Feb 18, 2017in International Journal of Ophthalmology1.33
· DOI :10.18240/IJO.2017.02.04
Jong Rok Oh2
Estimated H-index: 2
(SCH: Soonchunhyang University),
Jung Woo Han3
Estimated H-index: 3
+ 2 AuthorsTae Kwann Park13
Estimated H-index: 13
AIM: To investigate the effects of high glucose levels and anti-vascular endothelial growth factor (VEGF) agents (bevacizumab, ranibizumab and aflibercept) on retinal pigment epithelium (RPE) cells. METHODS: ARPE-19 cells were cultured at different glucose levels (5.5 mmol/L, 25 mmol/L, and 75 mmol/L). Cell viability was evaluated by MTT assay at 3d after treatment with D-glucose. Cell migration ability was measured by wound healing assay at 3d. A cell death detection kit was used to assess apoptosis at 3 and 14d. Cell proliferation was assessed by EdU assay at 3d. The culture medium was treated with anti-VEGF agents at clinically relevant concentrations. The experiment was then repeated at a different glucose level. RESULTS: The viability and migration of ARPE-19 cells were significantly decreased in the presence of 75 mmol/L as compared to 5.5 mmol/L glucose. The percentage of TUNEL-positive cells was significantly increased and the proliferative potential was decreased with 75 mmol/L compared to 5.5 mmol/L glucose. There were no significant differences in the results between 25 mmol/L and 5.5 mmol/L glucose. In the presence of 75 mmol/L glucose, the groups treated with anti-VEGF showed decreased cell viability and proliferation and increased apoptosis. However, there were no significant differences between the anti-VEGF groups. CONCLUSION: High glucose level decreases the viability, wound healing ability, and proliferation of RPE cells, while increasing apoptosis. Furthermore, anti-VEGF agents interfered with the physiological functions of RPE cells under high-glucose conditions, accompanied by decreases in cell viability and proliferation.
#1Michael J. ElmanH-Index: 35
#2Allison R. AyalaH-Index: 6
Last. Thomas W. StoneH-Index: 12
view all 8 authors...
Objective To report the 3-year follow-up results within a previously reported randomized trial evaluating prompt versus deferred (for ≥24 weeks) focal/grid laser treatment in eyes treated with intravitreal 0.5 mg ranibizumab for diabetic macular edema (DME). Design Multicenter, randomized clinical trial. Participants Three hundred sixty-one participants with visual acuity of 20/32 to 20/320 (approximate Snellen equivalent) and DME involving the fovea. Methods Ranibizumab every 4 weeks until no l...
403 CitationsSource
#1Jean-François Korobelnik (French Institute of Health and Medical Research)H-Index: 39
#2Diana V. Do (UNMC: University of Nebraska Medical Center)H-Index: 52
Last. David M. BrownH-Index: 46
view all 23 authors...
Purpose A head-to-head comparison was performed between vascular endothelial growth factor blockade and laser for treatment of diabetic macular edema (DME). Design Two similarly designed, double-masked, randomized, phase 3 trials, VISTA DME and VIVID DME . Participants We included 872 patients (eyes) with type 1 or 2 diabetes mellitus who presented with DME with central involvement. Methods Eyes received either intravitreal aflibercept injection (IAI) 2 mg every 4 weeks (2q4), IAI 2 mg every 8 w...
465 CitationsSource
#1Alexa Klettner (CAU: University of Kiel)H-Index: 22
#2Nihat Tahmaz (CAU: University of Kiel)H-Index: 1
Last. Johann Roider (CAU: University of Kiel)H-Index: 26
view all 5 authors...
Background/aim Anti-VEGF treatment is the therapy of choice in age-related macular degeneration, and is also applied in diabetic macular oedema or retinal vein occlusion. Recently, the fusion protein, aflibercept, has been approved for therapeutic use. In this study, we investigate the effects of aflibercept on primary RPE cells. Methods Primary RPE cells were prepared from freshly slaughtered pigs’ eyes. The impact of aflibercept on cell viability was investigated with MTT and trypan blue exclu...
35 CitationsSource
Background/aims Vascular endothelial growth factor (VEGF) is a key factor in the pathogenesis of neovascular retinal diseases including age-related macular degeneration. VEGF inhibitors including ranibizumab, pegaptanib or bevacizumab improve retinal morphology and vision in many patients. The recently approved drug aflibercept (VEGF Trap-Eye/Eyelea, Regeneron, Tarrytown, New York, USA) offers a new therapy modality. We therefore tested for toxic and anti-proliferating effects of aflibercept. Me...
50 CitationsSource
To investigate the effects of the vascular endothelial growth factor–neutralizing agent aflibercept on primary cultures of human trabecular meshwork cells (hTMC), human scleral fibroblasts (hFibro), and a retinal pigment epithelial cell line (ARPE-19). Various concentrations of aflibercept were incubated with confluent cell cultures for 24 hours. Ranibizumab was used as an active control for comparison. Assays of cellular metabolism (MTT assay) and cell viability (calcein dye uptake) were perfor...
20 CitationsSource
#1You-Hsuan Chen (NTHU: National Tsing Hua University)H-Index: 7
#2Hsiu-Chuan Chou (UEW: University of Education, Winneba)H-Index: 16
Last. Hong-Lin Chan (NTHU: National Tsing Hua University)H-Index: 23
view all 6 authors...
Abstract Retinopathy has been observed in around quarter of diabetic patients. Diabetic retinopathy can result in poor vision and even blindness since high glucose has been evidenced to weaken retinal capillary leading to leakage of blood into the surrounding space. In the present study, a proteomics-based approach has been applied to analyze a model retinal pigmented epithelium cell line, ARPE-19, grown in mannitol-balanced 5.5 mM, 25 mM and 100 mM D-glucose culture media and used as a model fo...
23 CitationsSource
#1Allen C. HoH-Index: 44
#2Ingrid U. ScottH-Index: 96
Last. Franco M. RecchiaH-Index: 21
view all 7 authors...
Objective To review the evidence regarding the safety and efficacy of current anti–vascular endothelial growth factor (VEGF) pharmacotherapies for the treatment of diabetic macular edema (DME). Methods Literature searches last were conducted in September 2011, in PubMed with no date restrictions, limited to articles published in English, and in the Cochrane Library without a language limitation. The combined searches yielded 532 citations, of which 45 were deemed clinically relevant for the auth...
73 CitationsSource
#1Felix TreumerH-Index: 14
#2Alexa KlettnerH-Index: 22
Last. Jost HillenkampH-Index: 28
view all 8 authors...
Abstract The purpose of this study was to investigate release of matrix metalloproteinases (MMP) 2 and 9 during retinal pigment epithelium (RPE) wound healing after Selective Retina Therapy (SRT) with laser energy levels below and above the threshold of RPE cell death. Following exposure to SRT using a prototype pulsed Nd:YLF laser with energies of 80–180 mJ/cm 2 fresh porcine RPE-monolayers with Bruch's membrane and choroid were cultured in modified Ussing chambers which separate the apical (RP...
33 CitationsSource
#1Quan Dong Nguyen (JHUSOM: Johns Hopkins University School of Medicine)H-Index: 7
#2David M. Brown (Houston Methodist Hospital)H-Index: 42
Last. Jason S. Ehrlich (Genentech)H-Index: 17
view all 12 authors...
Purpose To evaluate the efficacy and safety of intravitreal ranibizumab in diabetic macular edema (DME) patients. Design Two parallel, methodologically identical, phase III, multicenter, double-masked, sham injection–controlled, randomized studies. Participants Adults with vision loss from DME (best-corrected visual acuity [BCVA], 20/40–20/320 Snellen equivalent) and central subfield thickness ≥275 μm on time-domain optical coherence tomography (OCT). Intervention Monthly intravitreal ranibizuma...
1,025 CitationsSource
#1Philip J. Rosenfeld (UM: University of Miami)H-Index: 84
#1Philip J. Rosenfeld (UM: University of Miami)H-Index: 1
Last. Barbara A. Blodi (UW: University of Wisconsin-Madison)H-Index: 43
view all 6 authors...
Purpose To investigate the cause of visual acuity (VA) loss in patients with neovascular age-related macular degeneration (AMD) receiving monthly ranibizumab injections in the pivotal ranibizumab phase III trials. Design Retrospective analysis. Participants The Minimally Classic/Occult Trial of the Anti-VEGF Antibody Ranibizumab In the treatment of Neovascular AMD (MARINA) and Anti-VEGF Antibody for the Treatment of Predominantly Classic Choroidal Neovascularization in AMD (ANCHOR) trials. Metho...
204 CitationsSource
Cited By8
#1Zhongyi Yang (NCU: Nanchang University)H-Index: 1
#2Hanying Hu (NCU: Nanchang University)H-Index: 1
Last. Zhi-peng You (NCU: Nanchang University)H-Index: 2
view all 6 authors...
BACKGROUND: Diabetic retinopathy (DR) is a common complication of diabetes. This study investigated the effect of miR-7 on the regulation of cell proliferation via the HoxB3 gene and PI3K/AKT/mTOR signaling pathways in DR. Methods:Human retinal pigment epithelial cell line (ARPE-19) cultured in normal medium (Control) and high glucose medium (25mM glucose, HG) was transfected with mimics NC (HG+ mimics NC), miR-7 mimics (HG+miR-7 mimics), inhibitor NC (HG+ inhibitor NC), and miR-inhibitor (HG+mi...
2 CitationsSource
#1Bobak Bahrami (USYD: University of Sydney)H-Index: 7
#2Weiyong Shen (USYD: University of Sydney)H-Index: 26
Last. Mark C Gillies (USYD: University of Sydney)H-Index: 58
view all 6 authors...
BACKGROUND: Retinal pigment epithelium (RPE) is known to secrete factors important for retinal homeostasis. How this secretome changes in diabetic eyes treated with anti-vascular endothelial growth factor (VEGF) inhibitors is unclear. METHODS: Diabetic conditions were simulated in vitro using ARPE-19 cell-line culture, with high glucose (25 mM) culture media, and hypoxia was chemically induced using cobalt chloride. Stress was assessed using cell viability assays as well as Western blots and enz...
9 CitationsSource
#1Sanjar Batirovich Madrakhimov (Soonchunhyang University Hospital)H-Index: 2
#2Jin Young Yang (Soonchunhyang University Hospital)H-Index: 3
Last. Tae Kwann Park (SCH: Soonchunhyang University)H-Index: 13
view all 4 authors...
This study assessed the role of mechanistic target of rapamycin (mTOR) pathway in the human adult retinal pigment epithelial (ARPE) cell response after laser photocoagulation (LP). The effect of mTOR inhibition on ARPE-19 cell was investigated by rapamycin treatment after LP. Cell viability and proliferation were explored using MTT and EdU assays, respectively. The expression of mTOR-related proteins and epithelial-mesenchymal transition (EMT) markers was verified by Western blot. Rapamycin reta...
2 CitationsSource
: Objective: To evaluate short term changes in anterior chamber depth (ACD), central corneal thickness (CCT), intraocular pressure (IOP), and corneal endothelial cell density (ECD) after intravitreal anti-vascular endothelial growth factor (VEGF) injections. Materials and methods: A total of 100 eyes from 100 patients, who were naive for intravitreal injection treatment, were included in this retrospective study. Patients who received only a single type of intravitreal anti-VEGF injections (beva...
4 CitationsSource
#1Patrick Merz (Heidelberg University)H-Index: 5
#2Nina Röckel (Heidelberg University)H-Index: 1
Last. I. Schmack (Goethe University Frankfurt)H-Index: 5
view all 5 authors...
Background Ingrowth of newly formed blood and lymph vessels (angiogenesis) from the limbus region into the cornea can be treated successfully by subconjunctival application of antiangiogenic agents. Currently, there are several angiogenesis inhibitors from various manufacturers available, such as vascular endothelial growth factor (VEGF) antibodies. The aim of the study was to investigate potential cytotoxic effects of two anti-VEGF agents, ranibizumab (Lucentis®) and bevacizumab (Avastin®) on t...
5 CitationsSource
#1Magda Gharbiya (Sapienza University of Rome)H-Index: 5
#1Magda Gharbiya (Sapienza University of Rome)H-Index: 6
Last. Alessandro Lambiase (Sapienza University of Rome)H-Index: 59
view all 9 authors...
: Intravitreal injection (IVT) of antivascular endothelial growth factor (anti-VEGF) agents is widely used for the treatment of retinal vascular diseases. Recently, the injection of anti-VEGF agents in the ocular anterior chamber has been proposed for the treatment of neovascular glaucoma and potential side effects on the corneal structures have been investigated with contrasting results. Increasing evidence has demonstrated that VEGF inhibition is associated with cellular apoptotic changes and ...
5 CitationsSource
#1Xu LuoH-Index: 1
#2Shengjie GuH-Index: 1
Last. Jianhong ZhangH-Index: 1
view all 4 authors...
The pathological superoxidative condition that retinal pigment epithelium (RPE) cells experience contributed to the advancement of age-related macular degeneration (AMD), which was accompanied by significant neovascularization. Therefore, the discovery of novel pharmacological candidates to ameliorate oxidative damage (H2O2) against RPE cells and inhibit the following angiogenesis simultaneously is urgently needed. Herein, we found that kinsenoside (Kin), an active component derived from Anoecto...
13 CitationsSource
#1De Cillà SH-Index: 1
#1Stefano De CillàH-Index: 14
Last. Elena GrossiniH-Index: 23
view all 12 authors...
Background/Aims: the anti-vascular endothelial growth factors (VEGF), Aflibercept and Ranibizumab, are used for the treatment of macular degeneration. Here we exa
11 CitationsSource