September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Effects of anti-VEGF antibodies to the viability, proliferation and wound healing activity of cultured human retinal pigment epithelial cells under high glucose stress
Author Affiliations & Notes
  • Tae Kwann Park
    Ophthalmology, Soonchunhyang Univ Hospital, Bucheon-si, Korea (the Republic of)
  • Jong Rok Oh
    Ophthalmology, Soonchunhyang Univ Hospital, Bucheon-si, Korea (the Republic of)
  • Young-Hoon Ohn
    Ophthalmology, Soonchunhyang Univ Hospital, Bucheon-si, Korea (the Republic of)
  • Footnotes
    Commercial Relationships   Tae Kwann Park, None; Jong Rok Oh, None; Young-Hoon Ohn, None
  • Footnotes
    Support  This work was supported by grants from Basic Science Research Programthrough the National Research Foundation of Korea (NRF) funded by the ministry of Education, Science, and Technology (grant number 2013R1A1A2009899; Seoul, South Korea).
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 2078. doi:
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    • Get Citation

      Tae Kwann Park, Jong Rok Oh, Young-Hoon Ohn; Effects of anti-VEGF antibodies to the viability, proliferation and wound healing activity of cultured human retinal pigment epithelial cells under high glucose stress. Invest. Ophthalmol. Vis. Sci. 2016;57(12):2078.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : To investigate the changes of RPE cell viability, proliferative potential, and wound healing activity at high glucose condition and evaluate the effects of anti-VEGF antibodies to the RPE cells under high glucose level.

Methods : ARPE-19 cells were cultured in DMEM/Ham’s F-12 (1:1) at different glucose level (5, 25, and 75 mM). Viability was evaluated in methyl thiazolyl tetrazolium (MTT) assay at 5 days after treatment with each glucose concentration. Migration ability was measured in a wound healing assay at 3 days. Cell Death Detection Kit was used to assess apoptosis at 3 days and 14days. And proliferative potential was assessed by EdU Imaging kit at 3days. Cultured cells were treated with anti-VEGF antibodies at clinically relevant concentrations (bevacizumab 250 µg/mL, ranibizumab 125 µg/mL, aflibercept 500 µg/mL). Then, experiment was repeated at different glucose level. Significance was evaluated with an unpaired, two-tailed Student t test. A P-value < 0.05 was considered to indicate significance.

Results : The viability and migration of ARPE-19 cells were significantly decreased in high glucose levels. (Viability; 87.6±2.6% at75mM with 5mM set as 100% p<0.05, Migration; 24.5±1.3 % at 5mM, 25.6±1.2 % at25mM, 13.7±1.3% at 75mM, p<0.05). At 14 days, the percentage of TUNEL positive cell was significantly increased in 75mM glucose level compared to 5mM (75mM :152.3±18.8% with 25mM set as 100%). Percentage of EdU positive cell was also significantly decreasd (75mM : 35.8±10.5 % with 5mM set as 100%, p<0.05). In 75mM glucose level, the groups treated with Anti-VEGF, especially showed to decrease of cell viability, proliferation and increase of apoptosis (p<0.05). However, there was no significant difference generally between the anti-VEGF groups.

Conclusions : High glucose concentration interfered RPE cell viability, wound healing activity, and proliferation. Further, anti-VEGF antibodies aggravated those functions of RPE cells on high glucose level. It may be consider that anti-VEGF long-term use in diabetic retinopathy patients can give a negative effect on RPE function.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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