July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Effects of Intravitreal Connective Tissue Growth Factor Neutralizing Antibody on the Epiretinal Membrane Formation; an Experimental Study
Author Affiliations & Notes
  • Narsis Daftarian
    Ocular Tissue Engineering Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Tehran, Iran (the Islamic Republic of)
    Ophthalmic Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Tehran, Iran (the Islamic Republic of)
  • Omolbanin Bayeghi
    Ophthalmic Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Tehran, Iran (the Islamic Republic of)
    Ocular Tissue Engineering Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Tehran, Iran (the Islamic Republic of)
  • Mozhgan Rezaei Kanavi
    Ocular Tissue Engineering Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Tehran, Iran (the Islamic Republic of)
    Ophthalmic Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Tehran, Iran (the Islamic Republic of)
  • Hamid Ahmadieh
    Ophthalmic Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Tehran, Iran (the Islamic Republic of)
    Ocular Tissue Engineering Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Tehran, Iran (the Islamic Republic of)
  • Footnotes
    Commercial Relationships   Narsis Daftarian, None; Omolbanin Bayeghi, None; Mozhgan Rezaei Kanavi, None; Hamid Ahmadieh, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 5812. doi:
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      Narsis Daftarian, Omolbanin Bayeghi, Mozhgan Rezaei Kanavi, Hamid Ahmadieh; Effects of Intravitreal Connective Tissue Growth Factor Neutralizing Antibody on the Epiretinal Membrane Formation; an Experimental Study. Invest. Ophthalmol. Vis. Sci. 2019;60(9):5812.

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

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Abstract

Purpose : Retinal pigment epithelial (RPE) cells have been suggested to play an important role in epiretinal membrane (ERM) formation. Connective tissue growth factor (CTGF) as a downstream mediator of TGF-β, released by RPE cells has been found in ERMs. We compared the quality and quantity of ERM formation two weeks after the single intravitreal injection of a safe dose of anti-CTGF antibody alone, anti-VEGF (bevacizumab) alone, and combination of anti-CTGF/ bevacizumab in a rabbit model of RPE cell induced ERM formation.

Methods : With adherence to the ARVO regulations, intravitreal injection of 0.3 ml SF6 was performed in the right eyes of 24 adult male albino rabbits to induce posterior vitreous detachment. Three days later, human RPE cells obtained from the eye bank, cultured by 3 passages were injected into the vitreous cavity over the optic disc (250,000 in 0.1 ml). Rabbits were categorized into four groups that received single intravitreal injection of 0.05 ml of 50µg/ml humanized anti-CTGF ( group A), 0.05 ml of 25mg/ml bevacizumab (group B), and 0.05 ml anti-CTGF plus 0.05 ml bevacizumab ( group C). No antibody injection was done in the control group (group D). After 2 weeks, animals were sacrificed and paraffin embedded anterior-posterior sections were prepared from the right globes. Masson-trichrome for collagen and immunostaining by humanized anti-alpha-smooth muscle for fibroblasts differentiated from RPE cells were performed. Masked measurements of epiretinal membrane thickness and fibroblast counts on 20x light and fluorescent images (respectively) of the epiretinal membranes were performed by using the ImageJ software.

Results : Means of ERM thicknesses and fibroblast cell counts in ERMs in three treatment groups (A, B, C) were significantly less than the control group (P<0.001, <0.05, <0.05, respectively). Means of fibroblast counts for groups A and C were also significantly less than group B (P<0.05).

Conclusions : Intravitreal injection of anti-CTGF antibody significantly reduced the RPE induced ERM formation via inhibiting CTGF as the mediator of the final steps of fibrosis in various healing and restorative pathways.

This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.

 

Masson trichrome staining. Top to bottom: Groups A, B, C, D. Left to right: 4x and 10x magnification.

Masson trichrome staining. Top to bottom: Groups A, B, C, D. Left to right: 4x and 10x magnification.

 

IHC for anti-alpha-SMA. Top to bottom: Groups A, B, C, D. Left to right: Red alpha-SMA, dapi, merge.

IHC for anti-alpha-SMA. Top to bottom: Groups A, B, C, D. Left to right: Red alpha-SMA, dapi, merge.

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