May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Anti-VEGF Treatment on Human Tenons’ Fibroblasts Scarring Activity in vitro
Author Affiliations & Notes
  • Q. Qin
    Glaucoma Investigations Research Unit, Centre for Eye Research Australia, Melbourne, Australia
    School of Medicine, Monash University, Clayton, Australia
  • N. Van Bergen
    Glaucoma Investigations Research Unit, Centre for Eye Research Australia, Melbourne, Australia
  • D. Liew
    Department of Medicine, St Vincent's Hospital, Melbourne, Australia
  • P. Van Wijngaarden
    Glaucoma Investigations Research Unit, Centre for Eye Research Australia, Melbourne, Australia
    Royal Victorian Eye and Ear Hospital, Melbourne, Australia
  • A. P. Wells
    Department of Ophthalmology, Wellington Hospital, Wellington, New Zealand
  • J. G. Crowston
    Glaucoma Investigations Research Unit, Centre for Eye Research Australia, Melbourne, Australia
  • Footnotes
    Commercial Relationships  Q. Qin, None; N. Van Bergen, None; D. Liew, None; P. Van Wijngaarden, None; A.P. Wells, None; J.G. Crowston, None.
  • Footnotes
    Support  ORIA, Glaucoma Australia, RVEEH
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 4541. doi:https://doi.org/
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    • Get Citation

      Q. Qin, N. Van Bergen, D. Liew, P. Van Wijngaarden, A. P. Wells, J. G. Crowston; Anti-VEGF Treatment on Human Tenons’ Fibroblasts Scarring Activity in vitro. Invest. Ophthalmol. Vis. Sci. 2008;49(13):4541. doi: https://doi.org/.

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

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Abstract

Purpose: : To assess the effects of Anti-VEGF bevacizumab (Avastin) on Human Tenons’ Fibroblast (HTF) healing activity in-vitro.

Methods: : Fibroblasts were routinely cultured in RPMI media and bevacizumab was administered at a concentration ranging from 0.025 to 12.5mg/ml. Fibroblast viability and cell death was assessed using MTT colorimetric assay, Lactate Dehydrogenase assay and live/dead assays. Fibroblast contractility was assessed using floating collagen gel assay. In addition, the mechanism of HTF cell death (post bevacizumab administration) was explored via Hoechst staining and Scanning Electron Microscopy (SEM).

Results: : Bevacizumab induced a dose dependent reduction of HTF cell numbers, (83% reduction SD +/-6%) at a concentration of 12.5mg/mL at 72 hours. The administration of bevacizumab caused a moderate inhibition (30% reduction SD +/-8%) of fibroblast gel contraction from baseline. Under serum-free conditions, bevacizumab induced fibroblast cell death at concentrations greater than 7.5mg/ml. Scanning Electron Microscopy revealed marked vacuolization in bevacizumab treated fibroblasts suggestive of cell death via autophagy.

Conclusions: : Bevacizumab (Avastin) disrupts fibroblast proliferation, inhibits contraction ability of fibroblasts and induces cell death suggestive of autophagy. Our study findings suggest anti-VEGF has a direct effect on HTF activity and viability in culture. This supports a possible role for bevacizumab in inhibiting post-operative scarring after glaucoma filtration surgery.

Keywords: wound healing • vascular endothelial growth factor • apoptosis/cell death 
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