Purchase this article with an account.
J. S. Ellis, D. J. Paull, A. Khalili, S. Dhingra, A. Kato, S. Brocchini, P. T. Khaw; Inflammatory Cell and Fibroblast Model to Investigate Candidate Anti-Scarring Therapies for Ocular Disease. Invest. Ophthalmol. Vis. Sci. 2009;50(13):5929.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Failure of glaucoma and retinal surgery is often associated with scar tissue formation mediated by an inflammatory response. Various cytokines and growth factors are believed to be involved. Microglia are a major source of inflammatory cytokines in the retina, as such they provide an ideal model of inflammation. Here we describe a preliminary model of inflammation to assess both anti-inflammatory and anti-scaring medicines and their delivery systems.
Microglia were activated with 1 µg ml-1 lipopolysacharide (LPS) for 17 hours to create inflammatory conditioned media (CM). RT-PCR was used to confirm microglial activation and change in mRNA expression of inflammatory mediators. Gel contraction assays using Human Tenon Fibroblasts (HTFs) were used as a model of wound healing to investigate the effect of inflammatory cytokine rich CM on the degree of gel contraction over 72 hours.
Pretreatment of microglia with LPS resulted in significant increased mRNA expression of TNF (56%), MMP-9 (93 %), IL-6 (not expressed in control) and IL-1β (60 %), with a decrease in TGFβ2 (48%), confirming the activation of microglia. Preliminary data using gel contraction assays with HTFs and CM showed significantly more contraction (P<0.002, 47 % greater than control) in the first 24 hours of contraction, the difference in contraction after 72 hours was insignificant.
Microglia conditioned media provides a means to investigate the effect of simulated inflammation on a variety of cell lines and to gain a clearer understanding of inflammatory processes in the eye. This model will continue to be developed with the aim of testing candidate anti-scarring molecules.
This PDF is available to Subscribers Only