June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
In Vitro Evaluation of REMOGEN® OMEGA for HLA-DRA and MMP-9 Modulation and Cytotoxicity to Human Ocular Surface Cells
Author Affiliations & Notes
  • Eric G Romanowski
    The Charles T. Campbell Ophthalmic Microbiology Laboratory, Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, United States
  • Nicholas A Stella
    The Charles T. Campbell Ophthalmic Microbiology Laboratory, Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, United States
  • Robert M Q Shanks
    The Charles T. Campbell Ophthalmic Microbiology Laboratory, Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, United States
  • Footnotes
    Commercial Relationships   Eric Romanowski TRB Chemedica, Code F (Financial Support); Nicholas Stella TRB Chemedica, Code F (Financial Support); Robert Shanks TRB Chemedica, Code F (Financial Support)
  • Footnotes
    Support  TRB Chemedica; NIH Core Grant EY08098, The Eye & Ear Foundation of Pittsburgh; RPB
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 956 – A0425. doi:
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    • Get Citation

      Eric G Romanowski, Nicholas A Stella, Robert M Q Shanks; In Vitro Evaluation of REMOGEN® OMEGA for HLA-DRA and MMP-9 Modulation and Cytotoxicity to Human Ocular Surface Cells. Invest. Ophthalmol. Vis. Sci. 2022;63(7):956 – A0425.

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

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Abstract

Purpose : REMOGEN® OMEGA (REM) (TRB Chemedica) is a topical, preservative-free emulsion of essential fatty acids (eicosapentaenoic acid and docosahexanoic acid), vitamin E, glycerol, polyacrylic acid, and polymers that is used for treating symptoms and signs of dry eye and/or ocular surface damage, due to diseases such as superficial keratitis, Sjögren syndrome or primary dry eye syndrome. The goals of the current study were to determine whether pretreatment with REM affects the production of HLA Class II histocompatibility antigen (HLA-DRA) and matrix metalloproteinase 9 (MMP-9) by human corneal limbal epithelial (HCLE) cells after stimulation with lipopolysaccharide (LPS) as well as the overall cytotoxicity to HCLE cells. HLA-DRA and MMP-9 have been implicated as biomarkers of ocular inflammation in dry eye disease.

Methods : For the cytotoxicity assays, HCLE cells were incubated for 6 h with a 2-fold dilution series of REM (50%-0.1953%) or a mock solution. Cells were assessed for cytotoxicity using Presto Blue (Invitrogen). For the ELISA assays, HCLE cells were pretreated for 24 h with 50% REM or a mock solution, washed, then treated with or without LPS (10 mg/ml). ELISA assays for HLA-DRA (MyBiosource.com) and MMP-9 (Life Technologies-ThermoFisher) were carried out using the manufacturers’ instructions. Statistical analysis (ANOVA with Dunnett’s and Tukey’s Post-test) was performed with GraphPad Prism.

Results : No significant differences in cytotoxicity were demonstrated among the mock treated and the REM concentrations at 6 h (n=4 for each concentration, p>0.05). REM pretreatment of HCLE cells stimulated with LPS significantly reduced MMP-9 production (85.1%; n=4, p<0.01) and HLA-DRA production (52.5%; n=5, p<0.001) compared to mock-pretreated cells.

Conclusions : After 24 h of pretreatment, REM significantly reduced MMP-9 and HLA-DRA production by the ocular surface HCLE cell line. Despite a high concentration up to 50% and extended exposure, there was no statistically significant loss of ocular surface cell viability caused by REM. REM appears to be nontoxic and anti-inflammatory to HCLE cells in vitro.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

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