June 2017
Volume 58, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2017
Metabolic activity of human corneal epithelial cells after exposure to artificial tear-like formulations with varying pH and osmolalities
Author Affiliations & Notes
  • Lakshman N Subbaraman
    CCLR, School of Optometry & Vision Science, University of Waterloo, Waterloo, Ontario, Canada
  • Yun Ha (Sammie) Hwang
    CCLR, School of Optometry & Vision Science, University of Waterloo, Waterloo, Ontario, Canada
  • Lucy Liu
    CCLR, School of Optometry & Vision Science, University of Waterloo, Waterloo, Ontario, Canada
  • David McCanna
    CCLR, School of Optometry & Vision Science, University of Waterloo, Waterloo, Ontario, Canada
  • Lyndon William Jones
    CCLR, School of Optometry & Vision Science, University of Waterloo, Waterloo, Ontario, Canada
  • Footnotes
    Commercial Relationships   Lakshman Subbaraman, None; Yun Ha (Sammie) Hwang, None; Lucy Liu, None; David McCanna, None; Lyndon Jones, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2017, Vol.58, 3936. doi:
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      Lakshman N Subbaraman, Yun Ha (Sammie) Hwang, Lucy Liu, David McCanna, Lyndon William Jones; Metabolic activity of human corneal epithelial cells after exposure to artificial tear-like formulations with varying pH and osmolalities. Invest. Ophthalmol. Vis. Sci. 2017;58(8):3936.

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

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Abstract

Purpose : Symptoms of ocular discomfort can be caused due to the disruption of optimal pH and osmolality levels in the tear film. The objective of this study was to assess the metabolic activity of human corneal epithelial cells (HCEC) after exposure to artificial tear solutions (ATS) with varying pH and osmolality levels using an in vitro model.

Methods : Primary and immortalized HCEC were exposed to an ATS composed of proteins and lipids at concentrations found in the natural tears. Various ATS were prepared that differed in pH levels (ranging between 1 and 12) and osmolality (levels ranging from 100 to 1000 mmol/kg). HCEC were then exposed to ATS of varying pH and osmolalities for three hours at 37 degrees C and 5% CO2. Subsequently, the ATS was removed and replaced with 1mL of a solution composed of growth media and 10% alamarBlue. After incubation for 4 hours at 37 degrees C and 5% CO2, the change in fluorescence was determined using a microplate reader set at 530/590 excitation/emission. Statistical significance of the difference between treatment groups (three replicates were used for each treatment) was determined using an analysis of variance. Pairwise multiple comparison procedures were performed using the Bonferroni post-hoc test, with significance set at 0.05.

Results : Primary and immortalized HCEC were metabolically active between pH ranges 5-7. Cells exposed to extremes of pH outside this range were significantly less metabolically active (p<0.05). Exposure to ATS with osmotic levels >500 mmol/kg caused significant reductions in the metabolic activity of immortalized HCEC (p<0.05) when compared to the control (300 mmol/kg).

Conclusions : In conditions such as dry eye there may be an excessive evaporation of water from tears, causing a hypertonic environment for the cells. Furthermore, some dry eye patients may have an elevated pH in their tears. This investigation showed the tolerability of HCEC when exposed to ATS of varying pH and osmolalities using the alamarBlue metabolic activity as an indicator of cell health.

This is an abstract that was submitted for the 2017 ARVO Annual Meeting, held in Baltimore, MD, May 7-11, 2017.

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