June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Qualitative asymmetric mapping of lysozyme deposited on various contact lens materials using confocal laser scanning microscopy
Author Affiliations & Notes
  • Jaya Dantam
    Centre for Contact Lens Research, School of Optometry & Vision Science, Waterloo, ON, Canada
  • Miriam Heynen
    Centre for Contact Lens Research, School of Optometry & Vision Science, Waterloo, ON, Canada
  • Claudia Yvette Dominici
    Centre for Contact Lens Research, School of Optometry & Vision Science, Waterloo, ON, Canada
  • Lakshman N Subbaraman
    Centre for Contact Lens Research, School of Optometry & Vision Science, Waterloo, ON, Canada
  • Chantal Coles-Brennan
    Johnson & Johnson Vision Care, Inc., Jacksonville, FL
  • Zohra Fadli
    Johnson & Johnson Vision Care, Inc., Jacksonville, FL
  • Lyndon William Jones
    Centre for Contact Lens Research, School of Optometry & Vision Science, Waterloo, ON, Canada
  • Footnotes
    Commercial Relationships Jaya Dantam, None; Miriam Heynen, None; Claudia Dominici, None; Lakshman Subbaraman, Johnson & Johnson Vision Care, Inc. (F); Chantal Coles-Brennan, Johnson & Johnson Vision Care, Inc. (E); Zohra Fadli, Johnson & Johnson Vision Care, Inc. (E); Lyndon Jones, Johnson & Johnson Vision Care, Inc. (F)
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 6091. doi:
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      Jaya Dantam, Miriam Heynen, Claudia Yvette Dominici, Lakshman N Subbaraman, Chantal Coles-Brennan, Zohra Fadli, Lyndon William Jones; Qualitative asymmetric mapping of lysozyme deposited on various contact lens materials using confocal laser scanning microscopy . Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):6091.

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

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Abstract

Purpose: Previous studies have demonstrated the uptake and distribution of lysozyme in hydrogel contact lens (CL) materials using the traditional vial incubation method. The purpose of this study was to qualitatively assess the asymmetric uptake of lysozyme by different CL materials using a confocal laser scanning microscopy (CLSM) technique.

Methods: Six hydrogel CL materials belonging to various FDA groups (group I: polymacon; group II: omafilcon A, nesofilcon A, nelfilcon A; group IV: etafilcon A with Polyvinylpyrrolidone, ocufilcon B) were incubated in an artificial tear solution comprising fluorescently-labelled lysozyme for 16 h at 37°C. Appropriate controls were included. During the incubation, a novel asymmetric condition of lens exposure to ATS was simulated by blocking the posterior surface of the CL with a custom made Teflon® mount. After incubation, the central 5 mm of each CL was punched out and mounted on a microscope slide. The uptake of lysozyme by different CL materials was evaluated using CLSM with argon laser at 488nm.CLs were optically sectioned every 2 µm intervals (z stack). The fluorescence intensity profile of each CL sample was calculated with ImageJ software, using the “Plot Z axis profile” module. The data were normalized to plot the relative fluorescence for each CL profile.

Results: Mapping of asymmetric lysozyme distribution revealed different patterns of lysozyme penetration into the CLs. All CLs demonstrated the uptake of fluorescently-labelled lysozyme. Polymacon (FDA group I) revealed a relatively higher concentration of lysozyme within the bulk, while the group II materials showed a homogeneous distribution of lysozyme from the anterior to posterior surface. For the group IV materials, lysozyme was deposited at the surface and bulk in a gradient pattern, with decreasing deposition towards the posterior surface.

Conclusions: A novel model to demonstrate asymmetric uptake of lysozyme by different CL materials has been developed. The location of lysozyme in the bulk and on the surface varied across different FDA CL groups. It is well established that etafilcon A CLs exhibit high levels of lysozyme, and this study demonstrates that it is present throughout the material and most of it is located in the bulk.

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