May 2003
Volume 44, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2003
Purified Bovine Ocular Mucin Has Virtually No Surface Activity
P.J. Anderton; S.T. Tragoulias; M. Ball; F. Miano; G.R. Dennis; T.J. Millar
Author Affiliations & Notes
  • P.J. Anderton
    School of Optometry and Vision Science, UNSW, Cooperative Research Centre for Eye Research and Technology, Sydney, Australia
  • S.T. Tragoulias
    University of New South Wales, Cooperative Research Centre for Eye Research and Technology, Sydney, Australia
  • M. Ball
    University of New South Wales, Cooperative Research Centre for Eye Research and Technology, Sydney, Australia
  • F. Miano
    Biophysics R & D Department, SIFI SpA, Lavinaio, Italy
  • G.R. Dennis
    School of Science Food and Horticulture, UWS, Cooperative Research Centre for Eye Research and Technology, Sydney, Australia
  • T.J. Millar
    School of Science Food and Horticulture, UWS, Cooperative Research Centre for Eye Research and Technology, Sydney, Australia
  • Footnotes
    Commercial Relationships  P.J. Anderton, None; S.T. Tragoulias, None; M. Ball, None; F. Miano, None; G.R. Dennis, None; T.J. Millar, None.
  • Footnotes
    Support  Australian Government Co-operative Research Program, CRC for Eye Research and Technology
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 2502. doi:
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      P.J. Anderton, S.T. Tragoulias, M. Ball, F. Miano, G.R. Dennis, T.J. Millar; Purified Bovine Ocular Mucin Has Virtually No Surface Activity . Invest. Ophthalmol. Vis. Sci. 2003;44(13):2502.

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Abstract

Abstract: : Purpose: It is commonly believed that ocular mucins are surface active in tears. We have further investigated this idea by purifying bovine ocular mucin and comparing its surface activity with commercially available bovine submaxillary mucin (BSM), purified BSM, and hyaluronic acid. Methods: Ocular mucin was extracted from fresh bovine conjunctiva using 6M urea, 0.1% SDS, and 10mM DTT at 4°C for 16h. The extract and commercial BSM were purified using a Pharmacia Superose 6 column (16x600mm) in 50mM Na phosphate buffer containing 1M NaCl and 3mM DTT at pH7.5. It was desalted into water using a Pharmacia Fast Desalt column (10x100mm) and freeze dried. Purified and unpurified mucins were checked for purity using SDS-PAGE. The different mucins and hyaluronic acid were applied to an air/phosphate buffered saline interface in a Langmuir trough (maximum surface area 80cm2). The surface pressure was measured using a Wilhelmy balance as the film was compressed and expanded. Results: Ocular mucin and purified BSM had no detectable bands below 200kD. Commercial BSM had multiple bands with strongly stained bands at ~60kD, 20kD and 10kD. Bovine ocular mucin (10mg/mL, 50µL) and hyaluronic acid (1mg/mL, 45µL) had no detectable surface activity. Purified BSM (1mg/mL, 20-1000µL) gave negligible surface activity, but at a higher concentration (5mg/mL, 200µL) showed a maximum surface activity of 25mN.m–1. Commercial BSM gave strong surface activity even with low concentrations and low volumes (1mg/mL, 20µL). Conclusions: By examining the surface activity of mucin and hyaluronic acid, there is strong evidence to suggest that neither ocular mucin nor hyaluronic acid have surface activity, and hence any contribution to the surface activity of tears must come from their interaction with other tear film components. These differ from commercial BSM which had marked surface activity. This indicates that unpurified BSM cannot be used as a substitute for ocular mucin when modelling the tear film.

Keywords: cornea: surface mucins • cornea: tears/tear film/dry eye • protein structure/function 
© 2003, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Permission to republish any abstract or part of an abstract in any form must be obtained in writing from the ARVO Office prior to publication.
Copyright © 2025 Association for Research in Vision and Ophthalmology.
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