Evaluation of ocular surface glycocalyx using lectin-conjugated fluorescein

Abstract

Hiroshi Mochizuki1, Masaki Fukui1,2, Shin Hatou1,2, Masakazu Yamada1, Kazuo Tsubota21Division for Vision Research, National Institute of Sensory Organs, National Tokyo Medical Center, Tokyo, Japan; 2Department of Ophthalmology, Keio University School of Medicine, Tokyo, JapanPurpose: A glycocalyx plays important roles in the ocular surface epithelium, but there is no direct simple method to evaluate ocular surface glycocalyx. We tested a wheat germ agglutinin conjugate of fluorescein (F-WGA) as a marker to demonstrate ocular surface glycocalyx in vivo.Methods: After a 5% F-WGA solution was applied to the eyes of eight healthy volunteers, fluorescent intensities of the central cornea and bulbar conjunctiva were measured by fluorophotometry. A 10% fluorescein-conjugated dextran solution served as the control. Changes in fluorescent intensities of the ocular surface following a challenge with 5% N-acetyl cysteine, a conventional mucolytic agent, were tested in the second experiment. Saline instillation served as a control.Results: The ocular surface was diffusely stained by F-WGA. Breakup of the precorneal tear film was not apparent, possibly because F-WGA was bound to the glycocalyx of the ocular surface epithelium. F-WGA fluorescent intensities were high in the superior, nasal, and inferior regions of the bulbar conjunctiva and low in the temporal conjunctiva and cornea. No such regional differences were observed with fluorescein-conjugated dextran. F-WGA fluorescent intensities decreased significantly with N-acetyl cysteine instillation, whereas they were not affected by saline instillation.Conclusion: The fluorophotometric method may be used to evaluate the glycocalyx quantitatively in the ocular surface in vivo.Keywords: cornea, fluorescein, glycocalyx, lectin, mucin