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(Investigative Ophthalmology and Visual Science. 2003;44:4796-4804.)
© 2003 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.02-1083
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Effect of Heparin II Domain of Fibronectin on Aqueous Outflow in Cultured Anterior Segments of Human Eyes

Amy J. Santas,1,2 Cindy Bahler,2,3 Jennifer A. Peterson,1 Mark S. Filla,1,4 Paul L. Kaufman,4 Ernst R. Tamm,5 Douglas H. Johnson,3 and Donna M. Pesciotta Peters1,4

1From the Departments of Pathology and Laboratory Medicine and 4Ophthalmology and Visual Sciences, University of Wisconsin, Madison, Wisconsin; the 3Department of Ophthalmology, Mayo Clinic, Rochester, Minnesota; and the 5Department of Anatomy, University Erlangen-Nürnberg, Germany.

PURPOSE. To determine whether an integrin/syndecan-binding domain of fibronectin, called the heparin II (Hep II) domain, affects outflow facility in the human eye.

METHODS. Anterior segments of human eyes were placed in perfusion organ culture. One eye of each pair received the Hep II domain, and the fellow eye received DMEM or a heat-denatured Hep II domain. The Hep II domain was produced as a recombinant glutathione S-transferase (GST)-fusion protein. Microscopic changes were assessed.

RESULTS. Outflow facility in anterior segments treated with Hep II domain increased by 93% compared with that in anterior segments treated with DMEM. In contrast, facility in anterior segments treated with the heat-denatured Hep II domain showed very little change. Outflow facility remained high during Hep II domain perfusion and returned to baseline after removal of the protein. Electron microscopy revealed disruptions in the endothelial lining of Schlemm’s canal in anterior segments fixed during maximum effect and in anterior segments after facility had returned to baseline. Scattered disruptions of canal cells were noted in control anterior segments. Trabecular cells in other regions looked normal. Major changes in the extracellular matrix of the juxtacanalicular tissue were not observed. Repeated doses of the Hep II domain administered after facility returned to baseline increased facility in two of three anterior segments.

CONCLUSIONS. The Hep II domain of fibronectin increases outflow facility in the human anterior segment. This suggests that fibronectin-mediated interactions may have a role in modulating aqueous hydrodynamics. Such interactions may represent avenues of novel therapeutic interventions for glaucoma.




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