Exploration of Medicine
The potential role of high mobility group box 1 (HMGB-1) protein in the pathogenesis of keratoconus: a pilot immunofluorescence study
By Prem Chudgar, Sevnur Kömürlü Keceli, CHANDANA M. SOORANAHALLI and 3 others.
Abstract
Aim
This preliminary study aims to evaluate the expression of high mobility group box 1 (HMGB-1) in keratoconic corneas, normal corneas, and corneas from donors with Fuchs’ corneal dystrophy using immunohistochemistry.
Methods
Under a Loyola University Institutional Review Board (IRB) approved protocol (LU#219768), archived paraffin-embedded corneal tissues were obtained from the Loyola University Medical Center Department of Pathology. The study included three keratoconic corneas, three normal donor corneas, and three Fuchs’ dystrophy corneas. Tissue sections were deparaffinized in 100% xylene, rehydrated through a graded ethanol series, and subjected to antigen retrieval. Sections were stained with an HMGB-1 primary antibody, followed by a fluorescently conjugated anti-rabbit IgG secondary antibody and 4',6-diamidino-2-phenylindole (DAPI) nuclear counterstain. Immunofluorescence imaging and deconvolution analysis were performed using a Nikon Eclipse Ti2 microscope equipped with a digital camera. Two prespecified comparisons, keratoconus versus normal and Fuchs’ dystrophy versus normal, were evaluated using Welch’s independent-samples t tests with Holm adjustment for multiple testing. Statistical analyses were performed using IBM SPSS Statistics.