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Cellular and Molecular Bioengineering

Cellular and Molecular Bioengineering - Poster Session A

Poster H18 - Analysis of Scleral Macrophage Markers in a Mouse Glaucoma Model

Thursday, October 24, 2024

10:00 AM - 11:00 AM EST

Location: Exhibit Hall E, F & G

Presenting Author(s)

Maru Ando

Undergraduate researcher
Johns Hopkins University
Baltimore, Maryland, United States

Co-Author(s)

SC

Shiraj Crowdhury

Undergraduate researcher
Johns Hopkins University, United States
AM

Annie Mozzer

Research Technologist
Johns Hopkins University Center for Nanomedicine, United States
JJ

Joe Gerald Jesu Raj

Research Technologist
Johns Hopkins University Center for Nanomedicine, United States

Primary Investigator(s)

PI

Pitha Ian

Associate Professor
Department of Ophthalmology, Johns Hopkins Medicine, United States

Introduction: The sclera translates mechanical strain to the optic nerve head and scleral remodeling is a hallmark of glaucoma pathogenesis. Previous studies have shown that mouse and human sclera contain macrophage populations and selective macrophage depletion reduced scleral remodeling in myopia. Here we investigate scleral macrophage markers in a mouse glaucoma model.

Materials and

Methods: Ocular hypertension (OHT) was induced in C57BL/6J mice (3 months) by anterior chamber bead injection (BI) and intraocular pressure (IOP) was monitored serially in OHT and control eyes by rebound tonometry (TonoLab, TioLat, Inc., Helsinki, Finland) at 1, 3, 7, 14, and 21 days after injection. Glaucomatous eyes were only included if IOP elevation was >7 mmHg compared to fellow eye control at 1, 3, or 7 days after BI. Mice were euthanized, eyes enucleated and cryopreserved at 3, 7, and 28 days after BI. Scleral eye cups were dissected and cross-sections from the posterior to segments of the eye were processed for immunolabelling with macrophage markers (CD68, CD11b, and LYVE-1) were used to characterize macrophage populations within the peripapillary sclera (PPS) and scleral cross-sections of the mouse eye.

Results, Conclusions, and Discussions: CD68, CD11b, CD163, CD38, and LYVE-1 positive cells were detected in the PPS and peripheral sclera of non-glaucomatous mice. CD68, CD11b, and LYVE-1 positive cells were seen in the optic nerve, peripapillary sclera, and concentrated in the anterior and posterior sclera in the outer sclera layers and episcleral in non-glaucomatous mice. Following BI OHT for 3 days, 1 week, and 6 weeks, there was an observable increase in strongly positive CD68, CD11b, and LYVE-1 cells in PPS and peripheral sclera. Quantitative increase in CD68 and CD11b was also determined in three-day and six-week BI OHT eyes. This study shows multiple macrophage markers are present in peripheral and peripapillary region of sclera with an increase in three macrophage markers 3 and 28 days after IOP elevation. These results provide the basis for future studies to understand the role of scleral macrophages in glaucoma.

Acknowledgements (Optional): This work is supported by the Masason Foundation.