Analysis of 2-dimensional regional differences in the peripapillary scleral fibroblast cytoskeleton of normotensive and hypertensive mouse eyes

Abstract These studies aimed to study the mechanisms of glaucomatous peripapillary scleral (PPS) remodeling by investigating IOP-induced changes in fibroblast actin-collagen alignment and nuclear morphology in mouse PPS. Cryosections from the optic nerve heads (ONH) of eyes isolated 1- and 6-weeks a...

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Main Authors: Ann Mozzer, Joe Gerald Jesu Raj, Nushrat Yasmin, Edward Esposito, Ian Pitha
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-025-08251-4
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author Ann Mozzer
Joe Gerald Jesu Raj
Nushrat Yasmin
Edward Esposito
Ian Pitha
author_facet Ann Mozzer
Joe Gerald Jesu Raj
Nushrat Yasmin
Edward Esposito
Ian Pitha
author_sort Ann Mozzer
collection DOAJ
description Abstract These studies aimed to study the mechanisms of glaucomatous peripapillary scleral (PPS) remodeling by investigating IOP-induced changes in fibroblast actin-collagen alignment and nuclear morphology in mouse PPS. Cryosections from the optic nerve heads (ONH) of eyes isolated 1- and 6-weeks after bead-induced IOP elevation were imaged for nuclei, fibrillar actin (FA), and collagen (second harmonic generation, SHG). Two-dimensional nuclear morphology was analyzed using VAMPIRE machine-learning image analysis and FA-collagen alignment was determined by comparing vector fields of FA and SHG images. Nuclear morphology was regionally defined with the internal (pial) PPS (iPPS) containing nuclei with higher aspect ratios than the peripheral PPS (outer PPS, oPPS) and peripheral sclera. FA-collagen alignment was higher in the PPS than in the peripheral sclera (7.1 ± 2.5° versus 10.0 ± 1.4°, p = 0.05, n = 6). One and six weeks after BI, there were non-significant nuclear morphologic changes reflecting a transition to a rounder shape in all scleral regions and persistently reduced FA-collagen alignment in the PPS regions. This study therefore concludes that chronic IOP elevation is associated with persistent alterations in FA-collagen alignment that indicate sustained cellular responses to tissue stress.
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spelling doaj-art-db644f22d4dd4fac93d31440b184160f2025-08-20T04:01:24ZengNature PortfolioScientific Reports2045-23222025-07-0115111210.1038/s41598-025-08251-4Analysis of 2-dimensional regional differences in the peripapillary scleral fibroblast cytoskeleton of normotensive and hypertensive mouse eyesAnn Mozzer0Joe Gerald Jesu Raj1Nushrat Yasmin2Edward Esposito3Ian Pitha4Department of Ophthalmology, Center for Nanomedicine at the Wilmer Eye Institute, Johns Hopkins UniversityDepartment of Ophthalmology, Center for Nanomedicine at the Wilmer Eye Institute, Johns Hopkins UniversityDepartment of Ophthalmology, Center for Nanomedicine at the Wilmer Eye Institute, Johns Hopkins UniversityDepartment of Ophthalmology, Center for Nanomedicine at the Wilmer Eye Institute, Johns Hopkins UniversityDepartment of Ophthalmology, Center for Nanomedicine at the Wilmer Eye Institute, Johns Hopkins UniversityAbstract These studies aimed to study the mechanisms of glaucomatous peripapillary scleral (PPS) remodeling by investigating IOP-induced changes in fibroblast actin-collagen alignment and nuclear morphology in mouse PPS. Cryosections from the optic nerve heads (ONH) of eyes isolated 1- and 6-weeks after bead-induced IOP elevation were imaged for nuclei, fibrillar actin (FA), and collagen (second harmonic generation, SHG). Two-dimensional nuclear morphology was analyzed using VAMPIRE machine-learning image analysis and FA-collagen alignment was determined by comparing vector fields of FA and SHG images. Nuclear morphology was regionally defined with the internal (pial) PPS (iPPS) containing nuclei with higher aspect ratios than the peripheral PPS (outer PPS, oPPS) and peripheral sclera. FA-collagen alignment was higher in the PPS than in the peripheral sclera (7.1 ± 2.5° versus 10.0 ± 1.4°, p = 0.05, n = 6). One and six weeks after BI, there were non-significant nuclear morphologic changes reflecting a transition to a rounder shape in all scleral regions and persistently reduced FA-collagen alignment in the PPS regions. This study therefore concludes that chronic IOP elevation is associated with persistent alterations in FA-collagen alignment that indicate sustained cellular responses to tissue stress.https://doi.org/10.1038/s41598-025-08251-4
spellingShingle Ann Mozzer
Joe Gerald Jesu Raj
Nushrat Yasmin
Edward Esposito
Ian Pitha
Analysis of 2-dimensional regional differences in the peripapillary scleral fibroblast cytoskeleton of normotensive and hypertensive mouse eyes
Scientific Reports
title Analysis of 2-dimensional regional differences in the peripapillary scleral fibroblast cytoskeleton of normotensive and hypertensive mouse eyes
title_full Analysis of 2-dimensional regional differences in the peripapillary scleral fibroblast cytoskeleton of normotensive and hypertensive mouse eyes
title_fullStr Analysis of 2-dimensional regional differences in the peripapillary scleral fibroblast cytoskeleton of normotensive and hypertensive mouse eyes
title_full_unstemmed Analysis of 2-dimensional regional differences in the peripapillary scleral fibroblast cytoskeleton of normotensive and hypertensive mouse eyes
title_short Analysis of 2-dimensional regional differences in the peripapillary scleral fibroblast cytoskeleton of normotensive and hypertensive mouse eyes
title_sort analysis of 2 dimensional regional differences in the peripapillary scleral fibroblast cytoskeleton of normotensive and hypertensive mouse eyes
url https://doi.org/10.1038/s41598-025-08251-4
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