Universität zu Köln

Atta, Ghada, Schroedl, Falk ORCID: 0000-0002-8807-8982, Kaser-Eichberger, Alexandra ORCID: 0000-0002-6115-5036, Spitzer, Gabriel, Traweger, Andreas ORCID: 0000-0002-0220-4766, Heindl, Ludwig M. and Tempfer, Herbert (2021). Scleraxis expressing scleral cells respond to inflammatory stimulation. Histochem. Cell Biol., 156 (2). S. 123 - 133. NEW YORK: SPRINGER. ISSN 1432-119X

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Abstract

The sclera is an ocular tissue rich of collagenous extracellular matrix, which is built up and maintained by relatively few, still poorly characterized fibroblast-like cells. The aims of this study are to add to the characterization of scleral fibroblasts and to examine the reaction of these fibroblasts to inflammatory stimulation in an ex vivo organotypic model. Scleras of scleraxis-GFP (SCX-GFP) mice were analyzed using immunohistochemistry and qRT-PCR for the expression of the tendon cell associated marker genes scleraxis (SCX), mohawk and tenomodulin. In organotypic tissue culture, explanted scleras of adult scleraxis GFP reporter mice were exposed to 10 ng/ml recombinant interleukin 1-ss (IL1-ss) and IL1-ss in combination with dexamethasone. The tissue was then analyzed by immunofluorescence staining of the inflammation- and fibrosis-associated proteins IL6, COX-2, iNOS, connective tissue growth factor, MMP2, MMP3, and MMP13 as well as for collagen fibre degradation using a Collagen Hybridizing Peptide (CHP) binding assay. The mouse sclera displayed a strong expression of scleraxis promoter-driven GFP, indicating a tendon cell-like phenotype, as well as expression of scleraxis, tenomodulin and mohawk mRNA. Upon IL1-ss stimulation, SCX-GFP+ cells significantly upregulated the expression of all proteins analysed. Moreover, IL1-ss stimulation resulted in significant collagen degradation. Adding the corticosteroid dexamethasone significantly reduced the response to IL1-ss stimulation. Collagen degradation was significantly enhanced in the IL1-ss group. Dexamethasone demonstrated a significant rescue effect. This work provides insights into the characteristics of scleral cells and establishes an ex vivo model of scleral inflammation.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Atta, Ghada UNSPECIFIED UNSPECIFIED UNSPECIFIED Schroedl, Falk UNSPECIFIED orcid.org/0000-0002-8807-8982 UNSPECIFIED Kaser-Eichberger, Alexandra UNSPECIFIED orcid.org/0000-0002-6115-5036 UNSPECIFIED Spitzer, Gabriel UNSPECIFIED UNSPECIFIED UNSPECIFIED Traweger, Andreas UNSPECIFIED orcid.org/0000-0002-0220-4766 UNSPECIFIED Heindl, Ludwig M. UNSPECIFIED UNSPECIFIED UNSPECIFIED Tempfer, Herbert UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-579163
DOI:	10.1007/s00418-021-01985-y
Journal or Publication Title:	Histochem. Cell Biol.
Volume:	156
Number:	2
Page Range:	S. 123 - 133
Date:	2021
Publisher:	SPRINGER
Place of Publication:	NEW YORK
ISSN:	1432-119X
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language Cell Biology; Microscopy Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/57916