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Diverse effects of Perlecan-HS on cell and matrix regulation: Tendon destabilization in Hspg2 Exon 3 null HS deficient mice reveals essential homeostatic roles for HS in cellular regulation and tendon functionality

, , , ,
1 Raymond Purves Bone and Joint Lab, Kolling Institute of Medical Research, University of Sydney, Sydney, Australia
2 Murray Maxwell Biomechanics Laboratory, Royal North Shore Hospital, University of Sydney, St. Leonards, New South Wales, Australia
3 Surgical Skills Laboratory, Australian School of Advanced Medicine, Macquarie University, Sydney, New South Wales, Australia
4 Sydney Medical School, Northern, University of Sydney, Sydney, Australia
5 Graduate School of Biomedical Engineering, University of New South Wales, St. Leonards, New South Wales, Australia
DOI
10.7287/peerj.preprints.26644v1
Published
Accepted
Subject Areas
Biochemistry, Anatomy and Physiology
Keywords
Tendon, collagen, fibroblast growth factor, extracellular matrix, proteoglycan, heparan sulphate, tenocyte, biomechanics
Copyright
© 2018 Shu et al.
Licence
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ Preprints) and either DOI or URL of the article must be cited.
Cite this article
Shu CC, Smith MM, Appleyard RC, Little CB, Melrose J. 2018. Diverse effects of Perlecan-HS on cell and matrix regulation: Tendon destabilization in Hspg2 Exon 3 null HS deficient mice reveals essential homeostatic roles for HS in cellular regulation and tendon functionality. PeerJ Preprints 6:e26644v1

Abstract

The aim of this study was to determine the regulatory role of the perlecan HS side chains on cell and matrix homeostasis in tail and Achilles tendons in 3 and 12 week old Hspg2 exon 3 null HS deficient (Hspg2∆3-/∆3- ) and C57 BL/6 Wild Type (WT) mice. Tendons were biomechanically tested (ultimate tensile stress [UTS], tensile modulus [TM]) and glycosaminoglycan (GAG) and collagen (hydroxyproline) compositional analyses undertaken. Monolayer cultures of Hspg2∆3-/∆3- tenocytes stimulated with FGF-2 showed elevated Adamts4, Mmp2, 3, 13 gene expression compared to WT mice. Col1A1, Vcan, Bgn, Dcn, Lum, Hspg2, Ltbp1, Ltbp2, Eln and Fbn1 showed no major differences between genotypes. Type VI collagen and perlecan were immunolocalised in tail tendon and collagen fibrils imaged using transmission electron microscopy (TEM). The amplified catabolic phenotype of Hspg2∆3-/∆3- mice may account for the age-dependent decline in GAG observed in tail tendon and changes in UTS/TM biomechanics. Collagen fibril diameter increased in WT but decreased in Hspg2∆3-/∆3- tail tendons over 3 to 12 weeks. Achilles tenotomy resulted in changes in tendon material properties in both genotypes but, Hspg2∆3-/∆3- mice had a slower recovery of UTS after tenotomy. HS deficiency in Hspg2∆3-/∆3- tendon impaired tenocyte repair responses to FGF-2 and led to deleterious changes in tendon organization which was reflected in changes in their material properties. .

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This is a submission to PeerJ for review.

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Competing Interests

The authors declare that they have no competing interests.

Author Contributions

Cindy C Shu performed the experiments, prepared figures and/or tables, authored or reviewed drafts of the paper, approved the final draft.

Margaret M Smith conceived and designed the experiments, analyzed the data, approved the final draft, biostatistics, advice on qRTPCR.

Richard C Appleyard conceived and designed the experiments, performed the experiments, contributed reagents/materials/analysis tools, approved the final draft.

Christopher B Little conceived and designed the experiments, analyzed the data, authored or reviewed drafts of the paper, approved the final draft.

James Melrose conceived and designed the experiments, analyzed the data, contributed reagents/materials/analysis tools, prepared figures and/or tables, authored or reviewed drafts of the paper, approved the final draft.

Animal Ethics

The following information was supplied relating to ethical approvals (i.e., approving body and any reference numbers):

University of Sydney Animal Care and Ethics Research Committee approved all animal procedures conducted in this study.

Data Deposition

The following information was supplied regarding data availability:

Raw data is stored in the form of an electronic lab book on the Sydney University server. This is a large file specific data for particular figures in the submitted manuscript can be supplied upon request.

Funding

This work was supported by NHMRC Project Grant 1004032. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Diverse effects of Perlecan-HS on cell and matrix regulation: Tendon destabilization in Hspg2 Exon 3 null HS deficient mice reveals essential homeostatic roles for HS in cellular regulation and tendon functionality
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