Increased Collagen Crosslinking in Stiff Clubfoot Tissue: Implications for the Improvement of Therapeutic Strategies

Congenital clubfoot is a complex musculoskeletal deformity, in which a stiff, contracted tissue forms in the medial part of the foot. Fibrotic changes are associated with increased collagen deposition and lysyl oxidase (LOX)-mediated crosslinking, which impair collagen degradation and increase the t...

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Autores principales:	Jarmila Knitlova, Martina Doubkova, Adam Eckhardt, Martin Ostadal, Jana Musilkova, Lucie Bacakova, Tomas Novotny
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	relapsed clubfoot congenital idiopathic <i>Talipes equinovarus</i> collagen contraction crosslinking beta-aminopropionitrile (BAPN) Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/a6d2094ae7d443bbaed9ca3c597d843f
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spelling	oai:doaj.org-article:a6d2094ae7d443bbaed9ca3c597d843f2021-11-11T17:19:04ZIncreased Collagen Crosslinking in Stiff Clubfoot Tissue: Implications for the Improvement of Therapeutic Strategies10.3390/ijms2221119031422-00671661-6596https://doaj.org/article/a6d2094ae7d443bbaed9ca3c597d843f2021-11-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/21/11903https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Congenital clubfoot is a complex musculoskeletal deformity, in which a stiff, contracted tissue forms in the medial part of the foot. Fibrotic changes are associated with increased collagen deposition and lysyl oxidase (LOX)-mediated crosslinking, which impair collagen degradation and increase the tissue stiffness. First, we studied collagen deposition, as well as the expression of collagen and the amount of pyridinoline and deoxypyridinoline crosslinks in the tissue of relapsed clubfoot by immunohistochemistry, real-time PCR, and enzyme-linked immunosorbent assay (ELISA). We then isolated fibroblast-like cells from the contracted tissue to study the potential inhibition of these processes in vitro. We assessed the effects of a LOX inhibitor, β-aminopropionitrile (BAPN), on the cells by a hydroxyproline assay, ELISA, and Second Harmonic Generation imaging. We also evaluated the cell-mediated contraction of extracellular matrix in 3D cell-populated collagen gels. For the first time, we have confirmed significantly increased crosslinking and excessive collagen type I deposition in the clubfoot-contracted tissue. We successfully reduced these processes in vitro in a dose-dependent manner with 10–40 µg/mL of BAPN, and we observed an increasing trend in the inhibition of the cell-mediated contraction of collagen gels. The in vitro inhibitory effects indicate that BAPN has good potential for the treatment of relapsed and resistant clubfeet.Jarmila KnitlovaMartina DoubkovaAdam EckhardtMartin OstadalJana MusilkovaLucie BacakovaTomas NovotnyMDPI AGarticlerelapsed clubfootcongenital idiopathic <i>Talipes equinovarus</i>collagencontractioncrosslinkingbeta-aminopropionitrile (BAPN)Biology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 11903, p 11903 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	relapsed clubfoot congenital idiopathic <i>Talipes equinovarus</i> collagen contraction crosslinking beta-aminopropionitrile (BAPN) Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	relapsed clubfoot congenital idiopathic <i>Talipes equinovarus</i> collagen contraction crosslinking beta-aminopropionitrile (BAPN) Biology (General) QH301-705.5 Chemistry QD1-999 Jarmila Knitlova Martina Doubkova Adam Eckhardt Martin Ostadal Jana Musilkova Lucie Bacakova Tomas Novotny Increased Collagen Crosslinking in Stiff Clubfoot Tissue: Implications for the Improvement of Therapeutic Strategies
description	Congenital clubfoot is a complex musculoskeletal deformity, in which a stiff, contracted tissue forms in the medial part of the foot. Fibrotic changes are associated with increased collagen deposition and lysyl oxidase (LOX)-mediated crosslinking, which impair collagen degradation and increase the tissue stiffness. First, we studied collagen deposition, as well as the expression of collagen and the amount of pyridinoline and deoxypyridinoline crosslinks in the tissue of relapsed clubfoot by immunohistochemistry, real-time PCR, and enzyme-linked immunosorbent assay (ELISA). We then isolated fibroblast-like cells from the contracted tissue to study the potential inhibition of these processes in vitro. We assessed the effects of a LOX inhibitor, β-aminopropionitrile (BAPN), on the cells by a hydroxyproline assay, ELISA, and Second Harmonic Generation imaging. We also evaluated the cell-mediated contraction of extracellular matrix in 3D cell-populated collagen gels. For the first time, we have confirmed significantly increased crosslinking and excessive collagen type I deposition in the clubfoot-contracted tissue. We successfully reduced these processes in vitro in a dose-dependent manner with 10–40 µg/mL of BAPN, and we observed an increasing trend in the inhibition of the cell-mediated contraction of collagen gels. The in vitro inhibitory effects indicate that BAPN has good potential for the treatment of relapsed and resistant clubfeet.
format	article
author	Jarmila Knitlova Martina Doubkova Adam Eckhardt Martin Ostadal Jana Musilkova Lucie Bacakova Tomas Novotny
author_facet	Jarmila Knitlova Martina Doubkova Adam Eckhardt Martin Ostadal Jana Musilkova Lucie Bacakova Tomas Novotny
author_sort	Jarmila Knitlova
title	Increased Collagen Crosslinking in Stiff Clubfoot Tissue: Implications for the Improvement of Therapeutic Strategies
title_short	Increased Collagen Crosslinking in Stiff Clubfoot Tissue: Implications for the Improvement of Therapeutic Strategies
title_full	Increased Collagen Crosslinking in Stiff Clubfoot Tissue: Implications for the Improvement of Therapeutic Strategies
title_fullStr	Increased Collagen Crosslinking in Stiff Clubfoot Tissue: Implications for the Improvement of Therapeutic Strategies
title_full_unstemmed	Increased Collagen Crosslinking in Stiff Clubfoot Tissue: Implications for the Improvement of Therapeutic Strategies
title_sort	increased collagen crosslinking in stiff clubfoot tissue: implications for the improvement of therapeutic strategies
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/a6d2094ae7d443bbaed9ca3c597d843f
work_keys_str_mv	AT jarmilaknitlova increasedcollagencrosslinkinginstiffclubfoottissueimplicationsfortheimprovementoftherapeuticstrategies AT martinadoubkova increasedcollagencrosslinkinginstiffclubfoottissueimplicationsfortheimprovementoftherapeuticstrategies AT adameckhardt increasedcollagencrosslinkinginstiffclubfoottissueimplicationsfortheimprovementoftherapeuticstrategies AT martinostadal increasedcollagencrosslinkinginstiffclubfoottissueimplicationsfortheimprovementoftherapeuticstrategies AT janamusilkova increasedcollagencrosslinkinginstiffclubfoottissueimplicationsfortheimprovementoftherapeuticstrategies AT luciebacakova increasedcollagencrosslinkinginstiffclubfoottissueimplicationsfortheimprovementoftherapeuticstrategies AT tomasnovotny increasedcollagencrosslinkinginstiffclubfoottissueimplicationsfortheimprovementoftherapeuticstrategies
_version_	1718432137197125632

Increased Collagen Crosslinking in Stiff Clubfoot Tissue: Implications for the Improvement of Therapeutic Strategies

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