Testing Hypoxia in Pig Meniscal Culture: Biological Role of the Vascular-Related Factors in the Differentiation and Viability of Neonatal Meniscus

Menisci play an essential role in shock absorption, joint stability, load resistance and its transmission thanks to their conformation. Adult menisci can be divided in three zones based on the vascularization: an avascular inner zone with no blood supply, a fully vascularized outer zone, and an inte...

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Autores principales:	Barbara Canciani, Valentina Rafaela Herrera Millar, Margherita Pallaoro, Lucia Aidos, Federica Cirillo, Luigi Anastasia, Giuseppe Maria Peretti, Silvia Clotilde Modina, Laura Mangiavini, Alessia Di Giancamillo
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	hypoxia meniscus pig fibro-chondrocytes differentiation HIF-1α Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/fa69d239ad16447d8ee77d07887e6481
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spelling	oai:doaj.org-article:fa69d239ad16447d8ee77d07887e64812021-11-25T17:56:53ZTesting Hypoxia in Pig Meniscal Culture: Biological Role of the Vascular-Related Factors in the Differentiation and Viability of Neonatal Meniscus10.3390/ijms2222124651422-00671661-6596https://doaj.org/article/fa69d239ad16447d8ee77d07887e64812021-11-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/22/12465https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Menisci play an essential role in shock absorption, joint stability, load resistance and its transmission thanks to their conformation. Adult menisci can be divided in three zones based on the vascularization: an avascular inner zone with no blood supply, a fully vascularized outer zone, and an intermediate zone. This organization, in addition to the incomplete knowledge about meniscal biology, composition, and gene expression, makes meniscal regeneration still one of the major challenges both in orthopedics and in tissue engineering. To overcome this issue, we aimed to investigate the role of hypoxia in the differentiation of the three anatomical areas of newborn piglet menisci (anterior horn (A), central body (C), and posterior horn (P)) and its effects on vascular factors. After sample collection, menisci were divided in A, C, P, and they were cultured in vitro under hypoxic (1% O<sub>2</sub>) and normoxic (21% O<sub>2</sub>) conditions at four different experimental time points (T0 = day of explant; T7 = day 7; T10 = day 10; T14 = day 14); samples were then evaluated through immune, histological, and molecular analyses, cell morpho-functional characteristics; with particular focus on matrix composition and expression of vascular factors. It was observed that hypoxia retained the initial phenotype of cells and induced extracellular matrix production resembling a mature tissue. Hypoxia also modulated the expression of angiogenic factors, especially in the early phase of the study. Thus, we observed that hypoxia contributes to the fibro-chondrogenic differentiation with the involvement of angiogenic factors, especially in the posterior horn, which corresponds to the predominant weight-bearing portion.Barbara CancianiValentina Rafaela Herrera MillarMargherita PallaoroLucia AidosFederica CirilloLuigi AnastasiaGiuseppe Maria PerettiSilvia Clotilde ModinaLaura MangiaviniAlessia Di GiancamilloMDPI AGarticlehypoxiameniscuspigfibro-chondrocytesdifferentiationHIF-1αBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 12465, p 12465 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	hypoxia meniscus pig fibro-chondrocytes differentiation HIF-1α Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	hypoxia meniscus pig fibro-chondrocytes differentiation HIF-1α Biology (General) QH301-705.5 Chemistry QD1-999 Barbara Canciani Valentina Rafaela Herrera Millar Margherita Pallaoro Lucia Aidos Federica Cirillo Luigi Anastasia Giuseppe Maria Peretti Silvia Clotilde Modina Laura Mangiavini Alessia Di Giancamillo Testing Hypoxia in Pig Meniscal Culture: Biological Role of the Vascular-Related Factors in the Differentiation and Viability of Neonatal Meniscus
description	Menisci play an essential role in shock absorption, joint stability, load resistance and its transmission thanks to their conformation. Adult menisci can be divided in three zones based on the vascularization: an avascular inner zone with no blood supply, a fully vascularized outer zone, and an intermediate zone. This organization, in addition to the incomplete knowledge about meniscal biology, composition, and gene expression, makes meniscal regeneration still one of the major challenges both in orthopedics and in tissue engineering. To overcome this issue, we aimed to investigate the role of hypoxia in the differentiation of the three anatomical areas of newborn piglet menisci (anterior horn (A), central body (C), and posterior horn (P)) and its effects on vascular factors. After sample collection, menisci were divided in A, C, P, and they were cultured in vitro under hypoxic (1% O<sub>2</sub>) and normoxic (21% O<sub>2</sub>) conditions at four different experimental time points (T0 = day of explant; T7 = day 7; T10 = day 10; T14 = day 14); samples were then evaluated through immune, histological, and molecular analyses, cell morpho-functional characteristics; with particular focus on matrix composition and expression of vascular factors. It was observed that hypoxia retained the initial phenotype of cells and induced extracellular matrix production resembling a mature tissue. Hypoxia also modulated the expression of angiogenic factors, especially in the early phase of the study. Thus, we observed that hypoxia contributes to the fibro-chondrogenic differentiation with the involvement of angiogenic factors, especially in the posterior horn, which corresponds to the predominant weight-bearing portion.
format	article
author	Barbara Canciani Valentina Rafaela Herrera Millar Margherita Pallaoro Lucia Aidos Federica Cirillo Luigi Anastasia Giuseppe Maria Peretti Silvia Clotilde Modina Laura Mangiavini Alessia Di Giancamillo
author_facet	Barbara Canciani Valentina Rafaela Herrera Millar Margherita Pallaoro Lucia Aidos Federica Cirillo Luigi Anastasia Giuseppe Maria Peretti Silvia Clotilde Modina Laura Mangiavini Alessia Di Giancamillo
author_sort	Barbara Canciani
title	Testing Hypoxia in Pig Meniscal Culture: Biological Role of the Vascular-Related Factors in the Differentiation and Viability of Neonatal Meniscus
title_short	Testing Hypoxia in Pig Meniscal Culture: Biological Role of the Vascular-Related Factors in the Differentiation and Viability of Neonatal Meniscus
title_full	Testing Hypoxia in Pig Meniscal Culture: Biological Role of the Vascular-Related Factors in the Differentiation and Viability of Neonatal Meniscus
title_fullStr	Testing Hypoxia in Pig Meniscal Culture: Biological Role of the Vascular-Related Factors in the Differentiation and Viability of Neonatal Meniscus
title_full_unstemmed	Testing Hypoxia in Pig Meniscal Culture: Biological Role of the Vascular-Related Factors in the Differentiation and Viability of Neonatal Meniscus
title_sort	testing hypoxia in pig meniscal culture: biological role of the vascular-related factors in the differentiation and viability of neonatal meniscus
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/fa69d239ad16447d8ee77d07887e6481
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Testing Hypoxia in Pig Meniscal Culture: Biological Role of the Vascular-Related Factors in the Differentiation and Viability of Neonatal Meniscus

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