Introduction: Meniscus is a complex tissue made of a vascular outer zone and an avascular inner zone with chondrocytes existing under physiological conditions of sustained hypoxia. Using a hypoxic chamber to provide controlled hypoxia, this study was performed to determine whether persistent hypoxia enhances the differentiation of menisci tissue harvested from neonatal pigs. Materials and Methods: Meniscal bodies were in vitro cultured under hypoxia or normoxia conditions until 14 days. Samples were analysed at 0 (T0), 7 (T1), 10 (T2) and 14 (T3) days by immunofluorescence and RT-PCR [Type II Collagen (COL2A1), SRY-Box Transcription Factor 9 (SOX-9), Type I Collagen (COL1A1), Hypoxia Inducible Factor 1 Subunit Alpha (HIF-1α) and the vascular markers, the Vascular Endothelial Growth Factor A (VEGF) and Platelet And Endothelial Cell Adhesion Molecule 1 (CD31)]. Results: A maturation shifting was observed by immunofluorescence and RT-PCR analysis in hypoxic treatment characterized by 1) up-regulation of COL2A1 at T2 (P <0.05); 2) reduction in SOX9 expression (not significant); 3) greater increase of VEGF at T3 (P <0.001); 4) down-regulation of CD31 with a peak growth at T2 and a significantly decrease at T3 (P<0.05). Conclusion: This study shows that hypoxia can be considered as a booster to induce the differentiation of neonatal fibroblast-like meniscal cells into a mature fibro-cartilaginous phenotype by acting on angiogenesis markers: hypoxia induces VEGF, which is a chondrocyte survival factor during development, and it is also required for postnatal differentiation of chondrocytes. These results open considerably opportunities in the field of meniscus tissue engineering.

Hypoxia of neonatal meniscus: maturation towards a fibro-chondrocytes phenotype through the modulation of vascular markers / V. Herrera Millar, B. Canciani, F. Biancoli, M. Pallaoro, L. Mangiavini, L. Anastasia, F. Cirillo, G. Peretti, S. Modina, A. Di Giancamillo. ((Intervento presentato al 33. convegno Congress of the European Association of Veterinary Anatomists : July, 28th - 30th tenutosi a Ghent nel 2021.

Hypoxia of neonatal meniscus: maturation towards a fibro-chondrocytes phenotype through the modulation of vascular markers

V. Herrera Millar
Primo
;
B. Canciani
Secondo
;
M. Pallaoro;L. Mangiavini;L. Anastasia;F. Cirillo;G. Peretti;S. Modina
Penultimo
;
A. Di Giancamillo
Ultimo
2021

Abstract

Introduction: Meniscus is a complex tissue made of a vascular outer zone and an avascular inner zone with chondrocytes existing under physiological conditions of sustained hypoxia. Using a hypoxic chamber to provide controlled hypoxia, this study was performed to determine whether persistent hypoxia enhances the differentiation of menisci tissue harvested from neonatal pigs. Materials and Methods: Meniscal bodies were in vitro cultured under hypoxia or normoxia conditions until 14 days. Samples were analysed at 0 (T0), 7 (T1), 10 (T2) and 14 (T3) days by immunofluorescence and RT-PCR [Type II Collagen (COL2A1), SRY-Box Transcription Factor 9 (SOX-9), Type I Collagen (COL1A1), Hypoxia Inducible Factor 1 Subunit Alpha (HIF-1α) and the vascular markers, the Vascular Endothelial Growth Factor A (VEGF) and Platelet And Endothelial Cell Adhesion Molecule 1 (CD31)]. Results: A maturation shifting was observed by immunofluorescence and RT-PCR analysis in hypoxic treatment characterized by 1) up-regulation of COL2A1 at T2 (P <0.05); 2) reduction in SOX9 expression (not significant); 3) greater increase of VEGF at T3 (P <0.001); 4) down-regulation of CD31 with a peak growth at T2 and a significantly decrease at T3 (P<0.05). Conclusion: This study shows that hypoxia can be considered as a booster to induce the differentiation of neonatal fibroblast-like meniscal cells into a mature fibro-cartilaginous phenotype by acting on angiogenesis markers: hypoxia induces VEGF, which is a chondrocyte survival factor during development, and it is also required for postnatal differentiation of chondrocytes. These results open considerably opportunities in the field of meniscus tissue engineering.
Meniscus, hypoxia, in vitro culture, angiogenic factors, tissue engineering
Settore VET/01 - Anatomia degli Animali Domestici
Settore MED/33 - Malattie Apparato Locomotore
European Association of Veterinary Anatomis
https://eava2020.ugent.be/
Hypoxia of neonatal meniscus: maturation towards a fibro-chondrocytes phenotype through the modulation of vascular markers / V. Herrera Millar, B. Canciani, F. Biancoli, M. Pallaoro, L. Mangiavini, L. Anastasia, F. Cirillo, G. Peretti, S. Modina, A. Di Giancamillo. ((Intervento presentato al 33. convegno Congress of the European Association of Veterinary Anatomists : July, 28th - 30th tenutosi a Ghent nel 2021.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/870044
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