Marco Govoni1, Claudio Muscari2, Francesca Bonafè3, Paolo Giovanni Morselli4, Marilisa Cortesi5, Dante Dallari6, Emanuele Giordano7. 1. Reconstructive Orthopaedic Surgery and Innovative Techniques - Musculoskeletal Tissue Bank, IRCCS Istituto Ortopedico Rizzoli, Bologna, Italy. Electronic address: marco.govoni@ior.it. 2. BioEngLab, Health Science and Technology - Interdepartmental Center for Industrial Research (HST-CIRI), University of Bologna, Cesena, Italy; Department of Biomedical and Neuromotor Sciences (DIBINEM), University of Bologna, Bologna, Italy. 3. Department of Biomedical and Neuromotor Sciences (DIBINEM), University of Bologna, Bologna, Italy. 4. Department of Experimental, Diagnostic and Specialty Medicine (DIMES), University of Bologna, Bologna, Italy. 5. Laboratory of Cellular and Molecular Engineering "Silvio Cavalcanti" - DEI, University of Bologna, Cesena, Italy. 6. Reconstructive Orthopaedic Surgery and Innovative Techniques - Musculoskeletal Tissue Bank, IRCCS Istituto Ortopedico Rizzoli, Bologna, Italy. 7. BioEngLab, Health Science and Technology - Interdepartmental Center for Industrial Research (HST-CIRI), University of Bologna, Cesena, Italy; Laboratory of Cellular and Molecular Engineering "Silvio Cavalcanti" - DEI, University of Bologna, Cesena, Italy.
Abstract
PURPOSE: The aim of this study was to evaluate the effects exerted over chondrogenic commitment of human adipose-derived mesenchymal stem cells (ADSCs) by a very low oxygen tension (<1% pO2). MATERIALS/ METHODS: Cell morphology, mRNA levels of chondrocyte-specific marker genes and the involvement of p38 MAPK signalling were monitored in human ADSCs under a very low oxygen tension. RESULTS: Cell morphology was significantly changed after two days of hypoxic preconditioning when they featured as elongated spindle-shaped cells. SRY-box containing gene 9, aggrecan and collagen type II mRNA levels were enhanced under severe hypoxic culture conditions. Moreover, the inhibition of p38 MAPK resulted in a substantial reduction in transcription of the above-mentioned specific genes, proving the pivotal role of this pathway in the transcriptional regulation of chondrogenesis. CONCLUSIONS: Here, we propose a protocol showing the early commitment of stem cells towards the chondrogenic phenotype in only 2 days of culture via a very low hypoxic environment, in the absence of growth factors added in the culture medium.
PURPOSE: The aim of this study was to evaluate the effects exerted over chondrogenic commitment of human adipose-derived mesenchymal stem cells (ADSCs) by a very low oxygen tension (<1% pO2). MATERIALS/ METHODS: Cell morphology, mRNA levels of chondrocyte-specific marker genes and the involvement of p38 MAPK signalling were monitored in human ADSCs under a very low oxygen tension. RESULTS: Cell morphology was significantly changed after two days of hypoxic preconditioning when they featured as elongated spindle-shaped cells. SRY-box containing gene 9, aggrecan and collagen type II mRNA levels were enhanced under severe hypoxic culture conditions. Moreover, the inhibition of p38 MAPK resulted in a substantial reduction in transcription of the above-mentioned specific genes, proving the pivotal role of this pathway in the transcriptional regulation of chondrogenesis. CONCLUSIONS: Here, we propose a protocol showing the early commitment of stem cells towards the chondrogenic phenotype in only 2 days of culture via a very low hypoxic environment, in the absence of growth factors added in the culture medium.
Authors: Erwin Pavel Lamparelli; Joseph Lovecchio; Maria Camilla Ciardulli; Valentina Giudice; Tina P Dale; Carmine Selleri; Nicholas Forsyth; Emanuele Giordano; Nicola Maffulli; Giovanna Della Porta Journal: Pharmaceutics Date: 2021-03-17 Impact factor: 6.321
Authors: Alice Pasini; Joseph Lovecchio; Marilisa Cortesi; Chiara Liverani; Chiara Spadazzi; Laura Mercatali; Toni Ibrahim; Emanuele Giordano Journal: Ann Biomed Eng Date: 2021-02-04 Impact factor: 3.934