Viviana Costa1, Lavinia Raimondi2, Alice Conigliaro3, Francesca Salamanna4, Valeria Carina2, Angela De Luca2, Daniele Bellavia2, Riccardo Alessandro5, Milena Fini4, Gianluca Giavaresi6. 1. Rizzoli Orthopedic Institute, Bologna, Italy; Rizzoli Orthopedic Institute, Innovative Technological Platforms for Tissue Engineering, Theranostic and Oncology, Palermo, Italy. Electronic address: viviana.costa@ior.it. 2. Rizzoli Orthopedic Institute, Bologna, Italy; Rizzoli Orthopedic Institute, Innovative Technological Platforms for Tissue Engineering, Theranostic and Oncology, Palermo, Italy. 3. Department of Cellular Biotechnology and Hematology, Sapienza University of Rome, Rome, Italy. 4. Rizzoli Orthopedic Institute, Laboratory of Preclinical and Surgical Studies, Bologna, Italy. 5. Department of Biopathology and Medical Biotechnologies, Section of Biology and Genetics, University of Palermo, Palermo, Italy; Institute of Biomedicine and Molecular Immunology, National Research Council, Palermo, Italy. 6. Rizzoli Orthopedic Institute, Bologna, Italy; Rizzoli Orthopedic Institute, Innovative Technological Platforms for Tissue Engineering, Theranostic and Oncology, Palermo, Italy; Rizzoli Orthopedic Institute, Laboratory of Preclinical and Surgical Studies, Bologna, Italy.
Abstract
BACKGROUND AIMS: During bone formation, angiogenesis and osteogenesis are regulated by hypoxia, which is able to induce blood vessel formation, as well as recruit and differentiate human mesenchymal stromal cells (hMSCs). The molecular mechanisms involved in HIF-1α response and hMSC differentiation during bone formation are still unclear. This study aimed to investigate the synergistic role of hypoxia and hypoxia-mimetic microRNA miR-675-5p in angiogenesis response and osteo-chondroblast commitment of hMSCs. METHODS: By using a suitable in vitro cell model of hMSCs (maintained in hypoxia or normoxia), the role of HIF-1α and miR-675-5p in angiogenesis and osteogenesis coupling was investigated, using fluorescence-activated cell sorting (FACS), gene expression and protein analysis. RESULTS: Hypoxia induced miR-675-5p expression and a hypoxia-angiogenic response, as demonstrated by increase in vascular endothelial growth factor messenger RNA and protein release. MiR-675-5p overexpression in normoxia promoted the down-regulation of MSC markers and the up-regulation of osteoblast and chondroblast markers, as demonstrated by FACS and protein analysis. Moreover, miR-675-5p depletion in a low-oxygen condition partially abolished the hypoxic response, including angiogenesis, and in particular restored the MSC phenotype, demonstrated by cytofluorimetric analysis. In addition, current preliminary data suggest that the expression of miR-675-5p during hypoxia plays an additive role in sustaining Wnt/β-catenin pathways and the related commitment of hMSCs during bone ossification. DISCUSSION: MiR-675-5p may trigger complex molecular mechanisms that promote hMSC osteoblastic differentiation through a dual strategy: increasing HIF-1α response and activating Wnt/β-catenin signaling.
BACKGROUND AIMS: During bone formation, angiogenesis and osteogenesis are regulated by hypoxia, which is able to induce blood vessel formation, as well as recruit and differentiate human mesenchymal stromal cells (hMSCs). The molecular mechanisms involved in HIF-1α response and hMSC differentiation during bone formation are still unclear. This study aimed to investigate the synergistic role of hypoxia and hypoxia-mimetic microRNA miR-675-5p in angiogenesis response and osteo-chondroblast commitment of hMSCs. METHODS: By using a suitable in vitro cell model of hMSCs (maintained in hypoxia or normoxia), the role of HIF-1α and miR-675-5p in angiogenesis and osteogenesis coupling was investigated, using fluorescence-activated cell sorting (FACS), gene expression and protein analysis. RESULTS:Hypoxia induced miR-675-5p expression and a hypoxia-angiogenic response, as demonstrated by increase in vascular endothelial growth factor messenger RNA and protein release. MiR-675-5p overexpression in normoxia promoted the down-regulation of MSC markers and the up-regulation of osteoblast and chondroblast markers, as demonstrated by FACS and protein analysis. Moreover, miR-675-5p depletion in a low-oxygen condition partially abolished the hypoxic response, including angiogenesis, and in particular restored the MSC phenotype, demonstrated by cytofluorimetric analysis. In addition, current preliminary data suggest that the expression of miR-675-5p during hypoxia plays an additive role in sustaining Wnt/β-catenin pathways and the related commitment of hMSCs during bone ossification. DISCUSSION: MiR-675-5p may trigger complex molecular mechanisms that promote hMSC osteoblastic differentiation through a dual strategy: increasing HIF-1α response and activating Wnt/β-catenin signaling.
Authors: Evan Paul Tracy; Virginia Stielberg; Gabrielle Rowe; Daniel Benson; Sara S Nunes; James B Hoying; Walter Lee Murfee; Amanda Jo LeBlanc Journal: Am J Physiol Heart Circ Physiol Date: 2022-02-18 Impact factor: 4.733
Authors: Nádia de Cássia Noronha; Amanda Mizukami; Carolina Caliári-Oliveira; Juçara Gastaldi Cominal; José Lucas M Rocha; Dimas Tadeu Covas; Kamilla Swiech; Kelen C R Malmegrim Journal: Stem Cell Res Ther Date: 2019-05-02 Impact factor: 6.832
Authors: Laura Saieva; Maria Magdalena Barreca; Chiara Zichittella; Maria Giulia Prado; Marco Tripodi; Riccardo Alessandro; Alice Conigliaro Journal: Int J Mol Sci Date: 2020-05-28 Impact factor: 5.923