Sun H Peck1,2,3, Justin R Bendigo1,2,3, John W Tobias4, George R Dodge2,3,5, Neil R Malhotra1,2, Robert L Mauck2,3,6, Lachlan J Smith1,2,3. 1. Department of Neurosurgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. 2. McKay Orthopaedic Research Laboratory, Department of Orthopaedic Surgery, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. 3. Translational Musculoskeletal Research Center, Corporal Michael J. Crescenz VA Medical Center, Philadelphia, PA, USA. 4. Penn Genomic Analysis Core, University of Pennsylvania, Philadelphia, PA, USA. 5. Department of Otorhinolaryngology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA. 6. Department of Bioengineering, School of Engineering and Applied Science, University of Pennsylvania, Philadelphia, PA, USA.
Abstract
OBJECTIVE: Skeletal tissues such as intervertebral disc and articular cartilage possess limited innate potential to regenerate, in part due to their avascularity and low cell density. Despite recent advances in mesenchymal stem cell (MSC)-based disc and cartilage regeneration, key challenges remain, including the sensitivity of these cells to in vivo microenvironmental stress such as low oxygen and limited nutrition. The objective of this study was to investigate whether preconditioning with hypoxia and/or transforming growth factor-β 3 (TGF-β3) can enhance MSC survival and extracellular matrix production in a low oxygen and nutrient-limited microenvironment. DESIGN: MSCs from multiple bovine donors were preconditioned in monolayer in normoxia or hypoxia, with or without TGF-β3, and the global effects on gene expression were examined using microarrays. Subsequently, the effects of preconditioning on MSC survival and extracellular matrix production were examined using low oxygen and nutrient-limited pellet culture experiments. RESULTS: Hypoxic preconditioning resulted in upregulation of genes associated with growth, cell-cell signaling, metabolism, and cell stress response pathways, and significantly enhanced MSC survival for all donors in low oxygen and nutrient-limited pellet culture. In contrast, TGF-β3 preconditioning diminished survival. The nature and magnitude of the effects of preconditioning with either hypoxia or TGF-β3 on glycosaminoglycan production were donor dependent. CONCLUSIONS: These results strongly support the use of hypoxic preconditioning to improve postimplantation MSC survival in avascular tissues such as disc and cartilage.
OBJECTIVE: Skeletal tissues such as intervertebral disc and articular cartilage possess limited innate potential to regenerate, in part due to their avascularity and low cell density. Despite recent advances in mesenchymal stem cell (MSC)-based disc and cartilage regeneration, key challenges remain, including the sensitivity of these cells to in vivo microenvironmental stress such as low oxygen and limited nutrition. The objective of this study was to investigate whether preconditioning with hypoxia and/or transforming growth factor-β 3 (TGF-β3) can enhance MSC survival and extracellular matrix production in a low oxygen and nutrient-limited microenvironment. DESIGN: MSCs from multiple bovine donors were preconditioned in monolayer in normoxia or hypoxia, with or without TGF-β3, and the global effects on gene expression were examined using microarrays. Subsequently, the effects of preconditioning on MSC survival and extracellular matrix production were examined using low oxygen and nutrient-limited pellet culture experiments. RESULTS: Hypoxic preconditioning resulted in upregulation of genes associated with growth, cell-cell signaling, metabolism, and cell stress response pathways, and significantly enhanced MSC survival for all donors in low oxygen and nutrient-limited pellet culture. In contrast, TGF-β3 preconditioning diminished survival. The nature and magnitude of the effects of preconditioning with either hypoxia or TGF-β3 on glycosaminoglycan production were donor dependent. CONCLUSIONS: These results strongly support the use of hypoxic preconditioning to improve postimplantation MSC survival in avascular tissues such as disc and cartilage.
Authors: Beatriz Ranera; Ana Rosa Remacha; Samuel Álvarez-Arguedas; Tomás Castiella; Francisco José Vázquez; Antonio Romero; Pilar Zaragoza; Inmaculada Martín-Burriel; Clementina Rodellar Journal: Vet J Date: 2012-07-06 Impact factor: 2.688
Authors: Aiqun Wei; Sylvia A Chung; Helen Tao; Helena Brisby; Zhen Lin; Bojiang Shen; David D F Ma; Ashish D Diwan Journal: Tissue Eng Part A Date: 2009-09 Impact factor: 3.845
Authors: Jana Müller; Karin Benz; Michael Ahlers; Christoph Gaissmaier; Jürgen Mollenhauer Journal: Cell Transplant Date: 2011-03-08 Impact factor: 4.064
Authors: John T Martin; Sarah E Gullbrand; Bhavana Mohanraj; Beth G Ashinsky; Dong Hwa Kim; Kensuke Ikuta; Dawn M Elliott; Lachlan J Smith; Robert L Mauck; Harvey E Smith Journal: Tissue Eng Part A Date: 2017-04-19 Impact factor: 3.845
Authors: Rebecca Williams; Ilyas M Khan; Kirsty Richardson; Larissa Nelson; Helen E McCarthy; Talal Analbelsi; Sim K Singhrao; Gary P Dowthwaite; Rhiannon E Jones; Duncan M Baird; Holly Lewis; Selwyn Roberts; Hannah M Shaw; Jayesh Dudhia; John Fairclough; Timothy Briggs; Charles W Archer Journal: PLoS One Date: 2010-10-14 Impact factor: 3.240
Authors: Brent M Bijonowski; Qin Fu; Xuegang Yuan; Jerome Irianto; Yan Li; Samuel C Grant; Teng Ma Journal: Biotechnol Bioeng Date: 2020-07-05 Impact factor: 4.530
Authors: Chenghao Zhang; Sarah E Gullbrand; Thomas P Schaer; Sophie Boorman; Dawn M Elliott; Weiliam Chen; George R Dodge; Robert L Mauck; Neil R Malhotra; Lachlan J Smith Journal: Tissue Eng Part A Date: 2020-08-07 Impact factor: 3.845