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Home / Equine Research Bank / BMC Vet Res / The effect of hypoxia on chondrogenesis of equine synovial m...
BMC veterinary research2019; 15(1); 201; doi: 10.1186/s12917-019-1954-1

The effect of hypoxia on chondrogenesis of equine synovial membrane-derived and bone marrow-derived mesenchymal stem cells.

Abstract: Joint injury is extremely common in equine athletes and post-traumatic osteoarthritis (PTOA), a progressive and debilitating disease, is estimated to affect 60% of horses in the USA. The limited potential for intrinsic healing of articular cartilage has prompted intense efforts to identify a cell-based repair strategy to prevent progression of PTOA. Mesenchymal stem cells (MSCs) have the potential to become an ideal source for cell-based treatment of cartilage lesions; however, full chondrogenic differentiation remains elusive. Due to the relatively low oxygen tension in articular cartilage, hypoxia has been proposed as a method of increasing MSC chondrogenesis. The objective of this study was to investigate the effect of hypoxic culture conditions on chondrogenesis in equine synovial membrane-derived MSCs (SM-MSCs) and bone marrow-derived MSCs (BM-MSCs). MSCs were isolated from synovial membrane and bone marrow collected from 5 horses. Flow cytometric analysis was used to assess cell surface marker expression including CD29, CD44, CD90, CD105, CD45, CD-79α, MHCI and MHCII. MSC pellets were cultured in normoxic (21% O) or in hypoxic (5% O) culture conditions for 28 days. Following the culture period, chondrogenesis was assessed by histology, biochemical analyses and gene expression of chondrogenic-related genes including ACAN, COL2b, SOX9, and COL10A1. Results: Both cell types expressed markers consistent with stemness including CD29, CD44, CD90, CD105, and MHCI and were negative for exclusion markers (CD45, CD79α, and MHCII). Although the majority of outcome variables of chondrogenic differentiation were not significantly different between cell types or culture conditions, COL10A1 expression, a marker of chondrocyte hypertrophy, was lowest in hypoxic SM-MSCs and was significantly lower in hypoxic SM-MSCs compared to hypoxic BM-MSCs. Conclusions: Hypoxic culture conditions do not appear to increase chondrogenesis of equine SM-MSCs or BM-MSCs; however, hypoxia may downregulate the hypertrophic marker COL10A1 in SM-MSCs.
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Publication Date: 2019-06-14 PubMed ID: 31200719PubMed Central: PMC6567476DOI: 10.1186/s12917-019-1954-1Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study explored the effect of low-oxygen conditions on cartilage regeneration in mesenchymal stem cells derived from horse synovial membranes and bone marrow. The researchers found that although reduced oxygen did not generally improve cartilage healing, it did lower expression of a marker linked to overgrowth of cartilage cells in synovial-derived stem cells.

Introduction and Objective

  • This study was conducted to find an effective cell-based therapy for the treatment of post-traumatic osteoarthritis in horses.
  • Post-traumatic osteoarthritis (PTOA) is a debilitating condition, affecting around 60% of horses in the United States.
  • The researchers specifically aimed to understand the impact of hypoxic (low oxygen) culture conditions on the chondrogenesis (the process by which cartilage is formed) in two types of horse-originating mesenchymal stem cells namely synovial membrane-derived MSCs (SM-MSCs) and bone marrow-derived MSCs (BM-MSCs).

Research Methodology

  • MSCs were collected from synovial membranes and bone marrow of 5 horses.
  • Cells was tested with flow cytometry to confirm stem cell properties, using a range of cell surface markers such as CD29, CD44, CD90, CD105, MHCI and excluding markers like CD45, CD-79α, MHCII.
  • The MSCs were placed in either normal (21% oxygen) or hypoxic (5% oxygen) conditions for 28 days.
  • After the culture process, chondrogenesis was assessed through histologic, biochemical analyses and gene expression of chondrogenic-related genes like ACAN, COL2b, SOX9, and COL10A1.

Research Findings

  • Both SM-MSCs and BM-MSCs showed similar surface marker expression and thusly confirmed their stemness properties.
  • Most of the outcomes related to chondrogenic differentiation were not significantly different between the cell types or the culture conditions.
  • However, the gene expression of COL10A1, a marker associated with chondrocyte hypertrophy (overgrowth of cartilage cells), was found to be lowest in SM-MSCs cultured under hypoxic conditions. This was statistically lower than the COL10A1 expression in hypoxic BM-MSCs.

Conclusions

  • The findings suggest that hypoxic conditions did not improve cartilage regeneration in horse-derived MSCs. However, hypoxia seemed to play a role in reducing the expression of a marker connected to overgrowth of cartilage cells in SM-MSCs.

Cite This Article

APA
Gale AL, Mammone RM, Dodson ME, Linardi RL, Ortved KF. (2019). The effect of hypoxia on chondrogenesis of equine synovial membrane-derived and bone marrow-derived mesenchymal stem cells. BMC Vet Res, 15(1), 201. https://doi.org/10.1186/s12917-019-1954-1

Publication

BMC veterinary research
ISSN: 1746-6148
NlmUniqueID: 101249759
Country: England
Language: English
Volume: 15
Issue: 1
Pages: 201
PII: 201

Researcher Affiliations

Gale, Alexis L
  • Department of Clinical Studies, New Bolton Center, School of Veterinary Medicine, University of Pennsylvania, Kennett Square, PA, USA.
Mammone, Renata M
  • Department of Clinical Studies, New Bolton Center, School of Veterinary Medicine, University of Pennsylvania, Kennett Square, PA, USA.
Dodson, Michael E
  • Department of Clinical Studies, New Bolton Center, School of Veterinary Medicine, University of Pennsylvania, Kennett Square, PA, USA.
Linardi, Renata L
  • Department of Clinical Studies, New Bolton Center, School of Veterinary Medicine, University of Pennsylvania, Kennett Square, PA, USA.
Ortved, Kyla F
  • Department of Clinical Studies, New Bolton Center, School of Veterinary Medicine, University of Pennsylvania, Kennett Square, PA, USA. kortved@vet.upenn.edu.

MeSH Terms

  • Animals
  • Bone Marrow Cells / cytology
  • Cell Culture Techniques / methods
  • Cell Culture Techniques / veterinary
  • Cell Hypoxia
  • Cells, Cultured
  • Chondrogenesis
  • Horses
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / metabolism
  • Oxygen / metabolism
  • Synovial Membrane / cytology

Grant Funding

  • T35 OD010919 / NIH HHS

Conflict of Interest Statement

The authors declare that they have no competing interests.

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