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American journal of veterinary research2010; 71(1); 103-114; doi: 10.2460/ajvr.71.1.103

Evaluation of early cellular influences of bone morphogenetic proteins 12 and 2 on equine superficial digital flexor tenocytes and bone marrow-derived mesenchymal stem cells in vitro.

Abstract: To evaluate early cellular influences of bone morphogenetic protein (BMP)12 and BMP2 on equine superficial digital flexor tenocytes (SDFTNs) and equine bone marrow-derived mesenchymal stem cells (BMDMSCs). Methods: 9 adult clinically normal horses. Methods: BMDMSCs and SDFTNs were cultured in monolayer, either untreated or transduced with adenovirus encoding green fluorescent protein, adenovirus encoding BMP12, or adenovirus encoding BMP2. Cytomorphologic, cytochemical, immunocytochemical, and reverse transcriptase-quantitative PCR (RT-qPCR) analyses were performed on days 3 and 6. Genetic profiling for effects of BMP12 was evaluated by use of an equine gene expression microarray on day 6. Results: BMDMSCs and SDFTNs had high BMP12 gene expression and remained viable and healthy for at least 6 days. Type l collagen immunocytochemical staining for SDFTNs and tenocyte-like morphology for SDFTNs and BMDMSCs were greatest in BMP12 cells. Cartilage oligomeric matrix protein, as determined via RT-qPCR assay, and chondroitin sulfate, as determined via gene expression microarray analysis, were upregulated relative to control groups in SDFTN-BMP12 cells. The BMDMSCs and SDFTNs became mineralized with BMP2, but not BMP12. Superficial digital flexor tenocytes responded to BMP12 with upregulation of genes relevant to tendon healing and without mineralization as seen with BMP2. Conclusions: Targeted equine SDFTNs may respond to BMP12 with improved tenocyte morphology and without mineralization, as seen with BMP2. Bone marrow-derived mesenchymal stem cells may be able to serve as a cell delivery method for BMP12.
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Publication Date: 2010-01-02 PubMed ID: 20043789PubMed Central: PMC4246500DOI: 10.2460/ajvr.71.1.103Google Scholar: Lookup
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  • Non-U.S. Gov't

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 examines the early cell responses of bone morphogenetic proteins (BMP12 and BMP2) on horse tendon cells (Superficial Digital Flexor Tenocytes) and bone marrow stem cells (Bone Marrow-Derived Mesenchymal Stem Cells) to support tendon healing.

Study Design

  • The researchers used bone marrow-derived mesenchymal stem cells (BMDMSCs) and the cells from the superficial digital flexor tendon (SDFTNs) of 9 healthy adult horses for their study.
  • These cells were then cultured and altered using adenoviruses that express green fluorescent protein, BMP12 or BMP2.
  • The cells were then analysed using cytomorphologic, cytochemical, immunocytochemical tests and reverse transcriptase-quantitative PCR (RT-qPCR) in a 6 day period.
  • An additional look at the effects of BMP12 on gene expression was assessed using an equine gene expression microarray on the sixth day.

Results

  • Results indicated that both BMDMSCs and SDFTNs consistently expressed high levels of BMP12 and remained healthy and viable over 6 days.
  • It was observed that the staining for type l collagen (a protein found in tendons) was greatest in BMP12 treated cells, while cartilage oligomeric matrix protein (another protein found in tendons) and chondroitin sulfate (an important component of cartilage) were upregulated in SDFTN-BMP12 cells.
  • Importantly, BMP2 resulted in cells becoming mineralized (hardened as would be seen in bone tissue), a process which did not occur with BMP12.
  • Overall, the results indicate that SDFTNs responded better to BMP12 treatment by augmenting gene expression relevant to tendon healing. This happened without the cells undergoing mineralization, which was observed in BMP2 treatments.

Conclusions

  • The findings suggest that bone morphogenetic protein 12 (BMP12) may improve tendon cell structure (tenocyte morphology) without causing them to harden (mineralization).
  • This research suggests that mesenchymal stem cells from bone marrow could be used to deliver BMP12 to promote tendon healing, offering potential therapeutic strategies for various tendon injuries in horses.

Cite This Article

APA
Murray SJ, Santangelo KS, Bertone AL. (2010). Evaluation of early cellular influences of bone morphogenetic proteins 12 and 2 on equine superficial digital flexor tenocytes and bone marrow-derived mesenchymal stem cells in vitro. Am J Vet Res, 71(1), 103-114. https://doi.org/10.2460/ajvr.71.1.103

Publication

American journal of veterinary research
ISSN: 0002-9645
NlmUniqueID: 0375011
Country: United States
Language: English
Volume: 71
Issue: 1
Pages: 103-114

Researcher Affiliations

Murray, Shannon J
  • Comparative Orthopedic Molecular Medicine and Applied Research Laboratory, College of Veterinary Medicine, The Ohio State University, Columbus, OH 43210, USA.
Santangelo, Kelly S
    Bertone, Alicia L

      MeSH Terms

      • Animals
      • Bone Marrow
      • Bone Morphogenetic Protein 2 / genetics
      • Bone Morphogenetic Protein 2 / metabolism
      • Bone Morphogenetic Proteins / genetics
      • Bone Morphogenetic Proteins / metabolism
      • Cell Differentiation
      • Cells, Cultured
      • Gene Expression Regulation
      • Gene Transfer Techniques
      • Horses
      • Mesenchymal Stem Cells / drug effects
      • Protein Array Analysis
      • Tendons / cytology

      Grant Funding

      • F32 AR053805 / NIAMS NIH HHS
      • F32AR053805 / NIAMS NIH HHS
      • KO-8AR4920101A2 / NIAMS NIH HHS

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