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Home / Equine Research Bank / Mol Biol Rep / Induction of tenogenic differentiation of equine adipose-der...
Molecular biology reports2020; 47(9); 6855-6862; doi: 10.1007/s11033-020-05742-7

Induction of tenogenic differentiation of equine adipose-derived mesenchymal stem cells by platelet-derived growth factor-BB and growth differentiation factor-6.

Abstract: Managing tendon healing process is complicated mainly due to the limited regeneration capacity of tendon tissue. Mesenchymal stem cells (MSCs) have potential applications in regenerative medicine and have been considered for tendon repair and regeneration. This study aimed to evaluate the capacity of equine adipose tissue-derived cells (eASCs) to differentiate into tenocytes in response to platelet-derived growth factor-BB (PDGF-BB) and growth differentiation factor-6 (GDF-6) in vitro. Frozen characterized eASCS of 3 mares were thawed and the cells were expanded in basic culture medium (DMEM supplemented with 10% FBS). The cells at passage 5 were treated for 14 days in different conditions including: (1) control group in basic culture medium (CM), (2) induction medium as IM (CM containing L-prolin, and ascorbic acid (AA)) supplemented with PDGF-BB (20 ng/ml), (3) IM supplemented with GDF-6 (20 ng/ml), and (4) IM supplemented with PDGF-BB and GDF-6. At the end of culture period (14th day), tenogenic differentiation was evaluated. Sirius Red staining was used to assess collagen production, and H&E was used for assessing cell morphology. mRNA levels of collagen type 1 (colI), scleraxis (SCX), and Mohawk (MKX), as tenogenic markers, were analyzed using real-time reverse-transcription polymerase chain reaction (qPCR). H&E staining showed a stretching and spindle shape (tenocyte-like) cells in all treated groups compared to unchanged from of cells in control groups. Also, Sirius red staining data showed a significant increase in collagen production in all treated groups compared with the control group. MKX expression was significantly increased in PDGF-BB and mixed groups and COLI expression was significantly increased only in PDGF-BB group. In conclusion, our results showed that PDGF-BB and GDF-6 combination could induce tenogenic differentiation in eASCs. These in vitro findings could be useful for cell therapy in equine regenerative medicine.
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Publication Date: 2020-09-01 PubMed ID: 32875433PubMed Central: 3875481DOI: 10.1007/s11033-020-05742-7Google Scholar: Lookup
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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 research is aimed at studying how mesenchymal stem cells derived from equine adipose tissue react to specific growth factors (platelet-derived growth factor-BB and growth differentiation factor-6) and how those reactions may be employed in the repair and regeneration of tendons in equine veterinary medicine.

Introduction and Research Objective

  • Equine tendons are often challenging to heal due to their limited regenerative capabilities. The study aimed at investigating new mechanisms to improve this reparative process.
  • The primary objective of the study was to assess how equine adipose tissue-derived cells (eASCs) responded to two specific growth factors. The hypothesis was that these growth factors could promote these cells to transform into tenocytes (tendon cells), thereby potentially assisting tendon regeneration.

Methods

  • The researchers used eASCs derived from three mares, and these cells were cultured in a basic medium composed of DMEM (a type of cell culture media) with added FBS (Fetal Bovine Serum- a common growth supplement).
  • The resultant cells were subjected to different conditions including: a control group with no additives, one group supplemented with platelet-derived growth factor-BB (PDGF-BB), one with growth differentiation factor-6 (GDF-6), and one with both PDGF-BB and GDF-6.
  • At the end of the 14-day culture period, assessments were made regarding the potential for tenogenic differentiation (transformation into tendon cells). This was evaluated using a few key indicators, including specific staining techniques to view collagen production and cell morphology.
  • Real-time reverse transcription polymerase chain reaction, or qPCR, was also employed to analyze the mRNA for specific markers of tenogenic differentiation, namely collagen type 1 (colI), scleraxis (SCX), and Mohawk (MKX). These are proteins whose expression is associated with tenocyte differentiation and could provide key insights into the treatment’s success.

Results and Conclusion

  • Analysis revealed that all treated groups developed more elongated, tenocyte-like cell structures and increased collagen production compared to the control group. These results indicate successful tenogenic differentiation. However, the group that was exposed to both growth factors showed the most pronounced results.
  • Further qPCR analysis indicated that the combined treatment of PDGF-BB and GDF-6 resulted in a significant increase in the expression of tenogenic markers, suggesting successfully induced differentiation into tenocytes.
  • The study therefore concluded that the combination of PDGF-BB and GDF-6 can indeed induce tenogenic differentiation of eASCs. These findings could potentially offer valuable insights for the implementation of cell therapy techniques in equine regenerative medicine, particularly concerning tendon repair.

Cite This Article

APA
Javanshir S, Younesi Soltani F, Dowlati G, Parham A, Naderi-Meshkin H. (2020). Induction of tenogenic differentiation of equine adipose-derived mesenchymal stem cells by platelet-derived growth factor-BB and growth differentiation factor-6. Mol Biol Rep, 47(9), 6855-6862. https://doi.org/10.1007/s11033-020-05742-7

Publication

Molecular biology reports
ISSN: 1573-4978
NlmUniqueID: 0403234
Country: Netherlands
Language: English
Volume: 47
Issue: 9
Pages: 6855-6862

Researcher Affiliations

Javanshir, Shabnam
  • Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.
Younesi Soltani, Fatemeh
  • Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.
Dowlati, Gholamreza
  • Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.
Parham, Abbas
  • Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran. parham@um.ac.ir.
  • Stem Cell Biology and Regenerative Medicine Research Group, Research Institute of Biotechnology, Ferdowsi University of Mashhad, Azadi Square, Mashhad, 9177948974, Iran. parham@um.ac.ir.
Naderi-Meshkin, Hojjat
  • Stem Cells and Regenerative Medicine Research Group, Iranian Academic Center for Education, Culture and Research (ACECR), Khorasan Razavi Branch, Mashhad, Iran.

MeSH Terms

  • Animals
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / metabolism
  • Becaplermin / pharmacology
  • Cell Differentiation / genetics
  • Collagen Type I / genetics
  • Collagen Type I / metabolism
  • Female
  • Gene Expression Regulation, Developmental / drug effects
  • Gene Expression Regulation, Developmental / genetics
  • Growth Differentiation Factor 6 / pharmacology
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism
  • Horses
  • Mesenchymal Stem Cells / metabolism
  • Real-Time Polymerase Chain Reaction
  • Tendons / cytology
  • Tendons / metabolism
  • Tissue Engineering / methods

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Citations

This article has been cited 3 times.
  1. Rehman A, Nigam A, Laino L, Russo D, Todisco C, Esposito G, Svolacchia F, Giuzio F, Desiderio V, Ferraro G. Mesenchymal Stem Cells in Soft Tissue Regenerative Medicine: A Comprehensive Review.. Medicina (Kaunas) 2023 Aug 10;59(8).
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  2. Chen Y, Jiang L, Lyu K, Lu J, Long L, Wang X, Liu T, Li S. A Promising Candidate in Tendon Healing Events-PDGF-BB.. Biomolecules 2022 Oct 20;12(10).
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  3. Younesi Soltani F, Javanshir S, Dowlati G, Parham A, Naderi-Meshkin H. Differentiation of human adipose-derived mesenchymal stem cells toward tenocyte by platelet-derived growth factor-BB and growth differentiation factor-6.. Cell Tissue Bank 2022 Jun;23(2):237-246.
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