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In vitro cellular & developmental biology. Animal2025; doi: 10.1007/s11626-025-01049-8

In vitro model of equine cartilage degradation; using cartilage pellets differentiated from bone marrow-derived mesenchymal stem cells.

Abstract: The self-renewal capacity of chondrocytes in osteoarthritis (OA) joints is limited, and mesenchymal stem cells (MSCs) are crucial in disease treatment. This study established an OA model from equine bone marrow-derived mesenchymal stem cells (eBMSCs). The eBMSCs were cultured and differentiated into chondrocytes to generate cartilage pellets, which were induced for 7 d with inflammatory cytokines, interleukin-1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) to mimic OA conditions. Treated culture medium was collected to estimate enzyme activity (MMP-2, MMP-3, and MMP-9) using zymography, and the cartilage pellets were collected to estimate both anabolic gene (COL2A1) and catabolic gene expression (MMP2, MMP3, and MMP9) using qRT-PCR. Cartilage degradation was observed when induced with IL-1β + TNF-α on cartilage pellets. IL-1β + TNF-α decreased the expression levels of COL2A1 and MMP2 genes, and enhanced their enzymatic activities, while Alcian blue-positive glycosaminoglycan in cartilage pellets induced by IL-1β + TNF-α groups decreased. These results suggested that IL-1β + TNF-α induced on cartilage pellets from eBMSCs could be used as an in vitro OA model in horses.
© 2025. The Society for In Vitro Biology.
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Publication Date: 2025-05-27 PubMed ID: 40425901PubMed Central: 9197312DOI: 10.1007/s11626-025-01049-8Google 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 study focused on creating an in vitro model of osteoarthritis in horses by differentiating equine bone marrow-derived mesenchymal stem cells (eBMSCs) into chondrocytes to generate cartilage pellets, which were then induced with specific cytokines to mimic the conditions of the disease.

Creation of Cartilage Pellets from eBMSCs

  • The researchers initiated the study by culturing eBMSCs and subsequently differentiating them into chondrocytes.
  • These chondrocytes were then used to develop cartilage pellets, which formed the basis of their osteoarthritis (OA) model.

Induction of OA Conditions in Cartilage Pellets

  • The research team exposed the cartilage pellets to the inflammatory cytokines interleukin-1 beta (IL-1β) and tumor necrosis factor-alpha (TNF-α).
  • This exposure lasted for 7 days and was intended to simulate the conditions of osteoarthritis within the cartilage pellets.

Assessment of Enzyme Activity and Gene Expression

  • After treating the cartilage pellets with the cytokines, the team then assessed enzyme activity by evaluating the levels of MMP-2, MMP-3, and MMP-9 proteins through a process called zymography.
  • They also examined the expression of both anabolic and catabolic genes within the cartilage pellets to further understand the impacts of the cytokine treatment.

Observations and Findings

  • The researchers observed cartilage degradation in the pellets that were induced with IL-1β and TNF-α, signalling the effective modelling of osteoarthritis.
  • They also found that the cytokine treatment decreased the expression levels of the COL2A1 and MMP2 genes but increased their enzymatic activities.
  • Additionally, the levels of Alcian blue-positive glycosaminoglycan, a component of cartilage, decreased in the cartilage pellets that were induced by the cytokines.

Conclusion

  • The results of the study suggest that this method of inducing osteoarthritis within cartilage pellets created from eBMSCs can serve as an in vitro model for studying the disease in horses.
  • Through this model, researchers may gain a better understanding of osteoarthritis and develop more effective treatments in the future.

Cite This Article

APA
Euppayo T, Siengdee P, Limlenglert P, Nganvongpanit K, Watanabe G, Kasashima Y, Arai K. (2025). In vitro model of equine cartilage degradation; using cartilage pellets differentiated from bone marrow-derived mesenchymal stem cells. In Vitro Cell Dev Biol Anim. https://doi.org/10.1007/s11626-025-01049-8

Publication

In vitro cellular & developmental biology. Animal
ISSN: 1543-706X
NlmUniqueID: 9418515
Country: Germany
Language: English

Researcher Affiliations

Euppayo, Thippaporn
  • Faculty of Veterinary Medicine, Maejo University, Chiang Mai, 50300, Thailand. thippaporn_eu@mju.ac.th.
Siengdee, Puntita
  • Program in Applied Biological Sciences: Environmental Health, Chulabhorn Graduate Institute, Kamphaeng Phet 6 Road, Laksi, Bangkok, 10210, Thailand.
Limlenglert, Pakorn
  • Faculty of Veterinary Medicine, Maejo University, Chiang Mai, 50300, Thailand.
Nganvongpanit, Korakot
  • Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai, 50100, Thailand.
Watanabe, Gen
  • Laboratory of Veterinary Reproduction, Cooperative Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Tokyo, 183-8509, Japan.
Kasashima, Yoshinori
  • Equine Research Institute, Japan Racing Association, Tochigi, 329-0412, Japan.
Arai, Katsuhiko
  • Department of Tissue Physiology, Tokyo University of Agriculture and Technology, Tokyo, 183-8509, Japan.

Conflict of Interest Statement

Declarations. Conflict of interest: The authors declare no competing interests.

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