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Journal of veterinary science2018; 19(6); 735-743; doi: 10.4142/jvs.2018.19.6.735

TGF-β1 upregulates the expression of hyaluronan synthase 2 and hyaluronan synthesis in culture models of equine articular chondrocytes.

Abstract: We investigated the effect of transforming growth factor beta 1 (TGF-β1) on equine hyaluronan synthase 2 () gene expression and hyaluronan (HA) synthesis in culture models of articular chondrocytes. Equine chondrocytes were treated with TGF-β1 at different concentrations and times in monolayer cultures. In three-dimensional cultures, chondrocyte-seeded gelatin scaffolds were cultured in chondrogenic media containing 10 ng/mL of TGF-β1. The amounts of HA in conditioned media and in scaffolds were determined by enzyme-linked immunosorbent assays. mRNA expression was analyzed by semi-quantitative reverse transcription polymerase chain reaction. The uronic acid content and DNA content of the scaffolds were measured by using colorimetric and Hoechst 33258 assays, respectively. Cell proliferation was evaluated by using the alamarBlue assay. Scanning electron microscopy (SEM), histology, and immunohistochemistry were used for microscopic analysis of the samples. The upregulation of mRNA levels by TGF-β1 stimulation was dose and time dependent. TGF-β1 was shown to enhance HA and uronic acid content in the scaffolds. Cell proliferation and DNA content were significantly lower in TGF-β1 treatments. SEM and histological results revealed the formation of a cartilaginous-like extracellular matrix in the TGF-β1-treated scaffolds. Together, our results suggest that TGF-β1 has a stimulatory effect on equine chondrocytes, enhancing HA synthesis and promoting cartilage matrix generation.
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Publication Date: 2018-07-25 PubMed ID: 30041292PubMed Central: PMC6265591DOI: 10.4142/jvs.2018.19.6.735Google 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.

The research investigates how the transforming growth factor beta 1 (TGF-β1) affects expression of the hyaluronan synthase 2 gene and hyaluronan synthesis in equine articular chondrocytes, cultivated in both monolayer and three-dimensional cultures. The scientists found that TGF-β1 promotes hyaluronan synthesis and enhances the formation of a cartilage-like matrix in the cultures.

Research Methods

  • The research team conducted their investigation by treating equine chondrocytes (a type of cartilage cell) with various concentrations and durations of TGF-β1.
  • They employed two types of culture models: monolayer cultures and three-dimensional cultures. For the three-dimensional cultures, they used gelatin scaffolds seeded with chondrocytes and cultivated in chondrogenic media containing 10 ng/mL of TGF-β1.
  • To determine the amount of hyaluronan in the media and the scaffolds, they used enzyme-linked immunosorbent assays.
  • The research team further investigated mRNA expression via semi-quantitative reverse transcription polymerase chain reaction.
  • They also measured the uronic acid and DNA content through respective colorimetric and Hoechst 33258 assays.
  • The team evaluated cell proliferation using the alamarBlue assay.
  • Finally, microscopic analysis was made possible using a scanning electron microscope and histological examination.

Findings

  • It was found that the upregulation of the mRNA expression due to TGF-β1 exposure was dependent on the dose and the duration of the treatment.
  • TGF-β1 was reported to enhance the hyaluronan and uronic acid content within the scaffolds.
  • Interestingly, cell proliferation and DNA content were observed to be lower in the presence of TGF-β1 treatment.
  • Scanning electron microscopy and histological results confirmed the formation of a cartilage-like extracellular matrix when the chondrocytes were treated with TGF-β1.
  • In conclusion, it was posited that TGF-β1 has a stimulating effect on equine chondrocytes which leads to enhanced hyaluronan synthesis and an increased generation of a cartilage matrix.

Cite This Article

APA
Ongchai S, Somnoo O, Kongdang P, Peansukmanee S, Tangyuenyong S. (2018). TGF-β1 upregulates the expression of hyaluronan synthase 2 and hyaluronan synthesis in culture models of equine articular chondrocytes. J Vet Sci, 19(6), 735-743. https://doi.org/10.4142/jvs.2018.19.6.735

Publication

Journal of veterinary science
ISSN: 1976-555X
NlmUniqueID: 100964185
Country: Korea (South)
Language: English
Volume: 19
Issue: 6
Pages: 735-743

Researcher Affiliations

Ongchai, Siriwan
  • Thailand Excellence Center for Tissue Engineering and Stem Cells, Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand.
Somnoo, Oraphan
  • Thailand Excellence Center for Tissue Engineering and Stem Cells, Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand.
Kongdang, Patiwat
  • Thailand Excellence Center for Tissue Engineering and Stem Cells, Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand.
Peansukmanee, Siriporn
  • Equine Clinic, Department of Companion Animal and Wildlife Clinic, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai 50100, Thailand.
Tangyuenyong, Siriwan
  • Equine Clinic, Department of Companion Animal and Wildlife Clinic, Faculty of Veterinary Medicine, Chiang Mai University, Chiang Mai 50100, Thailand.

MeSH Terms

  • Animals
  • Cells, Cultured
  • Chondrocytes / drug effects
  • Chondrocytes / metabolism
  • Dose-Response Relationship, Drug
  • Enzyme-Linked Immunosorbent Assay / veterinary
  • Horses
  • Hyaluronan Synthases / metabolism
  • Hyaluronic Acid / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction / veterinary
  • Transforming Growth Factor beta1 / pharmacology
  • Up-Regulation / drug effects
  • Uronic Acids / metabolism

Conflict of Interest Statement

The authors declare no conflicts of interest.

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Citations

This article has been cited 8 times.
  1. Park S, Bello A, Arai Y, Ahn J, Kim D, Cha KY, Baek I, Park H, Lee SH. Functional Duality of Chondrocyte Hypertrophy and Biomedical Application Trends in Osteoarthritis. Pharmaceutics 2021 Jul 26;13(8).
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