TGF-β1 upregulates the expression of hyaluronan synthase 2 and hyaluronan synthesis in culture models of equine articular chondrocytes
10.4142/jvs.2018.19.6.735
- Author:
Siriwan ONGCHAI
1
;
Oraphan SOMNOO
;
Patiwat KONGDANG
;
Siriporn PEANSUKMANEE
;
Siriwan TANGYUENYONG
Author Information
1. Thailand Excellence Center for Tissue Engineering and Stem Cells, Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand.
- Publication Type:Original Article
- Keywords:
chondrocytes;
horses;
hyaluronan;
hyaluronan synthases;
transforming growth factors
- MeSH:
Bisbenzimidazole;
Cartilage;
Cell Proliferation;
Chondrocytes;
Culture Media, Conditioned;
DNA;
Enzyme-Linked Immunosorbent Assay;
Extracellular Matrix;
Gelatin;
Gene Expression;
Horses;
Hyaluronic Acid;
Immunohistochemistry;
Microscopy, Electron, Scanning;
Polymerase Chain Reaction;
Reverse Transcription;
RNA, Messenger;
Transforming Growth Factor beta;
Transforming Growth Factors;
Up-Regulation
- From:Journal of Veterinary Science
2018;19(6):735-743
- CountryRepublic of Korea
- Language:English
-
Abstract:
We investigated the effect of transforming growth factor beta 1 (TGF-β1) on equine hyaluronan synthase 2 (HAS2) 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. HAS2 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 HAS2 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.
