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 (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.

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“…The production of HA is regulated by growth factors, hormones, inflammatory cytokines and mechanical load. Previous study showed that TGF‐β1 can upregulate HAS2 expression and HA synthesis in culture models of equine articular chondrocytes 17 . A study suggested that IL‐1β is a crucial factor in the fragmentation of HA in inflammatory joints.…”

The Effect of Autologous Protein Solution on the Inflammatory Cascade in Stimulated Equine Chondrocytes

“…TGF-β1 is an important growth factor involved in the development, growth and maintenance of articular cartilage, although its role in osteoarthritis has not been completely elucidated ( 22 ). Several studies have shown that TGF-β1 supports chondrogenic differentiation of stem cells ( 23 , 24 ) and upregulates hyaluronan synthesis in cultured equine chondrocytes ( 25 ). However, TGF-β1 has also been implicated in osteophyte formation and synovial fibrosis, mainly in murine studies ( 26 , 27 ).…”

CircHYBID Regulates Hyaluronan Metabolism in Chondrocytes by Promoting the Expression of the hsa-miR-29b-3p Target Gene TGF-β1

“…From the circRNA-miRNA-mRNA network, we found that TGF-β1 was a potential downstream target of hsa-miR-29b-3p ( Figure 6). TGF-β1 has been reported to be closely related to HA metabolism and OA pathogenesis (20). Furthermore, a previous study has described the interaction between hsa-miR-29b-3p and TGF-β1 (21).…”