Abstract
Various miRNAs have been reported to regulate the chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs); however, whether miR-134 plays a role in this biological process remains undetermined. In the present study, we first evaluated the chondrogenic differentiation of BMSCs by Alcian blue staining, and examined the miR-134 expression by quantitative real-time PCR (qRT-PCR) during this process. And miR-134 inhibitor was used to investigate the functions of miR-134 in chondrogenic differentiation of BMSCs by Alcian blue staining, qRT-PCR, and Western blot. Subsequently, the correlation between miR-134 and SMAD6 was assessed via bioinformatics analysis and dual-luciferase reporter assay. Finally, the role of SMAD6 in chondrogenic differentiation of BMSCs was also determined through Alcian blue staining, qRT-PCR, and Western blot. As results showed that miR-134 expression was significantly down-regulated during chondrogenic differentiation, and inhibition of miR-134 obviously promoted chondrogenic differentiation. Dual-luciferase reporter assay indicated that miR-134 could directly target the 3′-UTRs of SMAD6, inhibit miR-134 expression in BMSCs, and up-regulate SMAD6 expression. Moreover, we found that overexpression of SMAD6 significantly promoted chondrogenic differentiation, and that SMAD6-induced promotion of chondrogenic differentiation could be reversed by miR-134 mimics. In conclusion, our findings suggest that miR-134 may act as a negative regulator during chondrogenic differentiation of BMSCs by interacting with SMAD6.
Highlights
The damage to articular cartilage, primarily caused by trauma and excessive mechanical conditions, is the most common clinical disease [1,2]
Since glycosaminoglycan deposition is a critical indicator of cartilage extracellular matrix accumulation, Alcian blue staining was performed to assess chondrogenic differentiation of bone marrow mesenchymal stem cell (BMSC) by examining glycosaminoglycan accumulation at 0, 14, and 21 days
To determine whether miR-134 plays a role in the chondrogenic differentiation of BMSCs, quantitative real-time PCR assay was applied at 0, 14, and 21 days of chondrogenic differentiation to examine miR-134 expression
Summary
The damage to articular cartilage, primarily caused by trauma and excessive mechanical conditions, is the most common clinical disease [1,2]. Articular cartilage damage in severe cases probably leads to osteoarthritis (OA), which subsequently causes disability and chronic pain, producing a significant social and economic burden in the world [3,4,5]. It was reported that the economic costs of OA in the United States were estimated at 15 billion dollars in 2011 alone, making OA the second most expensive disease [6]. The conservative interventions of OA mainly include injection of intra-articular hyaluronic acid, and oral non-steroidal anti-inflammatory drugs; these treatments could only alleviate chronic pain [7,8,9]. It is essential to explore some new effective therapeutic measures for articular cartilage damage
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
