Overexpression Effects of miR‐424 and BMP2 on the Osteogenesis of Wharton’s Jelly‐Derived Stem Cells
Asghar Fallah, Mahdieh Alipour, Zahra Jamali, Akbar Farjadfar, Leila Roshangar, Minoo Partovi Nasr, Parisa Hashemi, Marziyeh Aghazadeh
Abstract
Recently, the translational application of noncoding RNAs is accelerated dramatically. In this regard, discovering therapeutic roles of microRNAs by developing synthetic RNA and vector‐based RNA is attracting attention. Here, we studied the effect of BMP2 and miR‐424 on the osteogenesis of Wharton’s jelly‐derived stem cells (WJSCs). For this purpose, human BMP2 and miR‐424 DNA codes were cloned in the third generation of lentiviral vectors and then used for HEK‐293T cell transfection. Lentiviral plasmids contained miR424, BMP‐2, miR424‐BMP2, green fluorescent protein (GFP) genes, and helper vectors. The recombinant lentiviral particles transduced the WJSCs, and the osteogenesis was evaluated by real‐time PCR, Western blot, Alizarin Red staining, and alkaline phosphatase enzyme activity. According to the results, there was a significant increase in the expression of the BMP2 gene and secretion of Osteocalcin protein in the group of miR424‐BMP2. Moreover, the amount of dye deposition in Alizarin Red staining and alkaline phosphatase activity was significantly higher in the mentioned group ( p < 0.05). Thus, the current study results clarify the efficacy of gene therapy by miR424‐BMP2 vectors for bone tissue engineering. These data could help guide the development of gene therapy‐based protocols for bone tissue engineering.