Overexpression Effects of miR-424 and BMP2 on the Osteogenesis of Wharton’s Jelly-Derived Stem Cells

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.

Effect of lentivirus‐mediatedBMP2from autologous tooth on the proliferative and osteogenic capacity of human periodontal ligament cells

“…In recent years, many studies [46][47][48] This study confirms that BMP2 has a significant ability to promote the proliferation and osteogenic differentiation of hPDLCs.…”

“…In recent years, many studies 46–48 have shown that BMP2 plays an important role in tissue engineering and can significantly promote the osteogenic differentiation of stem cells. On the basis of previous studies, this study selected hPDLCs to explore the role of BMP2 in promoting osteogenesis.…”

miRNA-Guided Regulation of Mesenchymal Stem Cells Derived from the Umbilical Cord: Paving the Way for Stem-Cell Based Regeneration and Therapy

“…miR-216a was found capable of reversing the inhibiting effects of dexamethasone on the osteogenic process, while its expression levels were shown to positively correlate with osteoblastic differentiation and bone formation in adipose tissue-derived mesenchymal stem cells (ATMSCs) and UCMSCs [35]. Furthermore, it was demonstrated that an ex vivo engineered vector combining miR-424 and the bone morphogenetic protein-2 (BMP2) encoding gene has the capacity to promote osteogenesis in WJMSCs, with miR-424 potentially leading to the overexpression of OCN and BMP2 protein levels, the latter of which is considered an important regulator of the osteogenic/osteoblastic differentiation, and ultimately bone development [26].…”

The Molecular Regulatory Mechanism in Multipotency and Differentiation of Wharton’s Jelly Stem Cells

“…Furthermore, RUNX2 can mediate the BMP-2-induced expression of OSX [ 113 ]. BMP2 is regulated by the downstream target genes WISP1 [ 114 ] and miR424 [ 115 ] of the Wnt signaling pathway, which enhances its induction of OSX. OSX is another important regulator of osteogenic differentiation.…”