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.
Highlights
The skeleton microstructure is made up of mineralized extracellular matrix and bone remodeling units, including osteocytes, osteoblasts, osteoclasts, and lining cells
The synthesized BMP2-miR424 genes were cloned under a strong human cytomegalovirus promoter (CMV)
We demonstrated that an ex vivo combination of coding gene and microRNA gene therapy increased the rate of osteogenesis
Summary
The skeleton microstructure is made up of mineralized extracellular matrix and bone remodeling units, including osteocytes, osteoblasts, osteoclasts, and lining cells. Osteoblastic differentiation of stem cells is regulated by different genes that are coding several secretive molecules and transcription factors, including bone morphogenetic proteins (BMPs), Wnt proteins, Indian hedgehog homolog (IHH), and fibroblast growth factors (FGFs). The role of this miRNA was evaluated in osteogenesis According to these studies, upregulation of miR-322 (424), miR-34a, and miR10a expression follows by the overexpression of runt-related transcription factor-2 (Runx-2), osterix (Osx), and osteocalcin (OCN). Upregulation of miR-322 (424), miR-34a, and miR10a expression follows by the overexpression of runt-related transcription factor-2 (Runx-2), osterix (Osx), and osteocalcin (OCN) These complicated signaling pathways regulate bone formation. MiR-322 is reported as a regulator in osteoblast differentiation Overexpression of this microRNA enhances the BMP-2 response in bone marrow stem cells [43]. This study provides more evidence for a combination of coding and noncoding gene therapy and helps develop innovative gene therapy studies using novel biomaterials and advanced bioprinting techniques in bone regeneration
Sign-in/Register to view highlights & summary 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.
