Carbon Monoxide-Releasing Molecule-3 Enhances Osteogenic Differentiation of Rat Bone Marrow Mesenchymal Stem Cells via miR-195-5p/Wnt3a Pathway

Abstract

Purpose Bone marrow-derived mesenchymal stem cells (BMSCs) are hopeful in promoting bone regeneration as their pluripotency in differentiation. Our previous study showed that carbon monoxide-releasing molecule-3 (CORM-3) increased the osteogenic differentiation of rat BMSCs in vitro. However, the mechanism remained unclear. MicroRNAs (miRNAs) play a very important role in modulating the osteogenic differentiation of BMSCs. Therefore, we researched the miRNAs involved in CORM-3-stimulated osteogenic differentiation. Methods The CORM-3-stimulated osteogenic differentiation of rat BMSCs was further studied in vivo. Based on the gene sequencing experiment, the rat BMSCs were transfected with miR-195-5p mimics and inhibitor, pcDNA3.1-Wnt3a and Wnt3a siRNA. The osteogenic differentiation of rat BMSCs was measured by quantitative real-time polymerase chain reaction, Western blot and alizarin red staining. Additionally, the targeting relationship between miR-195-5p and Wnt3a was confirmed by the dual-luciferase assay. Results MiR-195-5p was down-expressed during the CORM-3-stimulated osteogenic differentiation of rat BMSCs. CORM-3-stimulated osteogenic differentiation of rat BMSCs was inhibited with miR-195-5p overexpression, evidenced by significantly reduced mRNA and protein expressions of runt-related transcription factor 2 and osteopontin, and matrix mineralization demonstrated. On the contrary, the osteogenic differentiation was enhanced with inhibition of miR-195-5p. CORM-3-stimulated osteogenic differentiation of rat BMSCs was increased by overexpression of Wnt3a, while the opposite was observed in the Wnt3a-deficient cells. Moreover, the decreased osteogenic differentiation capacity by increased expression of miR-195-5p was rescued by Wnt3a overexpression, showing miR-195-5p directly targeted Wnt3a. Conclusion These results demonstrate that CORM-3 promoted osteogenic differentiation of rat BMSCs via miR-195-5p/Wnt3a, which bodes well for the application of CORM-3 in the treatment of periodontal disease and other bone-defect diseases.

Mir-195-5p targets Smad7 regulation of the Wnt/β-catenin pathway to promote osteogenic differentiation of vascular smooth muscle cells

“…Comparisons between the control and model group cells during the period of osteogenic induction by β-GP revealed increases in the expression of miR-195-5p, Wnt3a, and β-catenin over time and a corresponding reduction in the expression of Smad7. The findings of a recent study have similarly indicated that carbon monoxide-releasing molecule 3 promotes osteogenic differentiation of rat BMSCs via miR-195-5p/Wnt3a [ 28 ], whereas other studies have confirmed that miR-195-5p enhances the osteogenic differentiation of cells and is associated with Smad7 [ 29 , 30 ]. These findings accordingly persuaded us to focus our research on the effects of miR-195-5p and Smad7 on the osteogenic differentiation of VSMCs and the Wnt/β-catenin pathway.…”

Gingival Tissue MiRNA Expression Profiling and an Analysis of Periodontitis-Specific Circulating MiRNAs

“…MiR-195-5p has been linked to osteogenic differentiation via Wnt3a. [28,29]. This miRNA has demonstrated anti-inflammatory properties in inflammatory models such as atherosclerosis [30], while it has also been shown to have pro-inflammatory effects in osteoarthritis [31] and ulcerative colitis [32], whereas hsa-miR-146a-5p may promote differentiation in dental stem cells by targeting Traf6 via the NF-κB signaling pathway [33].…”

“…A subgroup of participants with diagnosed RA was formed from patients undergoing rheumatological treatment at the VU Hospital Santaros Clinics. In total, 91 participants diagnosed with RA, who met the ACR/EULAR 2010 rheumatoid arthritis classification criteria, were enrolled in the study and were referred for periodontal status evaluation [29]. A single, qualified researcher rheumatologist (E.P.)…”