MicroRNAs (miRNAs) are small non-coding endogenous RNAs which control tissue development by regulating gene expression. So, miRNAs could be used in regenerative medicine to get optimal function of heart and bone tissues. To check the role of miR-148b and miR-499a on osteogenesis and cardiogenesis, respectively, we overexpressed these miRNAs in bone-marrow derived mesenchymal stem cells (BM-MSCs). Here in, the sequences of hsa-miR-148b (-3p and -5p) as well as hsa-miR-499a (-3p and -5p) were designed and cloned to lentiviral vectors. After the successful insertion of desired transgenes, related lentiviral particles were produced. Then, characterized BM-MSCs were transduced with them and allowed to differentiate towards osteogenic and cardiogenic development. At the end of three weeks, treated cells were subjected to differential tests including cellular staining, immunostaining as well as Western blotting for expression of protein hall markers. For the first time we had shown that ectopic expression of miR-148b-3p and miR-499a-5p raised osteogenesis and cardiogenesis of BM-MSCs, respectively. Increased osteoblastogenesis was confirmed by enhancement in mineralization and alkaline phosphatase (ALP) activity in miR-148b-3p overexpressed cells. In support of these data, the expressions of ALP and collagen type I (COLI) were dramatically up-regulated as it was shown by immunostaining and Western blotting. Besides, the expression of cardiac troponin T (cTnT) and α-actinin proteins were promoted in miR-499a- 5p infected cells. Since lentivirally overexpression of miR-148b-3p and miR-499a-5p could drive differentiation of osteoblasts and cardiomyocytes, these molecules might be applicable candidates as therapeutic biomarkers to improve heart and bone development.

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