Synergistic effects of overexpression of BMP-2 and TGF-β3 on osteogenic differentiation of bone marrow mesenchymal stem cells

Abstract

Bone morphogenetic protein 2 (BMP-2) and transforming growth factor β (TGF-β) isoforms are important in advancing bone regeneration. The aim of the present study was to investigate the positive and reciprocal effect of TGF-β3, one of the three TGF-β isoforms, on BMP-2 in promoting osteogenic differentiation. Exogenous BMP-2 and TGF-β3 genes were separately, and in combination, overexpressed in rabbit bone marrow-derived mesenchymal stem cells (rBMSCs). Expression levels of BMP-2 and TGF-β3 were evaluated using reverse-transcription-polymerase chain reaction (RT-PCR) and Western blotting assays. Furthermore, the osteogenic differentiation capacities of BMSCs were assessed by measuring Alizarin Red S staining, an alkaline phosphatase activity assay, and quantification of the osteogenic-specific genes, Runt-related transcription factor 2 (Runx2) and Osterix (Osx). Using lentiviral-mediated transfection, robust co-transfection efficiency of >90% was achieved. RT-PCR and immunoblotting results indicated a marked elevated expression of BMP-2 and TGF-β3 in rBMSCs undergoing co-transfection, compared with transfection with BMP-2 or TGF-β3 alone, indicating that BMP-2 and TGF-β3 are synergistically expressed in rBMSCs. Furthermore, enhanced osteogenic differentiation was observed in rBMSCs co-transfected with BMP-2/TGF-β3. The present study successfully delivered BMP-2 together with TGF-β3 into rBMSCs with high efficiency for the first time. Furthermore, TGF-β3 overexpression was demonstrated to enhance the osteogenic efficacy of BMP-2 in rBMSCs, and vice versa. This provides a potential clinical therapeutic approach for regenerating the function of osseous tissue, and may present a promising strategy for bone defect healing.

Identification of genomic differences among peripheral arterial beds in atherosclerotic and healthy arteries

“…The association of femoral plaques with the presence of bone-related features was illustrated by an enrichment of genes involved in osteoblast differentiation and bone morphogenesis in these plaques. Examples include the high expression levels of transforming growth factor beta 3 (TGFB3) and bone morphogenetic protein 2 (BMP), which have a synergistic effect of osteogenic differentiation 46 . BMP2 kinase (BMP2K), which may impair osteoblast differentiation, had reduced expression in femoral plaques 47 .…”

BMP4 knockdown of NCSCs leads to aganglionosis in the middle embryonic stage

“…However, at present, there is a lack of a genetic mouse models that exhibits HD gene knockdown with unmistakeable implementation. NCSCs originate from cells of the neural crest that migrate in chains as they colonize the embryonic gut, eventually forming the myenteric and submucosal plexus (27)(28)(29). Failure of the neural crest cells to colonize the gut leads to aganglionosis in the sigmoid colon, a pathological condition called Hirschsprung's disease, also known as congenital megacolon, in humans (28,29).…”

“…NCSCs originate from cells of the neural crest that migrate in chains as they colonize the embryonic gut, eventually forming the myenteric and submucosal plexus (27)(28)(29). Failure of the neural crest cells to colonize the gut leads to aganglionosis in the sigmoid colon, a pathological condition called Hirschsprung's disease, also known as congenital megacolon, in humans (28,29). At present, the mechanism associated withthe signalling pathways that adjust NCSCs for migration to the intestinal tract and differentiation in the enteric nervous system remains unclear.…”

“…Therefore, an improved understanding of the signalling pathways regulated by NCSCs and associated with the mechanisms of action is important to investigate the pathogenesis of aganglionosis. BMPs contribute to the largest subgroup of the TGF-β super family and were originally identified by their ability to induce bone development (27,28,31). Additionally, BMPs are expressed in the nervous system through out its differentiation.…”

Relationship between heterotopic ossification and traumatic brain injury

“… [51] revealed that implantation of 0.5–115 μg of partially purified recombinant human BMP-2A resulted in cartilage by Day 7 and bone formation by Day 14. Several studies have shown that BMP2 or BMP4 in combination with other factors, like vascular endothelial growth factor (VEGF) and TGF-β, can greatly promote endochondral bone formation [52] , [53] , [54] . The levels of BMP and its receptor, as well as the expression of its messenger RNA (mRNA), are all upregulated after brain injury [55] , [56] .…”