A Novel Human TGF-β1 Fusion Protein in Combination with rhBMP-2 Increases Chondro-Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells

Abstract

Transforming growth factor-beta (TGF-β) is involved in processes related to the differentiation and maturation of osteoprogenitor cells into osteoblasts. Rat bone marrow (BM) cells were cultured in a collagen-gel containing 0.5% fetal bovine serum (FBS) for 10 days in the presence of rhTGF (recombinant human TGF)-β1-F2, a fusion protein engineered to include a high-affinity collagen-binding decapeptide derived from von Willebrand factor. Subsequently, cells were moderately expanded in medium with 10% FBS for 4 days and treated with a short pulse of rhBMP (recombinant human bone morphogenetic protein)-2 for 4 h. During the last 2 days, dexamethasone and β-glycerophosphate were added to potentiate osteoinduction. Concomitant with an up-regulation of cell proliferation, DNA synthesis levels were determined. Polymerase chain reaction was performed to reveal the possible stemness of these cells. Osteogenic differentiation was evaluated in terms of alkaline phosphatase activity and mineralized matrix formation as well as by mRNA expression of osteogenic marker genes. Moreover, cells were placed inside diffusion chambers and implanted subcutaneously into the backs of adult rats for 4 weeks. Histological study provided evidence of cartilage and bone-like tissue formation. This experimental procedure is capable of selecting cell populations from BM that, in the presence of rhTGF-β1-F2 and rhBMP-2, achieve skeletogenic potential in vitro and in vivo.

Glycitin regulates osteoblasts through TGF-β or AKT signaling pathways in bone marrow stem cells

“…These proteins have important roles in cell proliferation, differentiation, formation of cell matrix, the formation of tissues and organs, During bone formation, the combination of TGF-β and its ligand may inhibit the formation of osteoclasts and bone absorption, but may additionally facilitate the osteogenesis of osteoblasts (23). Recent research has shown that TGF-β1 may promote the osteogenic differentiation of hMSCs and induce the expression of osteogenesis genes, ALP, collagen type I and osteocalcins (24). Conversely, the addition of TGF-β3 may inhibit the expression of ALP, suggesting that the TGF-β signaling pathway has a different role in the osteogenic differentiation of hMSCs (25).…”

Soluble Factors on Stage to Direct Mesenchymal Stem Cells Fate

“…Moreover, in vitro TGFβ1 drives MSCs fate toward osteoblasts generation and inhibits adipogenic differentiation; accordingly, TGFβ1 induces the switching from adipogenesis to osteogenesis when added to an adipogenic medium, acting mainly on the SMAD/C/EBPs/PPARγ signaling (Choy and Derynck, 2003 ; van Zoelen et al, 2016 ). In other reports, during in vitro MSC expansion, TGFβ1 reduces the number of osteoprogenitor cells and limits their expansion inducing a rapid terminal differentiation, suggesting that TGFβ1 effects on MSCs might depend on the commitment state of the cells (Walsh et al, 2003 ; Claros et al, 2014 ). Furthermore, TGFβ1 is a key molecule in chondrogenesis by stabilizing SOX9 via the canonical SMAD or the non-canonical p38 pathways (Coricor and Serra, 2016 ; Dexheimer et al, 2016 ).…”

Safety and efficacy of the mesenchymal stem cell in feline eosinophilic keratitis treatment

“…To assess the multipotentiality, fAd-MSCs at passage 2 were differentiated long adipogenic, osteogenic, and chondrogenic lineages according to standard protocols, as previously reported [ 26 , 27 ]. Cells were cultured in specific induction medium for 21 days.…”