The optimal concentration of this reagent for MSC bone differentiation is approximately 10 nM, which corresponds to physiologic concentrations [80]. Organic phosphate released after enzymatic hydrolysis of beta glycerol phosphate plays

an important role in matrix mineralization. This free phosphate is usually applied in 5–10 mM concentrations for MSC bone differentiation [81]. Ascorbic acid is a cofactor in the hydroxylation of prolins and lysine moiety of collagen molecules and is an abundant protein in the ECM. This reagent is used in 50–500 μM concentrations [82]. In addition learn more to these osteogenic supplements, there are other osteogenic factors, including BMP-2 and bFGF. Bone formation up to 84% has been reported with the application of human bone marrow-derived MSCs with hydrogel and 10 μg/mL of BMP-2 in a rat calvarial defect model [83]. Another investigation reported significantly greater bone formation with BMP-2- and bFGF-treated human bone marrow-derived MSCs [84]. Platelet rich plasma (PRP) is another known source of various growth factors, namely, platelet-derived growth factor, transforming growth factor-β and vascular endothelial growth factor. The applicability of PRP for the repair of bony defects is well established [85], and several Buparlisib investigators have advocated the use of this product

in combination with MSCs [86] and [87]. Many scaffolds have been used in different MSC-based bone augmentation procedures. At present, no perfect scaffold/carrier for MSC transfer has been developed, and evidence regarding the subject is sparse. Hydroxy apatite (HA), β-tricalcium phosphate (β-TCP) or a mixture of the two are often used as MSC transfer scaffolds [88] and [89]. Several studies have suggested the application of fibrin glue for cell delivery because fibrin glue is a biocompatible tissue adhesive that stabilizes seeded cells and provides an equally distributed population of cells throughout Florfenicol the carrier [87] and [90]. In general,

it is believed that MSCs can be safely cultured in vitro without risk of spontaneous malignant transformation [91]. Stenderup and colleagues cultured several strains of hMSCs from bone marrow at various ages (i.e. aged 18–81 years) until the cells reached their maximal life span without any evidence of transformation [92]. Furthermore, there have been no reports of human trials demonstrating the formation of tumors with culture-expanded hMSCs [93]. On the other hand, concerns have been raised about the safety of MSCs for clinical use, with studies reporting the potential risk of in vitro expanded MSCs developing into tumors on transplantation. In mice, there have been some reports of sarcoma formation by cultured murine MSCs in vitro and in vivo [94], [95] and [96].