TAK‐778 enhances osteoblast differentiation of human bone marrow cells

Abstract

TAK-778 has been shown to induce bone growth in in vitro and in vivo models. However, there are no studies evaluating the effect of TAK-778 on human cells. Thus, the aim of this study was to investigate osteogenesis induced by TAK-778 on human bone marrow cells. Cells were cultured in 24-well culture plates at a cell density of 2 x 10(4) cells/well in culture medium containing TAK-778 (10(-7), 10(-6), and 10(-5) M, each) or vehicle. During the culture period, cells were incubated at 37 degrees C in a humidified atmosphere of 5% CO(2) and 95% air. For attachment evaluation, cells were cultured for 4 and 24 h. After 7, 14, and 21 days, cell proliferation, cell viability, total protein content, alkaline phosphatase (ALP) activity, and bone-like formation were evaluated. Data were compared by ANOVA and Duncan's multiple range test. TAK-778 did not affect cell attachment and viability. Cell number was reduced by TAK-778 in all time period evaluated in a dose-dependent way. The effect of TAK-778 on total protein content, ALP activity and bone-like formation was a dose-dependent increase. The present results suggest that initial cell events such as cell attachment are not affected by TAK-778 while events that indicate osteoblast differentiation including reduced cell proliferation, and increased both ALP activity and bone-like formation are enhanced by TAK-778 in a time and dose-dependent way. It means that TAK-778 could be a useful drug to enhance new bone formation in clinical situations that require rapid restoration of physiologic function, such as orthopedic and maxillofacial surgery.

Effects of arachidonic acid and docosahexaenoic acid on differentiation and mineralization of MC3T3‐E1 osteoblast‐like cells

“…21,22,32 Osteogenic supplemented medium contains ascorbic acid and ß-glycerophosphate to stimulate greater collagen production and cross-linking. 33 To enhance osteogenesis further, the synthetic glucocorticoid dexamethasone is often added.…”

“…The extent of mineralization depends on the osteoblastic cell type as well as culture conditions. 21,22,32,40 MC3T3-E1 murine osteoblast-like cells used in this study are considered immediate osteoblast precursors as these cells constitutively express high levels of ALP and calcify in basal conditions. 30,47 It has been shown by Igarishi et al that the ALP activity of control MC3T3-E1 cells increases after seeding, reaches a maximum on day 15, and decreases thereafter up to day 21.…”

“…The final phase of osteoblast development is characterized by 19,20 Once mineralization starts, ALP activity decreases significantly. 21,22 Watkins et al have demonstrated that MC3T3-E1 cells, a murine osteoblast-like cell line, exposed to EPA (n-3 PUFA) demonstrated increased ALP activity compared with cells exposed to AA (n-6 PUFA). 2 This observation might be due to modulation of prostaglandin synthesis.…”

“…23,25,28 Results from our laboratory have shown that AA (n-6 PUFA) and DHA (n-3 PUFA), at physiological concentrations (2.5-20 mg ml À1 ), inhibit osteoblast cell proliferation in a dose-dependent manner. 29 Since there is a reciprocal relationship between reduced proliferation and subsequent induction of cell differentiation in vitro, 20,21,30 the effects of AA and DHA on osteogenesis was investigated.…”

<b>Analysis of indicators of osteogenesis, cytotoxicity and genotoxicity of an experimental β-TCP compared to other bone substitutes

“…The methodology used was based on previous studies that also examined the behavior of bone cells in vitro (Beloti, Martins, Xavier, & Rosa, 2008;Oliva et al, 2009;Quan et al, 2013;Rosa & Beloti, 2003Simão et al, 2007).…”