Rosa Adalberto L, Beloti Marcio M
Department of Oral and Maxillofacial Surgery, School of Dentistry of Ribeirao Preto, University of Sao Paulo, Brazil.
J Cell Biochem. 2003 Aug 15;89(6):1148-53. doi: 10.1002/jcb.10582.
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.
TAK - 778已被证明在体外和体内模型中均可诱导骨生长。然而，尚无研究评估TAK - 778对人类细胞的影响。因此，本研究的目的是探讨TAK - 778对人骨髓细胞诱导的成骨作用。将细胞以2×10⁴个细胞/孔的细胞密度接种于24孔培养板中，培养基中含有TAK - 778（分别为10⁻⁷、10⁻⁶和10⁻⁵ M）或溶剂。在培养期间，细胞于37℃、5% CO₂和95%空气的湿润环境中孵育。为评估细胞贴壁情况，细胞培养4小时和24小时。培养7天、14天和21天后，评估细胞增殖、细胞活力、总蛋白含量、碱性磷酸酶（ALP）活性和类骨形成情况。数据采用方差分析和邓肯多重极差检验进行比较。TAK - 778不影响细胞贴壁和活力。在所有评估时间段内，TAK - 778均以剂量依赖性方式减少细胞数量。TAK - 778对总蛋白含量、ALP活性和类骨形成的影响呈剂量依赖性增加。目前的结果表明，诸如细胞贴壁等初始细胞事件不受TAK - 778影响，而指示成骨细胞分化的事件，包括细胞增殖减少以及ALP活性和类骨形成增加，均被TAK - 778以时间和剂量依赖性方式增强。这意味着在需要快速恢复生理功能的临床情况下，如骨科和颌面外科手术中，TAK - 778可能是一种促进新骨形成的有用药物。
