Bioactive metabolites have wide-ranging biological activities and are a potential source of future research and therapeutic tools. Here, we use nanovibrational stimulation to induce osteogenic differentiation of mesenchymal stem cells, in the absence of off-target, nonosteogenic differentiation. We show that this differentiation method, which does not rely on the addition of exogenous growth factors to culture media, provides an artifact-free approach to identifying bioactive metabolites that specifically and potently induce osteogenesis. We first identify a highly specific metabolite, cholesterol sulfate, an endogenous steroid. Next, a screen of other small molecules with a similar steroid scaffold identified fludrocortisone acetate with both specific and highly potent osteogenic-inducing activity. Further, we implicate cytoskeletal contractility as a measure of osteogenic potency and cell stiffness as a measure of specificity. These findings demonstrate that physical principles can be used to identify bioactive metabolites and then enable optimization of metabolite potency can be optimized by examining structure-function relationships.

生物活性代谢物具有广泛的生物活性，是未来研究和治疗工具的潜在来源。在这里，我们使用纳米振动刺激来诱导间充质干细胞的成骨分化，在没有脱靶、非成骨分化的情况下。我们表明，这种不依赖于向培养基中添加外源生长因子的分化方法提供了一种无伪影的方法来识别特异性和有效诱导成骨的生物活性代谢物。我们首先确定了一种高度特异性的代谢物，硫酸胆固醇，一种内源性类固醇。接下来，筛选了具有类似类固醇支架的其他小分子，确定了醋酸氟氢可的松具有特异性和高效的成骨诱导活性。更远，我们将细胞骨架收缩性作为成骨效力的衡量标准，将细胞刚度作为特异性的衡量标准。这些发现表明，物理原理可用于识别生物活性代谢物，然后可以通过检查结构-功能关系来优化代谢物效力。