Background

The treatment of bone defects has always been a problem for clinicians. In recent years, research on human bone mesenchymal stem cells (hBMSCs) has found that promoting their osteogenic differentiation could be a useful therapeutic strategy for bone healing. Previous studies have been reported that Withania somnifera Dunal inhibits osteoclastogenesis by inhibiting the NF-κB signaling pathway. Withanolide B is an active component of W. somnifera Dunal, but its role in osteogenic differentiation of hBMSCs remains unknown. Here, we performed a preliminary study on the role of Withanolide B in promoting osteogenic differentiation and its possible mechanism.

Methods

We investigated the effect of Withanolide B on osteogenic differentiation of hBMSCs in vitro and in vivo. The effect of Withanolide B on the activity of hBMSCs was verified by CCK-8 assay and quantitative Real-time polymerase chain reaction (qPCR) and Western blotting analysis were used to verify the effect of Withanolide B on osteogenic differentiation-specific genes and proteins. The effect of Withanolide B on ALP activity and mineral deposition was verified by ALP and ARS staining. We then used a rat tibial osteotomy model to observe the effect of Withanolide B on bone healing.

Results

Withanolide B is noncytotoxic to hBMSCs and can effectively promote their osteogenic differentiation. Moreover, we found that Withanolide B can regulate the osteogenic differentiation of hBMSCs through the ERK1/2 and Wnt/β-catenin signaling pathways. When inhibitors of the ERK1/2 and Wnt/β-catenin signaling pathways were used, the enhancement of osteogenic differentiation induced by Withanolide B was attenuated. Withanolide B also effectively promoted bone healing in the rat tibial osteotomy model.

Conclusions

Our results suggest that Withanolide B can promote the osteogenic differentiation of hBMSCs through the ERK1/2 and Wnt/β-catenin signaling pathways and can effectively promote bone defect healing.

中文翻译：

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Withanolide B通过ERK1 / 2和Wnt /β-catenin信号通路促进人骨髓间充质干细胞的成骨分化。

背景

骨缺损的治疗一直是临床医生面临的问题。近年来，对人骨间充质干细胞（hBMSCs）的研究发现，促进其成骨分化可能是一种有效的骨愈合治疗策略。已有研究报道，Withania Smnifera Dunal通过抑制NF-κB信号通路来抑制破骨细胞生成。Withanolide B是W. somnifera Dunal的活性成分，但其​​在hBMSCs成骨分化中的作用仍然未知。在这里，我们对Withanolide B在促进成骨分化中的作用及其可能的机制进行了初步研究。

方法

我们调查了Withanolide B在体外和体内对hBMSCs成骨分化的影响。通过CCK-8试验验证了Withanolide B对hBMSCs活性的影响，并通过定量实时聚合酶链反应（qPCR）和蛋白质印迹分析来验证Withanolide B对成骨分化特异性基因和蛋白质的影响。通过ALP和ARS染色验证了Withanolide B对ALP活性和矿物质沉积的影响。然后，我们使用大鼠胫骨截骨模型观察Withanolide B对骨愈合的影响。

结果

Withanolide B对hBMSC无细胞毒性，可有效促进其成骨分化。此外，我们发现Withanolide B可以通过ERK1 / 2和Wnt /β-catenin信号通路调节hBMSC的成骨分化。当使用ERK1 / 2和Wnt /β-catenin信号通路的抑制剂时，Withanolide B诱导的成骨分化增强作用减弱。在大鼠胫骨截骨模型中，Withanolide B还可以有效促进骨愈合。

结论

我们的结果表明，Withanolide B可以通过ERK1 / 2和Wnt /β-catenin信号通路促进hBMSC的成骨分化，并可以有效地促进骨缺损的愈合。