Background. Oral and maxillofacial bone loss is highly prevalent among populations, and nowadays, increased attention has been focused on dentin derivatives serving as desirable graft materials for bone regeneration. In this study, dentin-derived inorganic mineral (DIM) was fabricated with a high-temperature calcination technique and the effects of DIM on the osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMMSCs) and the bone formation were elucidated. Methods. The effects of DIM on BMMSC proliferation and apoptosis capacity were evaluated by CCK-8, flow cytometry, and EdU assays. Alkaline phosphatase (ALP) activity detection, ALP staining, alizarin red staining, and osteogenic marker expression analysis were performed to investigate the influence of DIM on the osteogenic differentiation of BMMSCs, as well as the relevant signal mechanisms. The model of critical-sized defects in the calvarium of rats was constructed for exploring the in vivo efficiency of DIM on bone regeneration. Results. Cell viability assays indicated that DIM had no cytotoxicity. BMMSCs cultured with DIM presented a higher level of osteogenic differentiation ability than those in the control group. The activation in ERK and p38 signals was detected in DIM-treated BMMSCs, and both pathways and osteogenic process were suppressed while using ERK inhibitor U0126 and p38 inhibitor SB203580, respectively. Furthermore, the animal experiments revealed that DIM could dramatically enhance new bone formation compared to the control group. Conclusion. DIM could promote BMMSC osteogenic differentiation via triggering the ERK and p38 MAPK signaling pathways and might be a novel predictable material for facilitating bone formation.

中文翻译：

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牙本质来源的无机矿物质促进骨髓来源的间充质干细胞的成骨：骨再生的潜在应用

背景。口腔颌面部骨丢失在人群中非常普遍，如今，越来越多的注意力集中在牙本质衍生物作为骨再生的理想移植材料上。本研究采用高温煅烧技术制备牙本质衍生无机矿物质（DIM），并阐明 DIM 对骨髓间充质干细胞（BMMSCs）成骨分化和骨形成的影响。方法。通过 CCK-8、流式细胞术和 EdU 分析评估 DIM 对 BMMSC 增殖和凋亡能力的影响。采用碱性磷酸酶（ALP）活性检测、ALP染色、茜素红染色、成骨标志物表达分析，探讨DIM对BMMSCs成骨分化的影响及相关信号机制。建立大鼠颅骨临界尺寸缺损模型，探索DIM在体内对骨再生的作用。结果. 细胞活力测定表明 DIM 没有细胞毒性。与对照组相比，用 DIM 培养的 BMMSCs 具有更高水平的成骨分化能力。在 DIM 处理的 BMMSCs 中检测到 ERK 和 p38 信号的激活，分别使用 ERK 抑制剂 U0126 和 p38 抑制剂 SB203580 抑制了通路和成骨过程。此外，动物实验表明，与对照组相比，DIM 可以显着增强新骨的形成。结论。DIM 可通过触发 ERK 和 p38 MAPK 信号通路促进 BMMSC 成骨分化，可能是促进骨形成的新型可预测材料。