Background

In recent years, magnesium (Mg) has been extensively studied for manufacturing biodegradable orthopedic devices. Besides other advantages, researches have shown that magnesium-based implants can stimulate osteogenesis thus accelerating orthopedic trauma recovery, but its molecular mechanism is not fully understood. Meanwhile, long non-coding RNA (lncRNA) has been found to play vital role in regulating osteogenic differentiation.

Objective

To explore the role of lncRNA in Mg²⁺ (magnesium ions)-induced osteogenesis.

Methods

The effect of Mg²⁺ on mBMSCs proliferation was detected by the CCK-8 assay. The optimum concentration of Mg²⁺ (7.5 mM) in promoting mBMSCs osteogenesis was determined by ALP staining and Alizarin red staining, western blot and RT-qPCR were performed to detect osteogenic markers expressions. The lncRNAs and mRNAs expression profiles of mBMSCs were assessed by RNA-Seq and processed by bioinformatics analysis. The selected lncRNAs expression level was validated by RT-qPCR.

Results

The effect of Mg²⁺ in promoting osteogenesis was confirmed and the optimum concentration was determined as 7.5 mM. The lncRNAs and mRNAs differentially expressed between 7.5 mM Mg²⁺-treated group and control group was detected and functional analysis revealed that their function were associated with osteogenesis. The ceRNA networks were constructed for H19 and Dubr that aberrantly expressed in two groups. The ceRNA networks of selected lncRNAs (H19 and Dubr) were constructed.

Conclusions

This study identified H19 and Dubr as osteogenic associated lncRNAs involved in Mg²⁺-induced osteogenesis, and they might play their roles through lncRNA-miRNA–mRNA axis.

中文翻译：

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通过RNA-seq和生物信息学分析Mg2+诱导的成骨的LncRNA表达谱

背景

近年来，镁（Mg）被广泛研究用于制造可生物降解的骨科器械。除了其他优点外，研究表明镁基植入物可以刺激成骨，从而加速骨科创伤恢复，但其分子机制尚不完全清楚。同时，长链非编码RNA（lncRNA）被发现在调节成骨分化中发挥着重要作用。

客观的

^{探讨lncRNA在Mg 2+} (镁离子)诱导的成骨中的作用。

方法

采用CCK-8法检测Mg ²⁺对mBMSCs增殖的影响。^{通过ALP染色和茜素红染色、Western blot和RT-qPCR检测成骨标志物的表达，确定Mg 2+}促进mBMSCs成骨的最佳浓度（7.5 mM）。通过 RNA-Seq 评估 mBMSC 的 lncRNA 和 mRNA 表达谱，并通过生物信息学分析进行处理。通过 RT-qPCR 验证所选 lncRNA 表达水平。

结果

^{证实了Mg 2+}的促进成骨作用，确定最佳浓度为7.5 mM。检测7.5 mM Mg ²⁺处理组和对照组之间差异表达的lncRNA和mRNA ，功能分析表明它们的功能与成骨相关。ceRNA 网络是针对在两组中异常表达的 H19 和 Dubr 构建的。构建了选定的 lncRNA（H19 和 Dubr）的 ceRNA 网络。

结论

^{本研究将H19和Dubr确定为参与Mg 2+}诱导成骨的成骨相关lncRNA ，它们可能通过lncRNA-miRNA-mRNA轴发挥作用。