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MiR-26b-5p/TET3 regulates the osteogenic differentiation of human bone mesenchymal stem cells and bone reconstruction in female rats with calvarial defects

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MicroRNAs (miRNAs) play critical roles in the osteogenic differentiation of human bone mesenchymal stem cells (hBMSCs), but the mechanism by which miRNAs indirectly modulate osteogenesis remains unclear. Here, we explored the mechanism by which miRNAs indirectly modulate gene expression through histone demethylases to promote bone regeneration.

Methods and results

Bioinformatics analysis was performed on hBMSCs after 7 days of osteogenic induction. The differentially expressed miRNAs were screened, and potential target mRNAs were identified. To determine the bioactivity and stemness of hBMSCs and their potential for bone repair, we performed wound healing, Cell Counting Kit-8 (CCK-8), real-time reverse transcription quantitative polymerase chain reaction (RT‒qPCR), alkaline phosphatase activity, alizarin red S (ARS) staining and radiological and histological analyses on SD rats with calvarial bone defects. Additionally, a dual-luciferase reporter assay was utilized to investigate the interaction between miR-26b-5p and ten-eleven translocation 3 (TET3) in human embryonic kidney 293T cells. The in vitro and in vivo results suggested that miR-26b-5p effectively promoted the migration, proliferation and osteogenic differentiation of hBMSCs, as well as the bone reconstruction of calvarial defects in SD rats. Mechanistically, miR-26b-5p bound to the 3’ untranslated region of TET3 mRNA to mediate gene silencing.


MiR-26b-5p downregulated the expression of TET3 to increase the osteogenic differentiation of hBMSCs and bone repair in rat calvarial defects. MiR-26b-5p/TET3 crosstalk might be useful in large-scale critical bone defects.

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We acknowledge the funding support from National Key Research and Development Program (Grant No. 2022YFA1104401 and 2022YFC2504201 to Z.F.), National Natural Science Foundation of China (Grant No. 82130028 to Z.F.), CAMS Innovation Fund for Medical Sciences (Grant No. 2019-I2M-5-031 to Z.F.), Innovation Research Team Project of Beijing Stomatological Hospital, Capital Medical University (Grant No. CXTD202204 to Z.F.) and Young Scientist Program of Beijing Stomatological Hospital, Capital Medical University (Grant No. YSP202113 to C.Z.).

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W.Y. wrote the first draft of the manuscript. W.Y. and C.Z. performed the material preparation, data collection and analysis. C.Z. and Z.F. contributed to the study conception and design. All authors reviewed the final manuscript.

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Correspondence to Chen Zhang or Zhipeng Fan.

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The animal study was approved by the Ethics Committee Agreement of Beijing Statistical Hospital (No. KQYY-202311-006).

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Ye, W., Zhang, C. & Fan, Z. MiR-26b-5p/TET3 regulates the osteogenic differentiation of human bone mesenchymal stem cells and bone reconstruction in female rats with calvarial defects. Mol Biol Rep 51, 632 (2024).

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