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. 2020 Nov 6;8:600641. doi: 10.3389/fcell.2020.600641

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Copyright © 2020 Yin, Tian, Yu, Yang, Su, Zhao, Wang, Zhang, Pei, Li, Chen, Zhang, Miao and Qian.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Bioengineered recombinant miR-129-5p inhibitor enhanced osteoblast differentiation and rescued postmenopausal osteoporosis. (A) miR-129-5p expression levels of MC3T3-E1 cells treated with recombinant miR-129-5p inhibitor, as detected by reverse transcriptase-polymerase chain reaction (RT-PCR; mean ± SD, ^∗∗P < 0.01). MSA: tRNA^Met fused Sephadex aptamer. nCAR/anti129: novel recombinant miR-129-5p inhibitor. (B) Alp and alizarin red staining of MC3T3-E1 cells treated with recombinant miR-129-5p inhibitor, as detected by Alp staining and alizarin red staining. Alp: results of Alp staining. alizarin red: results of alizarin red staining. (C) Osterix and Runx2 expression levels of MC3T3-E1 cells treated with recombinant miR-129-5p inhibitor, as detected by RT-PCR (mean ± SD, ^∗∗P < 0.01). (D,E) Tcf7 and Lef1 expression levels of MC3T3-E1 cells treated with recombinant miR-129-5p inhibitor, as detected by RT-PCR (mean ± SD, ^∗∗P < 0.01, ^∗∗∗P < 0.001). (F) TCF7/LEF1 activities of MC3T3-E1 cells treated with recombinant miR-129-5p inhibitor, as detected by luciferase reporter assay (mean ± SD, ^∗∗∗P < 0.001). (G) TCF4 expression levels of MC3T3-E1 cells treated with recombinant miR-129-5p inhibitor, as detected by RT-PCR and Western blot (mean ± SD, ^∗∗∗P < 0.001). (H) Representative images showing calvarial mineral apposition rate of C57BL/6 mice after OVX and recombinant miR-129-5p inhibitor treatment. Scale bar: 10 μm. BL (baseline): sacrifice before RNA treatment. Sham: sham OVX operation group. OVX: OVX group. Mock: transfection reagent control group. MSA: empty recombinant tRNA treated group. nCAR/anti129: novel recombinant miR-129-5p–inhibitor treated group. (I) Calvarial mineral apposition rates of C57BL/6 mice after OVX and recombinant miR-129-5p inhibitor treatment (mean ± SD, ^∗∗∗P < 0.001).