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Updated: Nov 17, 2025
Isolation and Cultivation of Mandibular Bone Marrow Mesenchymal Stem Cells in Rats
Published on: August 25, 2020
microRNA-935-modified bone marrow mesenchymal stem cells-derived exosomes enhance osteoblast proliferation and
Ying Zhang1, Xiangyang Cao1, Peifeng Li1
1Medical Center of Hip, Luoyang Orthopedic-Traumatological Hospital, Orthopedics Hospital of Henan Province, Luoyang 471002, Henan, PR China.
Aims:
The involvement of several microRNAs (miRNAs) in osteogenic differentiation has been indicated recently. Also, exosomes, derived from different cells, could shuttle specific miRNAs to other cell systems. Nevertheless, the effect and mechanism of microRNA-935 (miR-935)-containing exosomes in osteoblasts remain basically unclear. The current work was set to inspect the relevance of bone marrow mesenchymal stem cells (BMSCs)-derived exosomes (BMSC-exo) carrying miR-935 to osteoporotic rats.
Methods:
The extracted BMSCs and purchased osteoblasts were cultured, followed by exosome isolation and identification. After cell grouping, osteoblasts were co-cultured with BMSCs. CCK-8, alizarin red staining as well as ALP staining were performed to detect osteoblast proliferation and activity. The binding connection between miR-935 and signal transducer and activator of transcription 1 (STAT1) was measured by dual-luciferase reporter gene assays. The expression profiles of miR-935, STAT1 and osteoblast-related proteins were assessed by RT-qPCR and Western blot. A rat model with osteoporosis was induced, and the BMD, BV/TV, Tb.N, Tb.Th and Tb.Sp values in rat bone tissues were observed by Micro-CT.
Results:
BMSC-exo inhibited STAT1 levels by the delivery of miR-935 into osteoblasts, while STAT1 silencing promoted ALP activity in osteoblasts and mineralized nodules. STAT1 was identified as a target gene of miR-935. Moreover, in vivo experiments showed that in ovariectomized rats, silencing of miR-935 significantly reduced BMD, BV/TV, Tb.N, Tb.Th and increased Tb.Sp.
Conclusion:
BMSC-exo carry miR-935 to promote osteoblast proliferation and differentiation through targeting STAT1.
Insights
Bone marrow mesenchymal stem cell-derived exosomes carrying microRNA-935 promote osteoblast function by targeting STAT1. This finding offers potential therapeutic strategies for osteoporosis treatment.
Area of Science:
- Cell Biology
- Biochemistry
- Regenerative Medicine
Background:
- MicroRNAs (miRNAs) play a role in osteogenic differentiation.
- Exosomes can transport miRNAs between cells.
- The function of miR-935-containing exosomes in osteoblasts is not well understood.
Purpose of the Study:
- To investigate the role of bone marrow mesenchymal stem cell (BMSC)-derived exosomes (BMSC-exo) carrying microRNA-935 (miR-935) in osteoporotic rats.
- To elucidate the mechanism by which BMSC-exo influence osteoblasts.
Main Methods:
- Isolation and characterization of exosomes from BMSCs.
- Co-culture of osteoblasts with BMSCs and BMSC-exo.
- Assessment of osteoblast proliferation and activity using CCK-8, alizarin red, and ALP staining.
- Dual-luciferase reporter assays to confirm the interaction between miR-935 and STAT1.
- RT-qPCR and Western blot to analyze gene and protein expression.
- Micro-CT analysis of bone mineral density and microarchitecture in an osteoporosis rat model.
Main Results:
- BMSC-exo delivered miR-935 into osteoblasts, inhibiting STAT1 expression.
- STAT1 was identified as a direct target of miR-935.
- Silencing STAT1 enhanced osteoblast activity and mineralization.
- In vivo, miR-935 silencing in ovariectomized rats led to reduced bone mineral density and impaired bone microarchitecture.
Conclusions:
- BMSC-exo carrying miR-935 promote osteoblast proliferation and differentiation.
- The mechanism involves targeting STAT1.
- This study highlights the therapeutic potential of BMSC-exo loaded with miR-935 for treating osteoporosis.
