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Extracellular vesicles from hypoxia preconditioned bone marrow mesenchymal stem cell improve peri-implant
Xi-Meng Cao1, Ya-Qin Wu1, Ying-Yi Shen1
1Department of Prosthodontics, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai, China; National Center for Stomatology and National Clinical Research Center for Oral Diseases, Shanghai, China; Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology, Shanghai, China; Shanghai Engineering Research Center of Advanced Dental Technology and Materials, Shanghai, China.
Purpose:
Poor peri-implant osseointegration of dental implants in patients with type 2 diabetes has become a major clinical challenge in recent years. BMSC (bone marrow mesenchymal stem cell)-derived extracellular vesicles may play an important role in peri-implant osseointegration, but the mechanism remains unclear. Improving the therapeutic effect of BMSC-derived extracellular vesicles and exploring their potential mechanisms can help provide new treatment strategies for dental implants in patients with type 2 diabetes.
Methods:
The extracellular vesicles derived from hypoxia (Hypo-EVs) or normoxia (Nor-EVs) preconditioned bone marrow mesenchymal stem cells (BMSCs) were co-cultured with BMSCs and human umbilical vein endothelial cells (HUVECs). The effect of extracellular vesicles on BMSCs cell proliferation was detected by CCK-8 assay and EdU assay, and the effect on angiogenesis ability of HUVECs was detected by wound healing assay, transwell migration assay, tube formation assay, enzyme-linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. A diabetic rat dental implant model was also established and the effect of extracellular vesicles on implant osseointegration was evaluated through micro-CT scanning and histological analysis. The differentially expressed miRNAs between Hypo-EVs and Nor-EVs were identified by high-throughput miRNA sequencing. Subsequently, the target genes and their roles in regulating angiogenesis were predicted and analyzed by bioinformatics analysis and dual luciferase reporter assay.
Results:
In vitro experiments indicated that hypoxia preconditioning could elevate extracellular vesicle production and promote cell proliferation of BMSCs and angiogenesis of HUVECs. Moreover, Hypo-EVs promoted peri-implant osteogenesis in rats with diabetes. Further investigation revealed the vital involvement of the miR-106b-5p/HIF-1α axis in promoting peri-implant osseointegration under high glucose condition.
Conclusion:
Extracellular vesicles derived from hypoxia-preconditioned BMSCs could improve the peri-implant osseointegration in rats with diabetes by promoting cell proliferation and angiogenesis, and the miR-106b-5p/ HIF-1α axis could be the underlying mechanism.
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