Chang Maolin, Chen Qianrou, Wang Beike, Zhang Zhen, Han Guangli
State Key Laboratory Breeding Base of Basic Science of Stomatology (Hubei-MOST) & Key Laboratory of Oral Biomedicine Ministry of Education, School & Hospital of Stomatology, Wuhan University, Wuhan, China.
Orthodontic Department Division II, School & Hospital of Stomatology, Wuhan University, Wuhan, China.
Int J Stem Cells. 2023 May 30;16(2):202-214. doi: 10.15283/ijsc21170. Epub 2023 Feb 28.
To investigate the role of exosomes from periodontal ligament cells (PDLCs) in bone marrow mesenchymal stem cell (BMSC) migration.
Human PDLCs were applied cyclic tension stretching. Exosomes were extracted from cultured PDLCs by ultracentrifugation, then characterized for their size, morphology and protein markers by NTA, TEM and western blotting. The process that PKH26-labeled exosomes taken up by BMSCs was assessed by confocal microscope. BMSC migration was examined by Transwell assay. Exosomes derived from PDLCs were identified. Cyclic tension stretch application on PDLCs can enhance the migration ability of BMSCs through exosomes. The exosomal miRNA expression profiles of unstretched and stretched PDLCs were tested by miRNA microarray. Four miRNAs (miR-4633-5p, miR-30c-5p, miR-371a-3p and let-7b-3p) were upregulated and six (miR-4689, miR-8485, miR-4655-3p, miR-4672, miR-3180-5p and miR-4476) were downregulated in the exosomes after stretching. Sixteen hub proteins were found in the miRNA-mRNA network. Gene Ontology and KEGG pathway analyses demonstrated that the target genes of differentially expressed exosomal miRNAs closely related to the PI3K pathway and vesicle transmission.
The exosomes derived from cyclic tension-stretched PDLCs can promote the migration of BMSCs. Alternation of microRNA profiles provides a basis for further research on the regulatory function of the exosomal miRNAs of PDLCs during orthodontic tooth movement.
探讨牙周膜细胞（PDLCs）来源的外泌体在骨髓间充质干细胞（BMSCs）迁移中的作用。
对人PDLCs施加周期性拉伸张力。通过超速离心从培养的PDLCs中提取外泌体，然后通过纳米颗粒跟踪分析（NTA）、透射电子显微镜（TEM）和蛋白质免疫印迹法对其大小、形态和蛋白质标志物进行表征。用共聚焦显微镜评估PKH26标记的外泌体被BMSCs摄取的过程。通过Transwell实验检测BMSCs的迁移。鉴定了源自PDLCs的外泌体。对PDLCs施加周期性拉伸张力可通过外泌体增强BMSCs的迁移能力。通过miRNA芯片检测未拉伸和拉伸后的PDLCs外泌体的miRNA表达谱。拉伸后，外泌体中有4种miRNA（miR-4633-5p、miR-30c-5p、miR-371a-3p和let-7b-3p）上调，6种miRNA（miR-4689、miR-8485、miR-4655-3p、miR-4672、miR-3180-5p和miR-4476）下调。在miRNA-mRNA网络中发现了16个枢纽蛋白。基因本体论（Gene Ontology）和京都基因与基因组百科全书（KEGG）通路分析表明，差异表达的外泌体miRNA的靶基因与PI3K通路和囊泡运输密切相关。
周期性拉伸张力作用下的PDLCs来源的外泌体可促进BMSCs的迁移。miRNA谱的改变为进一步研究正畸牙齿移动过程中PDLCs外泌体miRNA的调控功能提供了基础。
