海南医科大学学报2025，Vol.31Issue(17)：1336-1343,8.DOI:10.13210/j.cnki.jhmu.20241017.002
PDLSC-Exo对牙周膜干细胞迁移和成纤维分化的作用研究
Research on the effect of PDLSC-Exo on migration and fibroblast differentiation of periodontal ligament stem cells
摘要
Abstract
Objective:To investigate the effects of exosomes derived from periodontal ligament stem cells(PDLSC-Exo)on cell migration and fibroblast differentiation of PDLSCs.Methods:PDLSCs were extracted using enzymatic digestion;PDLSC-Exo was isolated by ultracentrifugation and characterized by transmission electron microscope(TEM),nanoparticle track-ing and Western blot;following on treatment with 20 μg/mL PDLSC-Exo,cell viability and apoptosis were assessed using MTT and annexin V-FITC/PI double staining,respectively,while cell migration was evaluated by scratch assay;fibroblast differentia-tion related genes were detected by qRT-PCR,and TGF-β/SMAD3 signaling levels were detected by Western blot;cell scratch assay was also used to investigate the effect of PDLSC-Exo on cell migration after inhibiting SMAD3 phosphorylation.Results:PDLSCs were spindle-shaped cells under a light microscope,and had both osteogenic and adipogenic differentiation abilities;PDLSCs positively expressed the mesenchymal stem cell surface antigens(CD73,CD90,and CD105),and negatively expressed the hematopoietic stem cell surface antigens(CD31 and CD45);PDLSC-Exo exhibited the cup-plate-shaped vesicular structure under a TEM with a diameter of 30 nm-150 nm;PDLSC-Exo was positive for exosome-specific markers(Alix and CD63)and negative for endoplasmic reticulum protein(Calnexin);in the PDLSC-Exo treatment group,cell viability of PDLSCs was in-creased,cell apoptosis showed a tendency to decrease,the expressions of fibroblast differentiation related genes COL1 and FAP were increased,healing area rate in scratch assay showed a tendency to increase,and level of P-SMAD2/3(canonical transform-ing growth factor-β(TGF-β)signaling pathway)was upregulated;after inhibiting SMAD3 phosphorylation,cell migration ability of PDLSCs promoted by PDLSC-Exo was downregulated.Conclusion:PDLSC-Exo promote cell migration and fibroblast differ-entiation of PDLSCs,which may be related to upregulation of TGF-β/SMAD signaling pathway.关键词
牙周膜干细胞/外泌体/成纤维分化/细胞迁移Key words
Periodontal ligament stem cells/Exosomes/Fibroblast differentiation/Cell migration分类
医药卫生引用本文复制引用
张宁,胡月,时小婷,曹楠珏,刘克达,孟圆,王蔚..PDLSC-Exo对牙周膜干细胞迁移和成纤维分化的作用研究[J].海南医科大学学报,2025,31(17):1336-1343,8.基金项目
This study was supported by the National Natural Science Foundation of China(82271037) （82271037）
Liaoning Province Science and Technology Plan Project(2022-BS-123) （2022-BS-123）
Shenyang Science and Technology Plan Project(2400023067,21-173-9-34) 国家自然科学基金资助项目(82271037) （2400023067,21-173-9-34）
辽宁省科学技术计划项目(2022-BS-123) （2022-BS-123）
沈阳市科技计划项目(2400023067,21-173-9-34) （2400023067,21-173-9-34）
