摘要
目的:研究氧化石墨烯(graphene oxide,GO)对骨髓间充质干细胞(bone marrow-derived mesenchymal stem cells,BMSCs)迁移能力的影响及相关机制,为氧化石墨烯在骨组织工程中的应用提供实验依据。方法:利用静电纺丝技术制备掺杂不同浓度GO的聚乳酸-羟基乙酸共聚物[poly(lactic-co-glycolic acid),PLGA]纤维支架,利用扫描电镜观察支架形貌,利用CCK-8、Transwell小室、划痕实验以及定量PCR(quantitative real time polymerase chain reaction,qPCR)观察GO对大鼠BMSCs的细胞活性、迁移能力和相关基因表达的影响。结果:所制备的纤维支架具有多孔的空间结构,有利于细胞的迁移;随着GO浓度升高,纤维直径有所下降;0.25wt%和0.5wt%的GO掺杂表现出了良好的生物相容性,促进了BMSCs的迁移并且上调了PI3K、Akt、MMP2和MMP9的基因表达,当GO浓度上升至1.0wt%时,该促进作用有所下降。结论:GO可以通过激活PI3K/Akt/MMPs信号通路促进BMSCs的迁移。
Objective:The aim of the study was to evaluate the effects of graphene oxide(GO)on the migration ability of bone marrow-derived mesenchymal stem cells(BMSCs)and investigate the related mechanisms.Methods:Pure polylactic-co-glycolic acid(PLGA)and 0.25,0.5 or 1.0wt%GO-PLGA fibrous scaffolds were prepared by electrospinning.Morphology of fibers and BMSCs adhered were observed by scanning electron microscope.Cell viability was examined by CCK-8 assay.The migration of BMSCs was evaluated by Transwell and cell wound healing assays.1-,4-,and 7-day after cell seeding,the expressions of PI3K,Akt,MMP2 and MMP9 were detected by qPCR.Results:All four groups showed a porous 3D structure.Incorporation of 0.5,and 1.0wt%GO reduced the diameter of fibers(P<0.05).BMSCs successfully adhered to the fibers after 12h.Incorporation of 1.0wt%GO showed a slight cytotoxicity,especially at 7th day(P<0.05).Incorporation of 0.25,and 0.5wt%GO enhanced the migration of BMSCs and promoted the expression of PI3K,Akt,MMP2 and MMP9 in a concentration-related pattern,and the trend was reversed at the concentration of 1.0 wt%(P<0.05).Conclusion:GO can promote the migration of BMSCs by activating PI3K/Akt/MMPs signaling pathways in a suitable concentration range.
作者
郑尚
苏俭生
ZHENG Shang;SU Jian-sheng(Department of Prosthodontics,School and Hospital of Stomatology,Tongji University,Shanghai Engineering Research Center of Tooth Restoration and Regeneration,Shanghai 200072,China)
出处
《口腔颌面外科杂志》
CAS
2018年第6期309-313,共5页
Journal of Oral and Maxillofacial Surgery
基金
国家自然科学基金(81572114、81873715)
上海市科学技术委员会项目(18441902100、 17140903600)
