摘要
牙髓炎和根尖周炎是口腔科目前较为常见的2种疾病,现有的治疗方案主要包括根管治疗和牙髓血运重建,能有效控制住炎症进而保存患牙,然而同时也会导致牙髓组织的永久失活,发生结构故障和继发感染。近年来,结合干细胞和生物材料等的组织工程技术使牙髓再生的研究逐渐进入人们的视野,其中从恒牙或乳牙中分离的牙髓间充质干细胞(dental pulp stem cells,DPSCs),因其多向分化和高增殖等特性已成为牙本质或者髓样组织再生的重要的干细胞来源。但是由于干细胞不能高效募集到损伤区域,影响受损区的活细胞数量和存活时间,显著降低了其应用疗效,因此,需要提高牙髓干细胞的迁移和增殖能力,本研究旨在探讨微核糖核酸31(microRNA-31,miR-31)能否有效提高DPSCs的增殖迁移能力。通过组织块酶消化法从牙髓组织中成功分离培养了DPSCs,比较了分别取自健康牙与炎症牙的牙髓组织和DPSCs中miR-31水平的差异,利用实时荧光定量PCR(RT-qPCR)技术检测的结果显示,与健康牙比较,炎症牙来源的牙髓组织和DPSCs中miR-31表达水平明显降低(P<0.05)。干扰和过表达DPSCs中的miR-31表达,具体分为NC组、miR-31 agomir(过表达)组和miR-31 antagomir(抑制剂)3个组,RTq-PCR结果显示,其转染成功(P<0.001)。CCK-8、细胞划痕以及Transwell迁移结果表明,与对照组相比,过表达miR-31成功提高了牙髓干细胞的增殖和迁移能力(P<0.05),且进一步通过Western印迹结果发现,过表达miR-31后增殖关键蛋白质增殖抗原(Ki67)、增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)以及迁移关键蛋白质CXC趋化因子受体4型(CXC chemokine receptor type 4,CXCR4)、基质金属蛋白酶2(matrix metalloproteinase2,MMP2)的蛋白质水平均显著升高(P<0.05)。本研究表明,miR-31能有效提高DPSCs的增殖迁移能力,以期为DPSCs更好应用于再生医学提供了有力的理论支持。
Pulpitis and periapical inflammation are two common diseases in stomatology today.Existing treatment options primarily include root canal therapy and pulp revascularization,which can effectively control inflammation and preserve the affected tooth while also causing permanent deactivation of the pulp tissue,structural failure,and secondary infection.In recent years,research on dental pulp regeneration has progressively entered the public consciousness because of tissue engineering technology that combines stem cells and biomaterials.Due to their multi-differentiation and high proliferation,dental pulp stem cells(DPSCs)isolated from permanent or deciduous teeth have emerged as a significant stem cell source for dentin or pulp tissue regeneration.However,the number and survival time of live cells in the damaged area are impacted,which significantly limits the efficacy of stem cells since they are unable to efficiently be recruited to the injured area.The ability of DPSCs to migrate and multiply must therefore be enhanced.This study sought to determine if miR-31(miR-31)may significantly enhance the proliferative and migratory capacities of DPSCs.The tissue block enzyme digestion method was used to successfully separate and culture DPSCs from dental pulp tissues,and the miR-31 levels in dental pulp tissues and DPSCs from normal and inflammatory teeth were compared.The results of real-time fluorescence quantitative PCR(RT-qPCR)revealed that the expression level of miR-31 in dental pulp tissues and DPSCs from inflammatory teeth was significantly lower when compared to the control group(P<0.05).Interference and over-expression of miR-31 expressions in DPSCs were specifically divided into three groups:the NC group,the miR-31 agomir(over-expressed)group and the miR-31 antagomir(inhibitor)group.RTq-PCR results showed that the transfection was successful(P<0.001).The results of CCK-8,wound-healing,and Transwell migration experiments showed that overexpression of miR-31 successfully improved the proliferation and migration a
作者
王佳微
曹利利
丁雪婷
田峰
王春芳
WANG Jia-Wei;CAO Li-Li;DING Xue-Ting;TIAN Feng;WANG Chun-Fang(School and Hospital of Stomatology,Shanxi Medical University,Taiyuan 030012,Shanxi,China;Shanxi Province Key Laboratory of Oral Diseases Prevention and New Materials,Taiyuan 030012,Shanxi,China;Experimental Animal Center,Shanxi Medical University,Taiyuan 030012,Shanxi,China)
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2023年第11期1620-1629,共10页
Chinese Journal of Biochemistry and Molecular Biology
基金
中央引导地方科技发展资金项目(No.YDZJSX2022A056)
山西省基础研究计划项目(No.202203021212373)资助。
关键词
人牙髓间充质干细胞
微RNA
miR-31
细胞增殖和迁移
human dental pulp mesenchymal stem cells(HDPSCs)
microRNA(miRNA)
miR-31
cell proliferation and migration
