正畸牙移动中牙周血管内皮祖细胞在骨改建中的作用机制探讨

doi:10.19439/j.sjos.2019.06.011

摘要/Abstract

摘要： 目的探讨正畸牙移动中牙周血管内皮祖细胞在骨改建中的作用机制。方法首先建立实验性大鼠牙移动模型,分离和培养血管内皮祖细胞(endothelial progenitor cells EPCs),用10 μmol/L Brdu标记EPCs并通过尾静脉注入模型大鼠,观察EPCs在牙周组织的分布情况。将VEGF加入EPCs细胞培养液,MTT法测细胞增殖能力,显微镜下观察细胞黏附情况,Transwell实验观察细胞迁移能力。制作不同时间点模型大鼠的VEGF免疫组织化学染色切片,与显微镜下观察不同时间点牙周组织VEGF的表达情况。采用SPSS 20.0软件包对数据进行统计学分析。结果成功建立了大鼠牙移动模型,从心脏血中分离EPCs,显微镜下观察到有些为梭形,将Brdu标记的EPCs经尾静脉注入模型大鼠中,观察到随着时间的增加,荧光强度逐渐增加,在3 d的标本中荧光强度达到最强。各个时间点上颌第一、二磨牙之间的间隙均为实验组低于对照组,且具有显著性差异（P<0.05）。VEGF免疫组织化学染色结果显示,无论是张力侧还是压力侧,实验组表达量均高于对照组。在成骨细胞和破骨细胞,VEGF均有表达,14 d时达到最大值。EPCs增殖、黏附能力实验表明,VEGF促进EPCs增殖,使其黏附力增强。Transwell实验表明,VEGF促进EPCs的趋化作用,VEGF对EPCs的生物作用具有调控作用。结论 EPCs能够聚集于牙周组织,参与牙周组织骨改建。EPCs趋化到牙周组织后,通过VEGF等因子的相互调控作用,参与牙周组织的改建程,促进牙周组织修复和骨改建。

关键词: 正畸牙移动, 血管内皮祖细胞, 骨改建

Abstract: PURPOSE: To investigate the mechanism of endothelial progenitor cells(EPCs) bone remodeling during orthodontic tooth movement. METHODS: Experimental tooth movement model was established. EPCs were isolated, cultured, and labeled with 10 μmol/L Brdu and injected into rats through tail vein to observe the distribution in periodontal tissue. VEGF was added to EPCs culture medium, cell proliferation ability was measured by MTT assay, cell adhesion was observed under microscope. Transwell assay was used to observe cell migration ability, and VEGF immunohistochemical staining sections of model rats at different time points were made. The expression of VEGF in periodontal tissues at different time points was defected. All data were imputed into SPSS 20.0 software package for statistical analysis. RESULTS: A rat model of tooth movement was successfully established. EPCs were isolated from cardiac blood. Some spindle-shaped EPCs were observed under microscope and injected into model rats using Brdu-labeled EPCs. With the increase of time, the intensity of fluorescence gradually increased. In the 3d specimen, the fluorescence intensity reached the strongest. The gap between the first and second molars in the experimental group was lower than that in the control group at each time point, with significant difference(P<0.05). The results of VEGF immunohistochemical staining showed that both the tension side and the pressure side of the experimental group were significantly higher than the control group(P<0.05). The expression of VEGF in both osteoblasts and osteoclasts reached a maximum at 14 days. EPCs proliferation and adhesion experiments demonstrated that VEGF promoted proliferation of EPCs and enhanced their adhesion. Transwell experiments showed that VEGF promoted chemotaxis of EPCs.VEGF regulated the biological effects of EPCs. CONCLUSIONS: EPCs can be accumulated in periodontal tissues and participate in periodontal bone remodeling. After EPCs chemotizing to periodontal tissues, they participate in the remodeling of periodontal tissues through mutual regulation of VEGF and other factors, and promote periodontal tissue repair and bone remodeling.

Key words: Orthodontic tooth movement, Endothelial progenitor cells, Bone remodeling

中图分类号:

R783.5

陈婉红, 蔡世雄, 吴小榕. 正畸牙移动中牙周血管内皮祖细胞在骨改建中的作用机制探讨[J]. 上海口腔医学, 2019, 28(6): 610-615.

CHEN Wan-hong, CAI Shi-xiong, WU Xiao-rong. Study on the mechanism of endothelial progenitor cells in periodontal revascularization during orthodontic tooth movement[J]. Shanghai Journal of Stomatology, 2019, 28(6): 610-615.

参考文献

[1] 蒋自然. IL-17在大鼠不同牙周状态压力侧牙槽骨中的表达和作用机制的初探[D]. 济南: 山东大学, 2017.
[2] 顾继航, 王兰珠, 韩博, 等. 在正畸力作用下大鼠牙周膜张力侧Runx2与β-catenin的表达[J]. 空军医学杂志, 2016, 32(2): 127-129.
[3] d'Apuzzo F, Cappabianca S, Ciavarella D, et al. Biomarkers of periodontal tissue remodeling during orthodontic tooth movement in mice and men: overview and clinical relevance[J]. Sci World J, 2013, 2013: 105873.
[4] 邓伟民, 黄思敏. 重视血管形成、免疫系统及甲状旁腺激素对骨的作用机制研究[J]. 实用医学杂志, 2015, 31(22): 3637-3640.
[5] 沈培晓, 汪海娅. 内皮祖细胞及造血祖细胞在心血管疾病中的意义及研究进展[J]. 老年医学与保健, 2015, 21(1): 59-61.
[6] 谢安明, 王雅君, 崔丽珺. 血管内皮祖细胞与VEGF对增生性糖尿病视网膜病变新生血管形成的影响[J]. 西安交通大学学报(医学版), 2013, 34(2): 233-236.
[7] 王仁, 张凯, 李爽, 等. 载脂蛋白A损伤小鼠骨髓源性EPCs血管发生能力及其机制[J]. 生物化学与生物物理进展, 2013, 40(8): 757-765.
[8] 张百灵, 王照飞, 张航, 等. CXCR4基因过表达HUCB-late EPCs移植对大鼠后肢缺血组织血管新生的影响[J]. 中国动脉硬化杂志, 2017, 25(3): 238-243.
[9] 栗洪师, 陈新, 冯岩, 等. 实验性牙移动模型大鼠构建:牙齿移动后空口咀嚼时间变化规律[J]. 中国组织工程研究, 2015, 19(49): 7988-7993.
[10] 卫肖艳, 张莉, 林明, 等. 大鼠骨髓源性内皮祖细胞的分离培养与鉴定[J]. 中国组织工程研究, 2013, 17(14): 2570-2577.
[11] 刘珍珍. 脐血内皮祖细胞糖基化修饰及其在缺血性动物模型的应用[D]. 苏州: 苏州大学, 2013.
[12] 王丽亚. 骨髓内皮祖细胞治疗大鼠急性脑缺血的实验研究[D]. 长春: 吉林大学, 2014.
[13] Altabas V, Altabas K, Kirigin L.Endothelial progenitor cells (EPCs) in ageing and age-related diseases: how currently available treatment modalities affect EPC biology, atherosclerosis, and cardiovascular outcomes[J]. Mech Ageing Dev, 2016, 159: 49-62.
[14] 洪微. 血管内皮祖细胞促进骨髓间充质干细胞向成骨细胞分化的实验研究 [D]. 沈阳: 中国医科大学, 2014.
[15] 底煜, 陈晓隆, 王爱媛. CCR7和VEGF在缺氧时视网膜血管内皮细胞中的表达及意义[J]. 国际眼科杂志, 2015, 15(3): 403-406.
[16] 杨瑞瑞, 杜怡峰, 郭守刚. VEGF动员内源性EPCs促进MCAO/R小鼠神经功能修复[C]// 山东省2013年神经内科学学术会议暨中国神经免疫大会2013论文汇编. 济南, 2013.
[17] 刘红林, 刘志军, 陈小兵, 等. 脐带间充质干细胞在颅脑损伤模型鼠体内的迁徙与定位[J]. 中国组织工程研究, 2016, 20(1): 31-35.
[18] Gong M, Dong W, Shi Z, et al.Vascular endothelial growth factor gene polymorphisms and the risk of renal cell carcinoma: Evidence from eight case-control studies[J]. Oncotarget, 2017, 8(5): 8447-8458.
[19] 越杨, 陈卓, 谢冰, 等. 正畸牙移动中牙周组织VEGF的表达及在骨改建中的作用机制探讨[J]. 上海口腔医学, 2018, 27(1):18-21.