文章摘要
王 燕,朱琳琳,王思雨,张舒婷,孙 雯,李 琥,张瀚文,严 斌.正畸驱动的骨皮质切开术对巨噬细胞的作用及机制研究[J].南京医科大学学报,2018,(5):582~589
正畸驱动的骨皮质切开术对巨噬细胞的作用及机制研究
Effects of orthodontically drivencorticotomy on macrophages and its mechanism
投稿时间:2017-01-03  
DOI:10.7655/NYDXBNS20180503
中文关键词: 牙槽骨  骨皮质切开  正畸牙移动  炎症  巨噬细胞  骨密度
英文关键词: alveolar bone  corticotomy  orthodontic tooth movement  inflammation  macrophage  bone mineral density
基金项目:国家自然科学基金(81571005);江苏省高等学校自然科学研究重大项目(17KJA320003);江苏省“333高层次人才培养工程”(BRA2016511);江苏省高校优势学科建设工程资助项目
作者单位
王 燕 南京医科大学口腔疾病研究江苏省重点实验室江苏 南京 210029南京医科大学附属口腔医院正畸科江苏 南京 210029 
朱琳琳 南京医科大学口腔疾病研究江苏省重点实验室江苏 南京 210029南京医科大学附属口腔医院正畸科江苏 南京 210029 
王思雨 南京医科大学口腔疾病研究江苏省重点实验室江苏 南京 210029南京医科大学附属口腔医院正畸科江苏 南京 210029 
张舒婷 南京医科大学口腔疾病研究江苏省重点实验室江苏 南京 210029南京医科大学附属口腔医院正畸科江苏 南京 210029 
孙 雯 南京医科大学口腔疾病研究江苏省重点实验室江苏 南京 210029 
李 琥 南京医科大学附属口腔医院正畸科江苏 南京 210029 
张瀚文 南京医科大学基础医学院江苏 南京 211166 
严 斌 南京医科大学口腔疾病研究江苏省重点实验室江苏 南京 210029南京医科大学附属口腔医院正畸科江苏 南京 210029 
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中文摘要:
      目的:研究正畸牙周围骨皮质切开所诱导的局部微环境对巨噬细胞产生的生物学作用。方法:将60只大鼠随机分成骨皮质切开辅助正畸组和传统正畸组各30只,加力后分别于3、5、7、14、21 d处死实验动物并对颌骨进行Micro?CT扫描,三维重建后测量移动牙周围牙槽骨骨密度,并通过免疫组化染色技术,观察牙周组织中CD68、CD11b、CD86及CD163阳性细胞数。将112只8周龄成年雄性C57BL/6小鼠随机分成牙槽骨皮质切开组及对照组各56只,并分别于3、5、7、14、21、28、42 d处死实验动物并取新鲜的牙槽骨组织悬液与小鼠腹腔巨噬细胞共培养,RT?PCR检测CD11b、CD68、白介素(IL)?6、IL?1β、IL?10及精氨酸酶(Arg)?1的mRNA表达,Western blot检测信号通路相关蛋白p65以及STAT3磷酸化水平。结果:牙槽骨皮质切开初期,骨皮质切开辅助正畸组和传统正畸组大鼠牙槽骨密度均降低,且前者明显低于后者(P < 0.05),同时免疫组化染色显示,前者CD68、CD11b、CD86及CD163阳性细胞比率显著高于后者(P < 0.05)。与牙槽骨组织悬液共培养后,牙槽骨皮质切开组巨噬细胞表面标记物CD11b和CD68的表达量在共培养第3天时开始逐渐增多,在14 d左右达到峰值并显著高于对照组(P < 0.05);M1型巨噬细胞标记物IL?6及IL?1β及M2型巨噬细胞标记物IL?10及Arg?1表达量在14 d左右到达峰值且显著高于对照组(P < 0.05)。p65蛋白以及磷酸化的STAT3蛋白表达量分别于共培养第5、7天和第7、14天显著高于对照组(P < 0.05)。结论:正畸驱动的骨皮质切开术通过活化并促进巨噬细胞的分型进而影响牙槽骨密度并促进牙齿的移动,其中NF?κB及JAK?STAT信号通路的激活可能发挥重要作用。
英文摘要:
      Objective:To investigate the biological effects of corticotomy?induced micro?environment around orthodontic teeth on macrophages. Methods:A total of 60 male Wistar rats were randomly divided into the experimental group(TM+CO)and control group(TM)with each group of 30 rats. The rats in each group were sacrificed after 3,5,7,14 and 21 days for tissue collection and assessments. Micro?CT examination was performed to acquire bone mineral density(BMD)around the upper left first molars,to analyze the change of bone remodeling. Morphological changes of periodontal tissues and the expression of CD68,CD11b,CD86,CD163?positive cells were determined by immunohistochemistry. A total of 112 male C57BL/6 mice of eight?week old were randomly divided into the corticotomy group and the vehicle group. The mice in each group were sacrificed after 3,5,7,14,21,28 and 42 days for tissue collection and co?culture. The relative mRNA expression of CD11b,CD68,IL?6,IL?1β,IL?10 and Arg?1 were respectively evaluated by RT?PCR. The protein change of p65 and STAT3 phosphorylation were detected by Western blot. Results:The value of BMD in both groups decreased significantly at the early stage and it was more obvious in the TM+CO group(P < 0.05). Meanwhile,the immunohistochemistry data showed that the expressions of CD68,CD11b,CD86,CD163?positive cells in the TM+CO group were all significantly increased compared with those in the TM group at day 3(P < 0.05). The real?time PCR analysis showed that the mRNA expression levels of CD11b and CD68 in macrophages were significantly higher in the corticotomy group than those in the vehicle group(P < 0.05);they increased from day 3 to day 14 and then declined. Moreover,the mRNA expression levels of M1 marker,TNF?α,IL?1β and M2 marker,IL?10,Arg?1 were significantly higher in the corticotomy group compared with those in the vehicle group(P < 0.05);they increased up to day 14 and then declined. Western blot analysis showed that the level of p65 was upregulated after co?culture with condition medium of maxilla in the corticotomy group early on day 5 and day 7,while the level of phosphorylated STAT3 was upregulated on day 7 and day 14 when compared with those in the vehicle group(P < 0.05). Conclusion:The activation of NF?κB and JAK?STAT signaling pathways may play an important role in corticotomy induced infiltration and polarization of macrophages and thereby influenced bone density and consequent orthodontic tooth movement.
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