骨髓基质细胞对大鼠牙周组织缺损的的修复作用及相关机制

Study on the repairing effect of bone marrow stromal cells on rat periodontal tissue defects and related mechanisms

目的研究骨髓基质细胞(BMSCs)对大鼠牙周组织缺损的修复作用,并探讨其相关分子机制。方法选取45只12周龄无特定病原体(SPF)级雄性SD大鼠为研究对象,每只大鼠在下颌第一和第二磨牙出制备牙周缺损区域,然后将大鼠按照随机数字表法分为膨体聚四氟乙烯(e-PTFE)组、BME组和BMSCs+BME组3组,每组15只。ePTFE组大鼠牙周缺损区域经覆盖e-PTFE膜;BME组大鼠先在牙周缺损区域覆盖BME胶原膜,然后在覆盖e-PTFE膜;BMSCs+BME组大鼠先在牙周缺损区域覆盖携带BMSCs的BME胶原膜,然后在覆盖e-PTFE膜。治疗4周后,安乐死大鼠测量各组大鼠新生牙槽骨和牙骨质面积、新生牙周膜宽度、血清和牙周缺损组织白细胞介素(IL)-1β、肿瘤坏死因子(TNF)-α以及IL-6含量,牙周缺损组织M1/M2巨噬细胞标志物表达水平。结果e-PTFE组和BME组大鼠新生牙槽骨和牙骨质面积、新生牙周膜宽度、血清和牙周缺损组织IL-1β、TNF-α以及IL-6含量,以及牙周缺损组织CD16、iNOS、CD206和Arg1 mRNA表达差异无统计学意义(P>0.05);术后第4周,BMSCs+BME组大鼠体重、新生牙槽骨和牙骨质面积、新生牙周膜宽度以及牙周缺损区域组织CD206和Arg1 mRNA均显著高于e-PTFE组和BME组,而大鼠血清和牙周缺损区域组织IL-1β、TNF-α以及IL-6含量以及牙周缺损区域组织CD16和iNOS mRNA显著低于e-PTFE组和BME组,差异均有统计学意义(P<0.05)。结论骨髓基质细胞在大鼠牙周组织缺损部位可促进M2巨噬细胞极化,进而抑制牙周缺损组织炎症反应,促进牙周缺损组织修复。

Objective To study the repairing effect of bone marrow stromal cells(BMSCs)on rat periodontal tissue defects,and to explore its related molecular mechanisms.Methods Forty-five 12-week-old specific pathogen free(SPF)male SD rats were selected as the research objects,and each rat prepared periodontal defect areas in the first and second mandibular molars.Then the rats were divided into 3 groups according to the random number table method:expanded polytetrafluoroethylene(e-PTFE)group,BME group and BMSCs+BME group,each group 15 pieces.The periodontal defect area of the rats in the e-PTFE group was covered with e-PTFE membrane;the rats in the BME group first covered the periodontal defect area with BME collagen membrane and then covered with the e-PTFE membrane;the rats in the BMSCs+BME group were covered with the periodontal membrane first The defect area was covered with BME collagen membrane carrying BMSCs,and then covered with e-PTFE membrane.After 4 weeks of treatment,the area of new-born alveolar bone and cementum,the width of new-born periodontal ligament,the content of interleukin(IL)-1β,tumor necrosis factor(TNF)-αand IL-6 in serum and periodontal defect tissue of rats in each group,the expression levels of M1/M2 macrophage markers in peripheral defect tissues were measured.Results There was no significantly different between e-PTFE group and BME group on new alveolar bone and cementum area,new periodontal ligament width,serum and periodontal defect tissue IL-1β,TNF-αand IL-6 content,and the expression of CD16,iNOS,CD206 and Arg1 mRNA in the periodontal defect tissue(P>0.05).At the 4 th week after operation,the body weight,the area of new alveolar bone and cementum,the width of the new periodontal ligament and the CD206 and Arg1 mRNA in the tissues of the periodontal defect in the BMSCs+BME group were significantly higher than those in the e-PTFE group and the BME group,and serum and periodontal defect area tissue IL-1β,TNF-αand IL-6 content and periodontal defect area tissue CD16 and iNOS mRNA in the BMSCs+BME group were significantly lower than those in the e-PTFE group and the BME group,the differences were statistically significant(P<0.05).Conclusion Bone marrow stromal cells can promote the polarization of M2 macrophages in the periodontal tissue defect of rats,thereby inhibiting the inflammatory response of periodontal defect tissue and promoting the repair of periodontal defect tissue.