目的 :研究来源于脂多糖(LPS)诱导的牙周炎大鼠上颌骨的间充质干细胞(MSC)表达破骨细胞因子的变化。方法:注射脂多糖构建牙周炎大鼠模型,6周后,处死大鼠,利用Micro CT观察上颌骨骨吸收情况,HE染色观察牙周组织的变化,抗酒石酸磷酸酶(TRAP)染色观察上颌骨内破骨细胞的数量;体外分离和培养大鼠上颌骨来源的MSCs,荧光定量PCR(q PCR)检测两组间充质干细胞中M-CSF、OPG和RANKL m RNA表达量,并计算OPG与RANKL的比值。结果:Micro CT显示实验组大鼠上颌骨牙槽骨吸收明显;HE染色显示实验组切片中牙龈明显退缩,结合上皮根向增殖,并有炎性细胞浸润;TRAP染色显示实验组上颌骨内破骨细胞数量明显增多;荧光定量PCR显示实验组BMMSCs表达的M-CSF增加,表达的OPG和RANKL减少,但OPG与RANKL的比值减小,差异具有统计学意义。结论:来源于LPS诱导的牙周炎大鼠,其颌骨的间充质干细胞在炎症状态下,表达的与破骨相关因子上调,可能有增强破骨细胞的功能,能加速骨吸收进程。
Objective: The aim of the present study is to investigate the effect on osteoclasts related cytokines of maxilla bone mesenchymal stem cells in periodontitis model of rat induced by liposaceharide (LPS). Methods: Periodontal inflammation was induced by injections of LPS into rat maxillary gingiva at the palatal aspect of the molars. Animals were killed after 6 weeks, and linear and volumetric alveolar bone loss was measured by mieroeomputed tomography. The prevalence of inflammatory infiltrate and osteoclasts was assessed from hematoxylin and eosin or tartrate-resistant acid phosphatase (TRAP)-stained sections. And maxillae were dissected for MSCs isolation and culture, and M-CSF, OPG and RANKL mRNA levels of MSCs was examined by qPCR, and the ratio of OPG to RANKL was calculated. Results: Linear and volumetric analysis of maxillae by Micro CT indicated significant loss of bone with LPS administration. Histologic examination revealed destruction of the periodontal ligament, increased inflammatory infiltrate, and more TRAP-positive osteoclasts in the LPS group compared to controls. Treatment of LPS injection up-regulates the expression of M-CSF and RANKL, and down-regulates OPG and RANKL expression of MSCs against control group, and the ratio of OPG to RANKL is down- regulated. Conclusion: The expression of M-CSF, OPG and RANKL by BM-MSCs in the periodontitis maxilla induced by LPS, may enhance the maturation and differentiation of osteoclasts and accelerate the process of bone absorption.