摘要
目的 通过大鼠牙周炎模型来考察比较甲硝唑羧甲基壳聚糖凝胶(M/CMCS)等试药对牙周组织和全血白介素1β(IL-1β)的影响。方法 采用SD大鼠上颌第一磨牙牙龈缘细丝结扎并接种牙龈卟啉单胞菌(P.gingivalis)法造模。72只模型大鼠随机分为4大组,每组18只。MODEL组(不涂布任何药物),M/CMCS组(涂布甲硝唑羧甲基壳聚糖凝胶),PERIO组(涂布盐酸米诺环素软膏),MNZ组(涂布甲硝唑凝胶)。造模成功后每天在结扎牙龈缘处涂药治疗2次。在药物治疗开始后满1周,满3周和满5周的3个不同时间点,各组分别取6只大鼠采集试验样本。大鼠腹腔麻醉后采集龈沟液和全血样本,然后分离截取上颌骨及牙周组织块,一侧组织块分离牙周软组织和牙槽骨,供ELISA实验测定IL-1β浓度,对侧组织块用于IL-1β的免疫组化实验。结果 龈沟液和牙周软组织的IL-1β浓度结果表现相似,与MODEL组比较,M/CMCS组、PERIO组和MNZ组在第1周、第3周和第5周各相同时间点均明显降低(P<0.05);M/CMCS组和PERIO组在3个相同时间点较MNZ组低(P<0.05);而M/CMCS组在第5周时比PERIO组又进一步降低(P<0.05)。而牙槽骨的IL-1β浓度,在第5周时,M/CMCS组与PERIO组差别未见统计学意义(P>0.05)。全血IL-1β浓度值,与MODEL组比较,3个药物治疗组在第1,3,5周均明显下降(P<0.05)。IL-1β免疫组化的平均光密度(MOD)值,3个药物治疗组比MODEL组在3个不同时间点均降低(P<0.05);M/CMCS组和PERIO组在第1,3,5周相同时间点均明显低于MNZ组(P<0.05),在第5周时间点,M/CMCS组低于PERIO组(P<0.05)。形态学观察,3个药物治疗组牙周状况较MODEL组均有不同程度改善,其中M/CMCS组的牙周形态较完整,牙槽骨吸收程度最轻,明显优于PERIO组和MNZ组。结论 甲硝唑羧甲基壳聚糖凝胶对大鼠牙周炎有抗炎和抗牙槽骨吸收作用,其机制可能在于其能够降低大鼠牙周炎龈沟液、牙周组织和全血中IL-1β的表达。
Objective To investigate the change of interleukin 1β(IL-1β)in periodontal tissues and blood from the periodontitis rats treated by metronidazole carboxymethyl chitosan gel(M/CMCS) and other control drugs.Methods The experimental periodontitis models of SD rats were established by orthodontic ligatures fixed around the cervical margins of the bilateral maxillary first molars together with the local infection of P.gingivalis.Seventy-two periodontitis model rats were randomly divided into 4 groups with 18 rats in each as follows: MODEL group(without any drugs),M/CMCS group(coated with metronidazole carboxymethyl chitosan gel),PERIO group(coated with minocycline hydrochloride ointment) and MNZ group(coated with metronidazole gel).Once the periodontitis models were confirmed to be successful, then different drugs were coated locally at the sites of gingival margin twice a day according to the different groups.Six rats were taken from each group for the collection of experimental samples at the end of the 1 st week, the 3 rd week and the 5 th week respectively after the beginning of different treatments for the rats in 4 groups.The rats were anesthetized intraperitoneally firstly, and the gingival crevicular fluids and whole blood samples were collected, and then two sides of the maxillary and periodontal tissue block were separated and intercepted after the rats were euthanized.Periodontal soft tissues and alveolar bone were separated from one side of the tissue block to analysis IL-1 β concentrations by ELISA tests, and the other side was used for IL-1 β immunohistochemical experiments.Results The statistical analysis results of the IL-1β concentrations in gingival crevicular fluids and periodontal soft tissues had a similar performance.Compared with the MODEL group, the concentrations of IL-1β in gingival crevicular fluid in M/CMCS group, PERIO group and MNZ group decreased significantly at the same time points in the first week, the third week and the fifth week(P<0.05);the concentrations of IL-1β of M/CMCS group and PERIO group were lower than those of MNZ group at the same time points(P<0.05);and the concentrations of IL-1β in M/CMCS group was lower than that in the PERIO group at the 5 th week(P<0.05).But there was no significant difference between M/CMCS group and PERIO group(P>0.05) for the IL-1β in alveolar bones at the fifth week.Compared with MODEL group, the concentrations of IL-1β in whole blood decreased significantly in M/CMCS group, PERIO group and MNZ group at 1-,3-and 5-week time points(P<0.05).The mean optical density(MOD) values of IL-1β immunohistochemical of the periodontal tissue in the three drug treatment groups were significantly lower than those in the MODEL group at different stages(P<0.05).The MOD values in M/CMCS group and PERIO group were significantly lower than that of MNZ group at the same time point of the first, third and fifth week(P<0.05),and the MOD value in M/CMCS group was lower than that in PERIO group at the same time point of the fifth week(P<0.05).Histopathomorphological observation showed that the periodontal conditions of the three drug treatment groups were improved to varying degrees compared with those of the MODEL group.Among them, the periodontal morphology of the M/CMCS group was more complete, and the alveolar bone absorption was the least, which was significantly better than that of the PERIO group and the MNZ group.Conclusion Metronidazole carboxymethyl chitosan gel has the effects of anti-inflammatory and anti-resorption of alveolar bone on rat periodontitis, and its mechanism may be related to the reductions of IL-1β in gingival crevicular fluids, periodontal tissues and whole blood of rats with periodontitis.
作者
陈东
周焱祎
CHEN Dong;ZHOU Yanyi(Department of Stomatology,Tangshan Second Hospital,Tangshan 063000,China)
出处
《中国煤炭工业医学杂志》
2022年第5期453-461,共9页
Chinese Journal of Coal Industry Medicine
基金
河北省卫计委医学科学研究重点课题计划(编号:20150971)。
关键词
牙周炎
白介素1Β
甲硝唑羧甲基壳聚糖凝胶
Periodontitis
Interleukin-1β
Metronidazole carboxymethyl chitosan gel
作者简介
通讯作者:陈东。
