| Research background and purposePeriodontitis is an inflammatory disease caused by plaque biofilm and characterized by the destruction of periodontal tissue.Periodontal pocket is a pathologically deepened gingival sulcus.It is one of the most important pathological changes of periodontitis.In addition,periodontal pathogens and their metabolites can inhibit the expression of various junctional proteins between epithelial cells and damage the epithelial barrier function.Subgingival scaling is necessary for the treatment of periodontitis in clinical practice.Mechanical damage during the treatment leads to varying degrees of destruction of the epithelium and connective tissue in the pocket and bleeding,resulting in pain.Bacteria in the oral cavity can enter the blood circulation through the ulcer surface of the periodontal pocket and cause transient bacteremia.The periodontal epithelium plays an important role in the control of microbial infection and the protection of subepithelial tissues.Therefore,it is of great significance to find an adjuvant therapy for the treatment of damaged periodontal pocket tissue.Oral ulcer powder is a kind of topical powder,which is composed of indigo naturalis,dried alum and borneol.It has the effect of healing sore and relieving pain,and is mainly used for the treatment of oral ulcer.Many clinical studies have shown that oral ulcer powder can significantly relieve pain,shorten healing time,reduce attack frequency and prolong recurrence interval in patients with recurrent oral ulcer.Its main component,indigo naturalis,can increase the expression of tight junction proteins and promote the healing of mucosal and skin lesions,has a certain effect on the treatment of psoriasis and ulcerative colitis.And traditional Chinese medicine is cheap、easy to obtain and has a long history,less side effects.Although oral ulcer powder has been widely used in clinical practice,it has not been reported in the literature as an adjuvant drug for periodontitis.The aim of this study was to explore whether the drug could promote the healing of the epithelium in the periodontal pocket,repair the histological morphology of the inner wall of the periodontal pocket,reduce inflammatory response,enhance the epithelial barrier function,and promote epithelial closure,so as to be applied in the treatment of periodontitis.Materials and methods1.Establishment of a rat model of experimental periodontitis with epithelial damage10 male Wistar rats were randomly divided into normal group(n=5)and model group(n=5).The normal group did not receive any treatment,and the model group was established by ligating the maxillary first molar with suture(orthodontic ligation wire combined with silk suture)and stimulating with butyrate and lipopolysaccharide.Ten rats were sacrificed after 5 days for sampling.The left maxillary first molars of the normal group and the model group were taken to make paraffin sections,and HE staining was performed to observe the histological changes to verify the successful establishment of the model.2.The study of oral ulcer powder in repairing epithelial damage of experimental periodontitis in rats20 male Wistar rats were randomly divided into natural healing group and treatment group with 10 rats in each group.Periodontal epithelial injury model was established in 20 rats,and ligature wires were removed 5 days later.The natural healing group did not do any treatment to heal naturally.the treatment group was given oral ulcer powder in periodontal pocket of rats 3 times a day.On the 2nd and 4th day of administration,5 samples were killed in each group.Paraffin sections were made from the left maxillary first molar of the rat.HE staining was performed for histological observation and statistical analysis.Occludin immunohistochemical staining and immunofluorescence staining were used to detect the expression levels of occludin protein in the two groups.Analyze the effect of oral ulcer Powder on epithelial injury of experimental periodontitis in rats.3.The effect of oral ulcer powder on inflammation in the rat experimental periodontitis model with epithelial damageGingiva around the right maxillary first molar of rats in normal group,model group,natural healing group(4d)and treatment group(4d)were taken.The expression levels of IL-1β,IL-6 and TNF-α in the four groups were detected by qRT-PCR.Masson staining was performed on paraffin sections to observe collagen fibers.Analyze the effect of oral ulcer Powder on periodontal inflammation in model rats.Results1.There was no abnormality in the periodontal tissue in the normal group,while the rats in the model group showed obvious attachment loss,periodontal inflammation and alveolar bone resorption.HE staining showed epithelial ulcer in the periodontal pocket of the model group,and the rat experimental periodontitis model was successfully established.2.He staining observation and statistical analysis showed that at the 2nd day,re epithelialization of epithelial ulcers within the periodontal pocket was more obvious in the treatment group than in the natural healing group,with statistical difference;At the 4th day,the two groups basically healed with no statistical difference.Immunohistochemistry and immunofluorescence staining showed that the expression of occludin protein in the treatment group(4d)was higher than that in the natural healing group(4d),and the difference was statistically significant.3.The results of qRT-PCR showed that the expression of inflammatory factors IL-1β,IL-6,and TNF-α in the model group was significantly higher than that in the normal group,the treatment group(4d)and the natural healing group(4d).The expression of inflammatory factors in the treatment group(4d)was significantly lower than that in the natural healing group(4d),and the differences were statistically significant.Masson staining showed that the collagen fibers in the model group were proliferated and the collagen fiber bundles were seriously broken.The collagen fibers in the treatment group(4d)and the natural healing group(4d)were repaired,but the treatment group(4d)had less collagen fiber proliferation and more new capillary formation than the natural healing group(4d).Conclusion1.An effective rat model of experimental periodontitis with periodontal epithelial damage could be established by suture ligation,lipopolysaccharide and butyrate stimulation.This model can better reproduce the state of the periodontal pocket of the periodontitis patients than the previous periodontitis model.2.In vivo,oral ulcer powder could promote the healing of periodontal pocket epithelial ulcer,promote the expression of tight junction protein occludin,and enhance epithelial barrier function.3.In vivo,oral ulcer powder was found to reduce periodontal tissue inflammation. |
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