| Background and objectiveOral ulcer is one of the most common complaints by patients in oral mucosa diseases.The exact pathogenesis of oral ulcer is still unknown.The inducements included various factors,such as trauma,infection,immune disorders,heredity,and psychological factors and so on.Some scholars pointed that oral ulcer is a basic clinical lesion,not an independent disease.There is still a lack of gold standard of treatment protocols.Local symptomatic treatment is the main treatment at present for the sake of antiphlogosis,analgesia,preventing secondary infection and promoting healing.Mouthwash is one style of preparation.Traditional Chinese medicine(TCM)has a long history of healthcare in China.Chinese herbal medicine(CHM)is an important composition of TCM.Compared to a single herb,the compound CHM is not a simple overlay of multi herbs and pays more attention to holistic.Several studies have demonstrated that compound CHM was effective on oral ulcer.Honeysuckle,chrysanthemumim indici flos,artemisia scopariae herba,angelicae dahuricae radix and asarui radix et rhizoma have been utilizing as the common CHM.Our research group applied the method of steam distillation to extract the five herbs.We further adopt the uniform design to confirm their optimal ratios by three indexs:the antibacterial activity,the inhibition rate of biofilm formation and the toxicity of human gingival fibroblasts.According to the optimal ratios,we compounded the extracts of the five herbs to invent a new Chinese medicine mouthwash,and found that it can effectively inhibit the inflammatory of experimental periodontitis.The aim of this study was to analyse the effect of this mouthwash for oral ulcer and the oral toxicity.Methods1.Under anesthesia with 2%pentobarbital sodium,rats were treated vertically with a plastic tube(diameter:5mm,length:4cm)which contained cotton sheets and soaked 70μL 20%sodium hydroxide solution on the right cheek mucosa for 30 seconds to establish the traumatic oral ulcer.2.Sixty female rats were inducted the oral ulcers by the above-mention method.All animals were randomly divided into four groups:test group,chlorhexidine positive group,LMZ positive group and negative group.After inhalation anesthesia of diethyl ether,the applications of 20 μL corresponding liquids were administered to cover the surface of ulcers for 3 min twice daily.The diameter of ulcers were measured daily by electronic vernier caliper to calculate the area.HE staining sections were made for histological observation.3.Acute oral toxicity test:20 rats were randomly divided into 2 groups(5 rats per sex per group):test group and control group.Animals in test group were given the compound honeysuckle Chinese medicine mouthwash by gavage,while in control group were given sterile water.The dose of intragastric administration was 5000mg per kg body weight.Abnormal clinical signs,body weight and food consumption were recorded for nine days.Necropsies were performed at the ninth day and organ coefficients were calculated.4.Subchronic oral toxicity test:60 rats were randomly divided into test group and control group(15 rats per sex per group).Animals in test group were received a gavage of the compound honeysuckle Chinese medicine mouthwash once a day for 93 days,while in control group were given sterile water.The dose of intragastric administration was 1000mg per kg body weight,and was adjusted every week.After 93-day drug administration,animals continued to be observed without treatment for 28 days as recovery period.During the experimental period,all animals were observed once a day for mortality and any abnormal clinical signs.Individual animal body weight and food consumption were measured every week during drug administration and recovery period.Hematology,serum chemistry and necropsy were conducted at day 45,93 and 118.Organ coefficients were calculated,and HE staining sections of organs were made for histological observation.Results1.After one day of induction,a rounded oral ulcer in the right cheek mucosa was clinically visible with delineating erythematous border and covered faint yellowish pseudomembrane.The model of traumatic oral ulcer was successfully established.2.According to the clinical performance,the changes of area and histopathology of ulcers in the four group,the compound honeysuckle Chinese medicine mouthwash can shorten the healing time of traumatic oral ulcer compare with the negative group,which was similar to chlorhexidine positive group and LMZ positive group.3.In acute oral toxicity study,the animals in test group didn’t develop abnormal clinical signs.There were no statistic differences of body weight,food consumption and organ coefficients between test group and control group(P>0.05).No abnormal organs or tissues were found in necropsies.4.In subchronic oral toxicity study,no mortality and abnormal clinical signs were observed in all animals during the 121-day study.There were no statistical differences in animal weight or food consumption between test group and control group(P>0.05).The differences between test group and control group had statistical significance in several parameters of hematology and serum chemistry at different periods(P<0.05),but these changes were within normal physiological range.There were no significant differences in organ coefficients of the above organs between the test and control groups(P>0.05).No abnormal organs or tissues were found in the necropsies and histopathology.Conclusions1.Compound honeysuckle Chinese medicine mouthwash can promote the healing of traumatic oral ulcer model in rats.2.Compound honeysuckle Chinese medicine mouthwash has no acute oral toxicity.3.Compound honeysuckle Chinese medicine mouthwash has no obvious sbchronic oral toxicity... |
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