The Experimental Study Of Chinese Nutgall And Scutellaria Baicalensis Georgi Extracts Inhibiting The Toxic Effect Of LPS In Human Periodontal Ligament Cells

At present, root canal therapy is the first-choice and the most effective therapy for pupal and periapical diseases. As the most important procedure of root canal therapy, root canal irrigation can eliminate and control the bacteria and bacterial products from the root canal system. Chinese nutgall has antibacterial activities and styptic effect; Scutellaria Baicalensis Georgi has many beneficial pharmacological effects including antioxidant, anti-inflammatory and anticancer effects. This study was to prepare medical powder by extracting from Chinese nutgall and Scutellaria Baicalensis Georgi as a new irrigant solution, in order to investgate the effect of it inhibiting the toxic of LPS in human periodontal ligament cells and to provide the theoretical and experimental proof for a new irrigant solution.Objective:To investigate suppression effect of different concentration Chinese nutgall and Scutellaria Baicalensis Georgi extracts in human periodontal ligament cells (HPDLs) treated with lipopolysaccharide, providing the theoretical foundation for a new irrigant solution.Methods:Human periodontal ligament cells were obtained in vitro by explant method and the study consists of two parts: Part1MTT was used to detect suppression effect of different concentration Chinese nutgall and Scutellaria Baicalensis Georgi extracts on human periodontal ligament cells proliferation treated with lipopolysaccharide. Experiments were carried out with human periodontal ligament cells which were plated in96-well plates from the fifth passages and were divided into three groups at random. Blank control group only added DMEM;drug group added different concentration Chinese nutgall and Scutellaria Baicalensis Georgi extracts(12.5μg/ml,25μg/ml,50μg/ml) and100μg/ml LPS in medium; LPS group added100μg/ml LPS in medium. After24h,48,,2h,Cell viability was determined using the3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.Part2To investigate the effect of Chinese nutgall and Scutellaria Baicalensis Georgi extracts on pro-inflammatory factor in human periodontal ligament cells treated with lipopolysaccharide. Human periodontal ligament cells were studied at the fifth passages and were divided into three groups at random. Blank control group only added DMEM; drug group added different concentration Chinese nutgall and Scutellaria Baicalensis Georgi extracts (12.5μg/ml,25μg/ml,50μg/ml)and100μg/ml LPS in medium; LPS group added100μg/ml LPS in medium. After24h,48,72h, ELISA was applied to detect the TNF-aαlevel of different test group in the culture supernatant.Results:1. Compared with control group,100μg/ml LPS was able to inhibit the HPDLs proliferation ability remarkably at24h,48h and72h (p<0.05). As time went on,there was significant difference between LPS group and three drug groups under the stimulation of LPS at24h、48h (p<0.05). At72h, only25μg/ml drug group could improve the HPDLs proliferation ability significantly. The optimal effect was25μg/ml at24h. 2. The level of TNF-a of LPS group was apparently higher than the blank group (p<0.05). Lower level TNF-a were observed in three drug groups than LPS group (p<0.05),especially in the25μg/ml at24h.Conclusion:Chinese nutgall and Scutellaria Baicalensis Georgi extracts could effectively suppress the toxic effect of lipopolysacchride in HPDLs and also have protective role on HPDLs.