The Effects Of Luteolin And Hesperitin On The Proliferation And Differentiation In Human Periodontal Ligament Cell Population

ObjectiveThe aim of this study was to explore the potential beneficial effect of effectivecomponents contained in traditional Chinese medicine on treating periodontal diseaseby observing the influence of Luteolin and Hesperitin on the proliferation anddifferentiation of human periodontal ligament cell population (PDLP), viewing toprovide the theoretical basis for the rational medicine in the application of periodontaldisease.MethodsHuman periodontal ligament cell population was primarily cultured by cover slipcovering tissue culture method in vitro. Then observed the growth conditions ofPDLP under inverted phase contrast microscope. Moreover, the progenitor of humanperiodontal ligament cell population was identified by Immunohistochemicalmethods and the cell growth curve was determined by MTT assay. Besides, theinfluence of Luteolin and Hesperitin on the proliferation and differentiation of humanperiodontal ligament cell population were investigated by MTT assay, ALP Kit,Coomassie brilliant blue assay, Q-PCR.Results1. The periodontal ligament cell population was in good condition. The growth curvemodel of PDLP liked "S". Immunohistochemical analysis showed that the vimentinstaining was in positive, the certain was in negative. The osteogenic induction ofPDLP showed that: Mineralization nodule was formed and alizarin red staining wasred.2. The effects of Luteolin and Hesperitin on the proliferation of human periodontalligament cell population.The MTT assay results showed that Luteolin and Hesperitin (100μM,10μM,1μM,0.1μM,0.01μM) could increase the proliferation of PDLP，and there weresignificant differences when compared with control group（P<0.05） 3. The effects of Luteolin and Hesperitin on the differentiation of PDLP.The ALP Kit, Coomassie brilliant blue assay results showed that Luteolin andHesperitin (100μM,10μM,1μM,0.1μM,0.01μM) can increase the ALP and totalprotein of PDLP（P<0.05）. The Q-PCR results showed that Luteolin and Hesperitin(100μM,10μM,1μM,0.1μM,0.01μM) can increase the expression of ALP（P<0.05）, except Luteolin in0.01μM, and Luteolin(1μM,0.1μM) and Hesperitin(0.1μM,0.01μM) can increase the expression of RUNX2（P<0.05）.Conclusion1. PDLP can be successfully cultured by Tissue adherent method and can differentiateinto boon-like tissue.2. At proper concentration, Luteolin and Hesperitin can increase the proliferation anddifferentiation of PDLP.