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Abstract：
OBJECTIVE: To assess the efficacy of Phyllanthus emblica extract in alleviating halitosis and reducing the inflammatory response to halitosis-related bacteria.
METHODS: This investigation, using Phyllanthus emblica fruit extract (PE), involved four aspects. First, we evaluated the effect on growth and aggregation of halitosis-related bacteria, including Fusobacterium nucleatum, Porphyromonas gingivalis, and Solobacterium moorei, using a microdilution assay and scanning electron microscopy. Second, volatile sulfur compound (VSC) levels were measured on individuals with halitosis in randomized short-term (26 participants) and double-blind randomized long-term trials (18 participants in each group) after rinsing with PE for 3, 6, and 12 h, and 28 days. Third, we analyzed pro-inflammatory cytokine expression in TR146 cells using quantitative real-time PCR and enzyme-linked immunosorbent assays. Lastly, we assessed pro-inflammatory cytokine secretion and Toll-like receptor (TLR) 2 mRNA expression via the same experimental methods in a three-dimensional oral mucosal epithelial model (3D OMEM).
RESULTS: PE extract dose-dependently inhibited the growth of F. nucleatum (50% inhibition concentration [IC50]=0.079%), P. gingivalis (IC50=0.65%), and S. moorei (IC50=0.07%) and effectively prevented bacterial aggregation. Furthermore, VSC contents decreased significantly at 3, 6, and 12 h after rinsing with 5% PE compared with those in the control. Long-term use of mouthwash containing 5% PE for 28 days led to a significant decrease in VSC contents. PE attenuated the F. nucleatum- or P. gingivalis-stimulated mRNA expression and protein release of interleukin (IL)-6 and IL-8 in TR146 cells. It also suppressed IL-8 and prostaglandin E2 secretion and TLR2 mRNA expression in F. nucleatum-induced OMEMs.
CONCLUSIONS: Our findings support the use of PE in oral care products to alleviate halitosis and it may reduce inflammation.
METHODS: This investigation, using Phyllanthus emblica fruit extract (PE), involved four aspects. First, we evaluated the effect on growth and aggregation of halitosis-related bacteria, including Fusobacterium nucleatum, Porphyromonas gingivalis, and Solobacterium moorei, using a microdilution assay and scanning electron microscopy. Second, volatile sulfur compound (VSC) levels were measured on individuals with halitosis in randomized short-term (26 participants) and double-blind randomized long-term trials (18 participants in each group) after rinsing with PE for 3, 6, and 12 h, and 28 days. Third, we analyzed pro-inflammatory cytokine expression in TR146 cells using quantitative real-time PCR and enzyme-linked immunosorbent assays. Lastly, we assessed pro-inflammatory cytokine secretion and Toll-like receptor (TLR) 2 mRNA expression via the same experimental methods in a three-dimensional oral mucosal epithelial model (3D OMEM).
RESULTS: PE extract dose-dependently inhibited the growth of F. nucleatum (50% inhibition concentration [IC50]=0.079%), P. gingivalis (IC50=0.65%), and S. moorei (IC50=0.07%) and effectively prevented bacterial aggregation. Furthermore, VSC contents decreased significantly at 3, 6, and 12 h after rinsing with 5% PE compared with those in the control. Long-term use of mouthwash containing 5% PE for 28 days led to a significant decrease in VSC contents. PE attenuated the F. nucleatum- or P. gingivalis-stimulated mRNA expression and protein release of interleukin (IL)-6 and IL-8 in TR146 cells. It also suppressed IL-8 and prostaglandin E2 secretion and TLR2 mRNA expression in F. nucleatum-induced OMEMs.
CONCLUSIONS: Our findings support the use of PE in oral care products to alleviate halitosis and it may reduce inflammation.
摘要：
目的：评价余甘子提取物减轻口臭和减轻口臭相关细菌炎症反应的疗效。
方法：这项调查，使用余甘子果实提取物（PE），涉及四个方面。首先,我们评估了对口臭相关细菌生长和聚集的影响，包括具核梭杆菌,牙龈卟啉单胞菌,和Solobacteriummoorei,使用微量稀释测定和扫描电子显微镜。第二,在用PE冲洗3、6和12小时后，在随机短期（26名参与者）和双盲随机长期试验（每组18名参与者）中测量了口臭个体的挥发性硫化合物（VSC）水平。28天。第三，我们使用实时定量PCR和酶联免疫吸附试验分析了TR146细胞中促炎细胞因子的表达.最后,我们通过相同的实验方法在三维口腔粘膜上皮模型(3DOMEM)中评估促炎细胞因子分泌和Toll样受体(TLR)2mRNA表达.
结果：PE提取物剂量依赖性地抑制核仁F.的生长（50％抑制浓度[IC50]=0.079％），牙龈卟啉单胞菌（IC50=0.65%），和S.moorei（IC50=0.07％），并有效防止细菌聚集。此外，与对照相比，用5％PE冲洗后3、6和12小时VSC含量显着降低。长期使用含5%PE的漱口水28天导致VSC含量显著降低。PE减弱了TR146细胞中白细胞介素（IL）-6和IL-8的核或牙龈卟啉单胞菌刺激的mRNA表达和蛋白质释放。它还抑制了有核F.诱导的OMEMs中IL-8和前列腺素E2的分泌以及TLR2mRNA的表达。
结论：我们的发现支持在口腔护理产品中使用PE来减轻口臭，并且可以减轻炎症。
方法：这项调查，使用余甘子果实提取物（PE），涉及四个方面。首先,我们评估了对口臭相关细菌生长和聚集的影响，包括具核梭杆菌,牙龈卟啉单胞菌,和Solobacteriummoorei,使用微量稀释测定和扫描电子显微镜。第二,在用PE冲洗3、6和12小时后，在随机短期（26名参与者）和双盲随机长期试验（每组18名参与者）中测量了口臭个体的挥发性硫化合物（VSC）水平。28天。第三，我们使用实时定量PCR和酶联免疫吸附试验分析了TR146细胞中促炎细胞因子的表达.最后,我们通过相同的实验方法在三维口腔粘膜上皮模型(3DOMEM)中评估促炎细胞因子分泌和Toll样受体(TLR)2mRNA表达.
结果：PE提取物剂量依赖性地抑制核仁F.的生长（50％抑制浓度[IC50]=0.079％），牙龈卟啉单胞菌（IC50=0.65%），和S.moorei（IC50=0.07％），并有效防止细菌聚集。此外，与对照相比，用5％PE冲洗后3、6和12小时VSC含量显着降低。长期使用含5%PE的漱口水28天导致VSC含量显著降低。PE减弱了TR146细胞中白细胞介素（IL）-6和IL-8的核或牙龈卟啉单胞菌刺激的mRNA表达和蛋白质释放。它还抑制了有核F.诱导的OMEMs中IL-8和前列腺素E2的分泌以及TLR2mRNA的表达。
结论：我们的发现支持在口腔护理产品中使用PE来减轻口臭，并且可以减轻炎症。
