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Stimulating Nonshivering Thermogenesis in Cold Exposed Humans: Emphasis on the Action of Green Tea ExtractsGosselin, Chantal 10 January 2012 (has links)
It has been demonstrated that EGCG and caffeine, naturally present in green tea, have thermogenic properties in thermoneutral conditions. The purpose of this study was to quantify the effect of the combined ingestion of EGCG/caffeine on thermogenic responses during a 3h mild cold exposure. Eight healthy males (22± 1 y) were exposed in a randomized, cross over, single blinded fashion to the cold (liquid conditioned suit perfused with 15°C water), after ingesting either a placebo (CON) or an extract of 1600mg of EGCG and 600mg of caffeine (EXP). Thermic, metabolic and electromyographic measurements were monitored at baseline and during cold exposure. After 180min of cold exposure, shivering intensity was significantly reduced by ~32% in EXP condition compared to CON. Area under the curve calculations for total shivering intensity was also reduced by ~21% in EXP (457±99 %MVC.min) compared to CON (361±81 %MVC.min; p=0.007). In contrast, the total area under curve of VO2 was ~25% higher in EXP (33.3±5.5 L O2) compared to CON (25.3±5.1 L O2; p=0.03). Total Heat production (Hprod) also increased by about 11% in the EXP condition (1535±112 kJ) compared to control (1372 ±106 kJ; p=0.002). The decrease in shivering activity combined with an increase in VO2 and Hprod, following the ingestion of EGCG and caffeine in the cold, indicates that nonshivering thermogenesis pathways can be significantly stimulated in adult humans.
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Stimulating Nonshivering Thermogenesis in Cold Exposed Humans: Emphasis on the Action of Green Tea ExtractsGosselin, Chantal 10 January 2012 (has links)
It has been demonstrated that EGCG and caffeine, naturally present in green tea, have thermogenic properties in thermoneutral conditions. The purpose of this study was to quantify the effect of the combined ingestion of EGCG/caffeine on thermogenic responses during a 3h mild cold exposure. Eight healthy males (22± 1 y) were exposed in a randomized, cross over, single blinded fashion to the cold (liquid conditioned suit perfused with 15°C water), after ingesting either a placebo (CON) or an extract of 1600mg of EGCG and 600mg of caffeine (EXP). Thermic, metabolic and electromyographic measurements were monitored at baseline and during cold exposure. After 180min of cold exposure, shivering intensity was significantly reduced by ~32% in EXP condition compared to CON. Area under the curve calculations for total shivering intensity was also reduced by ~21% in EXP (457±99 %MVC.min) compared to CON (361±81 %MVC.min; p=0.007). In contrast, the total area under curve of VO2 was ~25% higher in EXP (33.3±5.5 L O2) compared to CON (25.3±5.1 L O2; p=0.03). Total Heat production (Hprod) also increased by about 11% in the EXP condition (1535±112 kJ) compared to control (1372 ±106 kJ; p=0.002). The decrease in shivering activity combined with an increase in VO2 and Hprod, following the ingestion of EGCG and caffeine in the cold, indicates that nonshivering thermogenesis pathways can be significantly stimulated in adult humans.
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Stimulating Nonshivering Thermogenesis in Cold Exposed Humans: Emphasis on the Action of Green Tea ExtractsGosselin, Chantal 10 January 2012 (has links)
It has been demonstrated that EGCG and caffeine, naturally present in green tea, have thermogenic properties in thermoneutral conditions. The purpose of this study was to quantify the effect of the combined ingestion of EGCG/caffeine on thermogenic responses during a 3h mild cold exposure. Eight healthy males (22± 1 y) were exposed in a randomized, cross over, single blinded fashion to the cold (liquid conditioned suit perfused with 15°C water), after ingesting either a placebo (CON) or an extract of 1600mg of EGCG and 600mg of caffeine (EXP). Thermic, metabolic and electromyographic measurements were monitored at baseline and during cold exposure. After 180min of cold exposure, shivering intensity was significantly reduced by ~32% in EXP condition compared to CON. Area under the curve calculations for total shivering intensity was also reduced by ~21% in EXP (457±99 %MVC.min) compared to CON (361±81 %MVC.min; p=0.007). In contrast, the total area under curve of VO2 was ~25% higher in EXP (33.3±5.5 L O2) compared to CON (25.3±5.1 L O2; p=0.03). Total Heat production (Hprod) also increased by about 11% in the EXP condition (1535±112 kJ) compared to control (1372 ±106 kJ; p=0.002). The decrease in shivering activity combined with an increase in VO2 and Hprod, following the ingestion of EGCG and caffeine in the cold, indicates that nonshivering thermogenesis pathways can be significantly stimulated in adult humans.
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Stimulating Nonshivering Thermogenesis in Cold Exposed Humans: Emphasis on the Action of Green Tea ExtractsGosselin, Chantal January 2012 (has links)
It has been demonstrated that EGCG and caffeine, naturally present in green tea, have thermogenic properties in thermoneutral conditions. The purpose of this study was to quantify the effect of the combined ingestion of EGCG/caffeine on thermogenic responses during a 3h mild cold exposure. Eight healthy males (22± 1 y) were exposed in a randomized, cross over, single blinded fashion to the cold (liquid conditioned suit perfused with 15°C water), after ingesting either a placebo (CON) or an extract of 1600mg of EGCG and 600mg of caffeine (EXP). Thermic, metabolic and electromyographic measurements were monitored at baseline and during cold exposure. After 180min of cold exposure, shivering intensity was significantly reduced by ~32% in EXP condition compared to CON. Area under the curve calculations for total shivering intensity was also reduced by ~21% in EXP (457±99 %MVC.min) compared to CON (361±81 %MVC.min; p=0.007). In contrast, the total area under curve of VO2 was ~25% higher in EXP (33.3±5.5 L O2) compared to CON (25.3±5.1 L O2; p=0.03). Total Heat production (Hprod) also increased by about 11% in the EXP condition (1535±112 kJ) compared to control (1372 ±106 kJ; p=0.002). The decrease in shivering activity combined with an increase in VO2 and Hprod, following the ingestion of EGCG and caffeine in the cold, indicates that nonshivering thermogenesis pathways can be significantly stimulated in adult humans.
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Prevention of abdominal aortic aneurysm progression by oral administration of green tea polyphenol in a rat model / 緑茶ポリフェノール経口投与によるラット腹部大動脈瘤進展抑制効果Setozaki, Shuji 23 March 2017 (has links)
京都大学 / 0048 / 新制・課程博士 / 博士(医学) / 甲第20236号 / 医博第4195号 / 新制||医||1019(附属図書館) / 京都大学大学院医学研究科医学専攻 / (主査)教授 木村 剛, 教授 松原 和夫, 教授 川村 孝 / 学位規則第4条第1項該当 / Doctor of Medical Science / Kyoto University / DFAM
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Efeito do curativo de demora com EGCG, derivada do chá verde, na lesão periapical em cães / Effect of intracanal dressing with EGCG, derived from green tea, in periapical lesions in dogsLiévana, Fernanda Souza 07 December 2018 (has links)
O chá verde vem sendo utilizado na prevenção e tratamento de variadas doenças infecciosas e imunoinflamatórias, por apresentar efeitos benéficos decorrentes da presença de Epigalocatequina-3-galato (EGCG). Esta catequina apresenta papel antiinflamatório, anti-oxidante, anti-microbiano e mineralizador e poderia ser utilizado no tratamento da doença periapical. O objetivo do presente trabalho foi avaliar o efeito uma pasta à base de EGCG utilizada como curativo de demora em lesões periapicais experimentalmente induzidas em cães. Um total de 80 raízes de pré-molares de cão com rizogênese completa e lesões periapicais experimentalmente induzidas, foram aleatoriamente divididos em 4 grupos submetidos à diferentes protocolos de tratamento endodôntico: 1) curativo de demora com pasta à base de EGCG; 2) curativo de demora com EGCG em veículo aquoso; 3) curativo de demora com pasta à base hidróxido de cálcio (Pasta Calen); 4) tratamento endodôntico em sessão única. Para avaliação da resposta tecidual, os dentes foram avaliados radiograficamente e histopatologicamente. As imagens radiográficas obtidas antes e 120 dias após o tratamento, foram fotografadas e digitalizadas para o programa Image J 1.28 para mensuração das áreas (mm²) radiolúcidas periapicais. Aos 120 dias após a obturação doa canais radiculares os animais foram eutanasiados e os espécimes foram processados histotecnicamente, corados com HE e avaliados com microscopia de luz convencional e de fluorescência (infiltrado inflamatório, espessamento do ligamento periodontal e reabsorção de tecidos mineralizados). Os resultados obtidos foram analisados estatisticamente utilizando os testes quiquadrado, Fisher, Anova e pós teste de Tukey, com nível de significância de 5% em todos os testes. De acordo com os resultados radiográficos, o percentual médio de redução da área radiolúcida (± erro padrão da média) foi semelhante (p>0,05) nos grupos EGCG em veículo aquoso (64,57%; ±7,514); pasta à base de EGCG (59,95%; ±8,023) e pasta à base de hidróxido de cálcio (58,65%; ±6,192). Todos foram superiores ao grupo tratado em sessão única (19,49%; ±2,881) (p<0,01). A análise histopatológica mostrou que os grupos que receberam aplicação de curativo de demora com EGCG ou hidróxido de cálcio, apresentaram reaparação da lesão periapical com semelhança em todos os parâmetros avaliados. Por outro lado, o tratamento em sessão única resultou em manutenção da lesão periapical, com maior espessura do ligamento periodontal (p<0,001), persistência de infiltrado inflamatório moderado ou severo (p<0,01) e presença de reabsorção óssea e cementária (p<0,0001). Foi possível concluir que a pasta à base de EGCG proporcionou o reparo de lesões periapicais, constituindo possível medicação intracanal alternativa / Green tea has been used in the prevention and treatment of various infectious and immunoinflammatory diseases, since it has beneficial effects due to the presence of Epigallocatechin-3-gallate (EGCG). This catequin present anti-inflammatory, antimicrobial, antioxidant and mineralizing role that could be usefull in the treatment of periapical disease. The aim of the present study was to evaluate the effect of the intracanal dressing with an EGCG paste-based im periapical lesions experimentaly induced in dogs. A total of 80 dog premolar roots with complete rhizogenesis and experimentally induced periapical lesions were randomly divided into 4 groups submitted to different endodontic treatment protocols:1) intracanal dressing with EGCG based paste; 2) intracanal dressing with EGCG in aqueous vehicle; 3) intracanal dressing with based paste on calcium hydroxide (Calen paste); 4) single session endodontic treatment. To evaluate the tissue response, the teeth were evaluated radiographically and histopathologically. Radiographic images are selected before 120 days after treatment, were photographed and scanned for the program Image J 1.28 for the measurement of periapical radiolucent areas (mm2). To 120 days after the root canal filling animals were euthanasied and the specimens processed, stained in the rotin HE and evaluated under conventional and fluorescence microscopy (inflammatory infiltrate, periodontal ligament space and mineralized tissue resorption). The results were compared statistically using chisquare, Fisher, Anova and Tukey post-test, with a significance level of 5% in all tests. According to the radiographic results, the mean percentage reduction of the radiolucent area (standard pattern of the mean) was similar (p> 0.05) in the EGCG groups in aqueous vehicle (64.57%; ± 7.514); EGCG based paste (59.95%, ± 8.023) and calcium hydroxide based paste (58.65%, ± 6.192). All groups were higher for the group treated in single session (19.49%, ± 2.881) (p <0.01). The histopathological analysis showed that the groups that received the prescription of intracanal dressings with EGCG or calcium hydroxide, resulted in periapical lesion repair and were similar in all the evaluated parameters. However, treatment in single session did not repair the periapical lesion resulting in greatest measure of the periodontal ligament (p <0.001), persistence of moderate or severe inflammatory infiltrate (p <0.01) and presence of bone and cementum resorption (p <0.0001). It is possible to conclude that EGCG paste-based allows the periapical lesions repair, constituting possible alternative intracanal medication
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The Preventive Effect of Oral EGCG in a Fetal Alcohol Spectrum Disorder Mouse ModelLong, Ling, Li, Yi, Wang, Yi D., He, Qing Y., Li, Mei, Cai, Xiao D., Peng, Kou, Li, Xiang P., Xie, Dan, Wen, Yan Ling, Yin, Deling, Peng, Ying 01 November 2010 (has links)
Background: Fetal alcohol spectrum disorder (FASD) is a challenging public health problem. Previous studies have found an association between FASD and oxidative stress. In the present study, we assessed the role of oxidative stress in ethanol-induced embryonic damage and the effect of (-)-epigallocatechin-3-gallate (EGCG), a powerful antioxidant extracted from green tea, on the development of FASD in a murine model.Methods: Pregnant female mice were given intraperitoneal ethanol (25%, 0.005 to 0.02 ml/g) on gestational day 8 (G8) to establish the FASD model. On G10.25, mice were sacrificed and embryos were collected and photographed to determine head length (HL), head width (HW), and crown rump length (CRL). For mice given EGCG, administration was through a feeding tube on G7 and G8 (dose: 200, 300, or 400 mg/kg/d, the total amount for a day was divided into 2 equal portions). G10.25 embryos were evaluated morphologically. Brain tissues of G9.25 embryos were used for RT-PCR and western blotting of neural marker genes and proteins and detection of oxidative stress indicators.Results: Administration of ethanol to pregnant mice on G8 led to the retardation of embryonic growth and down-regulation of neural marker genes. In addition, administration of ethanol (0.02 ml/g) led to the elevation of oxidative stress indicators [hydrogen peroxide (H2O2) and malondialdehyde (MDA)]. Administration of EGCG on G7 and G8 along with ethanol on G8 ameliorated the ethanol-induced growth retardation. Mice given EGCG (400 mg/kg/d) along with ethanol had embryo sizes and neural marker genes expression similar to the normal controls. Furthermore, EGCG (400 mg/kg on G7 and G8) inhibited the increase in H2O2 and MDA.Conclusions: In a murine model, oxidative stress appears to play an important role in ethanol-induced embryonic growth retardation. EGCG can prevent some of the embryonic injuries caused by ethanol.
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Formulação à base de um extrato do chá verde desenvolvida para uso bucal: avaliação da atividade antimicrobiana e da alteração de cor dental / Formulation based of green tea extract developed for oral use: Evaluation of antimicrobial activity and dental color changeVilela, Marina Moscardini 27 November 2015 (has links)
Atualmente o digluconato de clorexidina (CHX) constitui o agente antimicrobiano de uso bucal mais utilizado na Odontologia devido ao seu amplo espectro de ação contra bactérias, fungos e vírus e efeito residual. No entanto, este agente apresenta efeitos colaterais quando utilizado por longo período, podendo causar coloração extrínseca nos dentes e restaurações, alteração no paladar, sensibilidade na língua e descamação das mucosas. Estes efeitos adversos conduzem à pesquisa de novas formulações. Entretanto, até o momento, nenhum agente antimicrobiano substituiu a clorexidina. Assim, o objetivo deste estudo foi desenvolver uma nova formulação antimicrobiana à base de um extrato de planta derivado do chá verde, a epigalocatequina-3-galato (EGCG), avaliar sua ação antimicrobiana contra microrganismos cariogênicos, in vitro e in vivo, e a possibilidade de alteração de cor dental, in vitro. A atividade antimicrobiana in vitro contra S. mutans, S. sobrinus e L. casei foi avaliada por meio da determinação da concentração inibitória mínima (CIM) e concentração bactericida mínima (CBM), e, posteriormente sua concentração antimicrobiana de uso clínico foi determinada in vivo, após seu uso por crianças com alto risco e atividade da doença cárie. A concentração de microrganismos cariogênicos foi determinada na saliva das crianças, antes e após a realização do bochecho em diferentes concentrações, até se obter a máxima ação antimicrobiana. A clorexidina a 0,12% (Periogard®) e a água destilada foram utilizadas como controle positivo e negativo, respectivamente. A possível alteração de cor dental foi avaliada na coroa de dentes decíduos e permanentes humanos anteriores recém-extraídos, por meio de espectrofotometria (Easy Shade) e obtenção dos valores absolutos de L*, a* e b* (Sistema CIELab). Foi determinada a cor inicial e após a realização de 1, 5, 10 e 30 simulações de bochechos com cada solução testada durante 1 minuto. Os resultados dos ensaios de CIM e CBM in vitro mostraram que a formulação de EGCG inibiu o crescimento de todos os microrganismos cariogênicos avaliados. A CIM de EGCG contra S. mutans, S. sobrinus e para o L. casei foi obtida nas concentrações de 125, 750 e 750 μg/mL, respectivamente. A CBM de EGCG contra o S. mutans, S. sobrinus e para o L. casei foi verificada nas concentrações de 250, 1000 e 1000 μg/mL, respectivamente. A CBM em comum entre os 3 microrganimos foi utilizada como concentração inicial para a avaliação da atividade antimicrobiana in vivo. A partir desse valor a concentração foi sendo dobrada até atingir o valor de 4000 μg/mL, que foi considerada a concentração de EGCG de uso clínico que causou redução da microbiota salivar cariogênica (86,48%) que mais se assemelhou à clorexidina (89,89%). A simulação de bochechos com a formulação de EGCG desenvolvida (4000 μg/mL) causou alteração no valor absoluto das coordenadas L*, a* e b* em dentes decíduos e permanentes, de forma semelhante à causada pela solução de clorexidina. A alteração dos valores absolutos de L* e b* foram reversíveis após a realização de profilaxia dental. Com base nas metodologias e nos resultados obtidos no presente estudo pode-se concluir que a formulação desenvolvida, à base de EGCG, apresenta efeito antimicrobiano contra microrganismos cariogênicos, in vitro e in vivo, e causa alteração de cor dental reversível. / Currently the digluconate of chlorhexidine (CHX) is the antimicrobial agent for oral use most often used in dentistry due to its broad spectrum of activity against bacteria, fungi and viruses, and residual effect. However, this agent has side effects when used for long periods, and may cause extrinsic staining on teeth and restorations, change in taste, tenderness in the language and peeling of the mucous membranes. These adverse events lead to research into new formulations. However, to date, no antimicrobial agent has replaced the chlorhexidine. The objective of this study was to develop a new antimicrobial formulation based of green tea extract, epigallocatechin-3-gallate (EGCG), evaluate their antimicrobial activity against cariogenic microorganisms in vitro and in vivo, and possibility of dental color change in vitro. The in vitro antimicrobial activity against S. mutans, S. sobrinus e L. casei was evaluated by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), and subsequently its antimicrobial concentration was determined in clinical use, after its use for children at high risk and activity of caries. The concentration of cariogenic microorganisms was determined in the saliva of children before and after performing the mouthwash at different concentrations to obtain maximum antimicrobial activity. The chlorhexidine 0.12% (Periogard®) and distilled water were used as positive and negative controls, respectively. The possible dental color change was evaluated on the crown of deciduous and permanent teeth freshly extracted human anterior by means of spectrophotometry (Easy Shade) and obtaining the absolute values of L *, a * and b * (CIELab system). The color was determined in the beginning and after the completion of 1, 5, 10 and 30 mouthwash simulations for 1 minute of each solution tested. In vitro results of MIC and CBM tests showed that formulation EGCG inhibited the growth of all cariogenic microorganisms evaluated similarly to chlorhexidine. The MIC of EGCG against S. mutans, S. sobrinus and L. casei was observed at concentrations of 125, 750 and 750 μg/ml, respectively. EGCG MBC against S. mutans, S. sobrinus and L. casei was observed at concentrations of 250, 1000 and 1000 μg/ml, respectively. The common CBM between the three microbes was used as initial concentration for the evaluation of the antimicrobial activity in vivo. From this value the concentration was being folded up to the value of 4000 μg/mL, which was considered the concentration of EGCG clinical use which caused reduction in salivary microbiota cariogenic (86.48%) that most resembled the chlorhexidine (89.89%). The simulation of mouthwash with the developed formulation EGCG (4000 μg/ml) caused change in the absolute value of the coordinates L *, a * and b * in deciduous and permanent teeth, similar to that caused by chlorhexidine . The changes of the absolute values of L* and b* were reversible after performing dental prophylaxis. Accordance with the procedures and the results obtained in this study it can be concluded that the formulation developed, based on EGCG, has antimicrobial effect against cariogenic microorganisms in vitro and in vivo, and causes reversible change dental color.
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Formulação à base de um extrato do chá verde desenvolvida para uso bucal: avaliação da atividade antimicrobiana e da alteração de cor dental / Formulation based of green tea extract developed for oral use: Evaluation of antimicrobial activity and dental color changeMarina Moscardini Vilela 27 November 2015 (has links)
Atualmente o digluconato de clorexidina (CHX) constitui o agente antimicrobiano de uso bucal mais utilizado na Odontologia devido ao seu amplo espectro de ação contra bactérias, fungos e vírus e efeito residual. No entanto, este agente apresenta efeitos colaterais quando utilizado por longo período, podendo causar coloração extrínseca nos dentes e restaurações, alteração no paladar, sensibilidade na língua e descamação das mucosas. Estes efeitos adversos conduzem à pesquisa de novas formulações. Entretanto, até o momento, nenhum agente antimicrobiano substituiu a clorexidina. Assim, o objetivo deste estudo foi desenvolver uma nova formulação antimicrobiana à base de um extrato de planta derivado do chá verde, a epigalocatequina-3-galato (EGCG), avaliar sua ação antimicrobiana contra microrganismos cariogênicos, in vitro e in vivo, e a possibilidade de alteração de cor dental, in vitro. A atividade antimicrobiana in vitro contra S. mutans, S. sobrinus e L. casei foi avaliada por meio da determinação da concentração inibitória mínima (CIM) e concentração bactericida mínima (CBM), e, posteriormente sua concentração antimicrobiana de uso clínico foi determinada in vivo, após seu uso por crianças com alto risco e atividade da doença cárie. A concentração de microrganismos cariogênicos foi determinada na saliva das crianças, antes e após a realização do bochecho em diferentes concentrações, até se obter a máxima ação antimicrobiana. A clorexidina a 0,12% (Periogard®) e a água destilada foram utilizadas como controle positivo e negativo, respectivamente. A possível alteração de cor dental foi avaliada na coroa de dentes decíduos e permanentes humanos anteriores recém-extraídos, por meio de espectrofotometria (Easy Shade) e obtenção dos valores absolutos de L*, a* e b* (Sistema CIELab). Foi determinada a cor inicial e após a realização de 1, 5, 10 e 30 simulações de bochechos com cada solução testada durante 1 minuto. Os resultados dos ensaios de CIM e CBM in vitro mostraram que a formulação de EGCG inibiu o crescimento de todos os microrganismos cariogênicos avaliados. A CIM de EGCG contra S. mutans, S. sobrinus e para o L. casei foi obtida nas concentrações de 125, 750 e 750 μg/mL, respectivamente. A CBM de EGCG contra o S. mutans, S. sobrinus e para o L. casei foi verificada nas concentrações de 250, 1000 e 1000 μg/mL, respectivamente. A CBM em comum entre os 3 microrganimos foi utilizada como concentração inicial para a avaliação da atividade antimicrobiana in vivo. A partir desse valor a concentração foi sendo dobrada até atingir o valor de 4000 μg/mL, que foi considerada a concentração de EGCG de uso clínico que causou redução da microbiota salivar cariogênica (86,48%) que mais se assemelhou à clorexidina (89,89%). A simulação de bochechos com a formulação de EGCG desenvolvida (4000 μg/mL) causou alteração no valor absoluto das coordenadas L*, a* e b* em dentes decíduos e permanentes, de forma semelhante à causada pela solução de clorexidina. A alteração dos valores absolutos de L* e b* foram reversíveis após a realização de profilaxia dental. Com base nas metodologias e nos resultados obtidos no presente estudo pode-se concluir que a formulação desenvolvida, à base de EGCG, apresenta efeito antimicrobiano contra microrganismos cariogênicos, in vitro e in vivo, e causa alteração de cor dental reversível. / Currently the digluconate of chlorhexidine (CHX) is the antimicrobial agent for oral use most often used in dentistry due to its broad spectrum of activity against bacteria, fungi and viruses, and residual effect. However, this agent has side effects when used for long periods, and may cause extrinsic staining on teeth and restorations, change in taste, tenderness in the language and peeling of the mucous membranes. These adverse events lead to research into new formulations. However, to date, no antimicrobial agent has replaced the chlorhexidine. The objective of this study was to develop a new antimicrobial formulation based of green tea extract, epigallocatechin-3-gallate (EGCG), evaluate their antimicrobial activity against cariogenic microorganisms in vitro and in vivo, and possibility of dental color change in vitro. The in vitro antimicrobial activity against S. mutans, S. sobrinus e L. casei was evaluated by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), and subsequently its antimicrobial concentration was determined in clinical use, after its use for children at high risk and activity of caries. The concentration of cariogenic microorganisms was determined in the saliva of children before and after performing the mouthwash at different concentrations to obtain maximum antimicrobial activity. The chlorhexidine 0.12% (Periogard®) and distilled water were used as positive and negative controls, respectively. The possible dental color change was evaluated on the crown of deciduous and permanent teeth freshly extracted human anterior by means of spectrophotometry (Easy Shade) and obtaining the absolute values of L *, a * and b * (CIELab system). The color was determined in the beginning and after the completion of 1, 5, 10 and 30 mouthwash simulations for 1 minute of each solution tested. In vitro results of MIC and CBM tests showed that formulation EGCG inhibited the growth of all cariogenic microorganisms evaluated similarly to chlorhexidine. The MIC of EGCG against S. mutans, S. sobrinus and L. casei was observed at concentrations of 125, 750 and 750 μg/ml, respectively. EGCG MBC against S. mutans, S. sobrinus and L. casei was observed at concentrations of 250, 1000 and 1000 μg/ml, respectively. The common CBM between the three microbes was used as initial concentration for the evaluation of the antimicrobial activity in vivo. From this value the concentration was being folded up to the value of 4000 μg/mL, which was considered the concentration of EGCG clinical use which caused reduction in salivary microbiota cariogenic (86.48%) that most resembled the chlorhexidine (89.89%). The simulation of mouthwash with the developed formulation EGCG (4000 μg/ml) caused change in the absolute value of the coordinates L *, a * and b * in deciduous and permanent teeth, similar to that caused by chlorhexidine . The changes of the absolute values of L* and b* were reversible after performing dental prophylaxis. Accordance with the procedures and the results obtained in this study it can be concluded that the formulation developed, based on EGCG, has antimicrobial effect against cariogenic microorganisms in vitro and in vivo, and causes reversible change dental color.
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Encapsulamento de epigalocatequina-3-Galato (EGCG) em nanopartículas para uso tópico bucal: desenvolvimento, caracterização e determinação da atividade antimicrobiana in vitro / Encapsulation of epigallocatechin-3-gallate (EGCG) in nanoparticles for oral topical use: development, characterization and determination of antimicrobial activity in vitroAna Paula Dias Moreno 14 June 2017 (has links)
O uso de agentes químicos coadjuvantes da higienização bucal pode ser necessário para o controle da microbiota cariogênica de indivíduos com alto risco e atividade da doença cárie. Atualmente o agente antimicrobiano mais recomendado é o digluconato de clorexidina (CHX) devido ao seu amplo espectro de ação e efeito residual. Contudo, quando utilizado por longos períodos, este agente químico apresenta efeitos colaterais. Neste contexto, os polifenóis naturais, como a Epigalocatequina3galato (EGCG), derivada do chá verde, vêm sendo propostos como alternativa aos agentes antimicrobianos sintéticos. Entretanto, os polifenóis não apresentam estabilidade ao longo do tempo, podendo se oxidar rapidamente. Desta forma, o encapsulamento da EGCG em nanopartículas poderia aumentar a sua biodisponibilidade e estabilidade física e química, manter o efeito deste polifenol no tecido alvo e potencializar sua eficácia farmacológica. Assim, o presente estudo teve como objetivo desenvolver e caracterizar sistemas de encapsulamento de EGCG e avaliar, in vitro, sua atividade antimicrobiana frente a microorganismos cariogênicos. Inicialmente, foram preparadas nanopartículas poliméricas (NPP) e carreadores lipídicos nanoestruturados (CLN), que foram caracterizados e avaliados quanto à sua atividade antimicrobiana in vitro frente aos microorganismos Streptococcus mutans, Streptococcus sobrinus e Lactobacillus casei. Após a análise dos resultados microbiológicos, o CLN foi selecionado para o encapsulamento da EGCG (CLNEGCG), por apresentar atividade antimicrobiana frente à todos os microorganismos avaliados. O CLNEGCG foi preparado pelo método de emulsão e sonicação e caracterizado quanto ao diâmetro, índice de polidispersão (PdI), potencial zeta (PZ), eficiência de encapsulamento (EE), cristalinidade, capacidade de mucoadesão e morfologia. A atividade antimicrobiana in vitro da EGCG livre e encapsulada foi avaliada por meio da determinação da concentração inibitória mínima (CIM) e concentração bactericida mínima (CBM). O diâmetro, PdI e o PZ dos CLN foram 228nm, 0,216 e 36,53mV, respectivamente, sendo que o encapsulamento da EGCG não alterou significativamente estes parâmetros. O CLN apresentou forma esférica, estabilidade por 330 dias e propriedade mucoadesiva devido a presença de quitosana na superfície do CLN. Além disso, a quitosana favoreceu o encapsulamento da EGCG obtendose uma EE de ~96%. As concentrações inibitórias e bactericidas mínimas do CLNEGCG (33,75 a 67,5 µg/mL) foram menores do que as verificadas para a EGCG livre (250 a 2.000 µg/mL), comprovando o aumento do potencial antimicrobiano com o encapsulamento da EGCG em nanocarreadores híbridos. De acordo com esses resultados, o CLNEGCG, desenvolvido no presente trabalho, constitui um sistema com potencial para o uso tópico bucal, pois além de ser estável e apresentar propriedade de mucoadesão e morfologia adequada, apresentaram alta atividade antimicrobiana frente aos principais microorganismos envolvidos no processo carioso. / The use of oral hygiene adjuvants may be necessary to control the cariogenic microbiota of individuals with high risk and caries disease activity. Currently the most recommended antimicrobial agent is chlorhexidine digluconate (CHX) because of its broad spectrum of action and residual effect. However, this chemical has side effects. In this context, as an Epigallocatechin3gallate (EGCG), a derivative of green tea, as an alternative to synthetic antimicrobial agents. However, there is no stability over time and can oxidize rapidly. Thus, the encapsulation of EGCG in nanoparticles can increase their bioavailability and physical and chemical stability, maintain the effect of this polyphenol on the target tissue and potentiate its pharmacological efficacy. Thus, the present study aimed to develop and characterize EGCG encapsulation systems and to evaluate, in vitro, its antimicrobial activity against cariogenic microorganisms. Initially, polymer nanoparticles (NPP) and nanostructured lipid carriers (CLN) were prepared and evaluated for their in vitro antimicrobial activity against Streptococcus mutans, Streptococcus sobrinus and Lactobacillus casei microorganisms. After the analysis of the microbiological results, the CLN was selected for the encapsulation of EGCG (CLNEGCG), due to its higher antimicrobial activity. The CLNEGCG was developed by emulsion and sonication method and was characterized in relation to diameter, polydispersity index (PdI), zeta potential (PZ), encapsulation efficiency (EE), crystallinity, mucoadhesion capacity and morphology. The in vitro antimicrobial activity of EGCG and its ability to evaluate minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The diameter, PdI and PZ of the CLNs were 228nm, 0.216 and 36.53mV, respectively, and the EGCG encapsulation did not significantly alter these parameters. The CLN showed a spherical structure, stability for 330 days and a mucoadhesive property due to a presence of chitosan on the CLN surface. In addition, a chitosan favored EGCG encapsulation resulting in an EE of ~ 96%. The minimum inhibitory and bactericidal concentrations of CLGEGCG (33.75 to 67.5 µg / mL) were lower than those for free EGCG (250 to 2,000 µg / mL), with increased antimicrobial potential with EGCG encapsulation in hybrid nanocarriers. According to the results, the CLNEGCG, developed in the present study, constitutes a system with potential for oral topical use, besides being stable and possessing mucoadhesion properties, presented high antimicrobial activity against the main microorganisms involved in the carious process.
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