Grape Seed Extract Attenuates Demyelination in Experimental Autoimmune Encephalomyelitis Mice by Inhibiting Inflammatory Response of Immune Cells / 中国结合医学杂志

OBJECTIVE@#To examine the anti-inflammatory effect of grape seed extract (GSE) in animal and cellular models and explore its mechanism of action.@*METHODS@#This study determined the inhibitory effect of GSE on macrophage inflammation and Th1 and Th17 polarization in vitro. Based on the in vitro results, the effects and mechanisms of GSE on multiple sclerosis (MS)-experimental autoimmune encephalomyelitis (EAE) mice model were further explored. The C57BL/6 mice were intragastrically administered with 50 mg/kg of GSE once a day from the 3rd day to the 27th day after immunization. The activation of microglia, the polarization of Th1 and Th17 and the inflammatory factors such as tumor necrosis factor- α (TNF- α), interleukin-1 β (IL-1 β), IL-6, IL-12, IL-17 and interferon-γ (IFN-γ) secreted by them were detected in vitro and in vivo by flow cytometry, enzyme linked immunosorbent assay (ELISA), immunofluorescence staining and Western blot, respectively.@*RESULTS@#GSE reduced the secretion of TNF-α, IL-1 β and IL-6 in bone marrow-derived macrophages stimulated by lipopolysaccharide (P<0.01), inhibited the secretion of TNF-α, IL-1 β, IL-6, IL-12, IL-17 and IFN-γ in spleen cells of EAE mice immunized for 9 days (P<0.05 or P<0.01), and reduced the differentiation of Th1 and Th17 mediated by CD3 and CD28 factors (P<0.01). GSE significantly improved the clinical symptoms of EAE mice, and inhibited spinal cord demyelination and inflammatory cell infiltration. Peripherally, GSE downregulated the expression of toll-like-receptor 4 (TLR4) and Rho-associated kinase (ROCKII, P<0.05 or P<0.01), and inhibited the secretion of inflammatory factors (P<0.01 or P<0.05). In the central nervous system, GSE inhibited the infiltration of CD45+CD11b+ and CD45+CD4+ cells, and weakened the differentiation of Th1 and Th17 (P<0.05). Moreover, it reduced the secretion of inflammatory factors (P<0.01), and prevented the activation of microglia (P<0.05).@*CONCLUSION@#GSE had a beneficial effect on the pathogenesis and progression of EAE by inhibiting inflammatory response as a potential drug and strategy for the treatment of MS.

Efecto de los extractos fitoquímicos en la eliminación del barro dentinario en el tratamiento de conductos radiculares / Effects of phytochemical extracts on smear layer removal in root canal treatment

Introduction: Studies suggested that phytochemical products are considered potential solutions to smear layer removal due to their biological safety in cleaning root canal systems, lower toxicity, lower irritant effect and antimicrobial effect. Objective: The aim of the present study was to perform a systematic review to evaluate smear layer removal of root canal systems by different natural product solutions. Methods: systematic review. Search of the literature was performed on Pubmed, Scopus, Cochrane, Web of Science, Scielo and Google Scholar according the PRISMA protocol. Studies were included if they performed the experiments of smear layer removal on extracted permanent human teeth. Articles published in any language without restriction of year of publication were included in this review. The risk of bias assessment in the included studies was evaluated using the Joanna Briggs Institute Critical Appraisal Checklist for Quasi-Experimental Studies. The search strategy resulted in the retrieval of 6221 publications. After the eligibility criteria application, 8 articles were selected for analysis. Results: It was observed that some natural products solutions showed effects on smear layer removal, especially on coronal third. The phytochemical products such as Salvadora persica, chamomile alcohol extract and grape seed extract demonstrated good potential of smear layer removal, however, its effectiveness and clinical applicability are still unclear. Conclusions: Although the limitations of this study, it is possible to highlight those phytochemical products such as Salvadora persica, chamomile alcohol extract and grape seed extract that demonstrated good potential on SL removal(AU)

Introducción: Los productos fitoquímicos se consideran soluciones potenciales para la eliminación del barro dentinario debido a su seguridad biológica en la limpieza del sistema de conductos radiculares, menor toxicidad, menor efecto irritante y efecto antimicrobiano. Objetivo: Evaluar la eliminación del barro dentinario de los sistemas de conductos radiculares mediante diferentes soluciones de productos naturales. Métodos: La búsqueda de la literatura se realizó en Pubmed, Scopus, Cochrane, Web of Science, SciELO y Google Scholar, según el protocolo PRISMA. Los estudios se incluyeron si realizaron los experimentos de eliminación del barro dentinario en dientes humanos permanentes extraídos. En esta revisión se incluyeron artículos publicados en cualquier idioma, sin restricción de año de publicación. La evaluación del riesgo de sesgo en los estudios incluidos se evaluó mediante la lista de verificación de evaluación crítica del Instituto Joanna Briggs para estudios cuasiexperimentales. La estrategia de búsqueda resultó en la recuperación de 6221 publicaciones. Después de la aplicación de los criterios de elegibilidad, se seleccionaron 8 artículos para su análisis. Resultados: Se observó que algunas soluciones de productos naturales mostraron efectos sobre la eliminación del barro dentinario, especialmente en el tercio coronal. Los productos fitoquímicos como salvadora pérsica, extracto de alcohol de manzanilla y extracto de semilla de uva demostraron un buen potencial de eliminación del barro dentinario; sin embargo, su efectividad y aplicabilidad clínica aún no están claras. Conclusiones: A pesar de las limitaciones de este estudio, es posible destacar aquellos productos fitoquímicos como salvadora pérsica, extracto de alcohol de manzanilla y extracto de semilla de uva demostraron buen potencial de eliminación del barro dentinario(AU)

Influência do uso da proantocianidina em um adesivo autocondicionante: avaliação da resistência de união / Influence of the use of proanthocyanidinin a self-etching adhesive: evaluation of bond strength / Influenciadel uso de proantocianidinaem um adesivo autograbante: evaluación de la fuerza de unión

Introdução:Os sistemas adesivos possibilitama execução de restaurações estéticas e minimamente invasivas, sendo, portanto,objeto de pesquisas para contornar os problemas que se apresentam no procedimento restaurador.Objetivo:Avaliar in vitroa resistência de união de um sistema adesivo autocondicionante, e deste modificado com soluções extrativas de semente de uva.Metodologia:Duas soluções extrativas foram preparadas comextrato de semente de uva em pó dissolvido em acetona e etanol. A partir delas e de umadesivo,seis sistemas adesivos autocondicionantes experimentais foram preparados, diferindo quanto aosolvente utilizado eàsproporções entre adesivo puro e solução extrativa(7,5%, 15% e 30%). Setenta incisivos bovinos hígidos tiveram as raízes removidas com disco de carborundum e as faces vestibulares desgastadas comlixas d'água de granulação 120, 240, 600 e 1200 sob refrigeração até expor a dentina superficial. Os dentes foram distribuídos aleatoriamenteem sete grupos distintos: Controle; A7,5; A15; A30; E7,5; E15; e E30, contendo 10 elementos cada. A aplicação dos adesivos foi executada de acordo com as recomendações do fabricante do adesivo controle. A restauração foi realizada com uma matriz de silicone com dimensões 2mm de altura e 4mm de diâmetro e inserido o material restaurador em incremento único e fotopolimerizado por 40s. Após três meses armazenados em água destilada, os espécimes foram submetidos ao teste de resistência de união. Foi empregado ométodo estatísticoTeste Paramétrico Anova 1 Fator e pós-teste de Tamhane (p<0,05). Resultados:Os grupos A7,5, E7,5 e E30 não apresentaram diferença em relação ao grupo Controle; A15 e A30 mostraram desempenho estatisticamente semelhante entre si; e E15 não apresentou diferença estatística em relação aos outros adesivos.Conclusões:A adição de proantocianidina teve efeitos diferentes,dependendodos solventes e das concentrações utilizadas, mas sem alterar significativamente o desempenho do adesivo (AU).

Introduction:Adhesive systems make it possible to perform aestheticand minimally invasive restorations, being the subject of research to circumvent the problems that arise in the restorative procedure.Objective:Evaluate in vitrothe bond strength of a self-etching adhesive system,and modified with extractive grape seed solutions. Methodology:Two extractive solutions were prepared with powdered grape seed extract dissolved in acetone and ethanol. From them and an adhesive, six experimental self-etching adhesive systems were prepared, differing in terms of the solvent used and the proportions between pure adhesive and extractive solution(7.5%, 15% and 30%). Seventy healthy bovine incisors had their roots removed with carborundum disc and the vestibular faces were worn with sandpaper with granulation water 120, 240, 600 and 1200 under refrigeration until the superficial dentin was exposed. The teeth were randomly assigned to seven different groups: Control; A7.5; A15; A30; E7.5; E15; and E30, containing 10 elements each. The application of the adhesives was carried out according to the recommendations of the manufacturer of the control adhesive. The restoration was performed with a silicone matrix with dimensions 2mm high and 4mm indiameter and the restorative material was inserted in a single increment and light cured for 40s. After three months stored in distilled water, the specimens were submitted to the bond strength test. The statistical method Parametric Test Anova 1 Factor and Tamhane post-test (p<0.05) were used. Results:Groups A7.5, E7.5 and E30 showed no difference in relation to the Control group; A15 and A30 showed a statistically similar performance; and E15 showed no statistical difference in relation to the other adhesives. Conclusions:The addition of proanthocyanidin had different effects, depending on the solvents and concentrations used, but without significantly altering the performance ofthe adhesive (AU).

Introducción: Sistemas adhesivos permiten realizar restauraciones estéticas y mínimamente invasivas, siendo objeto de investigación para sortear problemas que surgen en elprocedimiento restaurador. Objetivo: Evaluar in vitrola fuerza de unión de un sistema adhesivoautograbante y modificado con soluciones extractivas de semilla de uva. Metodología: Se prepararon dos soluciones extractivas con extracto de semilla de uva en polvo disuelto en acetona y etanol. A partir de ellos y de un adhesivo, se prepararon seis sistemas experimentales de adhesivos autograbantes, que se diferencian en cuanto al solvente utilizado y las proporciones entre adhesivo puro y solución extractiva (7,5%, 15% y 30%). Setenta incisivos bovinos sanos fueron removidos con un disco de carborundo y las caras vestibulares fueron usadas com lija de agua de granulación 120, 240, 600 y 1200 bajo refrigeración hasta que la dentina superficial quedo expuesta. Los dientes se asignaron aleatoriamente a siete grupos diferentes: Control; A7,5; A15; A30; E7,5; E15; y E30, que contiene 10 elementos cada uno. La aplicación de los adhesivos se realizó siguiendo las recomendaciones del fabricante del adhesivo de control. La restauración se realizó con matriz de silicona con 2mm de altura y 4mm de diámetro y el material restaurador se insertó en un solo incremento y se fotopolimerizó durante 40s. Tres meses después, almacenados em agua destilada, las muestras se sometieron a la prueba de resistencia de la unión. Se utilizó el método estadístico Prueba Paramétrica Factor Anova 1 y post-prueba de Tamhane (p<0,05). Resultados: Los grupos A7,5, E7,5 y E30 no mostraron diferencias em relación con el grupo Control; A15 y A30 mostraron un desempeño estadísticamente similar; y E15 no mostró diferencia estadística en relación con los otros adhesivos. Conclusiones: La adición de proantocianidina tuvo diferentes efectos, dependiendo de los disolventes y concentraciones utilizadas, pero sin alterar significativamente el rendimiento del adhesivo (AU).

Effectiveness of grape seed extract-based intracanal dressings against Enterococcus Faecalis and its influence on dentin microhardness and bond strength of filling material / Eficácia de medicamentos intracanais à base de extrato de semente de uva contra o Enterococcus Faecalis e sua influência na micro-dureza da dentina e na resistência de união do material obturador

The aim of this study was to evaluate the effectiveness of grape seed extract (GSE)-based intracanal dressings against Enterococcus faecalis (E. faecalis) and its influence on dentin microhardness and bond strength of the filling material. The root canals of 126 human teeth were distributed into three test groups: antimicrobial activity (60 teeth), dentin microhardness (30 teeth) and bond strength (36 teeth). In all three groups, specimens were subdivided into six groups, according to intracanal dressing protocols: G1 ­ distilled water (DW); G2 ­ 2% chlorhexidine gel (CHX); G3 ­ calcium hydroxide (Ca[OH]2)+DW; G4 ­ GSE+DW; G5 ­ Ca(OH)2+CHX; G6 ­ GSE+CHX. The counting of colony-forming units (CFUs), the Vickers microhardness tester and the push-out test were performed to evaluate the antimicrobial activity, dentin microhardness and bond strength, respectively. Specific statistical analysis was performed for each evaluation (α=5%). The greatest bacterial reduction was observed in G5 (Ca[OH]2+CHX) and G6 (GSE+CHX) (p<0.05). There was no statistically significant difference among groups in the dentin microhardness evaluation (p<0.05). The highest bond strength in the immediate evaluation was observed in G4 (GSE+DW) and G6 (GSE+CHX), whereas the highest bond strength after 12 months of storage was observed in G2 (CHX), G3 (Ca[OH]2+DW), G4 (GSE+DW), and G6 (GSE+CHX) (p<0.05). After the storage period, bond strength was increased in G2 (CHX) and G3 (Ca[OH]2+DW), and remained unchanged in G4 (GSE+DW) and G6 (GSE+CHX) (p<0.05). GSE-based intracanal dressings have antimicrobial potential against E. faecalis, have no influence in dentin microhardness and preserve the high bond strength of filling materials for root dentin over time.

O objetivo deste estudo foi avaliar a eficácia de medicamentos intracanal à base de extrato de semente de uva (GSE) contra Enterococcus faecalis (E. faecalis) e sua influência na microdureza da dentina e na resistência de união do material de obturação. Os canais radiculares de 126 dentes humanos foram distribuídos em três grupos de teste: atividade antimicrobiana (60 dentes), microdureza da dentina (30 dentes) e resistência adesiva (36 dentes). Nos três grupos, as amostras foram subdivididas em seis grupos, de acordo com os protocolos de curativos intracanal: G1 ­ água destilada (DW); G2 ­ gel de clorexidina a 2% (CHX); G3 ­ hidróxido de cálcio (Ca[OH]2) +DW; G4 ­ GSE+DW; G5 ­ Ca(OH)2+CHX; G6 ­ GSE+CHX. A contagem de unidades formadoras de colônias (UFCs), o testador de microdureza Vickers e o teste push-out foram realizados para avaliar a atividade antimicrobiana, a microdureza da dentina e a resistência adesiva, respectivamente. Análise estatística específica foi realizada para cada avaliação (α=5%). A maior redução bacteriana foi observada no G5 (Ca[OH]2+CHX) e G6 (GSE+CHX) (p<0,05). Não houve diferença estatisticamente significativa entre os grupos na avaliação da microdureza da dentina (p<0,05). A maior resistência adesiva na avaliação imediata foi observada no G4 (GSE+DW) e G6 (GSE+CHX), enquanto a maior resistência adesiva após 12 meses de armazenamento foi observada no G2 (CHX), G3 (Ca[OH]2+DW), G4 (GSE+DW) e G6 (GSE+CHX) (p<0,05). Após o período de armazenamento, a resistência de união aumentou no G2 (CHX) e G3 (Ca[OH]2+DW), permanecendo inalterada no G4 (GSE+DW) e G6 (GSE+CHX) (p<0,05). Os medicamentos intracanal à base de GSE têm potencial antimicrobiano contra E. faecalis, não influenciam na microdureza da dentina e preservam a alta resistência adesiva dos materiais de obturação da dentina radicular ao longo do tempo.

Antimicrobial activity of intracanal dressings composed by natural products associated to chlorhexidine and its influence on dentinal colour change / Avaliação da ação antimicrobiana de medicações intracanais à base de produtos fitoterápicos associados à clorexidina e sua influência na alteração de cor da estrutura dentinária

Purpose: evaluate the antimicrobial activity of intracanal dressings and their influence on dentinal colour changes. Material and methods: eighty single-rooted human extracted teeth were decoronated and divided into eight groups (n=10) according to intracanal dressing protocols inserted into the root canals: G1­distilled water (DW); G2­2% chlorhexidine gel (CHX); G3­calcium hydroxide (Ca[OH]2)+DW; G4­grape seed extract (GSE)+DW; G5­ginger extract (GE)+DW; G6­Ca(OH)2+CHX; G7­GSE+CHX; and G8­GE+CHX. The antimicrobial activity was evaluated by colony-forming units (CFUs) counting and dentinal colour changes was evaluated by digital spectrophotometry. Data were statistically analysed by One-way ANOVA followed by Tukey´s post hoc test (antimicrobial evaluation) and non-parametric Wilcoxon followed by the Mann- Whitney-U test (colour change evaluation) (α=0.05). Results: the highest bacterial reduction was observed in groups 4, 6, 7 and 8, with no significant difference between them (p<0.05). Groups 4 and 7 showed the highest medians of dentinal colour change (p<0.05). Conclusion: the addition of CHX improved the antimicrobial activity of GE-based intracanal dressing, with no effect in GSE-based intracanal dressing; moreover, these protocols induced significant dentinal colour changes. (AU)

Objetivo: avaliar a atividade antimicrobiana de medicações intracanais e sua influência na alteração da cor dentinária. Materiais e métodos: oitenta dentes humanos extraídos unirradiculares foram seccionados e divididos em oito grupos (n = 10), de acordo com os protocolos de medicação intracanal inseridos nos canais radiculares: água destilada G1 (DW); G2-2% de gel de clorexidina (CHX); hidróxido de cálcio G3 ­ (Ca [OH] 2) + DW; extrato de semente de uva G4 (GSE) + DW; extrato de gengibre G5 (GE) + DW; G6- Ca (OH) 2 + CHX; G7 ­ GSE + CHX; e G8-GE + CHX. A atividade antimicrobiana foi avaliada por contagem de unidades formadoras de colônias (UFCs) e as alterações de cor dentinária foram avaliadas por espectrofotometria digital. Os dados foram analisados estatisticamente por ANOVA one-way, seguida pelo teste post hoc de Tukey (avaliação antimicrobiana) e Wilcoxon não paramétrico, seguido pelo teste de Mann- Whitney-U (avaliação da mudança de cor) (α = 0,05). Resultados: a maior redução bacteriana foi observada nos grupos 4, 6, 7 e 8, sem diferença significativa entre eles (p < 0,05). Os grupos 4 e 7 apresentaram as maiores medianas da alteração da cor dentinária (p < 0,05). Conclusão: a adição de CHX melhorou a atividade antimicrobiana da medicação intracanal baseado em GE, sem efeito na medicação intracanal baseado em GSE; além disso, esses protocolos induziram alterações significativas na cor dentinária.(AU)

Effects of grape seed proanthocyanidin B2 pretreatment on oxidative stress and renal tubular epithelial cell apoptosis after renal ischemia reperfusion in mice

Abstract Purpose To investigate the effects of grape seed proanthocyanidin B2 (GSPB2) preconditioning on oxidative stress and apoptosis of renal tubular epithelial cells in mice after renal ischemia-reperfusion (RIR). Methods Forty male ICR mice were randomly divided into 4 groups: Group A: mice were treated with right nephrectomy. Group B: right kidney was resected and the left renal vessel was clamped for 45 minutes. Group C: mice were intraperitoneally injected with GSPB2 before RIR established. Group D: mice were intraperitoneally injected with GSPB2 plus brusatol before RIR established. Creatinine and urea nitrogen of mice were determined. Pathological and morphological changes of kidney were checked. Expressions of Nrf-2, HO-1, cleaved-caspase3 were detected by Western-blot. Results Compared to Group B, morphology and pathological damages of renal tissue were less serious in Group C. Western-blot showed that expressions of Nrf-2 and HO-1 in Group C were obviously higher than those in Group B. The expression of cleaved-caspase3 in Group C was significantly lower than that in Group B. Conclusion GSPB2 preconditioning could attenuate renal oxidative stress injury and renal tubular epithelial cell apoptosis by up-regulating expressions of Nrf-2 and HO-1 and down-regulating the expression of cleaved-caspase-3, but the protective effect could be reversed by brusatol.

Effect of feeding grape seed Proanthocyanidin extract on production performance, metabolic and anti-oxidative status of dairy cattle / Efeito da alimentação do extrato de semente de uva Proanthocyanidin sobre o desempenho da produção e o status metabólico e antioxidante de gado leiteiro

Aims to investigate the effects of grape seed proanthocyanidin extract (GSPE) on production performance, metabolism, and anti-oxidative status of Holstein dairy cattle in early lactation. Forty-eight multiparous Holstein dairy cattle were assigned to four groups (CON, G20, G40 and G80) and supplied with 0, 20, 40, and 80mg GSPE/kg of body weight/day. G20 significantly increased milk yield compared with other groups. Milk protein and non-fat-solids were increased in G20, G40 and G80 groups compared with the control group only at the 7th day during the experiment. No significant difference was observed in milk fat and somatic cell count, nor on parameters of energy metabolism in blood, liver function and kidney function between the four groups. There was no significant difference in glutathione peroxidase, superoxide dismutase, total antioxidant capacity, and hydrogen peroxide between the groups; but the malondialdehyde content of G20 significantly increased at day 14 in comparison with CON, and tended to increase at the 28th day. In conclusion, feeding 20mg GSPE/kg of body weight/day was associated with a significant increase in milk yield without detrimental effects on liver or kidney function and with substantial energy metabolism and antioxidant parameters improvement in early lactation dairy cattle.(AU)

O presente trabalho visa investigar os efeitos do extrato de semente de uva Proanthocyanidin (GSPE) sobre o desempenho da produção, o metabolismo e o status antioxidante de gado leiteiro Holstein em lactação precoce. Quarenta e oito vacas leiteiras multíparas Holstein foram divididas em quatro grupos (CON, G20, G40 e G80) e receberam 0, 20, 40 e 80mg de GSPE/kg de peso corporal/dia, respectivamente. O G20 aumentou significativamente o rendimento do leite em comparação com os outros grupos. A proteína e os sólidos não gordurosos do leite foram aumentados nos grupos G20, G40 e G80 somente no sétimo dia durante a experiência. Não foi observada diferença significativa na gordura do leite e na contagem de células somáticas, bem como nos parâmetros de metabolismo energético no sangue, na função hepática e na função renal entre os grupos em relação ao grupo controle. Não houve diferença significativa na glutationa peroxidase, na dimutase de superóxido, na capacidade antioxidante total e no peróxido de hidrogênio entre os grupos, mas o conteúdo malondialdeído do G20 aumentou significativamente no dia 14 em comparação com o CON, e tendia a aumentar no dia 28. Em conclusão, a alimentação de 20mg de GSPE/kg de peso corporal/dia foi associada a um aumento significativo no rendimento do leite, sem efeitos nocivos sobre a função hepática ou a renal, com o metabolismo de energia substancial e a melhoria dos parâmetros antioxidantes de gado leiteiro no início da lactação.(AU)

Effect of grape seed extract-containing phosphoric acid formulations on bonding to enamel and dentin

Abstract The aim was to evaluate the effect of 2% grape seed extract (GSE) containing phosphoric acid (PhA) on the bond strength to enamel and dentin. The control group was 37% PhA. The following three PhA formulations with 2% GSE and 20% ethanol were obtained: GSE5 = 5% PhA; GSE10 = 10% PhA; and GSE20 = 20% PhA. The enamel and dentin surfaces of molars were etched with the acid solutions, followed by Scotchbond Multi-Purpose adhesive and composite resin application. The tensile bond strength (TBS) test evaluated the bond to enamel after 24 h, and the microtensile bond strength (μTBS) test evaluated the bond to dentin after 24 h and 12-month water storage. Etched enamel and dentin were observed by scanning electron microscopy (SEM) and atomic force microscopy (AFM), respectively. The TBS data were submitted to one-way ANOVA, while µTBS data were submitted to two-way ANOVA and Tukey's test (α = 0.05). The TBS (MPa) to enamel did not significantly differ among the control (48.1 ± 15.7), GSE5 (46.1 ± 9.6), GSE10 (49.8 ± 13.6) and GSE20 (44.1 ± 11.9) groups (p = 0.537). The µTBS (MPa) to dentin of the control (28.4 ± 14.4) and GSE20 (24.1 ± 8.1) groups were significantly higher than those of the GSE5 (16.8 ± 7.4) and GSE10 (17.5 ± 6.6) groups at 24 h (p < 0.006). After 12-month storage, only GSE5 (21.0 ± 7.8) and GSE10 (17.6 ± 8.0) did not show significantly decreased μTBS (p > 0.145). SEM micrographs showed a shallower enamel etching pattern for GSE5. AFM images showed the formation of collagenous globular structures for GSE5 and GSE10. The different acid solutions did not influence the TBS to enamel, and the µTBS to dentin was stable over time when dentin was etched with GSE5 and GSE10.

Effect of grape seed extract (GSE) on functional activity and mineralization of OD-21 and MDPC-23 cell lines

Abstract Recent studies on functional tissue regeneration have focused on substances that favor cell proliferation and differentiation, including the bioactive phenolic compounds present in grape seed extract (GSE). The aim of this investigation was to evaluate the stimulatory potential of GSE in the functional activity of undifferentiated pulp cells and odontoblast-like cells. OD-21 and MDPC-23 cell lines were cultivated in odontogenic medium until subconfluence, seeded in 24-well culture plates in a concentration of 2x104/well and divided into: 1) OD-21 without GSE; 2) OD-21+10 µg/mL of GSE; 3) MDPC-23 without GSE; 4) MDPC-23+10 µg/mL of GSE. Cell proliferation, in situ detection of alkaline phosphatase (ALP) and total protein content were assessed after 3, 7 and 10 days, and mineralization was evaluated after 14 days. The data were analyzed by ANOVA statistical tests set at a 5% level of significance. Results revealed that cell proliferation increased after 10 days, and protein content, after 7 days of culture in MDPC-23 cells. In situ ALP staining intensity was higher in undifferentiated pulp cells and odontoblast-like cells after 7 and 10 days, respectively. A discrete increase in MDPC-23 mineralization after GSE treatment was observed despite OD-21 cells presenting a decrease in mineralized nodule deposits. Data suggest that GSE favors functional activity of differentiated cells more broadly than undifferentiated cells (OD-21). More studies with different concentrations of GSE must be conducted to confirm its benefits to cells regarding dentin regeneration.

Effect of grape seed extract on bond strength of restorative material to bleached enamel

Objective: The aim of this study was to evaluate the effect of different concentrations of a natural antioxidant (grape seed extract) on the bond strength of the restorative material to the bleached enamel. Methods: Forty fragments of healthy bovine incisors were randomly divided into four groups (n = 10): Group I: no bleaching; Group II: Bleaching with 35% hydrogen peroxide (HP) and without post-treatment; Group III: Bleaching with 35% HP + 5% grape seed extract; and Group IV: Bleaching with 35% HP + 10% grape seed extract. The bond strength at the adhesive interface was evaluated using the shear test (MPa). The data were analyzed by the analysis of variance (ANOVA) and Tukey test ( =0.05%). The fracture types were also analyzed and classified into: adhesive, cohesive or mixed. Results: Only Group III (bleached + 5% grape seed extract) had a significant increase (p<0.001) in bond strength values when compared to Group II bleached, without post- reatment). All groups showed a predominance of the adhesive type of fracture. Conclusion: It could be concluded that tooth bleaching decreases the bond strength to bleached enamel and 5% grape seed extract applied after dental bleaching improves the bond strength between the restorative material and the bleached enamel.

Objetivo: Este trabalho teve por objetivo avaliar in vitro o efeito de um antioxidante natural (extrato de semente de uva) em diferentes concentrações, na resistência de união do material restaurador ao esmalte clareado. Métodos: Quarenta fragmentos de incisivos bovinos hígidos, foram divididos aleatoriamente em quatro grupos (n=10): Grupo I: sem clareamento; Grupo II: clareado com peróxido de hidrogênio 35% (PH) e sem pós-tratamento; GIII: clareado PH 35% + extrato de semente de uva 5%; e Grupo IV: clareado com PH 35% + extrato de semente de uva 10%. A resistência de união da interface esmalte/material restaurador foi avaliada por meio do teste de cisalhamento (MPa). Os dados foram analisados pela análise de variância (ANOVA) e testes de Tukey ( =0,05%). Os tipos de fratura também foram analisados e classificados em: adesiva, coesiva ou mista. Resultados: Apenas o Grupo III (clareado + extrato de semente de uva 5%) apresentou aumento estatisticamente significante (p<0,001). dos valores de resistência de união comparado ao Grupo II (clareado e sem pós-tratamento). Todos os grupos mostraram um predomínio do tipo de fratura adesiva. Conclusão: O clareamento dental diminui significativamente a força de adesão ao esmalte dental clareado, e o extrato de semente de uva 5% aplicado após o clareamento dental melhora a resistência de união entre o material restaurador e o esmalte clareado.

Antibacterial efficacy of synthetic and natural-derived novel endodontic irrigant solutions

Abstract The aim of this study was to compare the efficacy of grape seed extract (GSE), calcium hypochlorite [Ca(ClO)2], and sodium hypochlorite (NaOCl) irrigant solutions with rotary or reciprocating instrumentation for disinfection of root canals inoculated with Enterococcus faecalis. The mesiobuccal root canals of mandibular molars were prepared and inoculated with Enterococcus faecalis for 21 days. The roots were then randomly divided into the following eight experimental groups (n=11) according to the instrumentation technique and disinfection protocol: ProTaper Next or Reciproc R25 with sodium chloride (control group), 6% NaOCl, 6% Ca(ClO)2, or 50% GSE used for irrigation during instrumentation. The antimicrobial activity was determined on the basis of a reduction in colony-forming units (CFUs) counted on bacterial samples collected before and after root canal instrumentation and expressed as a percentage of reduction. Data were evaluated by two-way ANOVA followed by Tukey HSD post-hoc tests (p<0.05). No significant differences were observed in bacterial reduction between the ProTaper Next and Reciproc R25 systems (p>0.05), regardless of the irrigant solution used. Furthermore, all active solutions (6% NaOCl, 50% GSE, and 6% Ca(ClO)2) showed similar potential to reduce bacterial counts (p>0.05) and were significantly more effective than sodium chloride (control) (p<0.05). The results suggest that the GSE and Ca(ClO)2 have potential clinical application as irrigant solutions in endodontic therapy since they present bactericidal efficacy against Enterococcus faecalis.

Resumo O objetivo deste estudo foi comparar a eficácia do extrato de semente de uva (ESU), hipoclorito de cálcio [Ca(ClO)2] e hipoclorito de sódio (NaOCl) como soluções irrigadores quando utilizadas com instrumentos reciprocantes e rotatórios para desinfecção de canais radiculares infectados com Enterococcus faecalis. Raízes mesio-vestibulares de molares inferiores foram preparados e inoculados com E. faecalis por 21 dias. As raízes foram aleatoriamente divididas em 8 grupos (n=11) de acordo com a técnica de instrumentação e protocolo de irrigação: ProTaper Next ou Reciproc R25 associados com soro fisiológico (grupo controle), Ca(ClO)2 6%, NaOCl 6% ou ESU 50%. A atividade antimicrobiana foi determinada pela redução do número de Unidades Formadoras de Colonias (UFCs) coletadas antes e após a instrumentação e expressas em porcentagens de redução. Os dados foram analisados estatisticamente pelos testes ANOVA seguido pelo teste complementar de Tukey HSD (p<0,05). Não foi encontrado diferença estatisticamente significante na redução bacteriana entre os sistemas ProTaper Next e Reciproc R25 (p>0.05), independente da solução irrigadora usada. Além disso, todas as soluções ativas (NaOCl, ESU e Ca(ClO)2) mostraram similar potencial em reduzir a quantidade de bactérias (p>0.05) e foram significativamente mais efetivas que o soro fisiológico (p<0.05). Pode-se concluir que o ESU e o Ca(ClO)2 apresentam potencial para aplicação clínica como irrigantes endodônticos uma vez que apresentaram efetividade antimicrobiana contra o E. faecalis.

Avaliação da expressão de citocinas pró e anti-inflamatórias após estímulo celular com diferentes agentes irrigantes / Evaluation of the expression of pro and anti-inflammatory cytokines after cellular stimulation with different irrigating agents

O objetivo do trabalho foi investigar o efeito de diferentes soluções químicas auxiliares, em concentrações subcitotóxicas, na expressão de citocinas pró e anti-inflamatórias, quando em contato com células da linhagem de linfoma humano, diferenciadas em macrófagos, human macrophage-like (U937), através da adição de 125ng/mL de PMA. As concentrações, citotóxica e subcitotóxica, de cada solução, foram determinadas de acordo com padrão ISO, utilizando-se fibroblastos de camundongos (L929). As células L929 foram cultivadas em meio MEM completo e mantidas em placas de 96 poços. As células foram então colocadas em contato com as diluições das soluções químicas auxiliares (a partir das concentrações recomendadas para uso, NaOCL 5,25%; CHX 2%; Quitosana 0,2%; HEBP 18%; GSE de Vitis vinífera, 6,5%), por 24 horas a 37ºC em estufa de CO2. Em seguida, o meio de cultivo contendo 1mg/mL de MTT foi adicionado aos poços em triplicata, para avaliação da viabilidade celular e determinação das concentrações citotóxicas (30% de morte das células L929) e subcitotóxicas (15% de morte) por regressão linear por meio do programa GraphPad Prism versão 6.0. As concentrações subcitotóxicas foram colocadas em contato com as células human macrophage-like (U937) por 60 min. Os sobrenadantes da pré-incubação (controles sem substâncias químicas auxiliares), e incubação com as soluções químicas auxiliares em meio DMEM completo foram congelados em freezer ultrafrio (­ 80ºC) e avaliados para as concentrações de dezessete citocinas através do kit Bio-Plex Pro Human Th17 Cytokine Panel® pelo método Luminex®. Foi possível obter valores de expressão de 7 citocinas pró-inflamatórias: IL-1ß, TNFα, IL-6, IL-17A, IL-17F, IL-22 e IL-23. As substâncias com maior atividade citotóxica (padrão ISO para morte celular) foram em ordem de grandeza, a quitosana, a CHX, o NaOCl, o HEBP e o GSE de Vitis vinifera. As concentrações subcitotóxicas das soluções químicas auxiliares, CHX e NaOCl, induziram ao aumento da maioria das citocinas pró-inflamatórias, exceto para a IL-6. Quanto a IL-1ß, a quitosana e a CHX, estas foram as que mais induziram a expressão pelas células U937 (P<0,05),enquanto o NaOCl induziu maior expressão de TNFα (P>0,05). Dentre as citocinas relacionadas ao fenótipo Th17, a CHX foi a que mais induziu a expressão de IL-17A (P<0,05), IL-17F e IL-23, seguido do NaOCl(P<0,05). O NaOCl induziu maior expressão de IL22 seguido da CHX (p<0,05). Quanto a IL-6, todas as substâncias modularam a sua expressão pelas células U937(p>0.05). O HEBP atuou como um excelente modulador da expressão de citocinas pró-inflamatórias, induzindo a redução da expressão de todas as citocinas testadas (P<0,05). O GSE de Vitis vinifera, apesar de ser o menos citotóxico não apresentou diferença estatística para o controle em nenhuma das situações avaliadas, exceto para a redução da IL-6 (assim como o HEBP e a quitosana) (p<0,05). A quitosana comportou-se de forma similar ao GSE de Vitis vinífera (p>0,05) exceto para a IL-1ß. Conforme observado, as substâncias químicas auxiliares são capazes de induzir a expressão de citocinas pró-inflamatórias diversas. O HEBP foi o agente químico que melhor modulou a expressão de citocinas pró-inflamatórias.

The aim of this investigation was to analyze the effect of different chemical endodontic solutions, in subcytotoxic concentrations, on the expression of pro and anti-inflammatory cytokines when in contact with human lymphoma lineage differentiated into human macrophages, human macrophage-like (U937), by the addition of 125ng/ml PMA. The cytotoxic and subcytotoxic concentrations of each solution were determined according to ISO standard using mouse fibroblasts (L929). L929 cells were cultured in complete MEM medium and maintained in 96-well plates. Dilutions of the auxiliary chemical solutions, from the recommended concentrations for use: NaOCL 5,25%; CHX 2%; Chitosan 0.2%; Etidronic acid 18% and Grape seed extract, Vitis vinifera, 6.5%, were carried out in complete MEM medium, and placed in contact with the cells for 24 hours at 37ºC in CO2 cell incubator, in triplicate. Thereafter, the culture medium containing 1 mg/mL MTT was added to the wells. Cytotoxic concentrations (30% death of L929 cells) and subcytotoxic (15% death) were determined by linear regression using GraphPad Prism version 6.0. Subcytotoxic concentrations were obtained in complete DMEM medium and maintained in contact for 60 min. with U937 cells. Controls without chemical substances were also performed. Supernatants from the pre-incubation, incubation with the chemical solutions and complete DMEM were removed, frozen in ultra-cold freezer (-80ºC) and evaluated for the concentrations of 17 cytokines through the Bio-Plex Pro Human Th17 Cytokine Panel® kit by Luminex®. The substances with higher cytotoxic activity were in order of magnitude: chitosan, CHX, NaOCl, etidronic acid and Vitis vinifera extract. The values of 7 pro-inflammatory cytokines: IL-1ß, TNFα, IL-6, IL-17A, IL-17F, IL-22 and IL-23 were obtained. Subcytotoxic concentrations of the irrigant agents (chlorhexidine and NaOCl) generally induced the increase of most pro-inflammatory cytokines, except for IL-6. Chitosan and CHX were the agents that most induced the expression of IL-1ß by U937 cells, whereas NaOCl induced higher TNFα expression (P> 0,05). Among the cytokines related to the Th17 phenotype, CHX was the agent that induced the highest expression of IL-17A (P <0,05), IL-17F and IL-23, followed by NaOCl (P<0,05). NaOCl induced the highest expression of IL22 followed by CHX. All of the substances modulated the expression of IL-6 by U937 cells (P >0,05). HEBP was an excellent modulator of the expression of pro-inflammatory cytokines, inducing the reduction of the expression of all cytokines tested. Vitis vinifera extract, despite being the least cytotoxic agent, did not present statistical differences compared to the control for any of the evaluated cytokines, except for the reduction of IL-6 (as well as etidronic acid and chitosan) (P<0,05). Chitosan, except for IL-1 ß, was very similar to the Vitis vinifera (P>0,05) extract. As observed, the auxiliary substances (especially the irrigants) are capable of inducing the expression of various pro-inflammatory cytokines. Etidronic acid was the chemical that modulated the expression of pro-inflammatory cytokines.

Effect of chemical and natural irrigant solutions on microhardness of root dentin­ an in vitro study

Aim: Modifications in the mechanical properties of dentin may reduce the fracture resistance of tooth, especially after endodontic treatment. The aim of present study was to evaluate the effect of the irrigation with different root canal irrigants on the microhardness of root dentin. Methods: The coronal portion of 60 single-rooted bovine incisors was sectioned and the pulpal tissue removed using endodontic K-files. The roots were cut transversely to obtain 2 fragments, which were embedded in acrylic resin and randomly distributed into six groups (n=20) according to the irrigation protocol: distilled water (DW) (control); 2% chlorhexidine solution (CHX); 6% sodium hypochlorite (NaOCl); 6% calcium hypochlorite (Ca[OCl]2); QMix; and 6.5% grape seed extract solution (GSE). The solutions were kept in contact with the root dentin specimens for 30 min. After that, irrigation with 5 mL of DW was performed. The Vickers microhardness was determined by performing three indentations in all specimens, using 300-g load and 20-second dwell time. The first indentation was made 1.000 µm from the root canal entrance, and two other indentations were made at a distance of 200 µm from each other. The microhardness value for each specimen was obtained as the average of the results for the three indentations. Data were statistically analyzed using one-way ANOVA with 5% significance level. Results: All the tested irrigant solutions maintained the same microhardness level of the root dentin when compared to the control group, with no statistically significant differences between them (p<0.05) Conclusion: The tested irrigant solutions did not present ability to modify the microhardness of root dentin

Hepatic protective effect of grape seed proanthocyanidin extract against Gleevec-induced apoptosis, liver Injury and Ki67 alterations in rats

ABSTRACT Gleevec (imatinib) is an antineoplastic chemotherapeutic agent used in the treatment of many types of cancer. The current study was conducted to examine the possible modifying effects of grape seeds proanthocyandins extract (GSPE) against apoptosis, liver injury and Ki67 alterations induced by Gleevec in male rats. 40 male albino rats were equally divided into four groups (First and second groups were control and GSPE groups; third group was Gleevec group and fourth group was treated with Gleevec and GSPE). Gleevec induced elevations in P53 and depletion of Bcl2 levels in liver tissues were compared with the control group. Liver sections in rats treated with Gleevec exhibited marked cellular infiltrations, vacuolar degeneration hepatocytes, numerous apoptotic cells, and congestion in central and portal veins, as well as a significant increase in the proliferating of Ki67 after Gleevec injection as compared with control group. In contrast, treatment with Gleevec and GSPE showed a moderate to good degree of improvement in hepatocytes with a significant increase in Ki67, a decrease in P53 and an increase in Bcl2 levels in liver tissues compared to treatment with Gleevec. Therefore, Gleevec induces apoptosis, injury and Ki67 changes in rat liver, whereas GSPE modulates these alternations.

Mechanical properties, water sorption characteristics, and compound release of grape seed extract-incorporated resins

Abstract Objective This study evaluated the effect of grape seed extract (GSE) incorporation on the mechanical properties, water sorption, solubility, and GSE release from the experimental adhesive resins. Material and Methods An experimental comonomer mixture, consisting of 40% Bis-GMA, 30% Bis MP, 28% HEMA, 0.26% camphorquinone and 1% EDMAB, was used to prepare four GSE-incorporated adhesive resins at concentrations of 0.5, 1, 1.5, and 2 wt%. The neat resin without GSE was used as the control. Six resin beams (25 mm x 2 mm x 2 mm) per group were prepared for flexural strength and modulus of elasticity evaluations using a universal testing machine at a crosshead speed of 1 mm/min. Five disks (6 mm in diameter and 2 mm in thickness) per group were used for microhardness measurements using a Leitz micro-hardness tester with Leica Qgo software. Five disks (7 mm in diameter and 2 mm in thickness) per group were prepared and stored in deionized water for 28 days. Water sorption, solubility, and GSE release in deionized water were calculated for each GSE-incorporated adhesive at the end of 28th day. Data was evaluated using one-way ANOVA and Tukey multiple comparisons. Results Flexural strength, modulus of elasticity and microhardness of GSE-incorporated adhesive decreased significantly with incorporation of 1.5% of GSE (p<0.05). Addition of GSE had no effect on the water sorption of the adhesive resins (p=0.33). The solubility of the resin also increased significantly with incorporation of 1.5% of GSE (p<0.05). Quantities of GSE release increased with increased concentration of GSE in the adhesive resin. Conclusion Up to 1% of GSE can be incorporated into a dental adhesive resin without interfering with the mechanical properties or solubility of the resins.

Effects of grape seed extract on periodontal disease: an experimental study in rats

Abstract Natural compounds capable of modulating the host response have received considerable attention, and herbal products are suggested as adjunctive agents in periodontal disease treatment. Objective This study aimed to demonstrate the effect of grape seed extract (GSE) on periodontitis. Material and Methods Ligature induced periodontitis was created in 40 rats and they were assigned to four equal groups. One group was fed laboratory diet (group A) while three groups received GSE additionally. Silk ligatures were placed around the cervical area of the mandibular first molars for four weeks to induce periodontitis. The GSE groups were reallocated regarding GSE consumption as: for two weeks before ligation (group B; totally eight weeks), from ligation to two weeks after removal of the ligature (group C; totally six weeks), and for two weeks from ligature removal (group D; totally two weeks). Sections were assessed histologically and immunohistochemically. Inflammatory cell number (ICN), connective tissue attachment level (CAL), osteoclast density (OD), IL-10 and TGF-β stainings in gingival epithelium (GE), connective tissue (GC), and periodontal ligament (PL) were used as the study parameters. Results Lower ICN, higher CAL, and lower OD were observed in the GSE groups (p<0.05). IL-10 was more intensive in the GSE groups and in the GEs (p<0.05). Group B showed the highest IL-10 for PL (p<0.05). TGF-ß was higher in the GEs of all groups (p<0.017). Conclusions The results suggest anti-inflammatory activities of GSE, but further investigations are needed for clarification of these activities.

Neuroprotective Effects of Grape Seed Procyanidin Extract on Ischemia-Reperfusion Brain Injury / 中国医学科学杂志(英文版)

Objective Oxidative stress (OS) plays a crucial role in ischemic stroke. Grape seed procyanidin extract (GSPE) was reported to be a critical regulator of OS. We hypothesized that GSPE might also be protective in ischemia-reperfusion brain injury. This study aimed to explore whether GSPE administration can protect mice from ischemia-reperfusion brain injury.Methods Transient middle cerebral artery occlusion (MCAO) was conducted followed by reperfusion for 24 hours to make ischemia-reperfusion brain injury in mice that received GSPE (MCAOG, n=60) or normal saline (MCAONS, n=60). Sham-operated mice (GSPE group and normal saline group) were set as controls. The neurological severity score (NSS) was used to evaluate neural function impairment 1 hour, 24 hour, 3 days and 7 days after MCAO. Mice underwent brain T2WI imaging with a 3T animal MRI scanner 24 hours after reperfusion, and the stroke volume of brains were calculated according to abnormal signal intensity. Immunohistopathological analysis of brain tissues at 24 h after reperfusion was performed for neuronal nuclear antigen (NeuN), CD34, Bcl-2, and Bax. Glutathione peroxidation (GSH-Px) activity and the level of malonaldehyde (MDA) of brain tissue were also examined. The above indexes were compared among the groups statistically.Results Significant functional improvement was observed 24 hours after MCAO in MCAOG group compared to MCAONS group (P<0.05). MCAOG group had smaller cerebral stroke volume (22.46 ± 11.45 mmvs. 47.84±9.06 mm, P<0.05) than MCAONS group 24 hours after MCAO. More mature NeuN-immunoreactive neurons and more CD34-positive cells in peri-infarct zones were observed in brain tissue of MCAOG mice 24 h after MCAO than that of MCAONS mice (both P<0.05). MCAONS mice had significantly higher number of Bax-positive cells in brain tissue than MCAOG (P<0.05). The mean MDA level was significantly lower (P<0.05) and the GSH-Px activity was significantly higher (P<0.05) in brains of MCAOG mice compared to those of MCAONS mice.Conclusion GSPE administration protects mice from ischemia-reperfusion brain injury through attenuating oxidative stress and apoptosis, promoting angiogenesis, and activating antioxidant enzyme GSH-Px. GSPE may represent a new therapeutical direction for the treatment of ischemia-reperfusion brain injury.

Atividade antimicrobiana do extrato de semente de uva Vitis vinifera sobre biofilme de Enterococcus faecalis: análise com microscopia confocal / Antimicrobial effectiveness of grape seed extract against Enterococcus faecalis biofilm: confocal microscopy analysis

As limitações dos irrigantes químicos utilizados em endodontia têm incentivado novas pesquisas acerca de substâncias alternativas para serem utilizadas no tratamento endodôntico. Estudos anteriores sugerem que o extrato de semente de uva (Vitis vinifera), tem atividade antimicrobiana assim como é eficaz no combate a cepas de Enterococcus faecalis. No entanto, os efeitos deste novo irrigante em biofilmes microbianos ainda não foram elucidados. Dessa forma, o objetivo deste estudo foi avaliar e quantificar o grau de desinfecção da dentina contaminada por biofilme de E. faecalis após irrigação com extrato de semente de uva Vitis vinífera 6,5% por meio de análise microscopia confocal a laser. A clorexidina (CHX) 2% e o hipoclorito de sódio (NaOCl) 5,25% foram utilizados como irrigantes para comparação. Foram confeccionadas quarenta amostras de discos de dentina que foram contaminadas com E. faecalis e incubadas por um período de 21 dias para formação do biofilme. As amostras foram divididas em diferentes grupos de acordo com o irrigante testado após transcorrido o período de contaminação: grupo NaOCl, grupo CHX, grupo Vitis vinifera e grupo solução salina (controle). Após a irrigação, os discos de dentina foram corados com o corante fluorescente LIVE/DEAD BacLight e analisados por microscopia confocal à laser para determinar a proporção de células mortas no biofilme. A aquisição das imagens e quantificação do biofilme foi realizada utilizando-se o software LAS X. Uma distribuição normal dos dados foi confirmada pelo teste Shapiro- Wilk (p>0,05). Por esse motivo, a análise estatística foi realizada utilizando-se One-way análise de variância. Comparações Post hoc pair-wise foram realizadas utilizando-se o teste Tukey para múltiplas comparações (P<0.05). O grupo no qual o NaOCl foi utilizado como irrigante apresentou maior quantidade de células bacterianas mortas do que os dos outros irrigantes (P<0,05). Já o grupo da Vitis vinifera demonstrou maior morte bacteriana quando comparada a CHX e ao grupo controle (P<0,05). Não foram observadas diferenças entre a CHX e o grupo controle (P>0,05). A irrigação com o extrato de semente de uva, vitis vinífera, apresentou atividade antimicrobiana sobre o biofilme de E. faecalis sendo esta inferior a apresenta pelo NaOCl e superior a apresentada pela CHX.

The limitations of chemical irrigants used in endodontics have encouraged further research on alternative substances to be used in endodontic treatment. Previous studies have suggested that grape seed extract (Vitis vinifera), an irrigant of natural origin, is effective against strains of Enterococcus faecalis. The aim of this study was to evaluate and quantify the degree of disinfection of dentin contaminated by E. faecalis biofilm after irrigation with Vitis vinifera grape seed extract 6,5% and to compare it with chlorhexidine (CHX) 2% and Sodium hypochlorite (NaOCl) 5,25% by confocal microscopy laser. Forty samples of dentin disks that will be contaminated with E. faecalis and incubated for a period of 21 days for biofilm formation were prepared. The samples were divided into different groups according to the irrigant tested after the period of contamination: NaOCl group, CHX group, Vitis vinifera group and control group . After irrigation, the dentin disks were stained with the fluorescent dye LIVE / DEAD BacLight and analyzed by laser confocal microscopy to determine the proportion of dead cells in the biofilm. The acquisition of the images and quantification of the biofilm was performed using the software LAS X. Normal distribution of the data was confirmed by Shapiro- Wilk test (p>0.05). Therefore, statistical analysis was performed using One-way analysis of variance. Post hoc pair-wise comparisons were performed using Tukey's test for multiple comparisons (P<0.05). NaOCl presented a higher amount of bacterial cells killed than the other irrigators (P <0.05). Irrigation with grape seed extract, Vitis vinifera, showed antimicrobial activity on the E. faecalis biofilm, being lower than that presented by NaOCl and higher than that presented by CHX.

[Effect of combination of topical ointment of aqueous extract of grape seed [Vitis Vinifera] and Eucerin on wound healing in diabetic rats]

Background and Objective: Diabetic mellitus is one of the main health problems of diabetic subjects skin wound. This study was done to evaluate the effect of topical ointment produced from aqueous extract of grape seed [Vitis Vinifera] plus Eucerin on wound healing in diabetic rats

Methods: In this experimental study, 48 male Wistar rats were randomly allocated into control, first, second and third experimental groups. Animals in control group were received only buffer citrate, interaperitonally. Diabetes was induced by 55 mg/kg/bw of Streptozotocin in three experimental groups. Skin wound [2x3 cm[2]] was created in left side of the lumbar region in three experimental groups. Animals in experimental gourp 1 did not received any treatment. The wounds of animals in experimental group 2 were treated with topical ointment of Eucerin twice a day. The wounds of animals in experimental group 3 were treated with topical ointment of aqueous extract of grape seed plus Eucerin with a ratio of 3:1 for 21 days twice a day. Wound healing process was evaluated using macroscopic and histological method

Results: The mean area and percentage of wound after treatment was significantly reduced in experimental group 2 and 3 in compare to experimental group 1 [P<0.05]. The percentage of improvement wound healing was significantly increased in experimental groups 2 and 3 in compare to experimental group 1 [P<0.05]. Synthesis of collagen fibers was increased in experimental group 3 in compare to experimental group 2 and 1

Conclusion: Topical application of aqueous extract of grape seed with ointment of Eucerin accelerates skin wound healing in diabetic rats

Grape Seed Procyanidin Extract Reduces Arsenic-Induced Renal Inflammatory Injury in Male Mice / 生物医学与环境科学（英文）

The aim of the present study is to evaluate the ability and mechanism by which grape seed procyanidin extract (GSPE) relieves arsenic trioxide (As2O3)-induced renal inflammatory injury. Therefore, male Kunming mice were treated with As2O3 and/or GSPE by gavage for 5 weeks. Mice were then sacrificed and inflammatory cytokines of kidneys were examined by ELISA, whereas the expression levels of molecules involved in the nuclear factor (NF)-κB signaling pathway were evaluated by both qRT-PCR and Western blot. Our results indicate that GSPE prevents As2O3-mediated renal inflammatory injury by inhibiting activation of the NF-κB signaling pathway and inflammatory cytokine production, while promoting expression of anti-inflammatory cytokines.