Evaluation of Cytotoxic effect of Moringa peregrina seeds on Oral Cancer, CAL 27, Cell Line and Red Blood Cells Hemolysis / Evaluación del efecto citotóxico de las semillas de Moringa peregrina en el cáncer oral, CAL 27, líneas celulares y hemólisis de glóbulos rojos

Background: Moringa peregrina Forssk is a well-known plant in ethnomedicine due to its widespread uses in various diseases like cough, wound healing, rhinitis, fever, and detoxification. The plant seeds contain compounds that are cytotoxic to many cancer cells. During the therapeutic use of plants via the oral route, some compounds present in the plants may be cytotoxic to normal cell lines and red blood cells. Objective: This study was the first report of investigation of the cytotoxic profile on oral cancer, CAL 27, cell line, and hemolytic activities on human erythrocytes of Moringa peregrina seeds ethanolic extract (MPSE). Methods: MPSE was screened for its cytotoxic effect against oral cancer, CAL 27, cell line using 3-(4, 5-dimethylthiazol-2-yl)-2, 5,-diphenyltetrazolium bromide (MTT) assay. The toxicity of MPSE on human erythrocytes was determined by in vitro hemolytic assay. Results: MPSE showed significant anti-proliferative activity against oral cancer, CAL 27 cell line at lower concentrations with half maximal inhibitory concentration (IC50) value of 21.03 µg/mL. At 1,000 µg/ml of MPSE, the maximum hemolysis was found to be 14.3% which is within safer limit. Conclusions: This study revealed a potential anti-oral cancer of MPSE and provided a baseline for its potential use in oral cancer treatment with minimum hemolytic effect on human RBCs.

La Moringa peregrina Forssk es una planta muy conocida en etnomedicina debido a sus usos generalizados en diversas enfermedades como la tos, la cicatrización de heridas, la rinitis, la fiebre y la desintoxicación. Las semillas de la planta contienen compuestos citotóxicos para muchas células cancerosas. Durante el uso terapéutico de las plantas por vía oral, algunos compuestos presentes en ellas pueden ser citotóxicos para las líneas celulares normales y los glóbulos rojos. Objetivo: Este estudio fue el primer informe de investigación del perfil citotóxico sobre el cáncer oral, CAL 27, línea celular, y las actividades hemolíticas en eritrocitos humanos del extracto etanólico de semillas de Moringa peregrina (MPSE). Métodos: Se examinó el efecto citotóxico del MPSE contra la línea celular de cáncer oral CAL 27 mediante el ensayo con bromuro de 3-(4, 5-dimetiltiazol-2-il)-2, 5,-difeniltetrazolio (MTT). La toxicidad del MPSE sobre los eritrocitos humanos se determinó mediante un ensayo hemolítico in vitro. Resultados: MPSE mostró una actividad antiproliferativa significativa contra el cáncer oral, línea celular CAL 27 a concentraciones más bajas con un valor de concentración inhibitoria media máxima (IC50) de 21,03 µg/mL. A 1.000 µg/ml de MPSE, la hemólisis máxima fue del 14,3%, lo que está dentro del límite de seguridad. Conclusiones: Este estudio reveló un potencial anticancerígeno oral de MPSE y proporcionó una base para su uso potencial en el tratamiento del cáncer oral con un efecto hemolítico mínimo en los glóbulos rojos humanos.

Grape seed extract inhibits apoptosis of rat growth plate chondrocytes and promotes tibial bone growth / 中国组织工程研究

BACKGROUND:Grape seed extract can inhibit chondrocyte apoptosis and aging,and improve osteoarthritis.However,the effects of grape seed extract on the apoptosis of chondrocytes in the growth plate and tibial growth are still unclear. OBJECTIVE:To investigate the effect of grape seed extract on interleukin-1β-induced apoptosis in rat growth plate cells and on tibial bone growth. METHODS:(1)Cell experiment:Growth plate chondrocytes from Sprague-Dawley rats were isolated,cultured,and identified.The cells were then randomly divided into control group,model group,grape seed extract group,miR-138-5p NC group and miR-138-5p inhibitor group.In the model group,20 ng/mL interleukin-1β was used to induce apoptosis in rat growth plate chondrocytes.In the grape seed extract group,20 ng/mL interleukin 1β was added along with 10 μmol/L grape seed extract solution for 48 hours.Cells in the miR-138-5p NC and inhibitor groups were transfected with 5 nmol/L miR-138-5p NC and 5 nmol/L miR-138-5p inhibitor for 12 hours,respectively,followed by addition of 20 ng/mL interleukin-1β.qRT-PCR was used to detect miR-138-5p and caspase-3 expression.Luciferase reporter assay was used to analyze the relationship between miR-138-5p and caspase-3 targeting.Cell counting kit-8 was used to detect cell proliferation activity.Flow cytometry was used to detect cell apoptosis.Caspase-3 and Bcl-2 protein expressions were detected by western blot.(2)Animal experiment:The animals were divided into normal control group,grape seed extract group and miR-138-5p inhibitor group.The effects of grape seed extract on epiphyseal closure and tibial growth of the tibial plateau in rats were observed. RESULTS AND CONCLUSION:miR-138-5p had a targeting relationship with caspase-3.Compared with the control group,cell proliferation was significantly reduced,apoptosis was significantly increased(P＜0.01),miR-138-5p,Bcl-2 expression was reduced(P＜0.01),and caspase-3 expression was increased(P＜0.01)in the model group.Compared with the mod group,the grape seed extract group showed a significant increase in cell proliferation,a significant decrease in apoptosis(P＜0.01),an increase in miR-138-5p and Bcl-2 expression(P＜0.01)and a decrease in caspase-3 expression(P＜0.01).Compared with the grape seed extract group,the miR-138-5p inhibitor group showed a significant decrease in cell proliferation,a significant increase in apoptosis(P＜0.01),a decrease in miR-138-5p and Bcl-2 expression(P＜0.05 or P＜0.01),and an increase in caspase-3 expression(P＜0.05).Intragastric administration of grape seed extract could delay epiphyseal closure of the tibial plateau and promote tibial bone growth in rats,whereas miR-138-5p inhibitor intervention inhibited the effect of grape seed extract on tibial bone growth in rats.To conclude,grape seed extract can inhibit apoptosis of rat growth plate chondrocytes through regulating the miR-138-5p/caspase-3 pathway,improve epiphyseal closure of rat tibial plateau and promote tibial bone growth.

Grape seed extract inhibits apoptosis in growth plate chondrocytes and promotes tibial growth in rats / 中国组织工程研究

BACKGROUND:Grape seed extract has been shown to be effective in inhibiting the growth of androgen-dependent tumors(e.g.,breast cancer),and thus grape seed extract could theoretically inhibit epiphyseal closure induced by estrogen in late adolescence. OBJECTIVE:To screen the effects of grape seed extract on apoptosis of growth plate chondrocytes and epiphyseal closure in rats. METHODS:(1)In vitro experiment:Growth plate chondrocytes from rat large tibia and femur at logarithmic growth stage were obtained and cultured in groups:normal control group,model control group(adding 17β-estradiol to induce apoptosis),positive control group(adding letrozole and 17β-estradiol),grape seed extract group(adding 17β-estradiol and 10 μg/mL grape seed extract),Caspase-9 inhibitor group(adding 17β-estradiol and Caspase-9 inhibitor),Caspase-9 agonist group(adding 17β-estradiol and Caspase-9 agonist).Cell apoptosis was detected by flow cytometry after 48 hours of culture.(2)In vivo experiment:Thirty 3-month-old Sprague-Dawley rats were randomly divided into model control group,positive control group and low-,medium-and high-dose groups,with five rats in each group.All rats were injected subcutaneously with 17β-estradiol(3 times per week)to establish epiphyseal closure models,followed by intragastric administration of letrozole in positive control group and 0.05,0.2 and 0.8 g/kg grape seed extract in low-,medium-and high-dose groups,respectively,once a day until over 2/3 of the epiphyseal plate in the model control group was closed.The length of the tibia was then observed.Another 18 Sprague-Dawley rats were randomly divided into model control group,positive control group,and medium-dose group,with 6 rats in each group,treated as above for 1.5 continuous months.The expression of Caspase-9 protein in rat growth plate cartilage was detected by western blot. RESULTS AND CONCLUSION:(1)In vitro experiment:17β-estradiol could induce apoptosis in growth plate chondrocytes,and letrozole,grape seed extract,and caspase-9 inhibitors could all inhibit apoptosis in growth plate chondrocytes.(2)In vivo experiment:When more than 2/3 of the epiphyseal plate in the model control group was closed,the number of rats with epiphysis closure in the positive control and medium-dose groups was less than that in the model control group(P＜0.05),and the tibial length was longer than that in the model control group(P＜0.05),and the Caspase-9 protein expression in the tibial growth plate was lower than that in the model control group(P＜0.05).To conclude,the appropriate dose of grape seed extract can effectively inhibit the apoptosis of growth plate chondrocytes and delay epiphyseal closure,which has the potential to promote bone growth.

Low concentrations of grape seed extract maintain osteoblast morphology, cell adhesion, and mineralization

Abstract The increase in life expectancy has led to a higher incidence of osteoporosis, characterized by an imbalance in bone remodeling. Several drugs are used for its treatment, but most promote undesirable side effects. The present investigation evaluated the effects of two low concentrations of grape seed extract (GSE) rich in proanthocyanidins on MC3T3-E1 osteoblastic cells. The cells were cultured in an osteogenic medium and divided into control (C), 0.1 µg/mL GSE (GSE0.1), and 1.0 µg/mL GSE (GSE1.0) groups to evaluate cell morphology, adhesion, and proliferation, in situ alkaline phosphatase (ALP) detection, mineralization and immunolocalization of osteopontin (OPN). The data obtained were analyzed by statistical tests for a significance of 5%. Cell morphology was maintained with both GSE concentrations, whereas cell adhesion significantly increased within three days in all groups. Cell proliferation increased significantly at seven days of culture, followed by a significant decrease in all experimental periods, with no statistical difference among them. In situ detection of ALP and mineralization increased with time, but within each period, no statistical differences among groups were observed. The expression of osteopontin was distributed regularly with more intensity after 24 hours in the GSE0.1 group. After three days, OPN expression was more intense in the control group, followed by GSE0.1 and GSE1.0 groups. Data obtained suggest that low concentrations of GSE do not affect the morphology and may stimulate the functional activity of osteoblastic cells.

Resumo O aumento da expectativa de vida tem levado a uma maior incidência de osteoporose, caracterizada por um desequilíbrio na remodelação óssea. Vários medicamentos são utilizados para o seu tratamento, contudo, a maioria promove efeitos colaterais indesejáveis. A presente investigação avaliou os efeitos de duas baixas concentrações de extrato de semente de uva (GSE) rico em proantocianidinas em células osteoblásticas MC3T3-E1. As células foram cultivadas em meio osteogênico e divididas em grupos controle (C), 0,1 µg/mL de GSE (GSE0.1) e 1,0 µg/mL de GSE (GSE1.0) para avaliar morfologia, adesão e proliferação celular, detecção in situ de fosfatase alcalina (ALP), mineralização e imunolocalização da proteína osteopontina (OPN). Os dados obtidos foram analisados por testes estatísticos para um nível de significância de 5%. A proliferação celular aumentou significativamente aos sete dias de cultura, seguido de uma diminuição significativa em todos os períodos experimentais, sem diferença estatística entre eles. A detecção in situ de ALP e mineralização aumentou com o tempo, mas dentro de cada período não foram observadas diferenças estatísticas entre os grupos. A morfologia celular foi mantida com ambas as concentrações de GSE, enquanto a adesão celular aumentou significativamente aos três dias em todos os grupos. A expressão de osteopontina distribuiu-se regularmente com maior intensidade após 24 horas no grupo GSE0.1. Após três dias, a expressão de OPN foi mais intensa no grupo controle, seguida pelos grupos GSE0.1 e GSE1.0. Os dados obtidos sugerem que baixas concentrações de GSE não afetam a morfologia e podem estimular a atividade funcional das células osteoblásticas.

Effects of Aqueous Seed Extracts of Sphenostylis stenocarpa on the Reproductive Indices of Male Rats

Effects of aqueous seed extracts of Sphenostylis stenocarpa on the reproductive indices of male rats were investigated. A total of 104 adult rats were used for the experiment, and were divided into 4 groups (group A – D) and replicated in triplicate. Group A served as the normal control, while groups B, C and D received three graded doses (800mg/kg, 1200mg/kg and 1600mg/kg) of the extracts, respectively, by oral intubation. The gonad characteristics, sperm parameters and hormonal analyses of the male rats were determined using standard procedures. These were ascertained prior to the commencement of treatment, and on weekly basis. Data were analyzed statistically using SPSS and R software at 95% confidence interval. An overall dose and time dependent showed significant differences in the mean weekly gonad characteristics of the male rats in the treatment groups when compared with the control. There was a significant reduction (p < 0.05) in the body weights of the male rats, but a significant increase (P < 0.05) in the testes weights, gonad somatic index, sperm count and sperm motility in the rats. The gonadal hormone testosterone, responded to the plant extracts, while follicle-stimulating and luteinizing hormones were largely undetected. There were significant increases in the testosterone levels of all the treated rats. Conclusively, aqueous seed extracts of Sphenostylis stenocarpa seems to possess ability to enhance reproductive health in male rats.

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.

Analysis of Mechanism of Antidepressant Effect of Sophora flavescens Seed Extract / 中国实验方剂学杂志

ObjectiveTo explore the antidepressant effect of Sophora flavescens seed extract and its molecular mechanism. MethodA mouse depression model was established by intraperitoneal injection of lipopolysaccharide（LPS）， and normal group， model group， fluoxetine group（2.5 mg·kg-1）， and S. flavescens seed low， medium and high dose groups（200， 400， 800 mg·kg-1） were set up for 7 d of consecutive gavage. Then the antidepressant effect of S. flavescens seed extract was evaluated by using open field test， elevated plus maze test and forced swimming test. Pathological morphological changes in the hippocampal tissue was observed by hematoxylin-eosin（HE） staining. Protein expression levels of G1/S-specific cyclin D1（Cyclin D1）， Wnt1， β-catenin and phosphorylated glycogen synthase kinase-3β（p-GSK-3β） in mouse brain tissues were detected by Western blot. Hippocampal cell apoptosis was detected by terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate（dUTP） nick end labeling（TUNEL）. ResultThe results of mouse behavioral experiments showed that compared with the normal group， the speed of movement in the open field and the distance of movement in the central area of the open field， and the time spent on the open arms of the elevated plus maze were significantly reduced in the model group（P<0.01）， while immobility time in the forced swimming test was significantly increased（P<0.05）. Compared with the model group， the S. flavescens seed medium and high dose groups had increased speed of movement in the open field test and time spent on the open arms of the elevated plus maze test（P<0.05， P<0.01）， and decreased immobility time in the forced swimming test（P<0.05）， the distance of movement in the central area of the open field test increased in the high dose group（P<0.05）. HE staining results showed that compared with the normal group， the hippocampal neuron structure of mice in the model group was damaged. Compared with the model group， after treatment of S. flavescens seed extract， the pathological state of the mouse hippocampal neuron structure was alleviated， and the neurons increased， were neatly arranged， and the cytoplasm was clear. Western blot results showed that the protein expression levels of Wnt1 and β-catenin in mouse brain tissue were significantly decreased（P<0.01）， while the protein expression levels of Cyclin D1 and p-GSK-3β were significantly increased（P<0.01） after LPS injection. Compared with the model group， protein expression levels of Wnt1 and β-catenin in brain tissue of S. flavescens seed medium and high dose groups were significantly increased（P<0.01）， while the protein expression levels of Cyclin D1 and p-GSK-3β were significantly decreased（P<0.01）. TUNEL staining results showed that the hippocampal cell apoptosis rate in the model group was significantly increased compared with that of the normal group（P<0.01）， while the hippocampal cell apoptosis rate in the S. flavescens seed medium and high dose groups was significantly decreased compared with that of the model group（P<0.01）. ConclusionS. flavescens seed extract can effectively improve the severity of depression in LPS-induced depressed mice， and its molecular mechanism is related to the regulation of neuroinflammation and hippocampal neuronal apoptosis mediated by Wnt/β-catenin signaling pathway.

Efecto de distintas Concentraciones de un Gel Experimental a Base de Extracto Etanólico de Semillas de Uva (Vitis vinifera L) sobre la Resistencia de Adhesión de Resina Nanohíbrida

El objetivo del estudio es determinar el efecto de distintas concentraciones de un gel experimental a base de extracto etanólico de semillas de uva (Vitis vinifera L) sobre la resistencia a la adhesión de resina nanohíbrida. Se obtuvo el extracto etanólico a base de Semillas de Uva (EESU) por maceración en alcohol al 70 % durante 45 días. Se reconstituyó el principio activo de las semillas de uva en forma de polvo mezclando 200 ml de agua ultrapura, 2 g. de Carbopol 940, 20 g. de glicerol, 20g. de propilenglicol, 1g. de benzoato de sodio, 1.5 de trietanolamina, obteniendo un gel con un pH de 14, en concentraciones de 5 %, 10 % y 15 %. La sustancia madre fue sometida a cromatografía en capa fina HPLC para identificar sus componentes químicos hallando compuestos fenólicos (ácido Gálico), taninos y en menor cantidad flavonoides. No existieron diferencias significativas entre los grupos control y experimentales cuya tracción se realizó inmediatamente al tratamiento de aclaramiento dental, a los 7 días y 14 días. El extracto etanólico de semillas de uva (Vitis Vinífera L) en concentraciones de 5 %, 10 % y 15 % tuvieron el mismo efecto sobre la resistencia a la adhesión de resina nanohíbrida.

The objective of the study is to determine the effect of different concentrations of an experimental gel based on ethanolic extract of grape seeds (Vitis vinifera L) on the resistance to adhesion of nanohybrid resin The ethanolic extract based on Grape Seeds (EESU) by maceration in 70 % alcohol for 45 days. The active principle of grape seeds was reconstituted in powder form by mixing 200 ml of ultrapure water, 2 g. of Carbopol 940, 20 g. glycerol, 20g. of propylene glycol, 1g. of sodium benzoate, 1.5 of triethanolamine, obtaining a gel with a pH of 14, in concentrations of 5 %, 10 % and 15 %. The base substance was subjected to HPLC thin-layer chromatography to identify its chemical components, finding phenolic compounds (Gallic acid), tannins and, to a lesser extent, flavonoids. There were no significant differences between the control and experimental groups whose traction was performed immediately after dental whitening treatment, at 7 days and 14 days. The ethanolic extract of grape seeds (Vitis vinifera L) in concentrations of 5 %, 10 % and 15 % had the same effect on the resistance to the adhesion of nanohybrid resin.

Use of two kinds of antioxidants to restore the bond strength of bleached enamel / 华西口腔医学杂志

OBJECTIVES@#This work aimed to evaluate the ability of two kinds of antioxidants, namely, grape-seed extract and sodium ascorbate, in restoring bond strength at the resin-enamel interface after bleaching.@*METHODS@#Ten groups of samples with 15 teeth per group were prepared for shear-bond-strength test at the resin-enamel interface after bleaching. The groups were as follows: control; no antioxidant; 2.5%, 5%, 10%, or 15% grape-seed extract; and 2.5%, 5%, 10%, or 15% sodium ascorbate. The peak values of shear bond strength when resin was debonded from teeth and the failure modes under a microscope were recorded. Ten other groups of teeth with two teeth per group were prepared and treated in a similar approach before resin bonding. The samples were cut vertically to the bonding interface. The structures of the bonding interface were compared by scanning electron microscopy.@*RESULTS@#No statistically significant difference in shear bond strength was found among the no-antioxidant, 2.5% grape-seed extract, and 2.5%, 5%, or 10% sodium ascorbate groups (@*CONCLUSIONS@#Immediately after bleaching, the bond strength of dental enamel significantly decreased. Bond strength can be restored by 5% grape-seed extract or 15% sodium ascorbate in 5 min.

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)

Melinjo Seed Extract / 日本補完代替医療学会誌

Melinjo (Gnetum gnemon L.) is distributed in Southeast Asia and its fruits, seeds and leafs are commonly eaten in Indonesia. It was found that melinjo seed extract contains some resveratrol dimers (gnetin C, gnemonoside A and gnemonoside D). Melinjo seed extract and gnetin C have been reported to show beneficial effects on several diseases, such as hyperuricemia, atherosclerosis, fatty liver, diabetes, dementia, cancer, skin aging and periodontitis etc．Melinjo seed extract is one of the desired functional food materials in Japan.

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.

Protective effects of Urtica dioica L. seed extract on liver tissue injury and antioxidant capacity in irradiated rats

Radiotherapy is often used for the treatment of cancer. However, it causes some side effects in patients. This study aimed to determine the hepatoprotective effects of Urtica dioica L. seed-extract (UDSE) in radiation-induced liver injury. Thirty-two male rats were randomly divided into 4 groups (n=8): control(C) group: no action was taken; radiation (R) group: irradiation was administrated at 5Gy single-fraction, radiation with UDSE(R+UDSE) group: irradiation was administrated at 5 Gy single-fraction and animals were fed pellets with 30 mL UDSE/kg; UDSE group: animals were fed pellets with 30 mL UDSE/kg. All of the experiments were performed in all of the groups over 10 days. Malondialdehyde (MDA) and reduced-glutathione (GSH) levels and superoxide-dismutase (SOD), catalase (CAT), glutathione-peroxidase (GSH-Px), aspartate-transaminase (AST), and alanine-aminotransferase (ALT) activities were determined. Histopathological findings were also evaluated in liver tissues. SOD, CAT and GSH-Px activities and GSH levels in the serum and liver were significantly increased, while MDA levels decreased in the R+UDSE group compared with the R group (P<0.05). Moreover, AST and ALT serum activities in the R+UDSE group were lower than those in the R group (P<0.05). In addition, radiation induced degenerative/necrotic changes in the R group were significantly compensated in the R+UDSE group. The results showed that radiation increased oxidative stress and decreased antioxidant capacity, as well as degeneration in the liver. However, UDSE attenuated these degenerative changes.

In vitro penetration activity of ionic liquid-Gnetum gnemon seed extract loaded solid lipid nanoparticles

Gnetum gnemon L. (Melinjo) seeds have high trans-resveratrol, which was known to have poor skin absorption ability.This study was performed to evaluate the skin absorption of ionic liquid-melinjo seed extract (IL-MSE) loaded solidlipid nanoparticles (SLN). The SLNs was formulated with glyceryl monostearate as the lipid ingredient, ceteareth-25,ceteareth-6, stearyl alcohol, an aqueous phase, and 10% melinjo seed extract. The SLNs were prepared by modificationof high-pressure homogenization. The polydispersity index (PDI), zeta potential, entrapment efficiency (EE), particlesize (PS), and morphological scanning electron microscopy (SEM) were evaluated. The in vitro penetration of IL-MSESLN was carried out by using the Franz cell diffusion method. The SLNs formulations showed an average of particlessize about 794 nm, and with high lipid and surfactant, the content could contributing to high entrapment efficiencyto almost 89%. Even though the polydispersity index of SLN was 1.229, and zeta potential measurement was 62.56mV. Up to 45% of the trans-resveratrol penetrated through the skin after 8 hours of the test run. The IL-MSE loadedto SLNs could improve the absorption of trans-resveratrol through the skin.

Effect of Trigonella foenum Extract and ZiO2 Nanoparticles on Some Pathogenic Fungi and Bacteria

Aim: The current venture, were made to evaluate the inhibitory effect of Trigonella foenum seed Extract and ZiO2 Nanoparticles on some selected species of Fungi and Bacteria. Materials and Methods: two bacterial species included Pseudomonas aeruginosa and Staphylococcus aureus and three fungal species which is Cryptococcus neoformans, Candidda albicans and Chaetomium were used to evaluate the antibacterial activity of Trigonella foenum Extract and ZiO2 Nanoparticles. Results: This study showed that the Zirconium Oxide (ZiO2) nanoparticles have antifungal and antibacterial activities on the isolates of Cryptococcus neoformans, Candida alicans and Staphylococcus aureus, respectively. While the antimicrobial activity of Zirconium Oxide nanoparticles on the Chaetomium and Pseudomonas aeruginosa was negative. All tested fungi and bacterial isolates were found to be sensitive to Trigonella foenum seed extract, the results of the compination of the ZiO2 Nanoparticle and the Trigonella foenum seed extract were poisitive for all tested fungi isolates and bacterial isolates. The XRD analysis was done for Zirconium Oxide nanoparticles and the result showed that the biocrystallization on the surface of the Zirconium Oxide manoparticles. The average partides size was about (29.8) nm. Conclusions: This investigation conclude that the use of Trigonella foenum seed Extract has the effect of killing all bacteria and fungi under study, result indicate the Trigonella foenun seed Extract best antibacterial efficacy than the ZiO2 together (AU)

The effect of date palm seed extract as a new potential radioprotector in gamma-irradiated mice

Objective: Date palm seed extract (DPSE) has various compounds revealing antioxidant features. This study aimed to evaluate the radioprotective effect of DPSE in total body gamma irradiation. Materials and Methods: At first, chemical characteristics of DPSE were analyzed by ultraviolet, visible and Fourier transform infrared spectroscopy. Then, the toxicity of DPSE was assessed. For this purpose, 60 mice were divided into five groups, and each of the groups were injected by the doses of 100, 200, 300, 400, and 500 mg/kg, respectively. At the termination of the experiment, mortality rate and weight loss of all mice were evaluated over a period of 30 days. Finally, the radioprotective effect of DPSE was evaluated by dividing 36 mice into three groups: control, test, and placebo and then were irradiated by Cobalt-60. Results: According to the findings, there was no mortality due to DPSE. Furthermore, for the maximum dose of 500 mg/kg, the number of mice surviving at the termination of the experiment with and without injection of DPSE was reported as 83% and 41%, respectively. In addition, a significant difference was obtained between radiated mice with and without DPSE injection (P = 0.035). Conclusion: The findings showed that DPSE injected into mice before irradiation has no toxicity and could protect mice from lethal effects of total body irradiation. The use of DPSE as a new radioprotector agent in the human needs further studies, particularly clinical trials

Radioprotective effect of grape seed extract against gamma irradiation in mouse bone marrow cells

Introduction: Ionizing radiations produce free radicals which are often responsible for DNA damage or cell death. Grape seed extract (GSE) is a natural compound having an antioxidant that protects DNA, lipids, and proteins from free radical damages. In this study, radioprotective effect of the GSE has been investigated in mouse bone marrow cells using micronucleus test. Materials and Methods: Four groups of mice were investigated in this study: Mice in Group 1 were subjected to injection of distilled water with no irradiation. Mice in Group 2 were exposed to 3 Gy gamma radiation after the injection of distillated water. Mice in Group 3 were injected with 200 mg/kg of the GSE without any irradiation. In another group, mice were exposed to three gray gamma irradiation after the injection of GSE. Animals were killed, and slides were prepared from the bone marrow cells 24 h after irradiation. The slides were stained with May Grunwald–Giemsa method and analyzed microscopically. The frequency of the micronucleated polychromatic erythrocytes (MnPCEs), micronucleated normochromatic erythrocyte (MnNCEs), and polychromatic erythrocyte/polychromatic erythrocyte + normochromatic erythrocyte (PCE/PCE + NCE) ratios was calculated. Results: Injection of GSE significantly decreased the frequency of MnPCEs (P < 0.0001) and MnNCEs (P < 0.05) and increased the ratio of PCE/PCE + NCE (P < 0.0001) compared to the irradiated control group. Discussion and Conclusions: GSE could reduce clastogenic and cytotoxic effects of gamma irradiation in mice bone marrow cells; therefore, it can be concluded that the GSE is a herbal compound with radioprotective effects against gamma irradiation. Free radical scavenging and the antioxidant effects of the GSE probably are responsible mechanisms for the GSE radioprotective effects

Trigonella foenum-graecum seed extract modulates expression of lipid metabolism-related genes in HepG2 cells

Objective: To investigate anti-dyslipidemic effects of hydroalcoholic fenugreek seed extracts, diosgenin, and 4-OH-Ile on HepG2 cell line. Methods: HepG2 cells were treated with hydroalcoholic fenugreek seed extracts, diosgenin, 4-OH-Ile, and orlistat. IC

Chemical Composition and Antimicrobial Efficiency of Swietenia macrophylla Seed Extract on Clinical Wound Pathogens

Microbial wound infection prolonged the hospitalization and increase the cost for wound management. Silver is commonly used as antimicrobial wound dressing. However, it causes several adverse side effects. Hence, this study was aimed to evaluate the antimicrobial efficiency of Swietenia macrophylla seed extract on clinical wound pathogens. Besides, the bioactive constituents of the seed extract were also determined. S. macrophylla seeds were extracted with methanol by maceration method. The seed extract inhibited 5 test bacteria and 1 yeast on disc diffusion assay. The antibacterial activity was broad spectrum, as the extract inhibited both Gram positive and Gram negative bacteria. On kill curve analysis, the antibacterial activity of the seed extract was concentration-dependent, the increase of extract concentration resulted in more reduction of bacterial growth. The extract also caused 99.9% growth reduction of Bacillus subtilis relative to control. A total of 21 compounds were detected in gas chromatography-mass spectrometry analysis. The predominant compounds present in the extract were oleic acid (18.56%) and linoleic acid (17.72%). In conclusion, the methanolic extract of S. macrophylla seeds exhibited significant antimicrobial activity on clinical wound pathogens. Further investigations should be conducted to purify other bioactive compounds from the seeds of S. macrophylla.