In situ effect of a proanthocyanidin mouthrinse on dentin subjected to erosion

Abstract Proanthocyanidin has been shown to be efficient in inhibiting matrix metalloproteinases. Objective The aim of this in situ study was to evaluate the protective effect of Proanthocyanidin-based mouthrinses either with naturally acidic or with a neutral pH applied on dentin subjected to erosion. Methodology Eight volunteers wore one palatal device in two phases (7 days washout) with 16 samples per group (n=8). The groups under study were: First Phase/ G1 - 10% proanthocyanidin mouthrinse (pH 7.0, Experimental group 1 - Purified Grape Seeds Oligomeric Proanthocyanidins), G2 - 10% proanthocyanidin mouthrinse (pH 3.0, Experimental group 2 - Purified Grape Seeds Oligomeric Proanthocyanidins). Second Phase/ G3 - 0.12% chlorhexidine mouthrinse (pH 7.0, Positive control group), G4 - no previous treatment (Negative control group). Each device was subjected to 3 erosive cycles (5 minutes) per day for 5 days. Treatments with different mouthrinses were applied once after the second erosive challenge (5 minutes). Profilometry was used to quantify dentin loss (µm). Results Data were analyzed by repeated measures of ANOVA followed by Fisher's test (p<0.05). G1 (1.17±0.69) and G3 (1.22±0.25) showed significantly lower wear values with no statistical difference between them. G2 (2.99±1.15) and G4 (2.29±1.13) presented higher wear values with no significant differences between them. Conclusion The 10% proanthocyanidin mouthrinse (pH 7.0) could be a good strategy to reduce dentin wear progression.

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.

Chlorhexidine and proanthocyanidin enhance the long-term bond strength of resin-based endodontic sealer

Abstract The aim of this study was to evaluate the effects of proanthocyanidin (PA) and chlorhexidine (CHX) on the bond strength (BS), failure pattern, and resin-dentin interface morphology of the endodontic sealers EndoREZ and AH Plus after 24 h and 6 months of water storage. A total of 120 prepared bovine roots were divided into six groups: AH Plus, CHX+AH Plus, PA+AH Plus, EndoREZ, CHX+EndoREZ, and PA+EndoREZ. Dentin was treated for 1 or 5 min with 2% CHX or 15% PA, respectively. Roots were filled and stored in water for 24 h or 6 months (n = 10). Root slices were subjected to push-out test and scanning electron microscopy (SEM). Data were compared using two-way ANOVA and student's t-test (α = 5%). BS decreased over time for AH Plus and untreated EndoREZ (p < 0.05). At 24 h, AH Plus had higher BS than EndoREZ (p < 0.001), with no differences among treatments for both sealers (p > 0.05). At 6 months, EndoREZ had higher BS values for CHX and PA than control (p < 0.05). AH Plus had higher BS than EndoREZ (p < 0.001), while with CHX or PA, similar BS was observed in both sealers (p > 0.05). Cohesive and mixed failures were observed in all groups. SEM revealed sealer tags in the root dentin. In conclusion, BS decreased with time and AH Plus had higher BS than EndoREZ in untreated dentin; however, CHX or PA enhanced long-term BS of EndoREZ. Overall, dentin treatment affected failure pattern and resin-dentin interface morphology, particularly for EndoREZ.

Effectiveness of plant-derived proanthocyanidins on demineralization on enamel and dentin under artificial cariogenic challenge

Dental caries is considered a disease of high prevalence and a constant problem in public health. Proanthocyanidins (PAs) are substances that have been the target of recent studies aiming to control or treat caries. Objective The aim of this in vitro study was to evaluate the efficacy of a treatment with grape seed extract, under cariogenic challenge, to minimize or even prevent the onset of caries in the enamel and dentin. Material and Methods Blocks of enamel and dentin (6.0x6.0 mm) were obtained from bovine central incisors, polished, and selected by analysis of surface microhardness (SH). The blocks were randomly divided into 3 groups (n=15), according to the following treatments: GC (control), GSE (grape seed extract), GF (fluoride – 1,000 ppm). The blocks were subjected to 6 daily pH cycles for 8 days. Within the daily cycling, the specimens were stored in buffered solution. The blocks were then analyzed for perpendicular and surface hardness and polarized light microscopy. Results The means were subjected to statistical analysis using the ANOVA and Fisher's PLSD tests (p<0.05). For enamel SH, GF showed the highest hardness values. In the dentin, GF was also the one that showed higher hardness values, followed by GSE. Regarding the cross-sectional hardness values, all groups behaved similarly in both the enamel and dentin. The samples that were treated with GSE and fluoride (GF) showed statistically higher values than the control. Conclusion Based on the data obtained in this in vitro study, it is suggested that grape seed extract inhibits demineralization of artificial carious lesions in both the enamel and dentin, but in a different scale in each structure and in a smaller scale when compared to fluoride. .

Stryphnodendron adstringens (Mart. ) Coville (Fabaceae) proanthocyanidins quantitation by RP-HPLC

Stryphnodendron adstringens (Mart.) Coville (barbatimão) is a tree belonging to the Fabaceae family, and it is commonly found in the southeastern Brazilian cerrado. The stem bark of this tree is traditionally used as an anti-inflammatory and antiseptic agent to treat leucorrhea and diarrhea, as well as to promote wound healing (owing to the presence of proanthocyanidins). Proanthocyanidins were obtained from the ethanolic extract of S. adstringens stem bark and assessed by reversed phase-high performance liquid chromatography with an ultraviolet/diode array detector. The identified compounds included gallic acid, catechin, gallocatechin (GC), epigallocatechin, and epigallocatechin gallate (EGCG). The selected markers, GC and EGCG, were simultaneously used for chromatographic validation (linearity range: 30-330 ng, equivalent to 3-33 µg/mL; r>0.998). The method showed precision (intra-day relative standard deviation [RSD]: 1.72% for GC; 1.16% for EGCG; inter-day RSD: 1.74%-2.60% for both markers), accuracy, robustness, and selectivity. The limits of detection and quantitation were 0.29 µg/ml and 0.89 µg/ml for GC, and 0.88 µg/mL and 2.67 µg/mL for EGCG, respectively. In addition, S. obovatum was evaluated and showed an average of 12.2 µg/mL for GC (equivalent to 1.22% w/w) and 14.2 µg/mL for EGCG (equivalent to 1.42% w/w) in the ethanolic extract. The quantitative results were compared to those obtained for S. adstringens, which showed that the markers are present in both species.

Stryphnodendron adstringens (Mart.) Coville (barbatimão) é uma árvore da família Fabaceae encontrada no cerrado do sudeste do Brasil. As cascas do caule das espécies são tradicionalmente utilizadas como agente antiinflamatório e antisséptico para o tratamento de diarreia e leucorreia, bem como para promover a cicatrização de feridas, devido à presença de proantocianidinas. Estas substâncias obtidas a partir do extracto etanólico (EE) de cascas do caule de S. adstringens foram avaliadas por cromatografia líquida de alta eficiência de fase reversa. As substâncias identificadas foram ácido gálico, catequina, galocatequina (GC), epigalocatequina e galato de epigalocatequina (EGCG). Os marcadores selecionados, GC e EGCG, foram utilizados simultaneamente para validação cromatográfica (intervalo de linearidade 30-330 ng, equivalente a 3-33 µg/mL, r>0,998). O método mostrou precisão (intra-dia DPR 1,72% para GC; 1,16% para EGCG; inter-dia DPR 1,74%-2,60% para ambos os marcadores), exatidão, robustez e seletividade. Os limites de detecção e de quantificação de 0,29 µg/mL e 0,89 µg/mL para a GC, e 0,88 ng/mL e 2,67 µg/mL para EGCG, respectivamente. Outra espécie (S. obovatum) avaliada, mostrou valores médios 12,2 µg/mL de GC (equivalente a 1,22% p/p) e 14,2 µg/mL de EGCG (equivalente a 1,42% p/p) encontrados no EE. Os resultados quantitativos comparam-se aos de S. adstringens e mostram que tais marcadores estão presentes em ambas as espécies.

Antioxidant properties of a dry product from Vitis vinifera seeds (Leucoselect) in rat liver endoplasmic reticulum / Propiedades antioxidantes de un producto seco de semillas de Vitis vinifera (Leucoselect) en retículo endoplásmico de hígado de rata

Leucoselect is a commercial dry product obtained from grape seeds and enriched in procyanidins, which display antioxidant activity in virtue to their ability to scavenge oxygen free radicals and to chelate transition metal ions. The hypoxanthine/xanthine oxidase and Cu2+/ascorbate systems are capable of generating reactive oxygen species; the latter system can also promote non-specific binding of copper ions to proteins. Therefore, we assessed the ability of Leucoselect to inhibit oxidative phenomena elicited by both oxidative systems on rat liver microsomes: lipid peroxidation, oxidation of protein thiols, and inhibition of the cytochrome P450 system. The antioxidant activity of Leucoselect was a reflection of its ability to scavenge oxygen free radicals, chelate copper ions, and protect microsomal membranes through direct interaction. These mechanisms were displayed in a dependent manner with the type of biomolecule studied and also with the oxidative system employed, which is an interesting phenomenon to consider when evaluating the antioxidant activity of herbal products.

Leucoselect es un producto comercial seco obtenido de semillas de uva y enriquecido en procianidinas, las cuales presentan actividad antioxidante debido a su capacidad para atrapar radicales libres y quelar metales de transición. Los sistemas hipoxantina/xantina oxidasa y Cu2+/ascorbato generan especies reactivas del oxígeno; este último sistema también promueve la unión inespecífica de iones cobre a proteínas. Por lo tanto, evaluamos la capacidad de Leucoselect para inhibir los fenómenos oxidativos producidos por ambos sistemas oxidantes en microsomas hepáticos de rata: lipoperoxidación, oxidación de tioles proteicos e inhibición de la actividad del sistema citocromo P450. La actividad antioxidante de Leucoselect fue un reflejo de su capacidad de atrapar radicales libres del oxígeno, quelar iones cobre y proteger membranas microsómicas por interacción directa. Dichos mecanismos se manifestaron en forma dependiente del tipo de biomolécula estudiada y del sistema oxidante empleado, fenómeno interesante de considerar al evaluar la actividad antioxidante de preparados herbales.

In vitro studies on inhibitory effect of proanthocyanidins in modulation of neutrophils and macrophages.

The role of proanthocyanidins (PC), a novel flavonoid extracted from grape seeds was studied in vitro in the modulation of neutrophil and macrophage function. We attempted to assess the levels of non-enzymatic and enzymatic mediators in the presence or absence of PC in 4-phorbol-12--myristate-13-acetate (PMA)-stimulated neutrophils isolated from humans and rats, E. coli endotoxin-stimulated macrophages and macrophages isolated from E. coli endotoxin-induced experimental periodontitis in rats. Addition of PC at a concentration of 50 µg/ml effectively blocked the release of reactive oxygen species (ROS) and reactive nitrogen species (RNS) and exhibited a marked inhibition of myeloperoxidase (MPO) and lysosomal enzymes (p<0.001), as compared to PMA-stimulated neutrophils (human and rats) and neutrophils isolated from experimental periodontitis in rats. The levels of ROS, RNS and lysosomal enzymes were found to be elevated (p<0.001) and addition of PC significantly (p<0.001) reduced these levels as compared to those from E. coli endotoxin-stimulatedmacrophages from rats and macrophages isolated from experimental periodontitis in rats (p<0.001). Thus, the study demonstrated that PC decreased the levels of ROS and RNS and also inhibited the MPO and lysosomal enzymes activities in experimental periodontitis in rats. In addition, this study clearly indicated that PC could be developed as an effective antiinflammatory agent.