The Effect of Solutions Containing Extracts of Vochysia tucanorum Mart., Myrcia bellaCambess., Matricaria chamomilla L. and Malva sylvestris L. on Cariogenic Bacterial Species and Enamel Caries Development.

This study evaluated the effect of experimental solutions containing plant extracts on bacterial species and enamel caries prevention. Microcosm biofilm was produced from human saliva mixed with McBain saliva (0.2% sucrose) on bovine enamel for 5 days (3 days under anaerobiosis and 2 days under aerobiosis) at 37°C. From the 2nd day, the following treatments were applied (1 × 60 s/day): Vochysia tucanorum (10 mg/mL); Myrcia bella (5 mg/mL); Matricaria chamomilla (80 mg/mL); Malva sylvestris, fluoride, and xylitol (Malvatricin Plus®); 0.12% chlorhexidine (CHX, PerioGard®); and PBS (negative control). The medium pH was measured. Quantitative polymerase chain reaction was performed for the detection of Streptococcus mutans and Lactobacillus spp. Enamel demineralization was measured by spectral-domain optical coherence tomography. The data were compared by means of the Kruskal-Wallis/Dunn, two-way ANOVA/Bonferroni, and ANOVA/Tukey tests (p < 0.05). The pH decreased after sucrose exposure; only CHX reestablished pH >5.5 by the last day. CHX also eliminated Lactobacillusspp., but the other treatments did not differ significantly from PBS. Malvatricin Plus® and CHX eliminated S. mutans, but the other treatments did not differ from PBS. Similar results were seen concerning the reduction of lesion depth and reflectivity. The experimental natural-extract solutions were ineffective against cariogenic bacteria and in preventing the development of enamel caries.

In vitro effect of curcumin-mediated antimicrobial photodynamic therapy on fibroblasts: viability and cell signaling for apoptosis.

Although it was demonstrated that curcumin-mediated antimicrobial photodynamic therapy (aPDT) is effective for reducing the viability of microbial cells and the vitality of oral biofilms, the cytotoxicity of this therapeutic approach for host cells has not been yet elucidated. Hence, the aim of this study was to evaluate the cytotoxicity and apoptotic effects of curcumin-mediated aPDT on mouse fibroblasts. Cells were treated with 0.6 or 6 µmol.L-1 curcumin combined with 0.075 or 7.5 J.cm-2 LED at 455 nm. Cytotoxicity was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and crystal violet (CV) assays, while quantitative reverse transcriptase-PCR (qRT-PCR) was used to assess the expression of Bax, Bad, Bcl-2, VDAC-1, cytochrome C, and Fas-L genes for apoptosis. The differences between groups were detected by Kruskal-Wallis and post hoc Dunn's tests for MTT and CV assays and by ANOVA and post hoc Tukey test for qRT-PCR (P < 0.05). The effect of 0.6 µmol.L-1 curcumin plus 0.075 J.cm-2 LED (minimum parameter) did not differ statistically from control group; however, the combination of 0.6 µmol.L-1 curcumin plus 7.5 J.cm-2 LED reduced viable cells in 34%, while the combinations of 6 µmol.L-1 curcumin plus 0.075 and 7.5 J.cm-2 LED reduced viable cells in 47% and 99%, respectively. aPDT increased significantly the relative expression of Bax/Bcl-2, cytochrome C, VDAC-1, and Fas-L genes, without influence on the ratio Bad/Bcl-2. Therefore, curcumin-mediated aPDT activated Bcl-2 apoptosis signaling pathways in mouse fibroblasts regarding present conditions, reducing the viability of cells with the increase of curcumin concentrations and light energies.

Photobiomodulation therapy can change actin filaments of 3T3 mouse fibroblast.

The purpose of this study was to investigate the effects that photobiomodulation therapy might produce in cells, in particular, related to their structure. Thus, this paper presents the results of morphological changes in fibroblasts following low-intensity light illumination. Mouse fibroblasts were grown on glass coverslips on either 4 kPa or 16 kPa gels, to mimic normal tissue conditions. Cells were photo-irradiated with laser light at either 625 nm or 808 nm (total energies ranging from 34 to 47 J). Cells were fixed at 5 min, 1 h, or 24 h after photo-irradiation, stained for both actin filaments and the cell nucleus, and imaged by confocal microscopy. A non-light exposed group was also imaged. A detailed analysis of the images demonstrated that the total polymerized actin and number of actin filaments decrease, while the nucleus area increases in treated cells shortly after photo-irradiation, regardless of substrate and wavelength. This experiment indicated that photobiomodulation therapy could change the morphological properties of cells and affect their cytoskeleton. Further investigations are required to determine the specific mechanisms involved and how this phenomenon is related to the photobiomodulation therapy mechanisms of action.

Effect of commercial herbal toothpastes and mouth rinses on the prevention of enamel demineralization using a microcosm biofilm model.

This work evaluated the effects of commercial toothpastes and mouth rinses containing natural/herbal agents on biofilm viability, extracellular polysaccharide (EPS) production and on enamel demineralization in vitro. Microcosm biofilm was produced on bovine enamel for 5 days and treated daily with: Orgânico natural® (toothpaste/mouth rinse), Boni Natural Menta & Malaleuca® (toothpaste/mouth rinse), Propolis & Myrrh® (toothpaste), Colgate Total 12 Clean Mint® (toothpaste, positive control), Malvatricin® Plus (mouth rinse), PerioGard® (mouth rinse, positive control) or PBS (negative control). Tom's Propolis & Myrrh® and Colgate Total 12® toothpastes and Malvatricin® Plus and PerioGard® mouth rinses significantly reduced biofilm viability (p < 0.05). Only PerioGard® had significant effects on biofilm thickness and EPS. Despite the indication that Tom's Propolis & Myrrh® significantly reduced lesion depth, only Colgate Total 12® significantly reduced mineral loss. Malvatricin® Plus significantly reduced mineral loss and lesion depth, as did PerioGard®. Some herbal products, Malvatricin® Plus and Tom's Propolis & Myrrh®, showed anticaries effects.

Effect of hydroalcoholic extract of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on the viability and activity of microcosm biofilm and on enamel demineralization.

OBJECTIVES: The aim of this study was to assess the effect of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves hydroalcoholic extracts on viability and metabolism of a microcosm biofilm and on enamel demineralization prevention. METHODOLOGY: Microcosm biofilm was produced on bovine enamel using inoculum from pooled human saliva mixed with McBain saliva, under 0.2% sucrose exposure, for 14 days. The biofilm was daily-treated with the extracts for 1 min. At the end, it was analyzed with respect to viability by fluorescence, CFU counting and extracellular polysaccharides (phenol-sulphuric acid colorimetric assay) and lactic acid (enzymatic assay) production. The demineralization was measured by TMR. The data were compared using ANOVA or Kruskal-Wallis (p<0.05). RESULTS: M. urundeuva All. at 100, 10 and 0.1 µg/mL and Q. grandiflora Mart. at 100 and 0.1 µg/mL reduced biofilm viability similarly to positive control (chlorhexidine) and significantly more than the negative-vehicle control (35% ethanol). M. urundeuva at 1000, 100 and 0.1 µg/mL were able to reduce both lactobacilli and mutans streptococci CFU counting, while Q. grandiflora (1000 and 1.0 µg/mL) significantly reduced mutans streptococci CFU counting. On the other hand, the natural extracts were unable to significantly reduce extracellular polysaccharides and lactic acid productions neither the development of enamel carious lesions. CONCLUSIONS: The extracts showed antimicrobial properties on microcosm biofilm, however, they had no effect on biofilm metabolism and caries protection.

Hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on Streptococcus mutans biofilm and tooth demineralization.

OBJECTIVES: This study evaluated the effect of the hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves (alone or combined) on the viability of Streptococcus mutans biofilm and on the prevention of enamel demineralization. METHODS: Strain of S. mutans (ATCC 21175) was reactivated in BHI broth. Minimum inhibitory concentration, minimum bactericidal concentration, minimum inhibition biofilm concentration and minimum eradication biofilm concentration were determined in order to choose the concentrations to be tested under biofilm model. S. mutans biofilm (5 × 105 CFU/ml) was produced on bovine enamel, using McBain saliva under 0.2% sucrose exposure, for 3 days. The biofilm was daily treated with the extracts for 1 min. The biofilm viability was tested by fluorescence and the enamel demineralization was measured using TMR. RESULTS: Myracrodruon urundeuva All. (Isolated or combined) at the concentrationsc ≥0.625 mg/ml was able to reduce bacteria viability, while Qualea Grandflora Mart. alone had antimicrobial effect at 5 mg/ml only (p < 0.05). On the other hand, none of the extracts were able to reduce enamel demineralization. CONCLUSIONS: The hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves (isolated or combined) have antimicrobial action; however, they do not prevent enamel caries under S. mutans biofilm model.

In situ effect of a commercial CPP-ACP chewing gum on the human enamel initial erosion.

OBJECTIVE: This study evaluated the in situ rehardening effect of a commercial chewing gum containing casein phosphopeptide - amorphous calcium phosphate (CPP-ACP) on initial erosion lesions. METHODS: Seventy-two human enamel blocks, after selection (initial surface hardness - SHi) and in vitro short-term acidic exposure (cola drink for 3 min - SHd) were randomly assigned to three groups. The factors under study were treatment (3 levels: GI chewing gum with CPP-ACP, GII chewing gum without CPP-ACP and GIII control group without gum) and intraoral period (2 levels: 2 and 24h). Twelve volunteers wore intraoral palatal devices for 24h in 3 crossover phases. On each phase, after 2h the surface hardness was assessed (SHf1) and the blocks were reinserted and the devices were used for additional 22 h (SHf2). In phases of GI and GII volunteers chewed the respective gum during 30 min, for 4 times with an interval of 4h. Percentage of surface hardness recovery (%SHR) was calculated after 2 and 24 h. The data were analysed by repeated measures ANOVA and Tukey's test. RESULTS: Chewing gum with CPP-ACP (2h=50.0%<24h=95.9%) showed higher hardness recovery than chewing gum without CPP-ACP (2h=30.0%<24 h=71.1%) and control (2 h=15.7%<24 h=40.9%) (p<0.05). CONCLUSIONS: The results suggest that saliva increased hardness of softened enamel after the use of conventional chewing gum (GII) and this effect was enhanced by the prolonged intraoral period (24 h) and by the use of CPP-ACP chewing gum (GI). CLINICAL SIGNIFICANCE: Since chewing gum is an alternative to enhance salivary defenses after erosive challenges, CPP-ACP chewing gum might be a supplementary strategy to potentiate the mineral precipitation of initial erosion lesions.

Anacardic acid from brazilian cashew nut trees reduces dentine erosion.

The aim of this study was to analyze the effect of solutions containing saturated anacardic acid (AA) on dentine erosion in vitro. AA was chemically isolated from natural cashew nutshell liquid obtained by continuous extraction in a Soxhlet extractor and was fully saturated by catalytic hydrogenation. Matrix metalloproteinase 2 (MMP-2) activity, when exposed to buffers containing 100 µmol/l AA, was analyzed using zymography. Bovine root samples were subjected to erosive demineralization (Sprite Zero™, 4 × 90 s/day) and remineralization with artificial saliva between the erosive cycles for 5 days. The samples were treated as follows, after the first and the last acid exposure (1 min; n = 12/group): (1) 100 µmol/l epigallocatechin-3-gallate (EGCG) (positive control); (2) 0.05% NaF; (3) 100 µmol/l saturated AA; (4) saturated AA and EGCG; (5) saturated AA, EGCG and NaF; (6) untreated (negative control). Dentine erosion was measured using a contact profilometer. Two dentine samples from each group were analyzed using scanning electron microscopy. Saturated AA reduced the activity of MMP-2. ANOVA and Tukey's test revealed that all treatments significantly reduced dentine loss compared to the negative control (6.03 ± 0.98 µm). Solutions containing saturated AA (1.97 ± 1.02 µm) showed the greatest reduction in dentine erosion compared to the NaF (3.93 ± 1.54 µm) and EGCG (3.79 ± 0.83 µm) solutions. Therefore, it may be concluded that AA significantly reduces dentine erosion in vitro, possibly by acting as an MMP-2 inhibitor.

Low-level laser therapy prevents prodromal signal complications on saphenectomy post myocardial revascularization.

BACKGROUND AND OBJECTIVE: One of the most frequent treatments for ischemic heart disease is myocardial revascularization, often applying the saphenous vein as a coronary graft. However, postoperative complications may occur, such as saphenous dehiscence. According to the literature, low-level laser therapy (LLLT) has been used in the treatment of several inflammatory processes in patients. Recently, its uses have expanded to include LLLT preventive therapy and postoperative treatment. Despite our department's successful application of LLLT in the treatment of saphenectomy incisions, many colleagues are still uncertain as to laser therapy's benefits. Therefore, the study's purpose was to evaluate tissue repair of prodromal surgical incisions after the administration of LLLT. MATERIALS AND METHODS: The pilot study included 14 patients, divided into two groups. Both groups of patients received the traditional treatment; additionally, the Laser Group (n = 7) received diode laser treatment (λ = 780 nm, fluence = 19 J/cm(2), pulse = 25 mW, time = 30 sec, energy = 0.75 J, irradiance = 625 mW/cm(2), beam spot size 0.04 cm(2)), which was applied on the edges of the saphenectomy incision. The Control Group (n = 7) received conventional treatment exclusively. RESULTS: In the Laser Group: all seven patients showed significant improvement, whereas the Control Group had twice as many complications, including critical rates of incisional dehiscence. CONCLUSIONS: LLLT was valuable in preventing prodromal complications in saphenectomy post myocardial revascularization.

Microhardness and chemical analysis of high-viscous glass-ionomer cement after 10 years of clinical service as ART restorations.

OBJECTIVES: The aim of this study was to evaluate the Knoop microhardness (KHN) and chemical composition of high-viscous glass-ionomer cement (HVGIC) after 10 years of clinical service. METHODS: Six HVGIC samples were cut from 10-year ART restorations. The sections were embedded in acrylic moulds with their longitudinal profile exposed. KHN was determined by performing three sequences of five indentations at 10, 30, 50, 70 and 90 µm of HVGIC outer surface. For the control group (n=6), HVGIC specimens were stored in distilled water for 24 months. Hardness measurements were taken at days 7, 30, 60, 120, 180, 360, and 720. For chemical analysis using SEM-EDX, 10-year and control specimens were dehydrated and coated with carbon. Data were analysed using T-test and ANOVA/Tukey's test (p<0.05). RESULTS: A significant KHN increase was observed in the control group up to the 180-day period. From this point the values stabilized and no more significant differences were found between the 10-year and the control KHN values. No statistical differences were observed amongst the KHN from inner distances compared to the outer surface of the 10-year HVGIC specimens. In one 10-year specimen, SEM-images identified the transformation of HVGIC in an altered layer with no glass filler particles detectable, and raised Ca, K and P contents. CONCLUSIONS: KHN values of ten-year HVGIC specimens were similar to the control group values at 180-day storage period. Except for one 10-year specimen in that an altered layer could be seen, chemical composition was similar amongst the depths evaluated.

The erosive potential of 1% citric acid supplemented by different minerals: an in vitro study.

PURPOSE: The objective of the present in vitro study was to evaluate the effect of different minerals in combination with 1% citric acid on dental erosion. MATERIALS AND METHODS: Ninety enamel samples were randomly allocated to nine groups (G1: pure 1% citric acid solution [control]; G2: with 1 mM Ca; G3: with 0.047 mM F; G4: with 1 mM Fe; G5: with 1 mM P; G6: with 1 mM Ca and 0.047 mM F; G7: with 1 mM Ca and 1 mM P; G8: with 1 mM Fe and 0.047 mM F; G9: with 1 mM Ca, 1 mM P, 0.047 mM F and 1.0 mM Fe). The samples were subjected to six pH cycles, each consisting of immersion in pure or modified 1% citric acid (1 min) followed by storage in artificial saliva (59 min). Enamel wear was assessed using profilometry. RESULTS: Data were analysed using analysis of variance and Tukey test (P < 0.05). Enamel loss (mean + or - SD) amounted to between 0.87 + or - 0.30 and 1.74 + or - 0.74 microm but did not significantly differ among the groups. CONCLUSIONS: The modification of 1% citric acid with different minerals did not have a protective effect on enamel erosion.

Protective effect of green tea on dentin erosion and abrasion.

OBJECTIVE: This in situ study evaluated the protective effect of green tea on dentin erosion (ERO) and erosion-abrasion (ABR). MATERIAL AND METHODS: Ten volunteers wore intraoral palatal appliances with bovine dentin specimens subjected to ERO or ERO + toothbrushing abrasion performed immediately (ERO+I-ABR) or 30 min after erosion (ERO+30-min-ABR). During 2 experimental 5-day crossover phases, the volunteers rinsed with green tea or water (control, 1 min) between each erosive (5 min, cola drink) and abrasive challenge (30 s, toothbrushing), 4x/day. Dentin wear was measured by profilometry. RESULTS: The green tea reduced the dentin wear significantly for all conditions compared to control. ERO+I-ABR led to significantly higher wear than ERO, but it was not significantly different from ERO+30-min-ABR. ERO+30-min-ABR provoked significant higher wear than ERO, only for the placebo treatment. CONCLUSIONS: From the results of the present study, it may be concluded that green tea reduces the dentin wear under erosive/abrasive conditions.

Protective effect of green tea on dentin erosion and abrasion

OBJECTIVE: This in situ study evaluated the protective effect of green tea on dentin erosion (ERO) and erosion-abrasion (ABR). MATERIAL AND METHODS: Ten volunteers wore intraoral palatal appliances with bovine dentin specimens subjected to ERO or ERO + toothbrushing abrasion performed immediately (ERO+I-ABR) or 30 min after erosion (ERO+30-min-ABR). During 2 experimental 5-day crossover phases, the volunteers rinsed with green tea or water (control, 1 min) between each erosive (5 min, cola drink) and abrasive challenge (30 s, toothbrushing), 4x/day. Dentin wear was measured by profilometry. RESULTS: The green tea reduced the dentin wear significantly for all conditions compared to control. ERO+I-ABR led to significantly higher wear than ERO, but it was not significantly different from ERO+30-min-ABR. ERO+30-min-ABR provoked significant higher wear than ERO, only for the placebo treatment. CONCLUSIONS: From the results of the present study, it may be concluded that green tea reduces the dentin wear under erosive/abrasive conditions.

Chlorhexidine and green tea extract reduce dentin erosion and abrasion in situ.

OBJECTIVES: This in situ/ex vivo study aimed to analyse the impact of possible MMP-inhibitors (chlorhexidine and green tea extract) on dentin wear induced by erosion or erosion plus abrasion. METHODS: Twelve volunteers took part in this cross-over and double-blind study performed in 4 phases of each 5 days. Bovine dentin samples were worn in palatal appliances and subjected to extraoral erosion (4 times/day, Coca-Cola, 5 min) or erosion plus abrasion (2 times/day, fluoride-free toothpaste and electrical toothbrush, 15s/sample). Immediately after each erosion, the appliances were reinserted in the mouth and the oral cavity was rinsed for 60s with: 250 ppm F solution (SnF(2)/AmF, pH 4.5, Meridol-Gaba, Switzerland), 0.12% chlorhexidine digluconate (0.06% chlorhexidine, pH 6.0, Periogard-Colgate, Brazil), 0.61% green tea extract solution (OM24, 100% Camellia Sinensis leaf extract, catechin concentration: 30+/-3%, pH 7.0, Omnimedica, Switzerland) or deionized water (pH 6.0, control). Dentin loss was assessed by profilometry (microm). The data were analysed by two-way repeated measures ANOVA and Bonferroni post hoc test. RESULTS: There was a significant difference between the conditions (EroxEro+Abr, p<0.001) and among the solutions (p<0.001). All solutions (F: 1.42+/-0.34; 1.73+/-0.50, chlorhexidine: 1.15+/-0.26; 1.59+/-0.32, green tea: 1.06+/-0.30; 1.54+/-0.55) significantly reduced the dentin wear when compared to control (2.00+/-0.55; 2.41+/-0.83) for both conditions. There were not significant differences among green tea extract, chlorhexidine and F solutions. CONCLUSIONS: Thus, the possible MMP-inhibitors tested in this study seem to be a promising preventive measure to reduce dentin erosion-abrasion, but their mechanism of action needs to be investigated in further studies.

TiF(4) and NaF at pH 1.2 but not at pH 3.5 are able to reduce dentin erosion.

OBJECTIVE: This study aimed to analyse and compare the protective effect of buffered (pH 3.5) and native (pH 1.2) TiF(4) in comparison to NaF solutions of same pH on dentin erosion. DESIGN: Bovine samples were pretreated with 1.50% TiF(4) or 2.02% NaF (both 0.48M F) solutions, each with a pH of 1.2 and 3.5. The control group received no fluoride pretreatment. Ten samples in each group were eroded with HCl (pH 2.6) for 10x60s. Erosion was analysed by determination of calcium release into the acid. Additionally, the surface and the elemental surface composition were examined by scanning electron microscopy (two samples in each group) and X-ray energy-dispersive spectroscopy in fluoridated but not eroded samples (six samples in each group). Cumulative calcium release (nmol/mm(2)) was statistically analysed by repeated measures ANOVA and one-way ANOVA at t=10min. RESULTS: TiF(4) and NaF at pH 1.2 decreased calcium release significantly, while TiF(4) and NaF at pH 3.5 were not effective. Samples treated with TiF(4) at pH 1.2 showed a significant increase of Ti, while NaF pretreatment increased F concentration significantly. TiF(4) at pH 1.2 led to the formation of globular precipitates occluding dentinal tubules, which could not be observed on samples treated with TiF(4) at pH 3.5. NaF at pH 1.2 but not at pH 3.5 induced the formation of surface precipitates covering dentinal tubules. CONCLUSION: Dentin erosion can be significantly reduced by TiF(4) and NaF at pH 1.2, but not at pH 3.5.

Light cola drink is less erosive than the regular one: an in situ/ex vivo study.

OBJECTIVE: This in situ/ex vivo study assessed the erosive potential of a light cola drink when compared to a regular one. METHODS: During 2 experimental 14-days crossover phases, eight volunteers wore palatal devices with 2 human enamel blocks. The groups under study were: group light, erosive challenge with light cola drink and group regular, erosive challenge with regular cola drink. During 14 days, erosive challenges were performed extraorally 3X/day. In each challenge, the device was immersed in 150ml of light cola (group light) or regular cola (group regular) for 5min. Erosion was analysed by surface profilometry (microm) and surface microhardness change (%SMH). The data were statistically analyzed using paired t test (p<0.05). RESULTS: Group light (0.6+/-0.2microm) showed significantly lesser wear than group regular (3.1+/-1.0microm). There was no significant difference between the groups for the %SMH (group light -63.9+/-13.9 and group regular -78.5+/-12.7). CONCLUSIONS: The data suggest that the light cola drink is less erosive than the regular one.