Effect of sodium hexametaphosphate and quercetin, associated or not with fluoride, on dentin erosion in vitro.

OBJECTIVE: to investigate the ability of solutions containing sodium hexametaphosphate, fluoride and quercetin, alone or in association, to prevent dentin erosion and to inhibit matrix metalloproteinases -2 and -9 activity using in vitro protocols. DESIGN: Root dentin blocks (n = 96) were prepared and divided into 8 experimental groups (n = 12/group), according to the solutions to be tested: Placebo; 0.24% sodium fluoride (F); 1.0% sodium hexametaphosphate (HMP); 0.03% quercetin (QC); F+HMP; F+QC; HMP+QC; and F+HMP+QC. Erosive challenges were performed 4×/day for 5 days. Specimens were treated with the respective solutions for one minute, twice a day. Next, dentin loss (profilometry) and integrated hardness area in depth (KHN × µm) were determined. The antiproteolytic potential was assessed by gelatin zymography. Dentin erosion results (log10-transformed) were submitted to one-way ANOVA, followed by Tukey's test. Integrated hardness area in depth data (raw) were submitted to two-way, repeated-measures ANOVA, followed by Holm-Sidak's test (p<0.05). RESULTS: Dentin erosion was significantly lower for F+HMP+QC than for all other treatments. At the shallowest depths (5-30 µm), blocks treated with F+HMP+QC had the highest integrated hardness area in depth values. All treatments completely inhibited matrix metalloproteinases-2 activity, except for the group QC (77% inhibition). For matrix metalloproteinases-9, all HMP-containing solutions or F+QC promoted total antiproteolytic activity. CONCLUSION: The association of fluoride, sodium hexametaphosphate, and quercetin must be considered a valuable strategy for novel product formulation for home and professional use, considering its superior protective effects against dentin erosion and its antiproteolytic potential.

Fluoride and trimetaphosphate association as a novel approach for remineralization and antiproteolytic activity in dentin tissue.

OBJECTIVE: The study evaluated the effect of solutions containing fluoride (F) and/or sodium trimetaphosphate (TMP) and F/TMP on the inhibition of MMP-2 and MMP-9, and on dentin remineralization in vitro. DESIGN: Bovine root dentin blocks were prepared, and caries-like lesions were induced in two thirds of the surface. Blocks were then randomly divided into 13 groups/solutions (n = 10): Placebo; 0.3 %, 1 % and 3 % NaOH-hydrolyzed TMP; 0.3 %, 1 % and 3 % TMP; 250, 500 and 1100 ppm F; 250 ppm F + 0.3 % TMP; 500 ppm F + 1 % TMP and 1100 ppm F + 3 % TMP. One third of each specimen was treated with the respective solutions in pH-cycling. The mineral concentration (gHAp × cm-3 × µm) was determined by computed X-ray microtomography, and data submitted to ANOVA and Student-Newman-Keuls' test (p < 0.05). The ability of the solutions to inhibit MMP-2 and MMP-9 activity was assessed by zymography. RESULTS: F/TMP association led to less mineral loss in the deeper region of the lesion and reduced the depth of lesions when compared to its counterpart without TMP (p < 0.001). 3 % TMP (hydrolyzed or not), 500 ppm F and 1100 ppm F completely inhibited MMP-2 activity, while for MMP-9 such effects were only achieved by treatment with 1100 ppm F + 3 % TMP. CONCLUSION: Treatment with 1100 ppm F + 3 % TMP fully inhibits the gelatinolytic activity of MMPs-2 and - 9 and shows greater remineralizing capacity in artificial caries lesions in dentin. However, hydrolyzing TMP does not improve its anti-proteolytic activity and its remineralizing capacity.

Efeito de soluções contendo hexametafosfato de sódio e quercetina, associados ou não ao fluoreto, sobre o desgaste erosivo dentinário in vitro / Effect of sodium hexametaphosphate and quercetin, associated or not with fluoride, on dentin erosion in vitro

O objetivo do presente estudo foi investigar o efeito de soluções contendo fluoreto (F), hexametafosfato de sódio (HMP) e quercetina (QC), sozinhos ou em associação, sobre a erosão dentinária e sobre a inibição de metaloproteinases da matriz (MMPs) -2 e -9, em protocolos in vitro. Blocos de dentina radicular bovina (4 × 4 × 2 mm; n = 96), selecionados por dureza superficial, foram aleatoriamente divididos em 8 grupos experimentais (n = 12/grupo) e tratados 2×/dia (um minuto) com as seguintes soluções: (1) água deionizada (controle negativo); (2) 1100 ppm F ("F"); (3) 1,0% HMP ("HMP"); (4) 0,03% QC ("QC"); (5) F+HMP; (6) F+QC; (7) HMP+QC; e (8) F+HMP+QC. Os blocos foram submetidos a desafios erosivos 4×/dia, por 5 dias (exposição dinâmica a ácido cítrico 50 mmol.l-1 , pH 3,2, 90 s). Em seguida, foram analisados quanto à perda dentinária (perfilometria) e à perda de dureza integrada em profundidade (área sob a curva, ∆KHN). O potencial antiproteolítico das soluções contendo F, HMP e/ou QC foi analisado por zimografia. Os dados de perda dentinária (log10) foram submetidos a ANOVA um critério, seguido do teste de Tukey. Os resultados de ∆KHN (dados brutos) foram submetidos a ANOVA dois critérios, medidas repetidas, seguido do teste HolmSidak (p< 0,05). O menor desgaste erosivo foi observado no grupo F+HMP+QC. Nas menores profundidades (5-30 µm), os blocos tratados com a solução contendo F+HMP+QC apresentaram os maiores valores de ∆KHN. A análise zimográfica mostrou que todos os tratamentos promoveram atividade antiproteolítica total da MMP-2, com exceção da QC administrada sozinha (inibição de 77%). Para MMP-9, todas as soluções contendo HMP e a associação de F+QC apresentaram atividade antiproteolítica total. Conclui-se que a adição de HMP e QC a soluções contendo F levou a uma maior proteção contra a erosão dentinária, tanto em superfície (perda dentinária) quanto em relação ao conteúdo mineral do tecido remanescente (∆KHN), além de promover uma completa inibição da atividade de MMPs -2 e -9 in vitro(AU)

The aim of the present study was to investigate the effect of solutions containing fluoride (F), sodium hexametaphosphate (HMP) and quercetin (QC), alone or in association, on dentin erosion and on the inhibition of matrix metalloproteinases (MMPs) - 2 and -9, using in vitro protocols. Bovine root dentin blocks (4 × 4 × 2 mm; n = 96), selected by surface hardness, were randomly divided into 8 experimental groups (n = 12/group) and treated 2×/day (one minute) with the following solutions: (1) deionized water (negative control); (2) 1100 ppm F ("F"); (3) 1.0% HMP ("HMP"); (4) 0.03% QC ("QC"); (5) F+HMP; (6) F+QC; (7) HMP+QC; and (8) F+HMP+QC. Blocks were submitted to erosive challenges 4×/day for 5 days (dynamic exposure to 50 mmol.l-1 citric acid, pH 3.2, 90 s). They were then analyzed for dentin loss (profilometry) and integrated hardness loss in depth (area under the curve, ∆KHN). The antiproteolytic potential of solutions containing F, HMP and/or QC was analyzed by zymography. Dentin loss results (log10 transformed) were submitted to one-way ANOVA, followed by Tukey's test. ∆KHN data (raw) were submitted to two-way, repeated-measures ANOVA, followed by the Holm-Sidak test (p< 0.05). The lowest dentin erosive wear was promoted by F+HMP+QC. At the lowest depths (5-30 µm), blocks treated with F+HMP+QC showed the highest values of ∆KHN. Zymography analysis showed that all treatments completely inhibited MMP-2 activity, except for QC administered alone (77% inhibition). For MMP-9, all the solutions containing HMP or the association of F+QC promoted total antiproteolytic activity. It was concluded that the addition of HMP and QC to F solutions led to greater protection against dentin erosion, both at the surface (dentin loss) and in relation to the mineral content of the remaining tissue (∆KHN), in addition to promoting a complete inhibition of MMPs -2 and -9 activity in vitro(AU)

Effect of red wine or its polyphenols on induced apical periodontitis in rats.

AIM: To evaluate the effect of red wine consumption or its polyphenols on the inflammation/resorption processes associated with apical periodontitis in rats. METHODOLOGY: Thirty-two three-month-old Wistar rats had apical periodontitis induced in four first molars and were then arranged into four groups: control (C)-rats with apical periodontitis; wine (W)-rats with apical periodontitis receiving 4.28 ml/kg of red wine; resveratrol+quercetin (R+Q)-rats with apical periodontitis receiving 4.28 ml/kg of a solution containing 1.00 mg/L of quercetin and 0.86 mg/L of resveratrol and alcohol (ALC)-rats with apical periodontitis receiving the alcoholic dose contained in the wine. The oral gavage treatments were administered daily, from day 0 to day 45. On the 15th day, apical periodontitis was induced, and on the 45th day, the animals were euthanized. Histological, immunohistochemical (RANKL, OPG, TRAP, IL-10, TNF-⍺ and IL-1ß) and micro-computed tomography for bone resorption analysis were performed in the jaws. The Kruskal-Wallis with Dunn's test was performed for nonparametric data, and the anova with Tukey's test for parametric data, p < .05. RESULTS: The median score of the inflammatory process was significantly lower in the R+Q group (1) compared to the C (2) (p = .0305) and ALC (3) (p = .0003) groups, and not different from the W (1.5) group. The immunolabeling for OPG was significantly higher in the R+Q group (p = .0054) compared to all groups; the same was observed for IL-10 (p = .0185), different from groups C and ALC. The R+Q group had the lowest TRAP cell count (p < .0001), followed by the W group, both inferior to C and ALC groups. The lowest bone resorption value was in the R+Q group (0.50mm3  ± 0.21mm3 ), significantly lower (p = .0292) than the C group (0.88mm3  ± 0.10mm3 ). The W group (0.60 mm3  ± 0.25 mm3 ) and R+Q group had less bone resorption compared to the ALC group (0.97 mm3  ± 0.22 mm3 ), p = .0297 and p = .0042, respectively. CONCLUSION: Red wine administration to rats for 15 days before induction of apical periodontitis decreased inflammation, TRAP marking and periapical bone resorption compared to alcohol. Resveratrol-quercetin administration reduced the inflammatory process in apical periodontitis, periapical bone resorption, and altered the OPG, IL-10 and TRAP expression compared to C and ALC groups.

Excessive caffeine intake increases bone resorption associated with periapical periodontitis in rats.

AIM: To evaluate the effect of excessive caffeine intake on the inflammation/resorption processes associated with periapical periodontitis (PP) in rats. METHODOLOGY: Sixteen Wistar rats were used. Periapical periodontitis was induced in the four first molars in each animal. The animals were arranged into two groups: control (C)-rats with periapical periodontitis; and caffeine (CAF)-rats with periapical periodontitis under caffeine administration protocol. The CAF animals received 10 mg/100 g of body weight/day of caffeine via gavage starting fifteen days before PP induction and continuing for thirty more days until euthanasia. On the 30th day, the animals were euthanized and the jaws removed for microcomputed tomography, histological and immunohistochemical analysis for RANKL, OPG, TRAP, IL-10, TNF-⍺ and IL-1ß. The Mann-Whitney test was performed for nonparametric data, and Student's t test was performed for parametric data, using p < .05. RESULTS: There was no significant difference in the weight change between the groups. The median score of the inflammatory process was significantly greater in the CAF group (3) compared with the C group (2), p = .0256. Bone resorption was greater in the group consuming caffeine (1.08 ± 0.15 mm3 ) compared with the C group (0.88 ± 0.10 mm3 ), p = .0346. The immunolabelling for RANKL, TRAP and IL-1ß was significantly higher in the CAF group when compared to the control, p < .05. No differences were found for the OPG, IL-10 and TNF-⍺ immunolabelling. CONCLUSION: Excessive caffeine exposure via gavage in rats was able to exacerbate the volume of periapical bone destruction, and the inflammatory pattern deriving from periapical periodontitis altering the expression of RANKL, IL-1ß and TRAP.

Análise in vitro da capacidade de soluções contendo fluoreto e/ou hexametafosfato na remineralização da dentina / In vitro analysis of the capacity of solutions containing fluoride and/or hexametaphosphate on the remineralization of dentin

O objetivo do presente estudo foi avaliar a capacidade de soluções contendo HMP e F, sozinhos ou em associação, em induzir a remineralização dentinária em um protocolo in vitro. Blocos de dentina radicular bovina (4 × 6 cm, n = 100) foram preparados e submetidos à indução de lesões de cárie artificiais em dois terços da superfície; cada bloco foi utilizado como seu próprio controle. Em seguida, os blocos foram divididos em 10 grupos experimentais (n=10/grupo), de acordo com as soluções a serem testadas: (1) Placebo (sem F ou HMP); (2) 0,5% HMP; (3) 0,75% HMP; (4) 1% HMP; (5) 250 ppm F; (6) 500 ppm F; (7) 1100 ppm F; (8) 250 ppm F + 0,5% HMP; (9) 500 ppm F + 0,75% HMP e (10) 1100 ppm F + 1% HMP. Os blocos foram tratados por um minuto, duas vezes ao dia com as respectivas soluções, e submetidos a uma ciclagem de pH durante 7 dias. Em seguida, foram determinadas a porcentagem de recuperação da dureza de superfície (%RDS) e a área integrada da lesão de subsuperfície (ΔKHN). Os dados foram submetidos a ANOVA e teste de Fisher LSD (p<0.05). Uma relação dose-reposta foi observada entre as concentrações de F nas soluções sem HMP e as variáveis %RDS e ΔKHN; quanto às soluções contendo apenas HMP, uma relação dose-resposta foi observada somente para ΔKHN. Em relação à %RDS, os grupos placebo e 0,5% HMP, e os grupos 0,75% e 1% HMP não apresentaram diferenças significativas entre si. Quando associado ao F, o HMP aumentou a capacidade de remineralização da superfície e subsuperfície dentinária, visto que os grupos contendo F + HMP apresentaram resultados significativamente melhores em relação aos grupos contendo F sozinho. Em acréscimo, a solução contendo 250 ppm F + 0,5% HMP promoveu um efeito remineralizador semelhante à solução contendo 500 ppm F. Já em relação à ∆KHN, diferenças estatísticas foram observadas entre todos os grupos na área tratada, sem diferenças significativas quanto às áreas controle e desmineralizada. Os resultados permitem concluir que a adição de HMP às soluções fluoretadas potencializou o efeito destas sobre a remineralização das lesões artificiais de cárie em dentina, tanto na superfície quanto em profundidade(AU)

The present study aimed to investigate the ability of solutions containing HMP and F, alone or in association, in promoting dentin remineralization in an in vitro protocol. Bovine root dentin blocks (4 × 6 cm, n = 100) were prepared, and caries-like lesions were induced in two thirds of the surface; each block served as its own control. Then, blocks were divided into 10 experimental groups (n = 10 / group), according to the solutions to be tested: (1) Placebo (without F or HMP); (2) 0.5% HMP; (3) 0.75% HMP; (4) 1% HMP; (5) 250 ppm F; (6) 500 ppm F; (7) 1100 ppm F; (8) 250 ppm F + 0.5% HMP; (9) 500 ppm F + 0.75% HMP and (10) 1100 ppm F + 1% HMP. Specimens were treated for one minute, twice a day with the respective solutions, and subjected to a pH-cycling regime for 7 days. Next, the percentage of the superficial hardness recovery (%SHR) and integrated loss of subsurface hardness (ΔKHN) were determined. Data were submitted to ANOVA and Fisher LSD's test (p<0.05). A dose-response relationship was observed between F concentrations in solutions without HMP and the variables %SHR and ΔKHN; as for the solutions containing HMP alone, a dose-response relationship was only observed for ΔKHN. Regarding %SHR, no significant differences were observed Placebo and 0.5% HMP groups, nor between 0.75% and 1% HMP groups. When associated with F, HMP was shown to increase the remineralizing capacity of the solutions both at the surface and the subsurface of dentin specimens, since the groups containing F + HMP showed significantly superior results compared to groups containing F alone. In addition, the solution containing 250 ppm F + 0.5% HMP promoted a remineralizing effect similar to that containing 500 ppm F. Regarding ∆KHN, significant differences were observed among all groups in the treated area, while no significant differences were observed among the groups in the control and demineralized areas. The results allowed the conclusion that the addition of HMP to fluoridated solutions significantly enhanced their remineralizing potential on dentin artificial caries lesions, both at the surface and in depth(AU)

Effects of different alcohol concentrations on the development of apical periodontitis in rats.

AIM: To investigate the effect of different alcohol concentrations on the development of apical periodontitis (AP) in rats. METHODS: Forty Wistar rats were arranged into five groups: (C) - control rats receiving sterile water as the only liquid; (G5) - animals receiving an alcohol solution at 5%, (G10) - alcohol solution at 10%, (G15) - alcohol solution at 15%, and (G20) - alcohol solution at 20%. The alcoholic solution or water was given to the groups as the sole source of hydration throughout the 30 days of the experiment. AP was induced in the mandibular molars on the first day. In the end, the animals were euthanized for histopathological and IL-1b, RANKL, OPG, and TRAP analyses. The Kruskal-Wallis test was used for nonparametric data, and ANOVA followed by the Tukey test were performed for parametric data, p < 0.05. RESULTS: G15 and G20 had a greater chronic inflammatory infiltrate (Score 3) and AP size bigger (1.59 ±â€¯0.41 and 1.83 ±â€¯0.38, respectively) than the C, G5 and G10 (p < 0.05). No significant difference was found in the IL-1b analyses. The G15 and G20 showed the highest immunolabeling pattern for RANKL and the lowest for OPG. The G20 had greater TRAP cells per mm (4.70 ±â€¯0.99) compared to the C, G5, and G10 (p < 0.05). Furthermore, G15 presented 3.92 ±â€¯0.64 TRAP cells/mm, higher than C (p < 0.05). CONCLUSIONS: G5 and G10 did not exert a protective or aggravating effect on the AP development. However, G15 and G20 had a significant effect on the AP severity, exacerbating the inflammation and osteoclast markers.