Abstract Objective To assess whether bleaching gel volume influences chromatic changes, hydrogen peroxide (HP) diffusion, inflammation, and oxidative stress in the pulp tissue. Methodology A total of 60 bovine teeth were divided into four groups, according to bleaching gel volume (n=15): without gel (WG); V30 (30 µL of 35% HP); V60 (60 µL); and V120 (120 μL). HP diffusion analysis was performed in the first session (T1). Chromatic changes (ΔE, ΔE00, and WID) were assessed after the first (T1), second (T2), third (T3) sessions, and 15 d (T4) after the end of treatment. Moreover, 20 rats were randomly divided into four groups (n=10) and their upper first molars were treated with different gel volumes: control (no treatment); V2 (2 μL of 17.5% HP); V4 (4 μL); and V8 (8 μL). After 24 h, rats were euthanized and the specimens processed for histological and immunohistochemical (nitric oxide synthase) evaluation. Data were analyzed using the Wilcoxon and Mann-Whitney tests (p<0.05). Results In vitro (bovine teeth), chromatic changes were not influenced by bleaching gel volume, showing similar values in all groups and sessions, except for the control group (p<0.05). The V120 group had the highest HP diffusion values (p<0.05). In vivo (pulp tissue), the V4 and V8 groups showed the highest inflammatory infiltrate in the pulp and significant oxidative stress (p<0.05). Conclusion The adverse effects on the dental pulp related to HP diffusion, pulp inflammation, and oxidative stress depend on bleaching gel volume, while the bleaching effect is not proportional to the volume used.

Abstract Objectives: To evaluate the influence of photobiomodulation with infrared laser (IRL) in the rat pulp tissue after bleaching, considering the immunolabeling of interleukin (IL)-23 and hypoxia-inducible factor (HIF)-1α. Methodology: The right and left molars of forty rats were divided into groups: Control – with placebo gel and Bleached – with 35% hydrogen peroxide (H2O2). Half of the rats received one IRL application on both sides, establishing a split-mouth design, which resulted in 4 groups with 20 hemi-maxillae each: Control, Bleach, IRL, and Bleached-IRL. Rats (n=10) from each group were euthanized, at 2- and 30-days mark, and the pulp tissue was evaluated using inflammation and immunolabeling scores. Wilcoxon and Mann-Whitney statistical tests were performed (p<0.05). Results: At the 2-days mark, the Bleached group had severe inflammation and necrosis in the occlusal thirds of the pulp, and moderate to severe inflammation in cervical third, whereas the Bleached-IRL had mild to moderate inflammation (p<0.05). At the 30-days mark, there was no inflammation, but tertiary dentine formation in the bleached groups. Regarding IL-23, severe immunolabeling was observed in the Bleached group (p<0.05) at the 2-days mark; at the 30-days mark, there was a reduction in immunolabeling, in which the Bleached group had moderate and the Bleached-IRL group had mild immunolabeling (p>0.05). HIF-1α was more evident at the 2-days mark in the Bleached group, without significant difference with the Bleached-IRL (p>0.05). The difference was observed between the bleached and control groups, without immunolabeling (p<0.05); at the 30-days mark, the Bleached group had reduction in HIF-1α immunolabeling, while the Bleached-IRL had an increase; the difference remained between the bleached and the controls groups (p<0.05) Conclusion: Photobiomodulation using IRL minimized the inflammation and IL-23 immunolabeling in the pulp tissue of rats after dental bleaching, but did not influence significantly the HIF-1α immunolabeling.

Resumo Este estudo avaliou a biocompatibilidade, biomineralização e maturação das fibras de colágeno induzidas por Resorbable Tissue Replacement (RTR®; fosfato β-tricálcico [TCP]), Bioglass (BIOG; vidro bioativo) e DM Bone® (DMB; hidroxiapatita e β-TCP) in vivo. Sessenta e quatro tubos de polietileno com ou sem (grupo controle; GC) os materiais (n=8/grupo/período) foram implantados aleatoriamente em tecido subcutâneo de 16 ratos machos Wistar (quatro por rato), pesando entre 250 a 280g. Os ratos foram mortos após 7 e 30 dias (n=8), e as amostras foram removidas para análise da inflamação utilizando hematoxilina-eosina; avaliação da biomineralização utilizando coloração de von Kossa (VK) e luz polarizada (LP); e maturação das fibras colágenas, utilizando picrosirius red (PSR). Os dados não-paramétricos foram analisados pelos testes de Kruskal-Wallis e Dunn, e os paramétricos pelo teste de one-way ANOVA (p<0.05). Aos 7 dias, todos os grupos induziram inflamação moderada (p>0,05). Aos 30 dias, houve inflamação leve nos grupos BIOG e GC, e inflamação moderada nos grupos RTR e DMB, com diferença significativa entre os GC e RTR (p<0,05). A cápsula fibrosa foi espessa aos 7 dias, e predominantemente fina aos 30 dias em todos os grupos. Todos os materiais exibiram estruturas positivas para VK e LP. Fibras colágenas imaturas foram predominantes aos 7 e 30 dias em todos os grupos (p>0,05), embora o DMB exibiu fibras mais maduras do que o BIOG aos 30 dias (p<0,05). RTR, BIOG e DMB foram biocompatíveis, induzindo inflamação que reduziu com o tempo, e biomineralização no tecido subcutâneo de ratos. O DMB exibiu mais fibras colágenas maduras do que o BIOG em período mais longo.

Abstract This study evaluated the biocompatibility, biomineralization, and collagen fiber maturation induced by Resorbable Tissue Replacement (RTR®; β-tricalcium phosphate [TCP]), Bioglass (BIOG; bioactive glass), and DM Bone® (DMB; hydroxyapatite and β-TCP) in vivo. Sixty-four polyethylene tubes with or without (control group; CG) materials (n=8/group/period) were randomly implanted in the subcutaneous tissue of 16 male Wistar rats (four per rat), weighting 250 to 280 g. The rats were killed after 7 and 30 days (n=8), and the specimens were removed for analysis of inflammation using hematoxylin-eosin; biomineralization assay using von Kossa (VK) staining and polarized light (PL); and collagen fiber maturation using picrosirius red (PSR). Nonparametric data were statistically analyzed by Kruskal-Wallis and Dunn tests, and parametric data by one-way ANOVA test (p<0.05). At 7 days, all groups induced moderate inflammation (p>0.05). At 30 days, there was mild inflammation in the BIOG and CG, and moderate inflammation in the RTR and DMB groups, with a significant difference between the CG and RTR (p<0.05). The fibrous capsule was thick at 7 days and predominantly thin at 30 days in all groups. All materials exhibited structures that stained positively for VK and PL. Immature collagen fibers were predominant at 7 and 30 days in all groups (p>0.05), although DMB exhibited more mature fibers than BIOG at 30 days (p<0.05). RTR, BIOG, and DMB were biocompatible, inducing inflammation that reduced over time and biomineralization in the subcutaneous tissue of rats. DMB exhibited more mature collagen fibers than BIOG over a longer period.

Abstract Objectives This study evaluated if the use of a bioactive glass-ceramic-based gel, named Biosilicate (BS), before, after or mixed with bleaching gel, could influence the inflammation of the dental pulp tissue of rats’ molars undergoing dental bleaching with hydrogen peroxide (H2O2). Methodology The upper molars of Wistar rats (Rattus norvegicus, albinus) were divided into Ble: bleached (35% H2O2, 30-min); Ble-BS: bleached and followed by BS-based gel application (20 min); BS-Ble: BS-based gel application and then bleaching; BS/7d-Ble: BS-based gel applications for 7 days and then bleaching; Ble+BS: blend of H2O2 with BS-based gel (1:1, 30-min); and control: placebo gel. After 2 and 30 days (n=10), the rats were euthanized for histological evaluation. The Kruskal-Wallis and Dunn statistical tests were performed (P<0.05). Results At 2 days, the Ble and Ble-BS groups had significant alterations in the pulp tissue, with an area of necrosis. The groups with the application of BS-based gel before H2O2 had moderate inflammation and partial disorganization in the occlusal third of the coronary pulp and were significantly different from the Ble in the middle and cervical thirds (P<0.05). The most favorable results were observed in the Ble+BS, which was similar to the control in all thirds of the coronary pulp (P>0.05). At 30 days, the pulp tissue was organized and the bleached groups presented tertiary dentin deposition. The Ble group had the highest deposition of tertiary dentin, followed by the Ble-BS, and both were different from control (P<0.05). Conclusion A single BS-based gel application beforehand or BS-based gel blended with a bleaching gel minimize the pulp damage induced by dental bleaching.

Abstract Aim To evaluate the cytotoxicity, biocompatibility and mineralization capacity of BIO-C PULPO, and MTA. Methodology L929 fibroblasts were cultured and MTT assay was used to determine the material cytotoxicity on 6, 24, and 48 h. A total of 30 male rats (Wistar) aged between 4 and 6 months, weighing between 250 and 300 g were used. Polyethylene tubes containing BIO-C PULPO, MTA, and empty tubes were implanted into dorsal connective tissue. After the experimental periods (7, 15, 30, 60, and 90 days) the tubes were histologically analyzed using hematoxylin-eosin (H&E), immunolabeling of IL-1β and TNF-α, and von Kossa staining, or without staining for polarized light analysis. The average number of inflammatory cells was quantified; the mineralization assessment was determined by the area marked in μm2 and semiquantitative immunolabeling analyses of IL-1β and TNF-α were performed. Then, data underwent statistical analysis with a 5% significance level. Results It was observed that BIO-C PULPO and MTA presented cytocompatibility at 6, 24, and 48 similar or higher than control for all evaluated period. On periods 7 and 15 days, BIO-C PULPO was the material with the highest number of inflammatory cells (p<0.05). On periods 30, 60, and 90 days, BIO-C PULPO and MTA presented similar inflammatory reactions (p>0.05). No statistical differences were found between Control, BIO-C PULPO, and MTA for immunolabeling of IL-1β and TNF-α in the different periods of analysis (p<0.05). Positive von Kossa staining and birefringent structures under polarized light were observed in all analyzed periods in contact with both materials, but larger mineralization area was found with BIO-C PULPO on day 90 (p<0.05). Conclusion BIO-C PULPO was biocompatible and induced mineralization similar to MTA.

Resumo Novas formulações de agregado de trióxido mineral (MTA) são constantemente introduzidas no mercado, geralmente em forma de pó e líquido. O Biocerâmico (Bio-C) Reparador (Repair) é um material pronto para uso sugerido como substituto do MTA, mas suas propriedades precisam ser estudadas. Este estudo avaliou a citotoxicidade, biocompatibilidade e biomineralização do Bio-C Repair comparado ao MTA-High Plasticity (MTA-HP) e MTA branco da Angelus (MTA-Ang). Fibroblastos L929 foram expostos a extratos dos materiais (não diluído, ½ e ¼ diluições; 6, 24 e 48 h). Tubos de polietileno contendo os materiais ou vazios (controle) foram implantados no tecido subcutâneo de ratos. Após 7 e 30 dias (n=8), os espécimes foram removidos para análises (hematoxilina-eosina, von Kossa e luz polarizada). Os dados da citotoxicidade foram analisados estatisticamente pelo teste de two-way ANOVA, e os dados da biocompatibilidade pelos testes de Kruskal-Wallis e Dunn (p<0,05). As células expostas aos materiais apresentaram maior viabilidade celular na maior parte dos períodos, comparados com o controle (p<0,05). O extrato não diluído e ½ diluição do MTA-HP apresentaram maior citocompatibilidade do que Bio-C Repair às 6h, e com ¼ diluição às 24h (p<0,05); o MTA-Ang branco apresentou maior citocompatibilidade do que o Bio-C Repair na maior parte dos períodos (p<0,05). O extrato não diluído do MTA-Ang branco apresentou maior citocompatibilidade às 6 e 24 h comparado ao MTA-HP, e com ½ diluição às 24h (p<0,05). A citocompatibilidade dos materiais foi semelhante às 48 h para a maior parte das diluições (p>0,05). Aos 7 e 30 dias, os grupos apresentaram inflamação moderada e leve, respectivamente (p>0,05). Todos os materiais mostraram estruturas positivas para von Kossa e luz polarizada. Em conclusão, o Bio-C Repair teve citocompatibilidade semelhante aos materiais à base de MTA, é biocompatível e induz à biomineralização.

Abstract New mineral trioxide aggregate (MTA) formulations are constantly introduced in the market, usually in a powder-and-liquid form. Bioceramic (Bio-C) Repair is a ready-for-use material suggested as substitute for MTA, but its properties need to be studied. This study evaluated the cytotoxicity, biocompatibility and biomineralization of Bio-C Repair compared to MTA Repair High-Plasticity (MTA-HP) and white MTA-Angelus (MTA-Ang). L929 fibroblasts were exposed to material-extracted (undiluted, ½ and ¼ dilutions; 6, 24 and 48h). Polyethylene tubes with material or empty (control) were implanted in the subcutaneous tissue of rats. After 7 and 30 days (n=8), the specimens were removed for analysis (hematoxylin-eosin, von Kossa and polarized light). Cytotoxicity data were statistically analyzed by two-way ANOVA, and biocompatibility data by Kruskal-Wallis and Dunn tests (p<0.05). The cells exposed to the materials had greater viability at most of the periods compared with control (p<0.05). The undiluted and ½ dilutions of MTA-HP extract showed higher cytocompatibility than Bio-C Repair at 6 h and with the ¼ dilution at 24 h (p<0.05); the white MTA-Ang showed higher cytocompatibility than Bio-C Repair at most of periods (p<0.05). The undiluted white MTA-Ang extract had higher cytocompatibility at 6 and 24h than MTA-HP, and with ½ dilution at 24h (p<0.05). The materials’ cytocompatibility was similar at 48h for most dilutions (p>0.05). At 7 and 30 days, the groups had moderate and mild inflammation, respectively (p>0.05). All materials showed positive structures for von Kossa and polarized light. In conclusion, Bio-C Repair had similar cytocompatibility to MTA-based materials is biocompatible and induces biomineralization.

Abstract: This study evaluated the cytotoxicity and biocompatibility of a new bioceramic endodontic sealer (i.e., Sealer Plus BC) in comparison with those of MTA Fillapex and AH Plus. L929 fibroblasts were cultured and Alamar Blue was used to evaluate cell viability of diluted extracts (1:50, 1:100, and 1:200) from each sealer at 24 h. Polyethylene tubes that were filled with material or empty (as a control) were implanted in the subcutaneous tissue of rats. The rats were killed after 7 and 30 d (n = 8), and the tubes were removed for histological analysis. Parametric data was analyzed using a one-way ANOVA test, and nonparametric data was analyzed via the Kruskal–Wallis test followed by the Dunn test (p < 0.05). A reduction in cell viability was observed in the extracts that were more diluted for Sealer Plus BC when compared to that of Control and AH Plus (p < 0.05). However, the 1:50 dilution of the Sealer Plus BC was similar to that of the Control (p > 0.05). Conversely, more diluted extracts of MTA Fillapex (1:200) and AH Plus (1:100 and 1:200) were similar to the Control (p > 0.05). Histological analysis performed at 7 d did not indicate any significant difference between tissue response for all materials, and the fibrous capsule was thick (p > 0.05). At 30 d, Sealer Plus BC was similar to the Control (p > 0.05) and MTA Fillapex and AH Plus exhibited greater inflammation than the Control (p < 0.05). The fibrous capsule was thin for the Control and for most specimens of Sealer Plus BC and AH Plus. Thus, Sealer Plus BC is biocompatible when compared to MTA Fillapex and AH Plus, and it is less cytotoxic when less-diluted extracts are used.

Resumo O gel clareador à base de peróxido de hidrogênio (H2O2) causa danos ao tecido pulpar. Este estudo investigou a ação de um anti-inflamatório tópico, o Otosporin®, nos dentes de ratos clareados com a hipótese nula de que o Otosporin® não é capaz de minimizar a inflamação da polpa gerada pelo gel clareador. Os molares dos ratos foram divididos em grupos: ClA: clareado (H2O2 a 35% / aplicação única de 30 min); CLA-O: clareado seguido do Otosporin® (10 min); e controle: gel placebo. No segundo dia após a clareação dentária, os ratos foram mortos e suas maxilas foram processadas para análise de hematoxilina-eosina e imunohistoquímica para o fator de necrose tumoral alfa (TNF-a), interleucina (IL)-6 e IL-17. Os dados coletados foram submetidos aos testes estatísticos de Kruskal-Wallis e Dunn com um nível de significância de 5% (p<0,05). O grupo CLA apresentou inflamação moderada à severa no terço oclusal da polpa coronária, com áreas necróticas; e CLA-O, inflamação leve (p<0,05). Houve diferença significativa nos terços oclusal e médio da polpa coronária entre o grupo CLA com os grupos CLA-O e controle (p<0,05). Não houve diferença no terço cervical (p>0,05). O grupo CLA apresentou maior imunoexpressão para TNF-a comparado aos grupos CLA-O e controle (p<0,05), com imunoexpressão moderada e leve, respectivamente. Em relação a IL-6 e IL-17, o grupo CLA apresentou maior imunoexpressão comparado ao controle (p<0,05); o CLA-O foi semelhante ao controle (p>0,05). O anti-inflamatório tópico Otosporin® pode reduzir a inflamação pulpar após clareação em dentes de ratos.

Abstract Bleaching gel containing hydrogen peroxide (H2O2) cause damages in pulp tissue. This study investigated the action of a topical anti-inflammatory, the Otosporin®, in rats’ bleached teeth with the null hypothesis of which the Otosporin® is no able to minimize the pulp inflammation that bleaching gel generates. The rat’s molars were divided into groups: BLE: bleached (35% H2O2 concentration /single application of 30 min); BLE-O: bleached followed by Otosporin® (10 min); and control: placebo gel. In the second day after dental bleaching, the rats were killed, and the jaws were processed for hematoxylin-eosin and immunohistochemistry analysis for tumor necrosis factor alpha (TNF-α), interleukin (IL)-6 and IL-17. The data collected were subjected to Kruskal-Wallis and Dunn statistical tests with at a 5% level of significance (p<0.05). The BLE group had moderate to strong inflammation in the occlusal third of the coronary pulp, with necrotic areas; and BLE-O, mild inflammation (p<0.05). There was a significant difference in the occlusal and middle thirds of the coronary pulp between the BLE with BLE-O and control groups (p<0.05). There was no difference in the cervical third (p>0.05). The BLE group had a high immunoexpression of TNF-α than BLE-O and control groups (p<0.05), with moderate and mild immunoexpression, respectively. Regarding IL-6 and IL-17, the BLE group had higher immunoexpression than control (p<0.05); the BLE-O was similar to the control (p>0.05). The topical anti-inflammatory Otosporin® can reduce pulp inflammation after dental bleaching in the rat teeth.

Resumo O peróxido de hidrogênio (H2O2) é capaz de penetrar pelos tecidos dentários, alterando a coloração destes, e causar danos a polpa. Este estudo avaliou a penetração por esmalte e dentina, a alteração de cor e a reposta tecidual pulpar, provocadas pelo uso de duas concentrações de H2O2 em protocolos de clareação dentária de consultório. Discos de dentes bovinos em câmaras pulpares artificiais receberam géis clareadores para avaliação da penetração por esmalte e dentina e da alteração de cor, formando os grupos: BLU (H2O2 20% - 1x50 min, Whiteness HP Blue); MAX (H2O2 35% - 3x15 min, Whiteness HP Maxx); e Controle (gel placebo - 1x50 min). A penetração por esmalte e dentina foi quantificada baseada na reação do H2O2 com o corante violeta leucocristal, e a alteração de cor foi analisada pelo sistema CIELab. Vinte ratos Wistar foram divididos em dois grupos (BLU e MAX), e tiveram os molares direito superiores tratados com os mesmos protocolos do estudo in vitro; os molares superiores do lado esquerdo serviram de controle. Após 2 dias, os animais foram eutanasiados e as maxilas examinadas por microscopia de luz. Foram atribuídos escores ao infiltrado inflamatório (1, ausente; 2, leve; 3, moderado; 4 severo ou necrose). Os dados foram submetidos a testes estatísticos (=0,05). O grupo MAX apresentou maior penetração de H2O2 por esmalte e dentina (p<0,05). A alteração de cor foi semelhante nos grupos clareados (p>0,05), mas diferente quando comparados grupos clareados com controle (p<0,05). MAX apresentou inflamação severa nos terços superiores da polpa coronária, e BLU apresentou inflamação moderada (p<0,05). Assim, protocolo para procedimento clareador de consultório utilizando baixas concentrações de H2O2 deve ser de escolha na clínica, por reduzir a penetração por esmalte e dentina, causando menos danos à polpa, e proporcionar mesma eficiência clareadora.

Abstract Hydrogen peroxide (H2O2) penetrates into the dental hard tissues causing color alteration but also alterations in pulpal tissues. Hard-tissue penetration, color alteration and the pulp response alterations were evaluated for two in-office bleaching protocols with H2O2. For trans-enamel/dentin penetration and color alteration, discs of bovine teeth were attached to an artificial pulp chamber and bleached according to the groups: BLU (20% H2O2 - 1x50 min, Whiteness HP Blue); MAX (35% H2O2 - 3x15 min, Whiteness HP Maxx); Control (1x50 min, placebo). Trans-enamel/dentin penetration was quantified based on the reaction of H2O2 with leucocrystal violet and the color analyzed by CIELab System. Twenty Wistar rats were divided into two groups (BLU and MAX) and their maxillary right molars were treated according to the same protocols of the in vitro study; the maxillary left molars were used as controls. After 2 days, the animals were killed and their maxillae were examined by light microscopy. The inflammation of pulp tissue was scored according to the inflammatory infiltrate (1, absent; 2, mild; 3, moderate; 4, severe/necrosis). Data were analyzed by statistical tests (α=0.05). MAX showed higher trans-enamel/dentinal penetration of H2O2 (p<0.05). The color alteration was similar for both groups (p>0.05), and different when compared to Control group (p<0.05). MAX showed severe inflammation in the upper thirds of the coronal pulp, and BLU showed moderate inflammation (p<0.05). In-office bleaching protocols using lower concentrations of hydrogen peroxide should be preferred due to their reduced trans-enamel/dentinal penetration since they cause less pulp damage and provide same bleaching efficiency.

ABSTRACT Dental materials in general are tested in different animal models prior to the clinical use in humans, except for bleaching agents. Objectives To evaluate an experimental rat model for comparative studies of bleaching agents, by investigating the influence of different concentrations and application times of H2O2 gel in the pulp tissue during in-office bleaching of rats’ vital teeth. Material and Methods The right and left maxillary molars of 50 Wistar rats were bleached with 20% and 35% H2O2 gels, respectively, for 5, 10, 15, 30, or 45 min (n=10 rats/group). Ten animals were untreated (control). The rats were killed after 2 or 30 days, and the maxillae were examined by light microscopy. Inflammation was evaluated through histomorphometric analysis with inflammatory cell count in the coronal and radicular thirds of the pulp. Fibroblasts were also counted. Scores were attributed to odontoblastic layer and vascular changes. Tertiary dentin area and pulp chamber central area were measured histomorphometrically. Data were compared by analysis of variance and Kruskal-Wallis test (p<0.05). Results After 2 days, the amount of inflammatory cells increased in the coronal pulp occlusal third up to the 15-min application groups of each bleaching gel. In the groups exposed to each concentration for 30 and 45 min, the number of inflammatory cells decreased along with the appearance of necrotic areas. After 30 days, reduction on the pulp chamber central area and enlargement of the tertiary dentin area were observed, without the detection of inflammation areas. Conclusion The rat model of extracoronal bleaching showed to be adequate for studies of bleaching protocols, as it was possible to observe alterations in the pulp tissues and tooth structure caused by different concentrations and application periods of bleaching agents.

ABSTRACT Dental materials, in general, are tested in different animal models prior to their clinical use in humans, except for bleaching agents. Objectives To evaluate an experimental rat model for comparative studies of bleaching agents by investigating the influence of different concentrations and application times of H2O2 gel in the pulp tissue during in-office bleaching of rats’ vital teeth. Material and methods The right and left maxillary molars of 50 Wistar rats were bleached with 20% and 35% H2O2 gels, respectively, for 5, 10, 15, 30, or 45 min (n=10 rats/group). Ten animals (control) were untreated. The rats were killed after 2 or 30 days, and the maxillae were examined by light microscopy. Inflammation was evaluated by histomorphometric analysis with inflammatory cell counting in the coronal and radicular thirds of the pulp. The counting of fibroblasts was also performed. Scores were attributed to the odontoblastic layer and to vascular changes. The tertiary dentin area and the pulp chamber central area were histomorphometrically measured. Data were compared by the analysis of variance and the Kruskal-Wallis test (p<0.05). Results After 2 days, the amount of inflammatory cells increased in the occlusal third of the coronal pulp until the time of 15 min for both concentrations of bleaching gels. In 30 and 45 min groups of each concentration, the number of inflammatory cells decreased along with the appearance of necrotic areas. After 30 days, a reduction in the pulp chamber central area and an enlargement of tertiary dentin area were observed without the detection of inflammation areas. Conclusion The rat model of extra coronal bleaching showed to be adequate for bleaching protocols studies, as it was possible to observe alterations in the pulp tissues and in the tooth structure caused by different concentrations and periods of application of bleaching agents.