Final irrigation with bioglass solution in regenerative endodontic procedure induces tissue formation inside the root canals, collagen maturation, proliferation cell and presence of osteocalcin.

AIM: To evaluate the influence of an experimental solution of cobalt-doped F18 bioactive glass (F18Co) on tissue repair following regenerative endodontic procedure (REP) in rat molars. METHODOLOGY: The F18Co solution was prepared at a ratio of 1:5 F18Co powder to distilled water. The right or left upper first molars of 12 Wistar rats were used, where the pulps were exposed, removed, and irrigated with 2.5% sodium hypochlorite (NaOCl), followed by 17% ethylenediaminetetraacetic acid (EDTA) (5 min each). Subsequently, the molars were divided into two groups (n = 6): REP-SS and REP-F18Co, where they received a final irrigation (5 min) with saline solution (SS) or F18Co solution, respectively. Then, intracanal bleeding was induced, and the tooth was sealed. Untreated molars were used as controls (n = 3). At 21 days, the rats were euthanized, and the specimens were processed for analysis of mineralized tissue and soft tissue formation inside the root canal using haematoxylin-eosin. The presence and maturation of collagen were evaluated by Masson's trichrome and picrosirius red staining. Immunolabelling analyses of proliferating cell nuclear antigen (PCNA) and osteocalcin (OCN) were performed. The data were submitted to the Mann-Whitney U-test (p < .05). RESULTS: There was a similar formation of mineralized tissue in thickness and length in REP-SS and REP-F18Co groups (p > .05). Regarding the presence of newly formed soft tissue, most specimens of the REP-F18Co had tissue formation up to the cervical third of the canal, whilst the REP-SS specimens showed formation up to the middle third (p < .05), and there was higher maturation of collagen in REP-F18Co (p < .05). The number of PCNA-positive cells found in the apical third of the root canal was significantly higher in the F18Co group, as well as the OCN immunolabelling, which was severe in most specimens of REP-F18Co, and low in most specimens of REP-SS. CONCLUSION: The final irrigation with F18Co bioactive glass solution in REP did not influence mineralized tissue formation but induced soft tissue formation inside the root canals, with higher collagen maturation, and an increase in PCNA-positive cells and OCN immunolabelling.

Influence of dental bleaching on the pulp tissue: A systematic review of in vivo studies.

BACKGROUND: Although several studies indicate the harmful effects of bleaching on pulp tissue, the demand for this procedure using high concentrations of hydrogen peroxide (HP) is high. OBJECTIVES: To investigate the influence of bleaching on the pulp tissue. METHODS: Electronic searches were conducted (PubMed/MEDLINE, Scopus, Cochrane Library and grey literature) until February 2021. Only in vivo studies that evaluated the effects of HP and/or carbamide peroxide (CP) bleaching gels on the inflammatory response in the pulp tissue compared with a non-bleached group were included. Risk of bias was performed according to a modified Methodological Index for Non-Randomized Studies scale for human studies and the Systematic Review Centre for Laboratory Animal Experimentation's RoB tool for animal studies. Meta-analysis was unfeasible. RESULTS: Of the 1311 studies, 30 were eligible. Of these, 18 studies evaluated the inflammatory response in animal models. All these studies reported a moderate-to-strong inflammatory response in the superficial regions of pulp, characterized by cell disorganization and necrotic areas, particularly during the initial periods following exposure to 35%-38% HP, for 30-40 min. In the evaluation of human teeth across 11 studies, seven investigated inflammatory responses, with five observing significant inflammation in the pulp of bleached teeth. In terms of tertiary dentine deposition, 11 out of 12 studies noted its occurrence after bleaching with 35%-38% HP in long-term assessments. Additionally, three studies reported significant levels of osteocalcin/osteopontin at 2 or 10 days post-treatment. Other studies indicated an increase in pro-inflammatory cytokines ranging from immediately up to 10 days after bleaching. Studies using humans' teeth had a low risk of bias, whereas animal studies had a high risk of bias. DISCUSSION: Despite the heterogeneity in bleaching protocols among studies, High-concentrations of HP shows the potential to induce significant pulp damage. CONCLUSIONS: High-concentrations of bleaching gel increases inflammatory response and necrosis in the pulp tissue at short periods after bleaching, mainly in rat molars and in human incisors, in addition to greater hard tissue deposition over time. However, further well-described histological studies with long-term follow-up are encouraged due to the methodological limitations of these studies. REGISTRATION: PROSPERO (CRD42021230937).

Influence of systemic antibiotic therapy on the development and progression of induced apical periodontitis in Wistar rats.

The aim of this study was to investigate the influence of systemic antibiotic therapy on the development and progression of induced apical periodontitis (AP) in Wistar rats. Fifty-six rats were submitted to pulp exposure of the lower left first molar for the induction of AP. On the same day, intraperitoneal antibiotic therapy was administered once a day, for 15 days, until euthanasia. The groups were formed according to the different treatments (n = 8): C-control; GEN-treated with gentamicin (10 mg/Kg); AC-treated with amoxicillin (100 mg/Kg); MZ-treated with metronidazole (40 mg/Kg); AMP-treated with ampicillin (100 mg/Kg); AMC group-treated with amoxicillin + clavulanic acid (100 mg/kg); CLI-treated with clindamycin (60 mg/kg). After euthanasia, the jaws were collected and processed for (1) histological and histometric analysis using hematoxylin and eosin staining, (2) analysis of collagen fibers using Picrosirius Red staining and (3) bacteriological analysis using Brown-Brenn staining. The data were analyzed statistically (p < 0.05). AP induction was confirmed in all groups. The AMC group had the lower intensity of inflammatory infiltrate (p = 0.028) and less periapical bone resorption compared to control (p = 0.006). Regarding collagen maturation, PSR staining revealed a predominance of mature collagen fibers in all groups. The AC and AMC groups had the lower amount of mature fibers and the highest amount of immature fibers, compared to all other groups (p < 0.001). All groups showed bacterial contamination; however, the AC and AMC groups showed a lower extent of bacterial contamination compared to the control (p < 0.001). It can be concluded that systemic antibiotic therapy influences the development and progression of induced AP.

Influence of ethylenediaminetetraacetic acid irrigation on the regenerative endodontic procedure in an immature rat molar model.

AIM: To analyse the influence of ethylenediaminetetraacetic acid (EDTA) on the repair process in immature rat molars after a regenerative endodontic procedure (REP). METHODOLOGY: The lower first molars of 12 4-week-old Wistar rats underwent pulpectomy in the mesial root and were divided into the following groups: sodium hypochlorite (NaOCl; n = 6) - the mesial canals were irrigated with 2.5% NaOCl for 5 min, and NaOCl-EDTA (n = 6) - the canals were irrigated with 2.5% NaOCl, followed by 17% EDTA for 5 min each. After evoking bleeding using a size 10 K-file, the cavities were sealed. Three molars on the untreated side were randomly used as control (control-15 d; n = 3), and three molars from the other three rats untreated were used as immediate control (n = 3). After 15 days (NaOCl, NaOCl-EDTA and control-15 d groups) or immediately (control-immediate), the animals were euthanized, and the teeth were subjected to histologic evaluation of tissue regeneration and presence of collagen fibres. Mann-Whitney U-test was used (p < .05). RESULTS: The experimental groups had newly formed cementum-like tissue and increased root length and thickness. Half of the specimens in NaOCl-EDTA group showed apical foramen closure, whilst the NaOCl group had partial apical closure. The experimental groups showed inflammatory infiltrate extending mainly to the medium third of the root canal. These parameters were similar between experimental groups (p > .05). Newly formed connective tissue in the pulp space was significantly higher in the NaOCl-EDTA group than in NaOCl group (p < .05). Regarding the collagen fibres, the NaOCl-EDTA group had more collagen fibres in the root tip, but there was no significant difference compared to NaOCl group, and both groups showed greater amount of immature fibres in this area; in the centre of the apical third of root canal, there was equivalence between mature and immature fibres from both groups (p > .05). CONCLUSIONS: Ethylenediaminetetraacetic acid irrigation improved newly formed intracanal connective tissue after REP in immature molars of rats; however, EDTA did not influence cementum-like tissue formation, apical closure, inflammatory infiltrate and maturation of collagen fibres.

Influence of sodium hypochlorite on cyclic fatigue resistance of nickel-titanium instruments: A systematic review and meta-analysis of in vitro studies.

OBJECTIVE: This systematic review/meta-analysis investigated the influence of NaOCl on cyclic fatigue resistance of endodontic NiTi instruments. MATERIALS AND METHODS: A systematic search until July 2022 in PubMed/MEDLINE, Embase, Scopus, Web of Science, SciELO, Cochrane Library, and grey literature was conducted. According to the PECOS strategy, only in vitro studies evaluating the effects of NaOCl on the cyclic fatigue resistance of NiTi instruments were eligible. Cyclic fatigue resistance was the primary outcome. A modified Joanna Briggs Institute's Checklist was used for risk of bias assessment. RESULTS: Of the 2,445 records screened, 37 studies were included. Most studies used simulated canals made of stainless-steel block with severe to moderate curvatures. NaOCl concentration varied from 1-6%, mainly at 37 °C. Regarding fatigue resistance, 23 studies using 1.2% to 6% NaOCl showed a reduction in the resistance compared to the control groups, especially when pre-heated. Four meta-analyses were performed according to the tested NiTi systems. The meta-analyses indicated that the PTU F2 files had higher reduction of fatigue resistance after exposure to 5.25% NaOCl; no differences between NaOCl and no immersion were observed for Reciproc R25, WaveOne 25.08, and WaveOne Gold Primary files. Included studies had low risk of bias. CONCLUSION: NaOCl appears to reduce cyclic fatigue resistance of certain NiTi files, especially when they are pre-heated, particularly in conventional NiTi files compared to some heat-treated instruments. It is possible that the temperature of the solution may have a greater influence on resistance than NaOCl itself. Important to note that an overall tendency toward no significant influence was observed among various systems. CLINICAL RELEVANCE: Precautions are necessary when a pre-heated high-concentration NaOCl is used to enhance its properties during root canal preparation, mainly using conventional wire.

Effects of diabetes mellitus on dental pulp: A systematic review of in vivo and in vitro studies.

OBJECTIVES: This systematic review (PROSPERO CRD42021227711) evaluated the influence of diabetes mellitus (DM) on the response of the pulp tissue and in the pulp cells behaviour. MATERIALS AND METHODS: Searches in PubMed/MEDLINE, Embase, Web of Science and OpenGrey were performed until March 2022. Studies evaluating the effects of DM in the pulp tissue inflammation and in the cell behaviour were included, followed by risk of bias assessment (Methodological Index for Non-Randomized Studies and SYRCLE's RoB tools). The meta-analysis was unfeasible, and a narrative synthesis for each outcome was provided. RESULTS: Of the 615 studies, 21 were eligible, mainly with in vivo analysis (16 studies). The pulp inflammation (10 studies) was analysed mainly by haematoxylin-eosin stain; DM increased pulp inflammation/degeneration in 9 studies, especially after dental procedures. The cell viability (5 studies) was analysed mostly using MTT assay; DM and glycating agents decreased cellular viability in 3 studies. DM reduced collagen in all of three studies. There were controversial results regarding mineralization; however, increased alkaline phosphatase was reported in three of four studies. CONCLUSIONS: DM seems to increase inflammation/degeneration and mineralization in the pulp tissue while reducing cell proliferation. Further analyses in human pulp are important to provide stronger evidence.

Biocompatibility, Biomineralization, and Maturation of Collagen by RTR®, Bioglass and DM Bone® Materials.

This study evaluated the biocompatibility, biomineralization, and collagen fiber maturation induced by Resorbable Tissue Replacement (RTR®; ß-tricalcium phosphate [TCP]), Bioglass (BIOG; bioactive glass), and DM Bone® (DMB; hydroxyapatite and ß-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.

Biocompatibility, biomineralization, and maturation of collagen by rtr®, bioglass and dm bone® materials

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.

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.