Effect of photobiomodulation therapy on TGF-ß release from dentin, migration and viability of dental pulp stem cells in regenerative endodontics treatment: An ex vivo study.

BACKGROUND AND AIM: Regenerative endodontic procedures (REPs) are oriented by the principles of tissue engineering, incorporating dental pulp stem cells (DPSC), crucial growth factors like Transforming growth factor-ß (TGF-ß1), and scaffolds to facilitate the regeneration of dental pulp tissues. The present study aimed to investigate the effect of photobiomodulation (PBM) therapy, using an 808 nm diode laser on cellular modulation mechanisms in REPs. METHOD AND MATERIAL: A total of 108 human dentin discs obtained from intact single root teeth were randomly assigned into six groups (n = 8): 1. Positive control (EDTA), 2. PBM-1 (3 J/cm2), 3. PBM-2 (5 J/cm2), 4. EDTA+PBM-1, 5. EDTA+PBM-2, and 6. Negative control (NaOCl). Then, an extract solution was prepared from each disc and the concentration of released TGF-ß1 from the discs was measured using enzyme-linked immunosorbent assay (ELISA). Moreover, the extract solution was added to DPSC culture medium to evaluate cell viability and migration through MTT assay and scratch test, respectively. RESULT: The group exposed to PBM-1 showed the highest cell viability, while treatment with EDTA and EDTA+PBM-2 decreased cellular viability. Also, the PBM-treated groups showed significantly higher release of TGF-ß1 compared to the negative control. EDTA and EDTA+PBM-1 showed the highest release among all the groups. No significant difference was found between EDTA and EDTA+PBM-1, as well as between PBM-1 and PBM-2. Moreover, the PBM-1 group exhibited the highest migration after 24 h, which was significantly greater than other groups, except for the PBM-2 group. CONCLUSION: According to the obtained data, 808 nm mediated-PBM (3 J/cm2), both independently and in conjunction with EDTA, enhanced the release of TGF-ß1 from dentin and improved cell viability and migration of DPSCs. It seems that, PBM under the specific parameters employed in this study, could be an effective adjunctive therapy in REPs.

Efficacy of disinfection procedures performed prior to regenerative endodontic therapy: An integrative review.

The aim of this integrative review was to identify whether the disinfection procedures performed prior to regenerative endodontic treatment were effective on biofilm removal from the root canals. The research was based on PubMed, Latin American and Caribbean Health Sciences Literature (Lilacs) and Scientific Electronic Library Online (SciELO) databases. Four articles were selected; one of the studies was in vivo and the others ex vivo. Different disinfection procedures were studied, characterised mainly by the use of intracanal medication, highlighting the double antibiotic paste, triple antibiotic paste and calcium hydroxide paste. Disinfection ability was evaluated against Enterococcus faecalis and multispecies biofilms by using the fluorescence technique and colony forming unit counting, for 7 to 21 days. Double antibiotic paste and triple antibiotic paste demonstrated excellent antibiofilm activity, unlike CH paste that showed limited disinfection, even when associated with different antimicrobial agents. Triple antibiotic paste was the most effective medication against biofilm.

Microbiome Changes during Regenerative Endodontic Treatment Using Different Methods of Disinfection.

INTRODUCTION: The purpose of this study was to characterize qualitatively and quantitatively the changes in the endodontic microbiome, in teeth with necrotic pulp, open apexes, and apical periodontitis, with 3 antimicrobial protocols, undertaken in a multicenter clinical trial. METHODS: Microbiological samples were collected from 116 regenerative endodontic teeth, and 97 qualified for inclusion. The teeth were randomly divided into 3 treatment groups: apexification (APEX), regeneration (REGEN), and revascularization (REVASC), all in 2 appointments. The group variables in the first appointment irrigants, and second appointment irrigants and medicaments were as follows: APEX: 5.25%-6% NaOCl, 5.25%-6% NaOCl + 17% EDTA and calcium hydroxide; REGEN: 1.25% NaOCl, 17% EDTA, and 0.1 mg/mL triple antibiotic paste (TAP); and REVASC 5.25% NaOCl, saline, and 1 g/mL TAP, respectively. Sampling was done upon access (S0), after irrigation in the first appointment (S1), and after using medication and irrigation in the second appointment (S2). RESULTS: Quantitative polymerase chain reaction analysis of the 16S ribosomal RNA gene showed significant reduction in bacterial load from S0 to S2 in all groups; however, the APEX and REVASC groups had significantly less residual DNA than the REGEN group (P = .0045). The relative abundance of Bacteroidetes, Fusobacteria, Spirochaetes, and Synergistetes were reduced with the treatment rendered. However, relative abundance of Firmicutes and Actinobacteria was not changed, and that of Proteobacteria increased. LEfSe analysis showed that reduction in bacterial taxa was more in REVASC than APEX, which in turn was more than in REGEN. CONCLUSION: Enhanced antimicrobial protocols lead to better reduction in quantitative and qualitative parameters of the endodontic microflora.

Effect of Citric Acid Versus EDTA on Radiographic Root Development in Regenerative Endodontic Treatment: An Animal Study.

INTRODUCTION: Regenerative endodontic treatment was introduced with the aim of reinforcing the root and enabling further root development. In the process of dentin mineralization, bioactive molecules are entrapped in the dentin matrix, which can be released later by rinsing the dentin with chelating agents. This study aimed to compare the effects of citric acid and EDTA on radiographic root development in regenerative endodontic treatment in an animal model. METHODS: Premolars of 3 beagle dogs were used in this study. The teeth at 1 side of the jaw were irrigated with EDTA according to the regeneration protocol of the American Association of Endodontists. The teeth in the other quadrant were irrigated with citric acid in the second treatment session. Follow-up radiographs were obtained at 6 months, and the primary and final data were analyzed in SPSS Version 23 (IBM Corp, Armonk, NY) using the analysis of covariance. RESULTS: There was no significant difference between the 2 irrigating solutions regarding the increase in root length (P = .668); however, EDTA showed significantly superior efficacy in increasing the root dentin thickness compared with citric acid (P = .022). CONCLUSIONS: Within the limitations of this study, the use of citric acid versus EDTA was not superior on radiographic root development in regenerative endodontic treatment.

Procedimentos regenerativos endodônticos com clorexidina a 2% e hidróxido de cálcio: relato de caso / Endodontic regenerative procedures with 2% chlorhexidine and calcium hydroxide: case report

Introdução: Muitas substâncias têm sido utilizadas como irrigantes e curativos intracanal nos tratamentos de dentes com rizogênese incompleta, visando obter a regeneração pulpar. A clorexidina a 2% em associação com hidróxido de cálcio P.A. pode ser uma alternativa. Métodos: No presente caso clínico, o dente #45 com periodontite apical, em um paciente com 13 anos de idade, foi tratado com solução de clorexidina a 2% como irrigante. A clorexidina na forma de gel a 2% foi utilizada como auxiliar da instrumentação e, em associação com o hidróxido de cálcio, foi utilizada como curativo durante 14 dias. Após esse período, o canal foi irrigado com solução fisiológica e, sob anestesia alveolar mandibular, uma lima K #70 foi utilizada ultrapassando levemente o ápice, para causar sangramento. Após observar que o sangue dentro dos canais alcançou a porção cervical, o cimento biocerâmico MTA Branco foi colocado na porção cervical do canal. Resultados: A apicificação e o desenvolvimento da raiz em comprimento e largura puderam ser observados já no controle de três meses, e continuaram até a última revisão, no controle de um ano. Alterações de cor não foram observadas nese período de acompanhamento. Conclusões: A clorexidina associada ao hidróxido de cálcio pode ser uma alternativa aos procedimentos de regeneração pulpar (AU).

Introduction: Many substances have been used as irrigating solutions to promote pulp regeneration and root development in teeth with incomplete rhizogenesis. The use of 2% chlorhexidine as an irrigating solution in association with Calcium Hydroxide could be an alternative. Methods: In the case of the patient, a 13-year-old boy, here presented, tooth 45 with apical periodontitis was treated with 2% Chlorhexidine solution as irrigant adjuvant to instrumentation, in association with Calcium Hydroxide used as a dressing for 14 days. After this period, the canal was cleaned with a physiological solution and under mandibular alveolar anesthesia, a # 70 K file was used slightly over the apex to cause bleeding. After observing that blood inside the canals had reached the cervical portion. White MTA was placed in the cervical portion of the canal. Results: Induced apexification and development of increasing root length and width could already be observed in the three-month control exam, and this continued until the last review in the one-year control exam. No color changes were observed in this follow-up period. Conclusions: Chlorhexidine and calcium hydroxide could be used as an alternative to pulp regeneration procedures (AU).

Are single-visit regenerative endodontic procedures successful in treating non-vital, immature young permanent teeth?

Design Systematic review.Data sources PubMed and Scopus databases were searched independently by two authors from inception to July 2018 using keywords and index words combined using Boolean terms. Articles were restricted to English and were not excluded based on study design. Conference proceedings, grey literature, letters and commentaries were excluded.Data extraction and synthesis Demographic data (age, sex), study design, pulpal and periapical pre-treatment diagnosis, pre-operative radiographic findings, intra-operative disinfection protocol, intracanal coronal barrier, duration of follow-up, and clinical and/or radiographic and/or histological outcomes after completion of single-visit regenerative endodontic procedures (REPs) on non-vital, immature permanent teeth. Quality of included case reports were assessed according to the Joanna Briggs Institute Critical Appraisal Checklist. Risk of bias assessment of randomised controlled trials was done using Cochrane Collaboration's tool. The quality of the animal study was assessed by the SYRCLE risk of bias assessment tool.Results A total of five case reports, one randomised controlled trial and one animal study were included in the final analysis. All case reports were rated as high quality while the randomised controlled trial was rated moderate-to-low risk of bias. Case reports and the animal study favoured single-visit REPs, while the results from the randomised trial reported only 50% success. Within this limited pool of studies, there were differences in aetiology, location, disinfection protocol (different concentrations of NaOCl and EDTA, saline, chlorhexidine), follow-up time intervals and subjective reporting of radiographic healing, making syntheses challenging.Conclusions Single-visit REPs may offer benefits of reduced treatment time and visits compared to conventional two-visit REPs.

12-Year Follow-Up of Regenerative Endodontic Treatment of Immature Permanent Upper Incisors with Acute Abscess.

This case report discusses the endodontic treatment of a 7-year-old girl who suffered trauma (intrusion) to the immature upper central incisors secondary to a fall from a bicycle. Thirty days after the accident the patient was brought by her mother for clinical and radiographic assessment with a chief complaint of swelling and tenderness to percussion and palpation. Acute apical abscess associated with immature teeth were diagnosed. A decision was made to perform regenerative endodontic treatment. Access cavities were made and the root canals were disinfected by irrigation with 2.5% sodium hypochlorite. Final irrigation was performed with 17% EDTA. Due to pain and presence of secretions, 2% chlorhexidine gel was applied as an intracanal medicament. Seven days later, at the second visit, the root canals were once again disinfected and the canals of the right and left permanent upper central incisors were filled with double antibiotic paste (metronidazole/ciprofloxacin) and calcium hydroxide paste, respectively. Zinc oxide was mixed in both pastes. At the third visit, after 21 more days, the pastes were removed and the periapical areas were stimulated with a #80 K-file to encourage clot formation within the pulp cavities. A mineral trioxide aggregate (MTA) paste cervical plug was placed and the teeth were restored with glass ionomer cement. Radiographs and CBCT scans demonstrated complete root formations. The patient has been followed for 12 years, with evidence of clinical success throughout.

12-year follow-up of regenerative endodontic treatment of immature permanent upper incisors with acute abscess

Abstract This case report discusses the endodontic treatment of a 7-year-old girl who suffered trauma (intrusion) to the immature upper central incisors secondary to a fall from a bicycle. Thirty days after the accident the patient was brought by her mother for clinical and radiographic assessment with a chief complaint of swelling and tenderness to percussion and palpation. Acute apical abscess associated with immature teeth were diagnosed. A decision was made to perform regenerative endodontic treatment. Access cavities were made and the root canals were disinfected by irrigation with 2.5% sodium hypochlorite. Final irrigation was performed with 17% EDTA. Due to pain and presence of secretions, 2% chlorhexidine gel was applied as an intracanal medicament. Seven days later, at the second visit, the root canals were once again disinfected and the canals of the right and left permanent upper central incisors were filled with double antibiotic paste (metronidazole/ciprofloxacin) and calcium hydroxide paste, respectively. Zinc oxide was mixed in both pastes. At the third visit, after 21 more days, the pastes were removed and the periapical areas were stimulated with a #80 K-file to encourage clot formation within the pulp cavities. A mineral trioxide aggregate (MTA) paste cervical plug was placed and the teeth were restored with glass ionomer cement. Radiographs and CBCT scans demonstrated complete root formations. The patient has been followed for 12 years, with evidence of clinical success throughout.

Resumo Este relato de caso discute o tratamento endodôntico de uma menina de 7 anos que sofreu trauma nos incisivos centrais superiores imaturos, após queda de bicicleta. Trinta dias após o acidente, a paciente foi levada pela mãe para avaliação clínica e radiográfica com queixa principal de inchaço e sensibilidade à percussão e palpação. Foram diagnosticados abscessos apicais agudos em ambos os dentes. Foi decidido por realizar o tratamento endodôntico regenerativo. Após acessos coronários, os canais radiculares foram submetidos à protocolos de antissepsia com auxílio de irrigação com hipoclorito de sódio a 2,5% e EDTA 17%. Devido à dor e presença de secreções, foi utilizada medicação intracanal com clorexidina gel 2% por 7 dias. Na segunda sessão, os canais radiculares foram novamente submetidos aos procedimentos de antissepsia e os canais dos incisivos centrais superiores permanentes direito e esquerdo foram preenchidos com pasta antibiótica dupla (Metronidazol / Ciprofloxacina) e pasta de hidróxido de cálcio, respectivamente. Em ambas as pastas foi adicionado o óxido de zinco. Na terceira sessão, após mais 21 dias, as pastas foram removidas e as regiões periapicais foram estimuladas com lima K nº 80 para incentivar a formação de coágulos sanguíneos no interior das cavidades pulpares. Tampões cervicais de MTA foram adaptados e os dentes restaurados com cimento de ionômero de vidro resinoso. Radiografias e tomografias computadorizadas demonstraram formações radiculares completas. A paciente é acompanhada há 12 anos, com evidências clínicas e radiográficas de sucesso.

Effectiveness of Autologous Platelet Concentrates in Management of Young Immature Necrotic Permanent Teeth-A Systematic Review and Meta-Analysis.

The use of autologous platelet concentrates (APCs) in regenerative endodontic procedures is inconsistent and unclear. The aim of this meta-analysis was to evaluate the effectiveness of autologous platelet concentrates compared to traditional blood-clot regeneration for the management of young, immature, necrotic, permanent teeth. The digital databases MEDLINE, SCOPUS, CENTRAL, Web of Science, and EMBASE were searched to identify ten randomized clinical trials. The outcomes at postoperative follow-up, such as dentinal wall thickness (DWT), increase in root length (RL), calcific barrier formation (CB), apical closure (AC), vitality response (VR), and success rate (SR), were subjected to both qualitative synthesis and quantitative meta-analysis. The meta-analysis showed that APCs significantly improved apical closure (risk ratio (RR) = 1.17; 95% CI: 1.01, 1.37; p = 0.04) and response to vitality pulp tests (RR = 1.61; 95% CI: 1.03, 2.52; p = 0.04), whereas no significant effect was observed on root lengthening, dentin wall thickness, or success rate of immature, necrotic teeth treated with regenerative endodontics. APCs could be beneficial when treating young, immature, necrotic, permanent teeth regarding better apical closure and improved response to vitality tests.

Regenerative Endodontic Therapy with Platelet Rich Fibrin: Case Series.

Regenerative endodontic therapy (RET) provides a novel treatment modality for the immature teeth with pulp necrosis. The aim of this case series was to evaluate RET of immature permanent teeth using platelet rich fibrin (PRF) at 36-month follow-up periods. In the present case series, three immature maxillary incisors diagnosed with pulp necrosis and apical periodontitis were treated with RET. The root canals were irrigated with 1.5% sodium hypochlorite (NaOCl) and medicated with triple antibiotic paste(TAP). At the second visit, TAP was removed and root canals were conditioned with 17% EDTA. PRF was used as a scaffold. MTA was placed over PRF and the teeth were restored with composite resin. Periapical radiographs and cone beam computerized tomography(CBCT) were used to evaluate the healing. At the end of the 36-month follow-up periods, there was no response to pulp sensibility tests with cold and electric pulp tester, but all teeth showed decreased periapical lesions or evidence of healing.

Regenerative Endodontic Treatment as a Biologically Based Approach for Non-Surgical Retreatment of Immature Teeth.

INTRODUCTION: The goal of regenerative endodontic treatment (RET) is to regenerate the pulp-dentin complex in immature teeth with pulp necrosis. There is no consensus in using RET in previously treated teeth. This article reports on long-term outcomes of RET in previously treated immature teeth. METHODS: Previously treated immature teeth with signs/symptoms of failure were included. After local anesthesia and rubber dam isolation, the root canal filling material was removed without the use of chemical solvents. The root canals were irrigated with 6% sodium hypochlorite using an EndoVac (Kerr Endodontics, Culver City, CA), and a creamy mix of metronidazole and ciprofloxacin was placed in the canals for 4 weeks. At the second visit, the antibiotic dressing was removed using 17% EDTA. Bleeding was induced into the root canal space, and SynOss Putty (Collagen Matrix Inc, Oakland, NJ) was placed. Bioceramic putty was placed over the SynOss Putty, and the teeth were restored at a subsequent visit. RESULTS: Five teeth in 4 patients were included. The patients' ages ranged from 14-46 years. Recall examinations were performed from 20 to 72 months with an average of 54.4 months. The apical openings of the teeth ranged from 1-3.7 mm. All teeth were functional and asymptomatic at the recall visits. All periapical lesions resolved with partial or complete mineralization of the root canals, and all teeth showed partial to complete apical closure at the follow-up visits. CONCLUSIONS: RET can be a viable option for nonsurgical retreatment of immature teeth. This article adds a new perspective to the field of regenerative endodontics.

Efeito antibiofilme e citotoxicidade do hidróxido de cálcio associado ao óleo essencial de Melaleuca alternifolia / Antibiofilm effect and cytotoxicity of calcium hydroxide associated with the Melaleuca alternifolia essential oil

Objetivo: Avaliar e comparar os efeitos antimicrobiano e antibiofilme, e a citotoxicidade promovida pela associação do hidróxido de cálcio ao óleo essencial de Melaleuca alternifolia (MA), em diferentes concentrações, e ao propilenoglicol (PG). Métodos: As seguintes medicações compuseram os grupos experimentais: G1) HC/MA 1%; G2) HC/MA 5%; G3) HC/MA 10%; G4) HC/MA 20%; e G5) HC/PG. Solução salina 0,85% e meio DMEM serviram como controle nos testes antimicrobianos e de citotoxidade em fibroblastos do ligamento periodontal humano (FbLP), respectivamente. A atividade antimicrobiana (n = 12) foi avaliada por meio do teste de difusão em ágar. O efeito antibiofilme (n = 12) imediato das medicações foi avaliado por meio do teste de viabilidade bacteriana em biofilmes de 72 horas de E. faecalis, formados sobre discos de dentina e tratados por sete dias com as medicações. Após a coleta microbiológica do biofilme remanescente, os discos de dentina foram imersos em meio estéril e armazenados por mais sete dias, para a análise do efeito antibiofilme residual das medicações, quando nova coleta microbiológica foi realizada. A atividade metabólica de FbLP foi avaliada por meio do ensaio colorimétrico MTS (n = 9). Os valores médios dos halos de inibição, em mm, das unidades formadoras de colônia, e o percentual de atividade metabólica celular foram analisados pelos testes Kruskal-Wallis e post hoc Dunn (α = 5%). Resultados:Todas as medicações experimentais apresentaram superior ação antimicrobiana e antibiofilme comparadas ao controle, solução salina (p < 0,05), e mantiveram viáveis os FbLP, semelhante ao controle DMEM (p > 0,05). Conclusão: A associação do óleo essencial de Melaleuca alternifolia, nas concentrações de 1%, 5%, 10% e 20%, ao hidróxido de cálcio promoveu excelente ação antimicrobiana, antibiofilme e biocompatibilidade com fibroblastos, de forma semelhante à associação com propilenoglicol.

Aim:To evaluate and compare the antimicrobial and antibiofilm effect, as well as the cytotoxicity of calcium hydroxide (CH) associated with the Melaleuca alternifolia (MA)essential oil, in different concentrations, and with propylene glycol. Methods: The following medications composed the experimental groups: G1) CH/MA 1%; G2) CH/MA 5%; G3) CH/MA 10%; G4) CH/MA 20%; and G5) CH/PG. Saline solution and culture medium DMEM were used as a control in antimicrobial and cytotoxicity tests in human periodontal ligament fibroblasts (PDLF), respectively. The antimicrobial activity (n = 12) was evaluated by the disk-diffusion agar method. The immediate antibiofilm effect (n = 12) of the medications was evaluated for bacterial viability in 72 hours-biofilms of E. faecalis, formed on the dentin disc surface and treated for seven days with medications. After microbiological sampling of the remaining biofilm, the dentin discs were immersed in sterile culture medium and stored for another seven days, for analysis of the residual antibiofilm effect of the medications, when a new microbiological sampling was performed. PDLF viability was evaluated by MTS colorimetric assay (n = 9). The mean values of the inhibition halos, in mm, the colony forming units, and the metabolic cell activity percentage were analyzed by means of Kruskal-Wallis and post hoc Dunn (α = 5%) tests. Results:All of the experimental medications presented higher antimicrobial and antibiofilm effects, when compared to the saline solution control (p < 0.05), and maintained the PDLF feasible, similar to the DMEM control (p > 0.05). Conclusions:The association of the Melaleuca alternifolia essential oil, at concentrations of 1%, 5%, 10%, and 20%, with calcium hydroxide promoted an excellent antimicrobial and antibiofilm activity, and biocompatibility with fibroblasts, similarly to the association with propylene glycol.

Avaliação da atenção secundária em endodontia em um Centro de Especialidades Odontológicas (CEO) / Evaluation of secondary care in endodontics at a Dental Specialties Center (DSC)

Resumo Objetivou-se avaliar a atenção secundária em endodontia em um Centro de Especialidades Odontológicas (CEO) de Belo Horizonte (MG). A coleta de dados utilizou dois formulários: (1) sobre o tratamento endodôntico, preenchido pelos endodontistas; (2) sobre o tratamento restaurador, com dados dos prontuários. O programa SPSS 19.0 foi utilizado para analisar os resultados por meio de frequência e percentis. Foram finalizados 452 tratamentos endodônticos em pacientes adultos. Os pacientes apresentaram mediana de 39 anos de idade, a maioria era do sexo feminino (69,7%) e apresentou guia de referência da atenção básica (96,2%). A maior parte dos tratamentos endodônticos foi realizada em pré-molares superiores (23,7%), seguida de molares inferiores (22,3%), utilizando técnica mista (74,1%) e em única sessão (64,2%). O encaminhamento para o tratamento restaurador foi para o CEO em 81,2% dos casos e finalizado em 24,1% casos (n = 109). A contrarreferência após o tratamento restaurador ocorreu em 58,7% dos casos finalizados. É necessário um planejamento conjunto do tratamento odontológico entre a atenção básica e secundária e, dentro desta última, entre as especialidades, além de um adequado processo de referência e contrarreferência, visando garantir a integralidade do cuidado e a eficiência e eficácia do serviço.

Abstract This study aimed to evaluate secondary endodontic care at a Dental Specialties Center (DSC) in Belo Horizonte, MG. Data collection used two forms: (1) on endodontic treatment, completed by the endodontists (2) on the restorative treatment, with data from the medical records. The SPSS 22.0 program was used to analyze the results using frequency and percentiles. In total, 452 endodontic procedures were completed in adult patients. The patients had a median of 39 years of age, most were female (69.7%) and had a primary care referral order (96.2%). Most endodontic treatments were performed in upper premolars (23.7%) followed by lower molars (22.3%), using a mixed technique (74.1%) and in a single session (64.2%). The referral for restorative treatment was for the DSC in 81.2% of cases and finished in 24.1% (n = 109). The counter-referral following restorative treatment occurred in 58.7% of the completed cases. It is necessary to jointly plan the dental treatment between primary and secondary care and, within the latter, among the specialties, in addition to an adequate referral and counter-referral process aimed at ensuring comprehensive care and efficient and effective service.

Evaluation of secondary care in endodontics at a Dental Specialties Center (DSC). / Avaliação da atenção secundária em endodontia em um Centro de Especialidades Odontológicas (CEO).

This study aimed to evaluate secondary endodontic care at a Dental Specialties Center (DSC) in Belo Horizonte, MG. Data collection used two forms: (1) on endodontic treatment, completed by the endodontists (2) on the restorative treatment, with data from the medical records. The SPSS 22.0 program was used to analyze the results using frequency and percentiles. In total, 452 endodontic procedures were completed in adult patients. The patients had a median of 39 years of age, most were female (69.7%) and had a primary care referral order (96.2%). Most endodontic treatments were performed in upper premolars (23.7%) followed by lower molars (22.3%), using a mixed technique (74.1%) and in a single session (64.2%). The referral for restorative treatment was for the DSC in 81.2% of cases and finished in 24.1% (n = 109). The counter-referral following restorative treatment occurred in 58.7% of the completed cases. It is necessary to jointly plan the dental treatment between primary and secondary care and, within the latter, among the specialties, in addition to an adequate referral and counter-referral process aimed at ensuring comprehensive care and efficient and effective service.

Objetivou-se avaliar a atenção secundária em endodontia em um Centro de Especialidades Odontológicas (CEO) de Belo Horizonte (MG). A coleta de dados utilizou dois formulários: (1) sobre o tratamento endodôntico, preenchido pelos endodontistas; (2) sobre o tratamento restaurador, com dados dos prontuários. O programa SPSS 19.0  foi utilizado para analisar os resultados por meio de frequência e percentis. Foram finalizados 452 tratamentos endodônticos em pacientes adultos. Os pacientes apresentaram mediana de 39 anos de idade, a maioria era do sexo feminino (69,7%) e apresentou guia de referência da atenção básica (96,2%). A maior parte dos tratamentos endodônticos foi realizada em pré-molares superiores (23,7%), seguida de molares inferiores (22,3%), utilizando técnica mista (74,1%) e em única sessão (64,2%). O encaminhamento para o tratamento restaurador foi para o CEO em 81,2% dos casos e finalizado em 24,1%  casos (n = 109). A contrarreferência após o tratamento restaurador ocorreu em 58,7% dos casos finalizados. É necessário um planejamento conjunto do tratamento odontológico entre a atenção básica e secundária e, dentro desta última, entre as especialidades, além de um adequado processo de referência e contrarreferência, visando garantir a integralidade do cuidado e a eficiência e eficácia do serviço.

Effect of ethylenediaminetetraacetic acid irrigation on immune-inflammatory response in teeth submitted to regenerative endodontic therapy.

AIM: To analyse longitudinally the immune-inflammatory response in teeth of mice that underwent a regenerative protocol with or without the use of ethylenediaminetetraacetic acid (EDTA) to irrigate the root canal system. METHODOLOGY: First maxillary molars of mice were devitalized using size 10 and 15 files. Teeth were divided into the following groups: Empty - the canals were left empty; Blood Clot (BC) - the canals were filled with a blood clot; and EDTA + Blood - the canals were irrigated with 0.06 mL of 17% EDTA for 1 min and filled with a blood clot. Access cavities were restored with Coltosol® . Animals were sacrificed at 7, 14 or 21 days after the operative procedures, and teeth were collected. RNA was extracted, mRNA expression of the cytokines IGF, NGF, IL-1α, IL-10, TGF and VEGF was assessed using real-time PCR, and the anova Kruskal-Wallis test was used. RESULTS: IL-1 mRNA expression was significantly higher in the EDTA + BC group than in the Empty and BC groups at the 7th and 14th days of evaluation (P < 0.05). IL-10 mRNA expression was similar across the three groups at all time periods. TGF-ß mRNA expression in the EDTA + BC group was significantly higher on the 7th and 21st days than on the 14th (P < 0.05); at day 21, TGF-ß mRNA expression was similar between the BC and EDTA + BC groups but significantly higher than in the Empty group (P < 0.05). IGF mRNA expression was significantly higher in the EDTA + BC group than in the other groups at all time periods. VEGF mRNA expression remained unchanged throughout the experimental period in all groups (P > 0.05). NGF mRNA expression was similar amongst all groups at the 7th and 21st days (P > 0.05). At the 14th day, however, there was a significant increase in NGF mRNA expression in the EDTA + Blood group (P < 0.05) when compared with the expression in the other groups. CONCLUSION: EDTA promoted increased expression of factors that have the potential to improve the outcome of regenerative endodontic treatment.

Direct and Indirect Effect of Chlorhexidine on Survival of Stem Cells from the Apical Papilla and Its Neutralization.

INTRODUCTION: Several irrigants have been used for disinfection in regenerative endodontic procedures including chlorhexidine (CHX). In this context, the antibacterial properties of disinfectants are mainly in focus of research even though they may have an undesirable impact on the fate of stem cells. In this study, we hypothesized that CHX has both a direct effect when applied to stem cells of the apical papilla (SCAPs) and an indirect effect when SCAPs are exposed to dentin previously conditioned with CHX. METHODS: Cell toxicity was evaluated in vitro using the CellTox green fluorescence assay (Promega, Madison, WI) and CellTiter-Glo (Promega) after SCAPs were exposed directly to a dynamic concentration range of CHX; apical papilla explant cultures were stained with ApopTag (Merck Millipore, Billerica, MA) after culture with CHX. Furthermore, standardized slabs from human dentin were treated with CHX and consecutively rinsed in EDTA, L-α-lecithin (Sigma-Aldrich, St Louis, MO), or L-α-lecithin followed by EDTA. After that, SCAPs were cultured on the slabs for 5 days, and cellular viability was determined (indirect effect). Data were treated nonparametrically and analyzed using the Krukal-Wallis test (P ≤ .05). RESULTS: Direct exposure of SCAPs to CHX highly affected cell viability at concentrations above 10-3%, whereas lower concentrations had no adverse effect. During the initial 60 minutes, concentrations of 10-2% CHX or higher resulted in early pronounced toxicity with a maximum effect within 15 minutes after exposure. Likewise, CHX-conditioned dentin slabs were detrimental to SCAP survival; however, the deleterious effects were completely reversed by neutralization with L-α-lecithin. CONCLUSIONS: Chlorhexidine is toxic to SCAPs when applied directly or indirectly via conditioned dentin. If applied for a short time and neutralized by L-α-lecithin, it can be a gentle and cell-preserving disinfectant before endodontic regeneration.

Case Report: Single visit photo-activated disinfection in regenerative endodontics.

Background: Root canal disinfection is considered critical for achieving successful regenerative endodontic procedures. Photo-activated oral disinfection is a novel disinfection method that can help to achieve the goal of regenerative endodontics. This article reports the clinical and radiographic results after single visit regenerative endodontics using photo-activated oral disinfection. Methods: An 8.5-year-old girl complained of fractured upper right central incisor. Pulp necrosis was diagnosed on the basis of clinical findings. The root canal was irrigated with sodium hypochlorite solution (1.5%) followed by saline. Then, the canal was dried with paper points. A combination of a photosensitizer solution and low power laser light were applied. EDTA solution was used as a final irrigant. Bleeding was induced, followed by placement of collagen resorbable matrix and white mineral trioxide aggregate. Two days later, the tooth was sealed and restored with permanent filling. Results: Clinical findings revealed no pain on percussion or palpation tests. Radiographic examination revealed an increase in root length, an increase of apical root thickness and apical closure at the 12-month follow-up period. Conclusion: Regenerative endodontics using photo-activated oral disinfection achieved successful outcomes in the necrotic immature permanent tooth.

Regenerative surgical therapy for peri-implantitis using deproteinized bovine bone mineral with 10% collagen, enamel matrix derivative and Doxycycline-A prospective 3-year cohort study.

OBJECTIVES: There is limited evidence regarding the long-term efficacy of regenerative treatment for peri-implantitis. The aim of this study was to evaluate a combination therapy of deproteinized bovine bone mineral with 10% collagen (DBBMC), enamel matrix derivative (EMD) and Doxycycline in the regeneration of bone defects associated with peri-implantitis. METHODS: Thirty patients diagnosed with peri-implantitis (BoP/suppuration, probing depth greater than 4 mm, minimum radiographic bone loss of 20%, at least 2 years in function) were enrolled in the study. Clinical measurements included probing depths, recession, radiographic bone fill, gingival inflammation and bleeding on probing/suppuration. Following surgical access and debridement, the implant surfaces were decontaminated with 24% EDTA for 2 min, and the bone defects were filled with a combined mixture of DBBMC, EMD and Doxycycline powder. The defects were covered with connective tissue grafts where necessary. Clinical measurements were recorded after 12, 24 and 36 months. RESULTS: The mean probing depth and bone loss at the initial visit was 8.9 mm (±1.9) and 6.92 mm (±1.26), respectively. Both mean probing depth and bone loss reduced significantly from baseline to 3.55 mm (±0.50) and 2.85 mm (±0.73) at 12 months, 3.50 (±0.50) and 2.62 mm (±0.80) at 24 months and 3.50 mm (±0.50) and 2.60 mm (±0.73) at 36 months. 56.6% of the implants were considered successfully treated (according to Successful Treatment Outcome Criterion: PD < 5 mm, no further bone loss >10%, no BoP/suppuration, no recession >0.5 mm for anterior implants and >1.5 mm for posterior implants) after 36 months. CONCLUSION: Regenerative treatment of peri-implantitis using a combined mixture of DBBMC, EMD and Doxycycline achieved promising results. The benefits of this protocol incorporating EMD should be tested in randomized clinical trials.

Antibacterial Efficacy and Discoloration Potential of Endodontic Topical Antibiotics.

INTRODUCTION: The optimal concentration for the use of endodontic topical antibiotics is not known. The aims of this study were to determine the minimum bactericidal concentrations (MBCs) and minimum inhibitory concentrations (MICs) of metronidazole, ciprofloxacin, minocycline, Augmentin (GlaxoSmithKline, Research Triangle Park, NC), and tigecycline against common endodontic pathogens and to evaluate ex vivo the antibacterial efficacy and discoloration effect of triple antibiotic paste (TAP), Augmentin, and tigecycline at different concentrations using a slow-release hydrogel scaffold. METHODS: Using the Epsilometer test method (Etest; bioMérieux USA, St Louis, MO), MICs and MBCs of selected antibiotics were determined against Fusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus intermedius, and Enterococcus faecalis. Biofilms of these bacterial species were then grown in extracted single-rooted teeth anaerobically for 3 weeks. Root canals were filled with TAP, Augmentin, and tigecycline at concentrations of 1 or 0.1 mg/mL in a degradable hydrogel scaffold or pure TAP at 1 g/mL for 7 days. Coronal discoloration was evaluated spectrophotometrically at 1, 2, and 3 weeks after dressing. RESULTS: MIC/MBC data showed significant efficacy of tigecycline, Augmentin, and minocycline compared with the other antibiotics (P < .05). Significant differences were found when comparing the log10 colony-forming units of all experimental groups (P < .05). TAP at 1 g/mL had no bacterial growth but caused the greatest discoloration. Hydrogel mixtures with TAP, Augmentin, or tigecycline at 1 mg/mL significantly reduced bacterial growth and the number of positive samples compared with those at 0.1 mg/mL (P < .05) with minimal discoloration. CONCLUSIONS: TAP, Augmentin, and tigecycline in a hydrogel at 1 mg/mL reduced bacterial growth significantly with minimal color change.

Cell Homing for Pulp Tissue Engineering with Endogenous Dentin Matrix Proteins.

INTRODUCTION: Compelling evidence pinpoints that pulp tissue engineering after the transplantation of stem cells is possible. Although intriguing, severe problems regarding clinical feasibility remain. Cell homing has been proposed as a viable alternative in which dentin-derived growth factors in a conducive scaffold may attract resident cells to form pulplike tissue. In this study, an ectopic animal model for in situ dental pulp tissue engineering was developed to evaluate whether pulplike tissue formation in empty root canals after the attraction of stem cells was possible and whether this could be enhanced by dentin-derived growth factors. METHODS: Three types of fibrin (custom-made fibrin, fibrin sealant, and plasma rich in growth factors [PRGF]) as well as a self-assembling peptide were evaluated in vivo in a modified tooth root model using human teeth. Root canal dentin was conditioned with EDTA, tooth roots were filled with growth factor-laden scaffolds, and dental pulp stem cells in collagen were placed at the root tip. Constructs were implanted into immunocompromised mice for 4 weeks and subsequently analyzed histologically. Differential interference contrast and second harmonic generation imaging were performed for selected sections. RESULTS: For custom-made fibrin and fibrin sealant with dentin matrix proteins, migration into the roots and the formation of a pulplike tissue were observed, whereas the peptide-based scaffold appeared less suitable. PRGF supported tissue formation regardless of the addition of dentin matrix proteins. In the test groups with dentin matrix proteins and EDTA conditioning, differentiated odontoblastlike cells extended cellular processes into the dentinal tubules, which coincided with the deposition of the newly formed collagenous dentin matrix. CONCLUSIONS: This new cell homing model provides evidence that fibrin derivatives make applicable scaffolds and that dentin-derived proteins induce chemotaxis and pulplike tissue formation.