Use of platelet-rich fibrin for bone repair: a systematic review and meta-analysis of preclinical studies

Abstract This systematic review evaluated the potential utility of platelet-rich fibrin (PRF) in bone repair in animals. The question is: can the use of PRF in bone defects in healthy rats induce bone repair compared to clot? This systematic review was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (Prisma). The protocol was registered with Prospero (CRD [42020162319]). The literature search involved nine databases, including grey literature. All studies evaluated the bone defects created in rats filled with PRF and clots (control). Biomaterial evaluation was also performed in this study. The risk of bias was assessed using the Systematic Review Center for Laboratory Animal Experimentation (Syrcle) tool for animal studies. A meta-analysis of quantitative data was performed to estimate the effect of PRF on bone repair in rats. Heterogeneity among the studies was assessed using the I2 statistic. The literature search retrieved 685 studies, 10 of which fulfilled the eligibility criteria, and 4 were included in the quantitative assessment. Analysis of the risk of bias revealed that most studies had a high risk of bias in performance and detection. Meta-analysis yielded divergent results and the absence of a statistically significant effect: PRF with control (standardized mean difference 2.54, 95% confidence interval -0.80-5.89; p = 0.14). In general, study heterogeneity was high (I2 ≥ 75.0%). The quality of the studies that influenced the conclusion of the review was based on the PICO, the sources and form of the search, the study selection criteria, the form of evaluation of publication bias, the evaluation of the quality of the studies, and data extraction by two researchers. PRF did not provide significant benefits for bone repair, resulting in unpredictable effects.

Physicochemical properties and osteoclastogenesis for three premixed calcium silicate-based sealers post set

Abstract: Solubility, pH, ion release, cytotoxicity, and osteoclastogenesis inhibition in bone marrow-derived monocyte macrophages (BMMs) were evaluated in EndoSequence BC Sealer (END), Bio-C Sealer (BC), and Sealer Plus BC (SPBC). pH was determined after immersion of the sealers in deionized water (DW) and Minimum Essential Medium Alpha (α-MEM). Solubility was obtained by mass loss. Ion release was measured by using X-ray fluorescence spectroscopy (XRF). Cytotoxicity was evaluated by MTT assay. Inhibition of osteoclastogenesis was evaluated by tartrate-resistant acid phosphatase (TRAP). Data were analyzed using the t-test, ANOVA and Tukey/Dunnett's post-hoc tests (α = 0.05). END had the highest pH in DW (p < 0.05), and BC, in α-MEM (p < 0.05). Solubility in DW was the lowest for SPBC (p < 0.005). The highest calcium release was observed for BC in DW at 12 h (p < 0.05), and in α-MEM at 12 and 24 h (p < 0.05). The lowest toxicity was detected for END (p < 0.05). BC had the highest inhibitory effect on osteoclasts (p < 0.05). Overall, the highest solubility and pH values were found in DW. However, the calcium silicate-based sealer showed higher solubility than the ISO standards. Calcium release was the highest for BC. END showed the highest cell viability, and BC, the highest osteoclast inhibition.

Ozone disinfection for viruses with applications in healthcare environments: a scoping review

Abstract The aim of this scoping review was to provide sufficient information about the effectiveness of ozone gas in virus inactivation of surfaces and objects under different environmental conditions. The review was performed according to the list of PRISMA SrC recommendations and the JBI Manual for Evidence Synthesis for Scoping Reviews. The review was registered in Open Science Framework (OSF). EMBASE (Ovid), Lilacs, LIVIVO, MEDLINE (PubMed), SciELO, Scopus and Web of Science were primary sources, and "gray literature" was searched in OpenGray and OpenThesis. A study was included if it reported primary data on the effect of ozone gas application for vehicle-borne and airborne virus inactivation. No language or publication date restriction was applied. The search was conduct on July 1, 2020. A total of 16,120 studies were screened, and after exclusion of noneligible studies, fifteen studies fulfilled all selection criteria. Application of ozone gas varied in terms of concentration, ozone exposure period and the devices used to generate ozone gas. Twelve studies showed positive results for inactivation of different virus types, including bacteriophages, SARS-CoV-2 surrogates and other vehicle-borne viruses. Most of the studies were classified as unclear regarding sponsorship status. Although most of the population has not yet been vaccinated against COVID-19, disinfection of environments, surfaces, and objects is an essential prevention strategy to control the spread of this disease. The results of this Scoping Review demonstrate that ozone gas is promising for viral disinfection of surfaces.

Accuracy of the dental pulp sensibility test using cold spray for the diagnosis of pulp diseases: an observational clinical study

The dental pulp sensibility test is one of the main auxiliary resources for the diagnosis of pulp pathologies, and its accuracy is still debatable. This cross-sectional observational study evaluated the accuracy of the pulp sensibility test (PST) using cold spray (1,1,1,2-tetrafluoroethane) for the diagnosis of pulp diseases and determined the effect of individual and clinical variables on the reliability of this test. The paper was designed following the STROBE statement. Sixty patients with indications for primary endodontic treatment were selected and examined from August 2017 to July 2018. Data collection was performed through interviews, clinical/radiographic examinations and the PST. The results of the cold test, along with data on sex, age, the tooth type regarding the root number, and the presence of restorations and caries, as well as the recent consumption of analgesics, were recorded. The presence of bleeding within the pulp chamber was used as the gold standard to compare with the clinical diagnosis and to identify the true-positive, false-positive, true-negative, and false-negative responses. The accuracy of PST achieved in subgroups of individual and clinical variables was compared using the chi-square test with a significance level of 5% (p < 0.05). The PST with the use of cold spray showed a sensitivity of 0.88, a specificity of 1.00, a positive predictive value of 1.00, a negative predictive value of 0.86, and an accuracy of 0.93. The accuracy of the cold spray was not affected by individual or clinical variables. The PST with the use of cold spray is an accurate and reliable method for determining the diagnosis of pulp diseases, especially in cases of pulp vitality or irreversible pulpitis.

Effect of irrigation protocols on root canal wall after post preparation: a micro-CT and microhardness study

Abstract: The aim of this study was to investigate the effects of different post space irrigation protocols for removing residual filling material from dentin walls, by using microcomputed tomography (micro-CT), and the influence of these protocols on dentin microhardness. Bovine incisors (n = 35) were filled with the single-cone technique and MTA Fillapex (Angelus, Londrina, PR, Brazil). Post space preparation (PSP) was performed 7 days after filling, using the Odous Touch electrical system (Odous De Deus Ind. e Com., Belo Horizonte, MG, Brazil), followed by post space irrigation using manual irrigation, passive ultrasonic irrigation, or Easy Clean, together with 2.5% sodium hypochlorite (NaOCl), or with 2.5% NaOCl and 17% EDTA (NaOCl/EDTA). Micro-CT scans were performed at three time points. The residual filling material was evaluated at three levels: cervical, middle and apical. The Knoop test was measured with four indentations around the canal lumen at three dentin depths: X (100 μm), Y (200 μm) and Z (400 μm). Statistical analysis was performed using ANOVA (p < 0.05). The effects of the activation method (p < 0.001), and the root level (p = 0.013), as well as the interaction between the irrigant and the activation method (p = 0.041), led to different percentages of residual filling material. Lower amounts of residual filling material were observed at the cervical versus the middle and apical levels (p < 0.05). No significant differences were observed in dentin microhardness (p > 0.05). The best removal of the residual filling material was performed using the Easy Clean tip and NaOCl/EDTA, regardless of the activation methods.

Effect of ScLL and 15d-PGJ2 on viability and cytokine release in LPS-stimulated fibroblasts: an in vitro study

Abstract This study evaluated the effect of a cyclopentenone-type PG, 15-Deoxy-Δ12,14-PG J2 (15d-PGJ2), and lectin (ScLL) on the viability of human gingival fibroblasts (HGFs), and on IL-6 and TGFβ-1 release by these fibroblasts, stimulated with lipopolysaccharide (LPS). HGFs were stimulated with LPS 10 μg/ml and treated with 15d-PGJ2 1 and 2 μg/ml, and ScLL 2 and 5 μg/ml, for 1 and 3h, and then evaluated for viability by MTT assay. Supernatant was collected to detect IL-6 and TGFβ-1 release, by ELISA. Positive control was cells kept in Dulbecco's Modified Eagle's Medium, and negative control was those kept in LPS. Data were analyzed by ANOVA and Dunnett's test (α = 0.05). No significant difference was found in viability among experimental groups at 1h (p > 0.05). Percentage of ScLL 5 µg/ml viable cells was similar to that of positive control at evaluated periods (p > 0.05), whereas the other groups had lower levels than the positive control (p < 0.05). IL-6 release was statistically higher for ScLL 5 μg/ml and 15d-PGJ2 2 µg/ml at 1h, compared with the other treated groups and positive control (p < 0.05). No significant differences were found among the groups at 3h (p > 0.05), except for ScLL 2 µg/ml and 15d-PGJ2 1 µg/ml, which showed lower IL-6 release compared with that of negative control (p < 0.05). No significant difference was found among the groups for TGFβ-1 release (p > 0.05). Results indicated that ScLL 5 μg/ml did not interfere in viability, and ScLL 2 µg/ml and 15d-PGJ2 1 µg/ml demonstrated reduced IL-6 release. Tested substances had no effect on TGFβ-1 release.

Can intra-radicular cleaning protocols increase the retention of fiberglass posts? A systematic review

Abstract The presence of residues within the root canal after post-space preparation can influence the bond strength between resin cement and root dentin when using fiberglass posts (FGPs). Currently, there is no consensus in the literature regarding what is the best solution for the removal of debris after post-space preparation. This systematic review involved "in vitro" studies to investigate if cleaning methods of the root canal after post-space preparation can increase the retention of FGPs evaluated by the push-out test. Searches were carried out in PubMed (MEDLINE) and Scopus databases up to July2017. English language studies published from 2007 to July 2017 were selected. 475 studies were found, and 9 were included in this review. Information from the 9 studies were collected regarding the number of samples, storage method after extraction, root canal preparation, method of post-space preparation, endodontic sealer, resin cement, cleaning methods after post-space and presence of irrigant activation. Five studies presented the best results for the association of sodium hypochlorite (NaOCl) and ethylenediamine tetra-acetic acid (EDTA), while in the other 4 studies, the solutions that showed improved retention of FGPs were photon-induced photoacoustic streaming (PIPS), Qmix, Sikko and EDTA. The results showed heterogeneity in all comparisons due to a high variety of information about cleaning methods, different concentrations, application time, type of adhesive system and resin cements used. In conclusion, this review suggests that the use of NaOCl/EDTA results in the retention of FGPs and may thus be recommended as a post-space cleaning method influencing the luting procedure.

A 12-Year Retrospective Study of Avulsion Cases in a Public Brazilian Dental Trauma Service

Abstract This study assessed the epidemiological characteristics and management of the permanent teeth avulsion cases attended in a Brazilian dental trauma service from December 2005 to August 2016. A retrospective study was conducted of case records of 93 patients involving 139 avulsed teeth. Data included sex, age, trauma etiology, location of the accident, number and position of avulsed teeth, and presence and type of associated traumatic lesions. Management of the avulsed teeth was addressed as: time elapsed until teeth were retrieved from the accident's location; teeth's cleaning method and storage media; time elapsed until seeking treatment and replantation. The majority of the patients were children from 6-10 (31.2%) and 11-15 years old (26.9%). Male patients were more affected than female. Bicycle accident was the main etiological factor (31.2%). In 56 (60.2%) cases, traumatic lesions to neighboring teeth were present. In 55 (59.1%) cases, lesions to adjacent soft tissues were reported. In 82 (88.2%) cases, patients requested treatment at the same day of the accident. Sixty-four teeth (46.0%) were immediately retrieved and 28 (20.1%) were not found. Forty-two teeth (30.2%) were kept dry. Only one tooth (0.7%) was immediately replanted at the accident's site, while 51 teeth (36.7%) were not replanted. Numerous avulsed teeth were inappropriately managed and immediate replantation was not frequent. Public policies must be created to raise awareness towards the particularities of avulsion cases.

Resumo Este estudo avaliou as características epidemiológicas e de manejo dos casos de avulsão de dentes permanentes atendidos em um serviço de trauma dental de dezembro de 2005 a agosto de 2016. Foi realizado um estudo retrospectivo de 93 casos, envolvendo 139 dentes avulsionados. Os dados incluíram sexo, idade, etiologia do trauma, localização do acidente, número e posição dos dentes avulsionados e presença e tipo de lesões traumáticas associadas. O manejo dos dentes foi abordado de modo a analisar: o tempo decorrido até que os dentes fossem recuperados do local do acidente; Método de limpeza dos dentes e meios de armazenamento; Tempo decorrido até a busca por tratamento e reimplante dental. A maioria dos pacientes eram crianças de 6-10 (31,2%) e 11-15 anos (26,9%). Os pacientes do sexo masculino foram mais acometidos que do feminino. O acidente de bicicleta foi o principal fator etiológico (31,2%). Em 56 (60,2%) casos, ocorreram lesões traumáticas aos dentes vizinhos. Em 55 (59,1%) casos foram relatadas lesões de tecidos moles. Em 82 (88,2%) casos, os pacientes solicitaram tratamento no mesmo dia do acidente. Sessenta e quatro dentes (46,0%) foram imediatamente recuperados e 28 (20,1%) não foram encontrados. Quarenta e dois dentes (30,2%) foram mantidos secos. Apenas um dente (0,7%) foi imediatamente reimplantado no local do acidente, enquanto 51 dentes (36,7%) não foram reimplantados. Numerosos dentes avulsionados foram manejados de forma inadequada e o reimplante imediato não foi frequente. Devem ser criadas políticas públicas para a conscientização da população sobre as particularidades dos casos de avulsão dental.

Evaluation of coconut water neutralized by different agents on the viability of human fibroblasts: an in vitro study / Avaliação da água de coco neutralizada por diferentes agentes na viabilidade de fibroblastos humanos: estudo in vitro

Objective: This study evaluated four types of pH adjustment of the coconut water (CW) on viability of human fibroblasts (HFF). Material and method: Natural and industrialized CW were adjusted to pH 7.0 using: (1) Sodium Hidroxide (NaOH), (2) Sodium bicarbonate (NaHCO3), (3) Triethanolamine (C6H15NO3), (4) 2-Amino-2-Methil-1-Propanol (C4H11NO). Fibroblasts were plated at 2×104/ well in 96 well plates and maintained in the CW solutions for 2 h and 4 h. Positive control was represented by HFF maintained in DMEM and the negative control by tap water. Cell viability was analyzed by MTT formazan method. Data were analyzed by 3-way ANOVA followed by Tukey's and Dunnet's test. Result: There are no significant effect on the cell viability regarding type of CW, period of evaluation, and the interactions between CW and period of evaluation, CW and pH adjustment method, pH adjustment method and period of evaluation (p>0.05). Conclusion The product used for CW pH adjustment did not influenced HFF viability, thought there are a tendency of better performance in natural CW.

Objetivo: Avaliar a eficácia de quatro tipos de substâncias usadas para ajuste do pH da água de coco (AC) sobre a viabilidade de fibroblastos humanos (HFF). Material e método: O pH da AC natural e industrializada foi ajustado para pH 7,0 utilizando: (1) Hidróxido de Sódio (NaOH), (2) bicarbonato de sódio (NaHCO3), (3) Trietanolamina (C6H15NO3), (4) 2-Amino-2- Methil-1-propanol (C4H11NO). Células HFF foram plaqueadas em 2×104 células/poço em placas de 96 poços e mantidas nas diferentes soluções de AC acima durante 2 h e 4 h. Controle positivo foi representado por HFF mantidas em DMEM e o controle negativo por água da torneira. A viabilidade celular foi avaliada pelo método de MTT Formazan. Os dados foram analisados por 3-way ANOVA seguido pelo teste de Tukey e Dunnett. Resultado: A viabilidade celular não é influenciada pelo período de avaliação, e as interações entre AC e período de avaliação, AC e método de ajuste de pH, método de ajuste de pH e período de avaliação (p> 0,05). Conclusão: O produto utilizado para ajuste do pH não interfere na viabilidade de FH, embora, haja uma tendência de melhor desempenho em AC natural.

Biological Effects of a Root Conditioning Treatment on Periodontally Affected Teeth - An In Vitro Analysis

Abstract The aim of this study was to evaluated the root surfaces modifications resulted by application of different chemicals agents, and their influence on the fibrin network and fibroblasts attachment. From 96 anterior mandibular human extracted incisor teeth, 192 dentin blocks of buccal and lingual surface were obtained and randomly divided into 6 groups: Cont- control group, which received no treatment; Root surface scaling and root planing (Srp); Citric acid-Srp; EDTA-Srp; Tetracycline capsule-Srp; Tetracycline gel-Srp. After dentin treatments the specimens were analyzed as follows: 1) demineralization level and residues of the product by scanning electron microscopy (SEM); 2) adhesion of blood components after 20 min of surface treatment by SEM; 3) fibroblast attachment after 24 h by SEM; 4) cell metabolism after 24 h by MTT assay. Data were analyzed using Fisher Exact, One-way ANOVA test followed by Dunn's test, Tukey test and Dunnett test (α=0.05). Citric acid, EDTA and Tetracycline gel resulted in adequate demineralization with no completely smear layer and smear plug removal on root dentin surface. Tetracycline capsule produced great tetracycline residues with several demineralization areas. Tetracycline gel and EDTA groups presented more fibroblast fixation than other experimental groups. The highest mean blood clot adhesion score was observed in roots treated with tetracycline gel. EDTA and Tetracycline gel surface treatment removed the smear layer over dentin surface and promoted adhesion of fibrin network and fibroblast cells attachment.

Resumo O objetivo deste estudo foi avaliar modificações nas superfícies radiculares sofridas pela aplicação de diferentes agentes químicos, e sua influência sobre a rede de fibrina e adesão de fibroblastos. A partir de 96 incisivos inferiores humanos, 192 blocos de dentina das superfícies vestibular e lingual foram obtidos e divididos aleatoriamente em 6 grupos: Cont-grupo controle, não recebeu nenhum tratamento; Raspagem e alisamento radicular (RAR); Ácido cítrico-SRP; EDTA-SRP; Tetraciclina em cápsula-SRP; Tetraciclina gel-SRP. Após o tratamento da dentina as espécimes foram analisadas: 1, nível de desmineralização e resíduos do produto por microscopia eletrônica de varredura (MEV); 2, adesão dos componentes sanguineos após 20 min na tratamento de superfície por SEM; 3, adesão de fibroblastos após 24h por SEM; 4, o metabolismo celular após 24 h por ensaio MTT. Os dados foram analisados por Fisher Exact, teste one-way ANOVA, seguido pelo teste de Dunn, teste de Tukey e teste de Dunnett (α = 0,05). O ácido cítrico, EDTA e gel tetraciclina resultaram na adequada desmineralização sem remoção completa de camada de smear layer e smear plug sobre a superfície da dentina radicular. Cápsula de tetraciclina produziu grandes resíduos de tetraciclina com várias áreas de desmineralização. Os grupos Gel de tetraciclina e EDTA apresentaram maior adesão de fibroblastos do que os demais grupos experimentais. O maior score de adesão de coágulo sanguineo foi observado nas superfícies tratadas com gel de tetraciclina. EDTA e Gel de tetraciclina removeram a camada de smear layer sobre a superfície da dentina e promoveu adesão da rede de fibrina e de fibroblastos.

Effect of lectin (ScLL) on fibroblasts stimulated with LPS - an in vitro study

Abstract: The lectin (ScLL) extracted from the Synadenium carinatum plant has been evaluated as an immunomodulator in diseases such as asthma, neosporosis and leishmaniasis. However, it has not yet been evaluated in the oral cavity. This study evaluated the effect of ScLL on viability, proliferation and release of IL-10 in human gingival fibroblasts (HGF) stimulated with lipopolysaccharide (LPS). HGF were stimulated with LPS 1 µg/ml and treated with ScLL in concentrations of 10, 5 and 2 µg/ml for 1 and 5 h, and evaluated by flow cytometry for viability, apoptosis (initial/advanced) and necrosis. The supernatant was collected to detect release of IL-10 by ELISA. The proliferation was assessed with the BrdU assay. Positive control consisted of cells maintained in Dulbecco's Modified Eagles Medium (DMEM), and the negative control, of those kept in tap water. Data were analyzed by ANOVA and Dunnett's test (α = 0.05). No significant difference was found for ScLL concentrations regarding viability or initial and advanced apoptosis (p=0.455). All the groups, including the positive control, had a significantly lower necrosis parameter than negative control at 5 h (p < 0.001). No difference was found for proliferation among the experimental groups (p = 0.832). ScLL at 5 and 2 µg/ml resulted in a lower release of IL-10 than positive and negative controls at 5 h (p = 0.047). The results indicated that ScLL concentrations tested were not cytotoxic, and had no effect on proliferation and release of IL-10 parameters. A thorough understanding of ScLL, regarding its immunomodulatory potential, may open the door to new perspectives for dentistry.

Influence of Implant Surfaces on Osseointegration: A Histomorphometric and Implant Stability Study in Rabbits

Abstract: The aim of this study was to evaluate the stability and osseointegration of implant with different wettability using resonance frequency analysis (RFA) and histomorphometric analysis (bone implant contact, BIC; and bone area fraction occupied, BAFO) after 2 and 4 weeks in rabbit tibiae. Thirty-two Morse taper implants (length 7 mm, diameter 3.5 mm) were divided according to surface characteristics (n=8): Neo, sandblasted and dual acid-etched; and Aq, sandblasted followed by dual acid-etched and maintained in an isotonic solution of 0.9% sodium chloride. Sixteen New Zealand rabbits were used. Two implants of each group were installed in the right and left tibiae according to the experimental periods. The RFA (Ostell(r)) was obtained immediately and after the sacrifice (2 and 4 weeks). The bone/implant blocks were processed for histomorphometric analysis. Data were analyzed using two-way ANOVA followed by Tukey's test and Pearson's correlation for ISQ, BIC and BAFO parameters (p=0.05). No significant effect of implant, period of evaluation or interaction between implant and period of evaluation was found for BIC and BAFO values (p>0.05). Only period of evaluation had significant effect for RFA values at 4 weeks (p=0.001), and at 2 weeks (p<0.001). RFA values were significantly higher at the final period of evaluation compared with those obtained at early periods. There was a significant correlation between BIC values and BAFO values (p=0.009). Both implant surfaces, Aq and Neo, were able to produce similar implant bone integration when normal cortical bone instrumentation was performed.

Resumo: O objetivo deste estudo foi avaliar a estabilidade e osseointegração de implantes com superfícies com diferentes molhabilidades empregando análise de frequência de ressonância (RFA) e histomorfometria (contato implante ósseo, BIC, e fração de área óssea ocupada, BAFO), nos períodos de 2 e 4 semanas em tíbias de coelhos. Trinta e dois implantes cone Morse (comprimento 7mm, diâmetro 3,5 mm), foram divididos de acordo com tratamento de superfície (n = 8): Neo, superfície jateada e condicionada com ácido; e Aq, superfície jateada e condicionada com ácido e mantida em solução isotônica de cloreto de sódio a 0,9%. Dezesseis coelhos tipo Nova Zelândia foram utilizados neste estudo. Dois implantes de cada grupo foram instalados nas tíbias direita e esquerda de acordo com os períodos experimentais. Os valores de RFA (Ostell(r)) foram obtidos imediatamente e após o sacrifício (2 e 4 semanas). Os blocos ósseos/implante foram processados para análise histomorfométrica. Os dados foram analisados usando ANOVA fatorial seguido pelo teste de Tukey e também por meio de correlação de Pearson para os fatores RFA, BIC e BAFO (P=0,05). Nenhum efeito significativo dos fatores tipo de implante, período de avaliação e da interação entre o tipo de implante e período de avaliação foram observados para os valores de BIC e BAFO. Apenas o período de avaliação resultou em efeito significativo para valores RFA após 2 semanas (p=0,001), e 4 semanas (p<0,001). Os valores de RFA valores foram significativamente mais elevados no final do período de avaliação em comparação com os obtidos em inicialmente. Houve correlação significativa entre os valores BIC e BAFO (p=0,009). Ambas as superfícies de implantes, Aq e Neo, são capazes de produzir adequada integração osso/implante em condição normal de instrumentação do osso cortical.

Biological characterization of implant surfaces - in vitro study / Caracterização biológica de superfície de titânio - estudo in vitro

Objective: Evaluate the biological performance of titanium alloys grade IV under different surface treatments: sandblasting and double etching (Experimental surface 1; Exp1, NEODENT); surface with wettability increase (Experimental surface 2; Exp2, NEODENT) on response of preliminary differentiation and cell maturation. Material and method: Immortalized osteoblast cells were plated on Exp1 and Exp2 titanium discs. The polystyrene plate surface without disc was used as control group (C). Cell viability was assessed by measuring mitochondrial activity (MTT) at 4 and 24 h (n = 5), cell attachment was performed using trypan blue exclusion within 4 hours (n = 5), serum total protein and alkaline phosphatase normalization was performed at 4, 7 and 14 days (n = 5). Data were analyzed using one-way ANOVA and Tukey test. Result: The values of cell viability were: 4h: C - 0.32±0.01A; Exp1 - 0.34±0.08A; Exp2 - 0.29±0.03A. 24h: C - 0.43±0.02A; Exp1 - ; 0.39±0.01A; Exp2 - 0.37±0.03A. The cell adhesion counting was: C -85±10A; Exp1- 35±5B; Exp2& - 20±2B. The amounts of serum total protein were 4d: C - 40±2B; Exp1 - 120±10A; Exp2 -130±20A. 7d: C 38±2B; Exp1 - 75±4A; Exp2 -70±6A. 14 d: C - 100±3A; Exp1 - 130±5A; Exp2 - 137±9A. The values of alkaline phosphatase normalization were: 4d: C - 2.0±0.1C; Exp1 - 5.1±0.8B; Exp2 - 9.8±2.0A<. 7d: C -1.0±0.01C; Exp1 - 5.3±0.5A; Exp2 - 3.0±0.3B. 14 d: C - 4.1±0.3A; Exp1 - 4.4±0.8A; Exp2 - 2.2±0.2B. ...

Objetivo: Avaliar o desempenho biológico de ligas de titânio grau IV submetidos a diferentes tratamentos de superfície - jateamento e duplo ataque ácido (Superfície experimental 1; Exp1, NEODENT) e superfície com aumento na molhabilidade (Superfície experimental 2; Exp2, NEODENT) em resposta preliminar de diferenciação e maturação celular. Material e método: Foram plaqueados osteoblastos imortalizados sobre discos de titânio de Exp1 e Exp2 e como controle o poço da placa de cultura sem disco (C). Empregou-se ensaios de viabilidade celular (MTT) em 4 e 24 horas (n = 5), adesão celular em 4 horas (n = 5), dosagem de proteínas totais e fosfatase alcalina normalizada em 4, 7 e 14 dias (n = 5). Os dados foram analisados por ANOVA em fator único seguido de teste de Tukey. Resultado: Os valores de viabilidade celular foram: 4h: C - 0,32±0,01A; Exp1 - 0,34±0,08A; Exp2 - 0,29±0,03A. 24h: C - 0,43±0,02A; Exp1 - 0,39±0,01A; Exp2 - 0,37±0,03A. A contagem de adesão celular foi: C - 85±10A; Exp1 - 35±5B; Exp2 - 20±2B. Os valores de proteínas totais foram: 4d: C - 40±2B; Exp1 - 120±10A; Exp2 - 130±20A. 7d: C - 38±2B; Exp1 - 75±4A; Exp2 - 70±6A. 14 d: C - 100±3A; Exp1 - 130±5A; Exp2 - 137±9A. Os valores de fosfatase alcalina normalizada foram: 4d: C - 2,0±0,1C; Exp1 - 5,1±0,8B; Exp2 - 9,8±2,0A, 7d: C - 1,0±0,01C; Exp1 - 5,3±0,5A; Exp2 - 3,0±0,3B, 14 d: C - 4,1±0,3A; Exp1 - 4,4±0,8A; Exp2 - 2,2±0,2B. Letras diferentes representam ...

Avaliação da reação tecidual a cimentosendodônticos recém-manipulados e suarelação com o subtipo de macrófago M1 / Evaluation of tissue reaction to freshly mixed endodontic sealers and their relationship with the M1 macrophage phenotype

Esse estudo avaliou a resposta tecidualpara cimentos recém-manipulados (Sealer 26, ou SE26;e Epiphany, ou EPH) e a relação entre macrófagos M1e reparação tecidual. Material: foram utilizados 21 ratospara avaliar a resposta tecidual aos 7, 14 e 21 diaspós-implantação. Metade das seções histopatológicasfoi marcada com anticorpos anti-iNOS (M1 macrófagossubconjunto), e metade foi classificada de acordo coma intensidade inflamatória. Resultados: no dia 7, nãofoi encontrada diferença significativa na intensidade dareação inflamatória entre os cimentos. Nos dias 14 e21, SE26 mostrou uma reação inflamatória mais intensado que o EPH (p < 0,05). Nos dias 7 e 14, o númerode células iNOS+ foi significativamente maior do queem EPH SE26 (p < 0,05). Conclusões: SE26 foi maistóxico do que o EPH. O grau de inflamação observadaem EPH teve relação inversa com a quantidade de macrófagosM1 observados.

Effects of Titanium Surfaces on the Developmental Profile of Monocytes/Macrophages

Due to the critical role of monocytes/macrophages (Mϕ) in bone healing, this study evaluated the effects of bio-anodized, acid-etched, and machined titanium surfaces (Ti) on Mϕ behavior. Cells were separated from whole human blood from 10 patients, plated on Ti or polystyrene (control) surfaces, and cultured for 72 h. At 24, 48 and 72 h, cell viability, levels of IL1β, IL10, TNFα, TGFβ1 inflammatory mediators, and nitric oxide (NO) release were analyzed by mitochondrial colorimetric assay (MTT assay) and immunoenzymatic assays, respectively. Real-time PCR was used to verify the expression of TNFα and IL10 at 72 h. The data were subjected to a Kruskal-Wallis analysis. IL1β, TNFα and TGFβ1 release were not significantly different between the Ti surfaces (p>0.05). The presence of NO and IL10 was not detected in the samples. Cell viability did not differ between the samples cultivated on Ti and those cultivated on control surfaces, except at 24 h (p=0.0033). With respect to the mediators evaluated, the surface characteristics did not induce a typical Th1 or Th2 cytokine profile, although the cell morphology and topography were influenced by the Ti surface during the initial period.

Devido ao papel crítico dos monócitos / macrófagos (Mϕ) na cicatrização óssea, este estudo avaliou os efeitos de superficies de titânio (Ti) bio-anodizada, ataque ácido e usinada sobre o comportamento Mϕ. As células foram separadas a partir de sangue humano de 10 pacientes, plaqueadas em Ti ou superfícies de poliestireno (controle), e cultivadas durante 72 h. Às 24, 48 e 72 h, a viabilidade celular e IL1β, IL10, TNFα, TGFβ1 e liberação de óxido nítrico foram analisados por ensaio colorimétrico mitocondrial (MTT) e ensaios imunoenzimáticos, respectivamente. PCR em tempo real foi utilizado para verificar a expressão de TNFα e IL-10 às 72 h. Os dados foram submetidos a uma análise de Kruskal-Wallis. IL1β autorização, TNFα e TGFβ1, não foram significativamente diferentes entre as superfícies de Ti (p>0,05). A presença de NO e de IL-10 não foi detectada nas amostras. A viabilidade celular não diferiu entre as amostras cultivadas em Ti e aquelas cultivadas em superfícies de controle, exceto às 24 h (p=0,0033). No que diz respeito aos mediadores avaliados, as características da superfície, não induziu resposta típica de citocinas Th1 ou Th2, embora a morfologia da célula e a topografia foram influenciadas pela superfície de Ti, durante o período inicial.

Levels of Immunoglobulin A1 in Peri-Implant Fluid and Saliva from Patients with Mucositis: A Preliminary Study

There are no studies evaluating the possible use of immunoglobulin A1 (IgA1) as an early marker for peri-implant inflammation. The aim of this study was to evaluate the IgA1 levels in peri-implant sulcular fluid (PISF) and saliva of partially edentulous patients as an indicator of mucositis. Twenty-seven patients were examined to determine the peri-implant status based on probing depth and bleeding on probing. Saliva and PISF around dental implants were collected and the IgA1 levels were evaluated by Elisa assay. IgA1 in saliva and PISF of these patients were compared and their correlations with clinical parameters were evaluated. Differences in IgA1 levels in saliva (821.1 ± 290.6; 779.8 ± 401.5) and PISF (26.6 ± 20.7; 25.1 ± 20.5) of healthy and mucositis groups, respectively were not observed (p>0.05). Correlation between clinical parameters and IgA1 in saliva or PISF was not observed in healthy or mucositis groups (p=0.607; p=0.826, respectively). These results suggest that IgA1 cannot be used as an immunological marker of mucositis.

Não existem estudos que avaliem a utilização de imunoglobulina A1 (IgA1) como marcador precoce da inflamação peri-implantar. O objetivo deste estudo foi avaliar os níveis de IgA1 do fluido sulcular peri-implantar (PISF) e saliva de pacientes parcialmente desdentados como indicador da mucosite. Vinte e sete pacientes foram examinados para determinar a condição peri-implantar com base na profundidade de sondagem e sangramento à sondagem. Saliva e PISF ao redor de implantes dentários foram coletados e os níveis IgA1 foram avaliados pelo teste Elisa. IgA1 na saliva e PISF destes pacientes foram comparados e suas correlações com parâmetros clínicos foram avaliados. Não foram observadas diferenças nos níveis de IgA1 (821,1 ± 290,6; 779,8 ± 401,5) na saliva e PISF (26,6 ± 20,7; 25,1 ± 20,5) de grupos saudáveis e mucosite, respectivamente (p>0,05). Correlação entre os parâmetros clínicos e IgA1 na saliva ou PISF não foi observada em grupos saudáveis ou mucosite (p=0,607; p=0,826, respectivamente). Estes resultados demonstraram que IgA1 não pode ser utilizada como marcador imunológico da mucosite.

Avaliação histoquímica da permeabilidade dentinária após a utilização do edta como auxiliar na irrigação do canal radicular / Histochemical evaluation of dentin permeability after using edta as auxiliar irrigation of the root canal

O presente estudo avaliou a permeabilidade dentinária em toda a extensão do canal radicular após o uso do EDTA associado a diferentes concentrações de NaOCl. Foram utilizados cinquenta incisivos bovinos divididos em cinco grupos experimentais (n=10) de acordo com as soluções irrigantes empregadas: G1- NaOCl 1%; G2- NaOCl 5,25%; G3- NaOCl 1% + EDTA; G4- NaOCl 5,25% + EDTA; G5- Água destilada (grupo controle). Após o preparo biomecânico, as raízes foram submetidas ao método de coloração histoquímico com sulfato de cobre a 10% e ácido rubeânico a 1% para identificação da permeabilidade dentinária. Para a análise morfométrica as amostras foram seccionadas transversalmente, obtidas três secções de cada terço, desidratadas e clarificadas. Analisadas em microscopia óptica para quantificar a penetração dos íons cobre na dentina radicular. Após a análise estatística dos dados, os resultados indicaram que, houve diferença significativa entre os terços analisados seguindo uma ordem de efetividade do terço cervical maior que o médio e este maior que o apical. Entretanto, nenhuma diferença significante em relação à permeabilidade foi encontrada quando se comparou as diferentes soluções estudadas. Concluiu-se que o uso do EDTA associado com diferentes concentrações de hipoclorito de sódio, como irrigante final, não promoveu alteração na permeabilidade dentinária no canal radicular em relação as soluções usadas isoladamente.

This study evaluated the dentine permeability on the length of the root canal after the use of EDTA associated with different concentrations of NaOCl. Fifty bovine incisors were used divided into five experimental groups (n = 10) according to the irrigating solutions used: G1- 1% NaOCl; G2- 5.25% NaOCl; G3- 1% NaOCl + EDTA; G4- 5.25% NaOCl + EDTA; G5- distilled water (control group). After biomechanical preparation, the roots were subjected to histochemical staining method with 10% copper sulfate and then in 1% rubeanic acid alcohol solution for identification of dentin permeability. For the morphometric analysis the samples were cross sectioned, obtained three sections of each third, dehydrated and clarified. Analyzed with light microscopy to quantify the penetration of copper ions in the root dentin. After statistical analysis, the results indicated that there were significant differences among thirds analyzed by an order of effectiveness of the cervical third larger than the middle and this larger than the apical. However, no significant difference in relation to the permeability was found when comparing the different solutions studied. In conclusion, the use of EDTA associated with different concentrations of sodium hypochlorite, as final rinse, did not cause changes in dentin permeability in relation solutions used alone.

Soy milk as a storage medium to preserve human fibroblast cell viability: an in vitro study

Soy milk (SM) is widely consumed worldwide as a substitute for cow milk. It is a source of vitamins, carbohydrates and sugars, but its capacity to preserve cell viability has not been evaluated. The purpose of the present study was to investigate the efficacy of SM to maintain the viability of human fibroblasts at short periods compared with different cow milks. Human mouth fibroblasts were cultured and stored in the following media at room temperature: 10% Dulbecco's Modified Eagle Medium (DMEM) (positive control group); long shelf-life ultra-high temperature whole cow milk (WM); long shelf-life ultra-high temperature skim cow milk (SKM); powdered cow milk (PM); and soy milk (SM). After 5, 15, 30 and 45 min, cell viability was analyzed using the MTT assay. Data were analyzed statistically by the Kruskal-Wallis test with post-analysis using the Dunn's method (α=0.05). SKM showed the lowest capacity to maintain cell viability in all analyzed times (p<0.05). At 30 and 45 min, the absorbance levels in control group (DMEM) and SM were significantly higher than in SKM (p<0.05). Cell viability decreased along the time (5-45 min). The results indicate that SM can be used as a more adequate storage medium for avulsed teeth. SKM was not as effective in preserving cell viability as the cell culture medium and SM.

O leite de soja (LS) é largamente consumido em todo o mundo como substituto para o leite bovino. Este é uma fonte de vitaminas, carboidratos e açúcares, mas a sua capacidade para preservar a viabilidade celular não foi avaliada. A finalidade do estudo foi investigar a eficácia do LS em manter a viabilidade de fibroblastos humanos em períodos curtos em comparação com diferentes leites bovinos. Fibroblastos de boca humanos foram cultivados e armazenados nos seguintes meios à temperatura ambiente: 10% de meio Dulbecco's Modified Eagle (DMEM) (grupo controle positivo); leite bovino integral longa vida (LI); leite bovino desnatado longa vida - LD; leite em pó - LP; leite de soja - LS. Depois de 5, 15, 30 e 45 min, a viabilidade celular foi analisada usando o método de MTT. Os dados foram analisados estatisticamente pelo teste de Kruskal-Wallis e posteriormente usando o método de Dunn (α=0,05). O grupo LD apresentou a menor capacidade para manter a viabilidade celular em todos os tempos analisados (p<0,05). Aos 30 e aos 45 min, os níveis de absorbância no grupo controle (DMEM) e LS foram significativamente maiores que no grupo LD (p<0,05). A viabilidade celular diminuiu ao longo do tempo (5-45 min). Os resultados indicaram que LS pode ser usado como meio de armazenamento mais adequado para dentes avulsionados. LD não foi eficaz na preservação da viabilidade das células como o meio de cultura de células e o LS.

Cytotoxic response of two cell lines exposed in vitro to four endodontic sealers

Aim: To investigate the cytotoxicity of four endodontic sealers with different bases – Epiphany(EPH), AH Plus (AHP), Sealer 26 (S26) and Endofill (ENF) – on human foreskin fibroblasts(HFF) and mouse macrophages (J774/G8). Methods: Cells were placed in direct contact withfreshly prepared endodontic sealers in polypropylene tubes. The cells were incubated for 24, 48and 72 h. Cytotoxicity was assessed using the MTT assay (cell viability) and Griess reagent (NOrelease). Results: On the HFF cultures, EPH showed the lowest viability levels of all four sealersat 24 h (p<0.05), but over time (72h), EPH lessened its toxic levels in a similar pattern as the otherthree materials (p>0.05). The viability of all four sealers on the macrophage cultures showed nostatistically significant difference over time, except between EPH and AHP at 72 h (p<0.05).Although uniformity was not detected in macrophage and fibroblast release of NO in response tosealers over time, a trend of increased NO levels for EPH (p<0.05) was observed. Conclusions:The response pattern varied depending on time and type of cell line used for analysis, althoughthe results indicate a higher cytotoxicity for EPH in short-term tests.

Cytotoxicity of bovine and porcine collagen membranes in mononuclear cells

This study compared the cytotoxicity and the release of nitric oxide induced by collagen membranes in human mononuclear cells. Peripheral blood was collected from each patient and the separation of mononuclear cells was performed by Ficoll. Then, 2x10(5) cells were plated in 48-well culture plates under the membranes in triplicate. The polystyrene surface was used as negative control. Cell viability was assessed by measuring mitochondrial activity (MTT) at 4, 12 and 24 h, with dosage levels of nitrite by the Griess method for the same periods. Data had non-normal distribution and were analyzed by the Kruskal-Wallis test (p<0.05). Statistically significant differences (p<0.05) were observed between the membranes and the control in the experimental period, although there was a significant reduction in viability over time (p<0.01). At 4 and 12 h, the porcine membrane induced a higher release of nitrite compared with the control and bovine membrane, respectively (p<0.01), and this difference was maintained at 24 h (p<0.05). This in vitro study showed that the porcine collagen membrane induces an increased production of proinflammatory mediators by mononuclear cells in the first hours of contact, decreasing with time.

Neste estudo foi comparada a citoxicidade e a liberação de nitrito induzidos por membranas de colágeno bovino e suíno em células mononucleares humanas. Foram coletados sangue periférico de cada paciente, e realizada separação de mononucleares por gradiente de Ficoll. Um total de 2x10(5) células foram plaqueadas em placas de cultura de 48 poços sob as membranas, em triplicata. O poço sem membrana serviu como controle negativo. A viabilidade celular foi avaliada medindo a atividade mitocondrial (MTT) em 4,12 e 24 h, com dosagens dos níveis de nitrito pelo método de Griess nos mesmos períodos. As amostras não apresentaram distribuição normal, sendo realizado o teste de Kruskal-Wallis (p<0,05). Foram observadas diferenças estatisticamente significantes entre as membranas e o controle nos período analisados (p<0,05), embora tenha ocorrido redução da viabilidade em função do tempo (p<0,01). Em 4 e 12 h a membrana suína induziu maior liberação de nitrito comparado ao controle e à membrana bovina, respectivamente (p<0,01). Tal diferença foi mantida em 24 h (p<0,05). Este estudo in vitro demonstrou que a membrana colágena suína induz uma maior produção de mediador pró-inflamatório pelas células mononucleares nas primeiras horas de contato, diminuindo com o tempo.