Bending Resistance and Cyclic Fatigue Life of Reciproc, Unicone, and WaveOne Reciprocating Instruments.

INTRODUCTION: This study evaluated the bending resistance and cyclic fatigue life of a new single-file reciprocating instrument (Unicone; Medin, Nové Mesto na Morave, Czech Republic). Reciproc (VDW, Munich, Germany) and WaveOne (Dentsply Maillefer) instruments were used as references for comparison. METHODS: Flexibility was determined by 45° bending tests using a universal testing machine. The cyclic fatigue test was performed using a custom-made device. For this test, an artificial canal with a 60° angle and a 5-mm radius of curvature was used. Scanning electron microscopic analysis was performed to determine the mode of fracture and possible deformations at the helical shaft. Statistical analysis for the bending resistance test was performed using parametric methods (ie, 1-way analysis of variance). Post hoc pair-wise comparisons were performed using the Tukey test for multiple comparisons (P < .05). Weibull analysis was used to calculate the mean life, beta, and eta parameters. RESULTS: Reciproc presented significantly lower bending resistance than the other tested systems (P < .05), whereas no differences were observed between WaveOne and Unicone (P > .05). When mean life was compared among the brands, Reciproc lasted longer than WaveOne with a probability of 99.9%, longer than Unicone in the "RECIPROC ALL" mode with a probability of 99.9%, and longer than Unicone in the "WAVEONE ALL" mode with a probability of 99.9% (all statistically significant). Moreover, WaveOne lasted longer than Unicone in the "RECIPROC ALL" mode with a probability of 98.5% and longer than Unicone in the "WAVEONE ALL" mode with a probability of 99.8% (all statistically significant). Finally, Unicone in the "RECIPROC ALL" mode lasted longer than Unicone in the "WAVEONE ALL" mode with a probability of 95.3% (statistically significant). CONCLUSIONS: The new reciprocating instrument Unicone showed lower cyclic fatigue resistance compared with Reciproc R25 and WaveOne Primary files.

Long-Term Cytotoxicity, pH and Dissolution Rate of AH Plus and MTA Fillapex.

The aim of the present study was to verify the long-term cytotoxic effects of the MTA Fillapex and to compare them with AH Plus. Dissolution rate and pH were also evaluated. Human osteoblast cells were incubated with elutes of fresh specimens from AH Plus and MTA Fillapex, and with elutes of the same specimens for 4 successive weeks. Elute's pH was evaluated at each time point. A multiparametric cell viability assay was performed. For dissolution rate, ISO methodology was used. The results were analyzed by one-way analysis of variance, complemented with the Tukey post-test (p<0.05). No significant difference was found among the materials when fresh mixed (p>0.05). After 1 week, AH Plus became non-cytotoxic on all three evaluated parameters. Conversely, MTA Fillapex remained severely and mildly cytotoxic over the entire experimental period (p<0.05). The dissolution rate of AH Plus was significantly lower than MTA Fillapex at all time points (p>0.05). The pH of AH Plus was significantly lower than MTA Fillapex at the second and third week (p<0.05). In the other tested time points no statistical difference was observed. In conclusion, MTA Fillapex remained cytotoxic after 4 weeks and its cytotoxicity may be related to the high dissolution rate of this material.

Long-Term Cytotoxicity, pH and Dissolution Rate of AH Plus and MTA Fillapex

Abstract The aim of the present study was to verify the long-term cytotoxic effects of the MTA Fillapex and to compare them with AH Plus. Dissolution rate and pH were also evaluated. Human osteoblast cells were incubated with elutes of fresh specimens from AH Plus and MTA Fillapex, and with elutes of the same specimens for 4 successive weeks. Elute's pH was evaluated at each time point. A multiparametric cell viability assay was performed. For dissolution rate, ISO methodology was used. The results were analyzed by one-way analysis of variance, complemented with the Tukey post-test (p<0.05). No significant difference was found among the materials when fresh mixed (p>0.05). After 1 week, AH Plus became non-cytotoxic on all three evaluated parameters. Conversely, MTA Fillapex remained severely and mildly cytotoxic over the entire experimental period (p<0.05). The dissolution rate of AH Plus was significantly lower than MTA Fillapex at all time points (p>0.05). The pH of AH Plus was significantly lower than MTA Fillapex at the second and third week (p<0.05). In the other tested time points no statistical difference was observed. In conclusion, MTA Fillapex remained cytotoxic after 4 weeks and its cytotoxicity may be related to the high dissolution rate of this material.

Resumo O objetivo do presente estudo foi verificar os efeitos citotóxicos de longo prazo da MTA Fillapex e comparar com os do AH Plus. Solubilidade e pH também foram avaliados. Osteoblastos humanos foram incubados com elutos de amostras frescas de AH Plus e MTA Fillapex, e com elutos dos mesmos espécimes pelas 4 semanas seguintes. O pH foi avaliado a cada semana. Um ensaio multiparamétrico de viabilidade celular foi realizado. Para solubilidade foi utilizada metodologia ISO. Os resultados foram analisados por ANOVA, complementada com o pós-teste de Tukey (p<0,05). Nenhuma diferença significativa foi encontrada entre os materiais frescos quando avaliados em relação a citotoxicidade (p>0,05). Depois de uma semana, o AH Plus tornou-se não-citotóxico em todos os três parâmetros avaliados. Por outro lado, MTA Fillapex permaneceu citotóxico durante todo o período experimental (p<0,05). A solubilidade do AH Plus foi significativamente menor do MTA Fillapex para todos os períodos avaliados (p>0,05). O pH da AH Plus foi significativamente menor do que o MTA Fillapex na segunda e na terceira semana (p<0,05). Nos outros períodos testados não houve diferença estatística. Em conclusão, o MTA Fillapex permaneceu citotóxico após 4 semanas e a sua citotoxicidade pode estar relacionada à elevada solubilidade deste material.

Cytotoxicity and gelatinolytic activity of a new silicon-based endodontic sealer.

PURPOSE: To compare the cytotoxicity, gelatinolytic activity, and protein levels (MMP-2 and MMP-9) produced by 3T3 fibroblasts cells after stimulation with GuttaFlow 2 and AH Plus. METHODS: 3T3 fibroblasts were incubated with elutes of GuttaFlow 2 and AH Plus for 24 h. The cytotoxicity of tested materials was determined using the MTT and the LDH assay. Supernatants of cell cultures incubated with sealers were collected to determine the levels of MMP-2 and MMP-9 gelatinolytic activity by gelatin zymography. Cell lysates were used to determine MMP-2 and MMP-9 protein levels by Western Blot. Data were analyzed using ANOVA and Tukey test (P<0.05). RESULTS: AH Plus showed significantly less cell viability (mitochondrial activity of cells) than GuttaFlow 2 (P<0.01). Moreover, GuttaFlow 2 was noncytotoxic, showing no statistically significant difference in LDH leakage levels compared to the control group (P>0.05). Specific characterization of MMPs demonstrated that GuttaFlow 2 seemed not to affect MMP-2 levels compared with the control group, while AH Plus had elevated gelatinolytic activity and protein levels of MMP-2 as confirmed by quantitative measurements. No detectable gelatinolytic activity or protein levels of MMP-9 (92 kDa) was observed in any tested group. CONCLUSIONS: GuttaFlow 2 did not showed cytotoxic effects and did not induce MMP-2 or MMP-9 expression.

Knowledge of Dentists on the Management of Tooth Avulsion Injuries in Rio de Janeiro, Brazil.

PURPOSE: To evaluate dentists' professional experience and knowledge of emergency management of tooth avulsion injuries in Rio de Janeiro, Brazil. MATERIALS AND METHODS: A total of 100 questionnaires were evaluated. The first part of the questionnaire consisted of questions regarding personal information. The second part evaluated dentists' knowledge of emergency management in cases of dental avulsion. The responses for each question were counted and expressed as percentages. RESULTS: All dentists had a college degree or above. Only three dentists had a Master's or PhD degree. Most of the dentists (94.5%) considered time and storage media important for the prognosis of avulsed teeth. However, the dentists did not show consistent responses about the adequate time and ideal storage media to transport avulsed teeth. CONCLUSION: The study highlighted Brazilian dentists' need for continuing education in order to improve current knowledge in emergency management of avulsed teeth.

Reciprocating versus rotary systems for root filling removal: assessment of the apically extruded material.

INTRODUCTION: This study was designed to quantitatively evaluate the amount of apically extruded debris by comparing the ProTaper Universal Retreatment system (Dentsply Maillefer, Ballaigues, Switzerland) with 2 reciprocating single-file systems (Reciproc [VDW, Munich, Germany] and WaveOne [Dentsply Maillefer]) during endodontic retreatment. METHODS: Forty-five mandibular premolars with a single canal were prepared with the ProTaper Universal system and then obturated. The specimens were divided into 3 groups (n = 15) according to the system used for filling removal: ProTaper Universal Retreatment system associated with the ProTaper Universal system (until file F4 40/0.06]), Reciproc system (Reciproc R40 [40/0.06]), and WaveOne system (WaveOne Large [40/0.08]). Sodium hypochlorite was used as an irrigant, and the apically extruded debris was collected in glass vials and then dried. The mean weight of debris was assessed with a microbalance and statistically analyzed using 1-way analysis of variance and post hoc Tukey multiple comparison tests (P < .05). RESULTS: The ProTaper Universal Retreatment system produced significantly more debris compared with the Reciproc and WaveOne systems (P < .01). The reciprocating systems showed no significant difference between them (P > .05). CONCLUSIONS: Under the conditions of the present study, all systems caused apical debris extrusion. Reciprocating systems were associated with less debris extrusion when compared with a conventional rotary retreatment system.

Evaluation of gelatinases, tissue inhibitor of matrix metalloproteinase-2, and myeloperoxidase protein in healthy and inflamed human dental pulp tissue.

INTRODUCTION: The aim of this study was to compare the gelatinolytic activity of matrix metalloproteinase (MMP)-2 and MMP-9 and the expression of tissue inhibitor of matrix metalloproteinase (TIMP)-2 and myeloperoxidase protein (MPO) in clinically healthy human pulp and inflamed pulp tissue specimens. METHODS: Twenty dental pulps clinically diagnosed as inflammatory tissues and 20 healthy pulp tissues from enclosed third molars were harvested and evaluated. The gelatinolytic activity for MMP-2 and MMP-9 was assessed by using the zymography technique, TIMP-2 gene expression was evaluated using the enzyme-linked immunosorbent assay, and MPO was determined using the MPO assay. RESULTS: Data showed increased levels of MMP-9, active MMP-2, TIMP-2, and MPO in inflammatory pulp tissues compared with healthy tissues (P < .05). No statistical difference could be observed for pro-MMP-2 (P > .05). CONCLUSIONS: Although all samples were associated with MMP-2 expression, the active form of this MMP was observed only in inflamed pulps. Inflamed pulps showed an up-regulation of MMP-9, TIMP-2, and MPO.

Self-adjusting file cleaning-shaping-irrigation system improves root-filling bond strength.

INTRODUCTION: The aim of the present study was to assess the bond strength of root fillings in oval-shaped canals prepared with the self-adjusting file (SAF) system. METHODS: A careful specimen selection resulted in 2 equal groups, each consisting of 12 extracted mandibular canines with oval canals that had vital pulps before extraction. One group was subjected to the SAF protocol, and the other group underwent conventional protocol, which was the ProTaper system with syringe-needle irrigation. Full-strength sodium hypochlorite was used as an irrigant for both groups. The teeth were obturated in a standardized way, filled with a lentulo spiral as the root filling, and then prepared for micropush-out assessment by using root slices of 1-mm thickness. Loading was performed on a universal testing machine at a speed of 0.5 mm/min(-1). The Student's t test for pairwise comparisons was applied to assess the effect of each preparation technique on the push-out bond strength. RESULTS: All specimens showed measurable adhesive properties to root dentin. In addition, no premature failure occurred. The group-by-location interaction was significant (P = .0071); thus, the group comparisons were dependent on the canal third. Overall, the push-out bond strength was the highest in the coronal third and the lowest in the apical third. SAF-prepared specimens displayed significantly higher bond strengths (P = .0012, 0.51-5.9 MPa). CONCLUSIONS: The present study showed that SAF preparation markedly influenced root-filling push-out bond strength in oval-shaped canals. Further investigations are needed to provide a better understanding of the physicochemical modifications of the root dentin prepared with the SAF cleaning-shaping-irrigation system.

Biological monitoring of a xenomaterial for grafting: an evaluation in critical-size calvarial defects.

Our purpose was to evaluate the osteoconduction potential of mixed bovine bone (MBB) xenografts as an alternative for bone grafting of critical-size defects in the calvaria of rats. After surgery, in the time intervals of 1, 3, 6, and 9 months, rats were killed and their skulls collected, radiographed and histologically prepared for analysis. The data obtained from histological analysis reported that the particles of MBB did not promote an intense immunological response, evidencing its biocompatibility in rats. Our results clearly showed the interesting evidence that MBB was not completely reabsorbed at 9 months while a small amount of newly formed bone was deposited by osteoprogenitor cells bordering the defect. However, this discrete bone-forming stimulation was unable to regenerate the bone defect. Overall, our results suggest that the properties of MBB are not suitable for stimulating intense bone regeneration in critical bone defects in rats.

Rat forming incisor requires a rigorous ECM remodeling modulated by MMP/RECK balance.

Reversion-inducing-cysteine-rich protein with Kazal motifs (RECK) is a single membrane-anchored MMP-regulator and regulates matrix metalloproteinases (MMP) 2, 9 and 14. In turn, MMPs are endopeptidases that play a pivotal role in remodeling ECM. In this work, we decided to evaluate expression pattern of RECK in growing rat incisor during, specifically focusing out amelogenesis process. Based on different kinds of ameloblasts, our results showed that RECK expression was conducted by secretory and post-secretory ameloblasts. At the secretory phase, RECK was localized in the infra-nuclear region of the ameloblast, outer epithelium, near blood vessels, and in the stellate reticulum. From the transition to the maturation phases, RECK was strongly expressed by non-epithelial immuno-competent cells (macrophages and/or dendritic-like cells) in the papillary layer. From the transition to the maturation stage, RECK expression was increased. RECK mRNA was amplified by RT-PCR from whole enamel organ. Here, we verified the presence of RECK mRNA during all stages of amelogenesis. These events were governed by ameloblasts and by non-epithelial cells residents in the enamel organ. Concluding, we found differential expression of MMPs-2, -9 and RECK in the different phases of amelogenesis, suggesting that the tissue remodeling is rigorously controlled during dental mineralization.

Physicochemical characterization of two deproteinized bovine xenografts.

Calcium phosphate salts, or more specifically hydroxyapatite, are products of great interest in the fields of medical and dental science due to their biocompatibility and osteoconduction property. Deproteinized xenografts are primarily constituted of natural apatites, sintered or not. Variations in the industrial process may affect physicochemical properties and, therefore, the biological outcome. The purpose of this work was to characterize the physical and chemical properties of deproteinized xenogenic biomaterials, Bio-Oss (Geistlich Biomaterials, Wolhuser, Switzerland) and Gen-Ox (Baumer S.A., Brazil), widely used as bone grafts. Scanning electron microscopy, infrared region spectroscopy, X-ray diffraction, thermogravimetry and degradation analysis were conducted. The results show that both materials presented porous granules, composed of crystalline hydroxyapatite without apparent presence of other phases. Bio-Oss presented greater dissolution in Tris-HCl than Gen-Ox in the degradation test, possibly due to the low crystallinity and the presence of organic residues. In conclusion, both commercial materials are hydroxyapatite compounds, Bio-Oss being less crystalline than Gen-Ox and, therefore, more prone to degradation.

Physicochemical characterization of two deproteinized bovine xenografts

Calcium phosphate salts, or more specifically hydroxyapatite, are products of great interest in the fields of medical and dental science due to their biocompatibility and osteoconduction property. Deproteinized xenografts are primarily constituted of natural apatites, sintered or not. Variations in the industrial process may affect physicochemical properties and, therefore, the biological outcome. The purpose of this work was to characterize the physical and chemical properties of deproteinized xenogenic biomaterials, Bio-Oss (Geistlich Biomaterials, Wolhuser, Switzerland) and Gen-Ox (Baumer S.A., Brazil), widely used as bone grafts. Scanning electron microscopy, infrared region spectroscopy, X-ray diffraction, thermogravimetry and degradation analysis were conducted. The results show that both materials presented porous granules, composed of crystalline hydroxyapatite without apparent presence of other phases. Bio-Oss presented greater dissolution in Tris-HCl than Gen-Ox in the degradation test, possibly due to the low crystallinity and the presence of organic residues. In conclusion, both commercial materials are hydroxyapatite compounds, Bio-Oss being less crystalline than Gen-Ox and, therefore, more prone to degradation.

Expression of matrix metalloproteinases-2 and -9 and RECK during alveolar bone regeneration in rat.

MMPs are endopeptidases that play a pivotal role in ECM turnover. RECK is a single membrane-anchored MMP-regulator. Here, we evaluated the temporal and spatial expression of MMP-2, MMP-9, and RECK during alveolar bone regeneration. The maxillary central incisor of Wistar rats was extracted and the animals were killed at 1, 3, 7, 10, 14, 21, 28, and 42 days post-operatively (n = 3/period). The hemimaxillae were collected, demineralized and embedded in paraffin. Immunohistochemical analysis was performed by the immunoperoxidase technique with polyclonal antibodies. On day 1, polymorphonuclear cells in the blood clot presented mild immunolabeling for MMPs. During bone remodeling, osteoblasts facing new bone showed positive staining for gelatinases and RECK in all experimental periods. MMPs were also found in the connective tissue and endothelial cells. Our results show for the first time that inactive and/or active forms of MMP-2, MMP-9 and RECK are differentially expressed by osteogenic and connective cells during several events of alveolar bone regeneration. This may be important for the replacement of the blood clot by connective tissue, and in the formation, maturation and remodeling of new bone.

Expression of metalloproteinase 2 in the cell response to porous demineralized bovine bone matrix.

The purpose of the study was to analyze the involvement of metalloproteinase 2 (MMP-2) and macrophages in the tissue and cell response to the organic graft material produced from bovine cancellous bone. Thirty adult male white Wistar rats (Rattus norvegicus) received implants of blocks of demineralized bovine bone matrix between the fasciae of the quadriceps muscle. The specimens collected at 3, 7, 14, 21 and 28 days after implantation (n = 6/period). Sections of 6 microm thick were stained with hematoxylin and eosin and immunolabeled with anti-MMP-2 and anti-CD68 using standard avidin-biotin-peroxidase method. The tissue response to the material was initially mediated by polymorphonuclear neutrophils, evolving to a mononuclear inflammatory infiltrate with macrophages and few lymphocytes and plasma cells and presence of inflammatory multinucleated giant cells (GC) in contact with the material that exhibited signs of resorption. The number of cells immunolabeled to MMP-2 was highest at day 7 (103.2 +/- 39.1), but significantly decreased (F = 3.67; p = 0.044) until day 28 (45.9 +/- 13.1). CD68 immunostaining also significantly decreased (F = 6.75; p = 0.007) from day 7 (49.5 +/- 10.4) to day 28 (19.5 +/- 8.9). A positive and statistically significant correlation was observed between the evolutions of these two variables. The material had been almost completely resorbed at day 28. Among cells present at the granuloma, anti-MMP-2 immunostaining was predominant and more intense in macrophages, yet lightly immunolabeled multinucleated giant cells were found in close contact with the material. Thus, considering the experimental limitations of this study, we concluded that MMP-2 produced by macrophages participates in the resorption of demineralized bovine bone.