Effect of reciprocating instrumentation on chlorhexidine substantivity on human dentin: chemical analysis followed by confocal laser microscopy

The present research analyzed the reciprocating instrumentation associated to chlorhexidine (CHX) substantivity as its correlation with E. faecalis viability in ex vivo root canals. Eighty extracted single-rooted human teeth were used, being 40 to high-performance liquid chromatography (HPLC) and 40 to confocal laser scanning microscopy (CLSM). In both, teeth were decoronated and the cervical third was prepared. In the CLSM analysis, the root canals were inoculated with E. faecalis for 14 days. Samples were divided into 4 groups (n=10) according to instrumentation technique: no instrumentation and irrigation with distilled water (control); manual instrumentation (K-File); rotary instrumentation (ProTaper Next); and reciprocating instrumentation (Reciproc R25). Two percent chlorhexidine was applied as irrigating substance in experimental groups. Longitudinal grooves resulted in 2 halves root and 20 proof bodies in each group. Samples were divided by chance in two groups (n=10) and the outcomes were evaluated after two days and one week. The retained chlorhexidine and live cells after instrumentation techniques in each evaluation time was measured by HPLC and CLSM, respectively. Specific analysis was applied for experimental tests (p≤0.05). Both rotary as well as reciprocating techniques significantly reduced the amount of chlorhexidine on dentin in all observation periods (p<0.05). After evaluation times, all experimental groups presented lower live cells compared to control, but without statistically difference. Intragroup comparisons in times of evaluation showed no differences in instrumentation techniques, in chlorhexidine retention and number of live cells (p>0.05). Reciprocating instrumentation does not interfere on chlorhexidine substantivity.

Effect of human tooth bone graft materials on proliferation and differentiation of mice mononuclear macrophage RAW264.7 / 中国修复重建外科杂志

Objective: To investigate the effect of human tooth bone graft materials on the proliferation, differentiation, and morphology of macrophages, and to understand the biocompatibility and cytotoxicity of human tooth bone graft materials. Methods: Fresh human teeth were collected to prepare human tooth bone graft materials, the adhesion of mouse mononuclear macrophages RAW264.7 to human bone graft materials was observed under confocal microscopy. Scanning electron microscopy was used to observe the morphology of human tooth bone graft materials, OSTEONⅡ synthetic highly resorbable bone grafting materials, and untreated tooth powder (dental particles without preparation reagents). Different components of the extract were prepared in 4 groups: group A (DMEM medium containing 10% fetal bovine serum), group B (human tooth bone graft materials), group C (OSTEONⅡ synthetic highly resorbable bone grafting materials), group D (untreated tooth powder without preparation reagents). The 4 groups of extracts were co-cultured with the cells, and the cytotoxicity was qualitatively determined by observing the cell morphological changes by inverted microscope. The cell proliferation and differentiation results and cell relative proliferation rate were determined by MTT method to quantitatively determine cytotoxicity. The cell viability was detected by trypanosoma blue staining, and tumor necrosis factor α (TNF-α ) and interleukin 6 (IL-6) expressions were detected by ELISA. Results: Scanning electron microscopy showed that the surface of the human tooth bone graft material and the OSTEONⅡ synthetic highly resorbable bone grafting materials had a uniform pore structure, while the untreated tooth particle collagen fiber structure and the demineralized dentin layer collapsed without specific structure. Confocal microscopy showed that the cells grew well on human tooth bone graft materials. After co-culture with the extract, the morphology and quantity of cells in groups A, B, and C were normal, and the toxic reaction grades were all grade 0, while group D was grade 3 reaction. MTT test showed that the cytotoxicity of groups B and C was grade 0 or 1 at each time point, indicating that the materials were qualified. The cytotoxicity was grade 2 in group D at 1 day after culture, and was grade 4 at 3, 5, and 7 days. Combined with cell morphology analysis, the materials were unqualified. The trypanosoma blue staining showed that the number of cells in groups A, B, and C was significantly higher than that in group D at each time point ( P<0.05), but no significant difference was found among groups A, B, and C ( P<0.05). ELISA test showed that the levels of TNF-α and IL-6 in groups A, B, and C were significantly lower than those in group D ( P<0.05), but no significant difference was found among groups A, B, and C ( P<0.05). Conclusion: The human tooth bone graft materials is co-cultured with mice mononuclear macrophages without cytotoxicity. The extract has no significant effect on cell proliferation and differentiation, does not increase the expression of inflammatory factors, has good biocompatibility, and is expected to be used for clinical bone defect repair.

Caracterización estereológica de odontoblastos en pulpas dentarias humanas sanas y con pulpitis reversible / Stereological characterization of odontoblasts in normal healthy and reversible pulpitis in human dental pulps

La reacción y reparación dentinaria depende del número de odontoblastos. Los métodos para obtener estimaciones fiables sobre la cantidad de odontoblastos en la pulpa dental han sido subjetivos y sesgados, sobre todo al evaluar los cambios cuantitativos y la potencial capacidad reparativa en presencia de caries. El objetivo de este trabajo fue estimar y comparar cuantitativamente el número, densidad y volumen de odontoblastos en dientes sanos y con diagnóstico de pulpitis reversible producto de caries a través de herramientas estereológicas. Se utilizaron dientes premolares humanos obtenidos de exodoncias, divididos en un grupo sano y otro cariado. Fueron fijados y descalcificados con ácido nítrico al 5%. Siguiendo el protocolo del orientator se obtuvieron 5 secciones de 5 mm teñidas por H-E de cada diente. Se aplicó el recuento estereológico de los odontoblastos con el test multipropósito M42. Se estimaron las densidades de número (Nv), volumen (Vv) y superficie (Sv), y calcularon las Medias (±DE) por diente, y Medias (±EE) por grupo. Las diferencias entre grupos se analizaron mediante la prueba T, con un valor p 0,05 de significación estadística. En dientes sanos, la Media (±EE) para Nv de odontoblastos fue 0,409x105/mm3 (±0,018x105/mm3), para Vv 19,714% (±1,43%) y para Sv 21,016 mm2/mm3 (±1,425 mm2/mm3). En dientes cariados, la Nv fue 0,521x105 /mm3 (±0,023x105/mm3), la Vv 24,686% (±1,625%) y la Sv 23,203 mm2/mm3 (±1,364 mm2/mm3). Al comparar las Nv, los odontoblastos del grupo con caries aumentaron significativamente (p=0,0062), al igual que la Vv (p=0,0197). Existe un aumento del número de odontoblastos en los dientes con pulpitis reversible, lo que condicionaría su capacidad de respuesta. La metodología empleada puede ser aplicable para determinar el comportamiento pulpar y cuantificar variables de respuesta odontoblástica en tratamientos restauradores atraumáticos de manera imparcial y reproducible.

Dentinal reaction and repair depends on factors like the amount of odontoblasts. Methods for obtaining reliable estimates of the number of odontoblasts in the dental pulp have been subjective and biased, especially when assessing the potential quantitative changes and reparative capacity in the presence of cavities. The aim of this study was to estimate and quantitatively compare, through stereological tools, the number, density and volume of odontoblasts in healthy teeth, diagnosed with reversible pulpitis due to caries. We used human premolars obtained from extractions, divided into groups of healthy teeth and teeth with caries, fixed and decalcified in 5% nitric acid. Following the orientator protocol, five 5-µm-thick sections stained by HE were obtained from each tooth. Stereological counting for odontoblast with M42 multipurpose test was applied. Numerical density (Nv), volume density (Vv) and surface density (Sv) were estimated, and the mean (± SD) for each tooth, and Mean (± SE) per group were calculated. Differences between groups were analyzed by t test, with p 0.05 for statistical significance. In the healthy teeth group, the mean (± SE) for Nv was 0.409x105/mm3 (±0.018x105/mm3), Vv 19.714% (±1.43%) and to Sv 21.016 mm2/mm3 (±1.425 mm2/mm3) odontoblast cells. In the caries teeth group, the Nv was 0.521x105 /mm3 (±0.023x105/mm3), Vv 24.686% (±1.625%) and Sv 23.203 mm2/mm3 (±1.364 mm2/mm3). When comparing Nv, an increased in odontoblasts significantly (p = 0.0062), as well as Vv (p = 0.0197) in caries teeth group. There is an increased number of odontoblasts in teeth with reversible pulpitis, which would condition its responsiveness. The methodology can be applied to determine pulp behavior, and quantify variables of odontoblastic response in atraumatic restorative treatments in an impartial and reproducible form.

Radiodensity evaluation of dental impression materials in comparison to tooth structures

In the most recent decades, several developments have been made on impression materials' composition, but there are very few radiodensity studies in the literature. It is expected that an acceptable degree of radiodensity would enable the detection of small fragments left inside gingival sulcus or root canals. OBJECTIVE: The aim of this study was to determine the radiodensity of different impression materials, and to compare them to human and bovine enamel and dentin. MATERIAL AND METHODS: Twenty-five impression materials, from 5 classes, were studied: addition and condensation silicones, polyether, polysulfides and alginates. Five 1-mm-thick samples of each material and tooth structure were produced. Each sample was evaluated 3 times (N=15), being exposed to x-ray over a phosphor plate of Digora digital system, and radiodensity was obtained by the software Digora for Windows 2.5 Rev 0. An aluminum stepwedge served as a control. Data were subjected to Kruskal-Wallis and Dunn's method (α=0.05). RESULTS: Different materials and respective classes had a different behavior with respect to radiodensity. Polysulfides showed high values of radiodensity, comparable to human enamel (p>0.05), but not to bovine enamel (p<0.05). Human dentin was similar only to a heavy-body addition silicon material, but bovine dentin was similar to several materials. Generally, heavy-body materials showed higher radiodensity than light-body ones (p<0.05). CONCLUSION: Impression materials' radiodensity are influenced by composition, and almost all of them would present a difficult detection against enamel or dentin background in radiographic examinations.

Comparison of resin push-out strength to root dentin of bovine- and human-teeth.

Aim : To compare the push-out strength of bovine- and human-root dentin and, thus, evaluate the suitability of bovine-root dentin to substitute human-root dentin for bond strength testing. Materials and Methods : Ten single-rooted human-teeth and ten bovine incisors were prepared using a #3 bur of a fiber post system (12 mm long). The posts were duplicated with resin cement (Duolink). The root canals were treated with All Bond 2 adhesive system and the resin posts were cemented using Duolink. The specimens were cut perpendicular to their long axis, yielding disc-specimens with 1.5 mm thickness, which were submitted to a push-out test (1 mm/min). Ten bond strength values per group (n = 10) were used for statistical analysis (Student t test, a =.05). Results : Statistically significant differences were found for the bond strength values between bovine- (4.1 ± 1.3 MPa) and human-root dentin (8.6 ± 5.7 MPa) (P =.0001). Conclusion : The push-out strengths of bovine- and human-root dentin were statistically different.