RESUMEN
BACKGROUND: In the face of the COVID-19 pandemic, the Defence Science and Technology Laboratory (Dstl) and Defence Pathology combined to form the Defence Clinical Lab (DCL), an accredited (ISO/IEC 17025:2017) high-throughput SARS-CoV-2 PCR screening capability for military personnel. LABORATORY STRUCTURE AND RESOURCE: The DCL was modular in organisation, with laboratory modules and supporting functions combining to provide the accredited SARS-CoV-2 (envelope (E)-gene) PCR assay. The DCL was resourced by Dstl scientists and military clinicians and biomedical scientists. LABORATORY RESULTS: Over 12 months of operation, the DCL was open on 289 days and tested over 72 000 samples. Six hundred military SARS-CoV-2-positive results were reported with a median E-gene quantitation cycle (Cq) value of 30.44. The lowest Cq value for a positive result observed was 11.20. Only 64 samples (0.09%) were voided due to assay inhibition after processing started. CONCLUSIONS: Through a sustained effort and despite various operational issues, the collaboration between Dstl scientific expertise and Defence Pathology clinical expertise provided the UK military with an accredited high-throughput SARS-CoV-2 PCR test capability at the height of the COVID-19 pandemic. The DCL helped facilitate military training and operational deployments contributing to the maintenance of UK military capability. In offering a bespoke capability, including features such as testing samples in unit batches and oversight by military consultant microbiologists, the DCL provided additional benefits to the UK Ministry of Defence that were potentially not available from other SARS-CoV-2 PCR laboratories. The links between Dstl and Defence Pathology have also been strengthened, benefitting future research activities and operational responses.
RESUMEN
The aim of this study was to evaluate the effect of different materials used for dentin prophylaxis on the microtensile bond strengths (µTBS) of adhesively cemented indirect composite restorations. Sixty bovine incisors had the buccal surface ground with wet #600-grit silicon carbide abrasive paper to obtain a flat exposed superficial dentin and were submitted to different prophylaxis protocols, as follows: 3% hydrogen peroxide (HydP); 0.12% chlorhexidine (Chlo); sodium bicarbonate jet (SodB); 50-µm aluminum oxide air abrasion (AirA); pumice paste (PumP), and control group-water spray (Cont). After prophylaxis protocols a resin composite block (3.0 mm × 5.0 mm × 5.0 mm) was adhesively cemented using dual resin cement (Rely X ARC). After 24 hours of water storage, specimens were serially sectioned perpendicular to the bonded interface into 1-mm-thick slices. Each specimen was trimmed with a diamond bur to an hourglass shape with a cross-sectional area of approximately 1.0 mm(2) at the bonded area. Specimens were tested (µTBS) at 0.5 mm/min using a universal testing machine. Scanning electron microscopy was used to examine the effects of prophylaxis techniques on dentin. Bond strength data (MPa) were analyzed by one-way analysis of variance and failure mode by Fisher test (α=0.05). µTBS data, means (SD), were (different superscripted letters indicate statistically significant differences): AirA, 25.2 (7.2)(a); PumP, 24.1 (7.8)(a); Chlo, 21.5 (5.6)(a); Cont, 20.6 (8.1)(a); HydP(,) 15.5 (7.6)(b); and SodB(,) 11.5 (4.4)(c). The use of aluminum oxide air abrasion, pumice paste, and chlorhexidine before acid etching did not significantly affect µTBS to dentin; however, the use of hydrogen peroxide and sodium bicarbonate jet significantly reduced µTBS.
Asunto(s)
Recubrimiento Dental Adhesivo , Profilaxis Dental/métodos , Recubrimientos Dentinarios/química , Dentina/ultraestructura , Grabado Ácido Dental/métodos , Óxido de Aluminio/química , Animales , Antiinfecciosos Locales/química , Bisfenol A Glicidil Metacrilato/química , Bovinos , Cementación/métodos , Clorhexidina/química , Resinas Compuestas/química , Materiales Dentales/química , Análisis del Estrés Dental/instrumentación , Peróxido de Hidrógeno/química , Ensayo de Materiales , Microscopía Electrónica de Rastreo , Ácidos Fosfóricos/química , Polietilenglicoles/química , Ácidos Polimetacrílicos/química , Cementos de Resina/química , Silicatos/química , Bicarbonato de Sodio/química , Estrés Mecánico , Propiedades de Superficie , Factores de Tiempo , Agua/químicaRESUMEN
The in vivo preclinical pharmacodynamic profile of TD-5108, a selective 5-HT(4) receptor agonist with high intrinsic activity, was compared to that of the clinically studied gastrointestinal pro-kinetic agents, tegaserod, cisapride and mosapride. The activity of TD-5108 was evaluated in guinea pig colonic transit, rat oesophageal relaxation and dog gastrointestinal smooth muscle contractility models. Subcutaneous administration of TD-5108, tegaserod, cisapride and mosapride increased guinea pig colonic transit (rank order of potencies: TD-5108 > tegaserod > cisapride > mosapride). Following intravenous and intraduodenal dosing, TD-5108, tegaserod, cisapride and mosapride produced dose-dependent relaxation of the rat oesophagus. On a molar basis, TD-5108 was approximately twofold less potent than tegaserod following intravenous dosing but 6- or 86-fold more potent than cisapride or mosapride, respectively, and 9- or 18-fold more potent than tegaserod or cisapride, respectively, after intraduodenal administration. Orally dosed TD-5108 increased the contractility of the canine antrum, duodenum and jejunum with higher potency than tegaserod. The selective 5-HT(4) receptor agonist, TD-5108, demonstrates robust in vivo activity in the guinea pig, rat and dog gastrointestinal tracts.
Asunto(s)
Compuestos de Azabiciclo/farmacología , Tránsito Gastrointestinal/efectos de los fármacos , Agonistas del Receptor de Serotonina 5-HT4 , Agonistas de Receptores de Serotonina/farmacología , Administración Oral , Animales , Compuestos de Azabiciclo/administración & dosificación , Benzamidas/administración & dosificación , Benzamidas/farmacología , Cisaprida/administración & dosificación , Cisaprida/farmacología , Colon/efectos de los fármacos , Colon/metabolismo , Perros , Relación Dosis-Respuesta a Droga , Esófago/efectos de los fármacos , Esófago/metabolismo , Femenino , Cobayas , Indoles/administración & dosificación , Indoles/farmacología , Masculino , Morfolinas/administración & dosificación , Morfolinas/farmacología , Contracción Muscular/efectos de los fármacos , Músculo Liso/efectos de los fármacos , Músculo Liso/metabolismo , Ratas , Ratas Sprague-Dawley , Agonistas de Receptores de Serotonina/administración & dosificaciónRESUMEN
Microtensile bond strength values are influenced by specimen shape and attachment method to the gripping device during testing. We hypothesized that stress distribution inside the testing specimen is affected by microtensile specimen shape and attachment method. Rectangular, hourglass-, and dumbbell-shaped specimens, all with a 1 mm(2) cross-sectional testing region, were modeled as indirect ceramic restorations luted to dentin. Three specimen attachments were investigated: (1) posterior surface; (2) posterior, superior, and lateral surfaces; and (3) all surfaces. Qualitative and quantitative analyses were carried out according to von Mises' criteria. Stress analysis showed a direct correlation between attachment modes and stress distribution, with shear stresses observed in models with less surface attachment. Increasing the number of faces for specimen attachment to the metallic gripping device resulted in a more homogeneous and regular distribution of stress, with tensile stress concentrated at the adhesive interface. Dumbbell-shaped specimens showed improved stress distribution compared with rectangular and hourglass-shaped specimens.
Asunto(s)
Materiales Dentales/química , Análisis de Elementos Finitos , Bisfenol A Glicidil Metacrilato/química , Recubrimiento Dental Adhesivo/métodos , Porcelana Dental/química , Análisis del Estrés Dental/instrumentación , Dentina/ultraestructura , Elasticidad , Humanos , Imagenología Tridimensional , Compuestos de Litio/química , Ensayo de Materiales , Polietilenglicoles/química , Ácidos Polimetacrílicos/química , Cementos de Resina/química , Estrés Mecánico , Propiedades de Superficie , Resistencia a la TracciónRESUMEN
This study investigated the effect of three equivalent radiant exposure energy application sequences (EASs): high intensity power (HIP: 1,177 mW/cm2, 40 seconds), low intensity power (LOP: 573 mW/cm2, 71 seconds) and ramp (RMP: 650 mW/cm2, 5 seconds, then the irradiance increases to 1047 mW/cm2, 37 seconds) on the microtensile bond strength (MTBS) in both low and high C-factor cavity preparations. Thirty Paradigm blocks (Z100) were mounted in stone, with their top surfaces parallel to the mounting block. High C-factor (HC = 3.8) Class I cavity preparations were then prepared in 15 of the Paradigm blocks using a computer-controlled specimen former. Low C-factor (LC = 1.8) Class II cavity preparations were prepared in the remaining 15 blocks by sectioning them perpendicularly using a slow-speed diamond saw. Five samples, one from each experimental group (3 EASs x 2 C-factors), were prepared and stored in the dark for 48 hours in distilled water at 37 degrees C prior to MTBS testing on the third day and on the remainder of the samples (n = 30). Mode of fracture was determined with a stereomicroscope at 20-40x magnification. The findings of this study indicated that HC, in combination with HIP, had significantly lower bond strength (27.54 MPa) than LC with HIP (51.39 MPa). On the other hand, there was no significant difference between high and low C-factors with the other EASs (LOP or RMP). There was also no significant effect for EAS (HIP, RAMP and LOP), with C-factor (HC or LC) held constant. The HIP-HC group had the highest percentage of adhesive (20%) and mixed adhesive (50%) failures (70% total). Adhesive and mixed adhesive failures occurred in other groups, but with lower percentages (RMP-LC: 40% total) (LOP-HC: 40% total).
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Resinas Compuestas/efectos de la radiación , Recubrimiento Dental Adhesivo , Preparación de la Cavidad Dental/clasificación , Materiales Dentales/efectos de la radiación , Grabado Ácido Dental , Resinas Acrílicas/química , Adhesividad , Resinas Compuestas/química , Materiales Dentales/química , Humanos , Luz , Iluminación/instrumentación , Ensayo de Materiales , Poliuretanos/química , Dosis de Radiación , Cementos de Resina/química , Propiedades de Superficie , Temperatura , Resistencia a la Tracción , Factores de Tiempo , Agua/química , Circonio/química , Circonio/efectos de la radiaciónRESUMEN
The 5-HT4 receptor agonist and gastroprokinetic, tegaserod, possesses 5-HT2B receptor antagonist activity. However, the relevance of such activity is unclear. In this study, the 5-HT2B receptor antagonist and 5-HT4 agonist activities of tegaserod were investigated. Two piezoelectric crystals were implanted on the stomach fundus or oesophagus of anaesthetized Sprague-Dawley rats. Measurement of the transmission time of ultrasonic pulses between the implanted crystals provided a continuous record of inter-crystal distance, and thus of muscle length. In the stomach fundus, tegaserod (1 and 3 mg kg(-1)), administered subcutaneously (s.c.), inhibited the contractile response evoked by the 5-HT2B receptor agonist, BW 723C86 (0.01-1 mg kg(-1) intravenously (i.v.)). SB 206553 (1 mg kg(-1) s.c.), a selective 5-HT2B/2C receptor antagonist, also inhibited the BW 723C86-mediated responses. In the rat oesophagus, tegaserod (0.001-0.3 mg kg(-1) i.v. or 0.003-3 mg kg(-1) s.c.) increased inter-crystal distance, consistent with smooth muscle relaxation; the responses were inhibited by the 5-HT4 antagonist, piboserod (0.1 mg kg(-1) s.c.). Data from this in vivo rat study are consistent with tegaserod-induced 5-HT4 receptor-mediated oesophageal relaxation, and antagonism of 5-HT2B receptor-mediated stomach fundus contraction. The clinical relevance of the 5-HT2B receptor antagonism of tegaserod remains to be determined.
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Indoles/farmacología , Antagonistas del Receptor de Serotonina 5-HT2 , Agonistas del Receptor de Serotonina 5-HT4 , Agonistas de Receptores de Serotonina/farmacología , Animales , Esófago/efectos de los fármacos , Esófago/fisiología , Fundus Gástrico/efectos de los fármacos , Fundus Gástrico/fisiología , Fármacos Gastrointestinales/farmacología , Masculino , Contracción Muscular/efectos de los fármacos , Músculo Liso/efectos de los fármacos , Músculo Liso/fisiología , Ratas , Ratas Sprague-DawleyRESUMEN
1 Tegaserod (Zelnorm) is a potent 5-hydroxytryptamine4 (5-HT4) receptor agonist with clinical efficacy in disorders associated with reduced gastrointestinal motility and transit. The present study investigated the interaction of tegaserod with 5-HT2 receptors, and compared its potency in this respect to its 5-HT4 receptor agonist activity. 2 Tegaserod had significant binding affinity for human recombinant 5-HT2A, 5-HT2B and 5-HT2C receptors (pKi=7.5, 8.4 and 7.0, respectively). The 5-HT2B receptor-binding affinity of tegaserod was identical to that at human recombinant 5-HT4(c) receptors (mean pKi=8.4) in human embryonic kidney-293 (HEK-293) cells stably transfected with the human 5-HT4(c) receptor. 3 Tegaserod (0.1-3 microm) inhibited 5-HT-mediated contraction of the rat isolated stomach fundus potently (pA2=8.3), consistent with 5-HT(2B) receptor antagonist activity. Tegaserod produced, with similar potency, an elevation of adenosine 3',5' cyclic monophosphate in HEK-293 cells stably transfected with the human 5-HT4(c) receptor (mean pEC50=8.6), as well as 5-HT4) receptor-mediated relaxation of the rat isolated oesophagus (mean pEC50=8.2) and contraction of the guinea-pig isolated colon (mean pEC50=8.3). 4 Following subcutaneous administration, tegaserod (0.3 or 1 mg kg(-1)) inhibited contractions of the stomach fundus in anaesthetized rats in response to intravenous dosing of alpha-methyl 5-HT (0.03 mg kg(-1)) and BW 723C86 (0.3 mg kg(-1)), selective 5-HT2B receptor agonists. At similar doses, tegaserod (1 and 3 mg kg(-1) subcutaneously) evoked a 5-HT4 receptor-mediated increase in colonic transit in conscious guinea-pigs. 5 The data from this study indicate that tegaserod antagonizes 5-HT2B receptors at concentrations similar to those that activate 5-HT4 receptors. It remains to be determined whether this 5-HT2B receptor antagonist activity of tegaserod contributes to its clinical profile.
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Indoles/farmacología , Receptor de Serotonina 5-HT2B/efectos de los fármacos , Receptores de Serotonina 5-HT4/efectos de los fármacos , Antagonistas de la Serotonina/farmacología , Agonistas de Receptores de Serotonina/farmacología , Animales , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Células Cultivadas , Colon/efectos de los fármacos , AMP Cíclico/metabolismo , Esófago/efectos de los fármacos , Fundus Gástrico/efectos de los fármacos , Tránsito Gastrointestinal/efectos de los fármacos , Cobayas , Técnicas In Vitro , Masculino , Contracción Muscular/efectos de los fármacos , Músculo Liso/efectos de los fármacos , Presión , Unión Proteica , Ensayo de Unión Radioligante , Ratas , Ratas Sprague-DawleyRESUMEN
Crowns and large amalgams protect structurally compromised teeth to various degrees in different situations. The aim of this investigation was to evaluate the survival of teeth with these two types of restorations and the factors associated with better outcomes. Retrospective administrative and chart data were used. Survival was defined and modeled as: (1) receipt of no treatment and (2) receipt of no catastrophic treatment over five- and 10-year periods. Analyses included: Kaplan-Meier survival curves, Log-Rank tests, and Cox proportional hazards regression modeling. Crowns survived longer with no treatment and with no catastrophic treatment; however, mandibular large amalgams were least likely to have survived with no treatment, and maxillary large amalgams were least likely to have survived with no catastrophic treatment. Having no adjacent teeth also decreased survival. Crowns survived longer than large amalgams, but factors such as arch type and the presence of adjacent teeth contributed to the survival of large amalgams.
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Coronas , Amalgama Dental , Fracaso de la Restauración Dental , Restauración Dental Permanente/métodos , Diente Premolar , Femenino , Humanos , Estudios Longitudinales , Masculino , Diente Molar , Modelos de Riesgos Proporcionales , Retratamiento , Estudios Retrospectivos , Análisis de Supervivencia , Factores de TiempoRESUMEN
The cariostatic effect of resin-modified glass ionomer (RMGI) on secondary root caries is well-documented. However, this beneficial effect may be dependent upon the mode of cavity surface treatment. To investigate this relationship, we studied 4 cavity surface treatments prior to the placement of RMGI: no treatment (None), polyacrylic acid (PAA), phosphoric acid (H(3)PO(4)), and Scotchbond Multi-Purpose adhesive (SMP) as a control. Specimens were aged for two weeks in synthetic saliva, thermocycled, and subjected to an artificial caries challenge (pH 4.4). Polarized light microscopy (PLM) and microradiography (MRG) showed significantly less demineralization with the H(3)PO(4) cavity surface treatment as revealed by ANOVA and Tukey's multiple comparisons (p < or = 0.05). Dentin fluoride profiles determined by electron probe microanalysis (EPMA) supported PLM and MRG findings. It may be concluded that removal of the smear layer with phosphoric acid provides significantly enhanced resistance to secondary root caries formation adjacent to RMGI restorations.
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Grabado Ácido Dental , Preparación de la Cavidad Dental , Dentina/efectos de los fármacos , Cementos de Ionómero Vítreo/química , Caries Radicular/prevención & control , Capa de Barro Dentinario , Desmineralización Dental/prevención & control , Resinas Acrílicas/farmacología , Análisis de Varianza , Cariostáticos/uso terapéutico , Restauración Dental Permanente/métodos , Microanálisis por Sonda Electrónica , Fluoruros/uso terapéutico , Humanos , Microrradiografía , Microscopía Electrónica de Rastreo , Microscopía de Polarización , Diente Molar , Ácidos Fosfóricos/farmacología , Prevención Secundaria , Estadísticas no Paramétricas , Propiedades de SuperficieRESUMEN
OBJECTIVE: To determine the failure mode between dentin-adhesive resin-resin composite bonded joint produced with a chevron-notch short-bar (CNSB) and microtensile test methods. METHODS: Forty teeth were randomly selected for microtensile and forty for CNSB specimen fabrication and stored in 0.5% chloramine T at 37 degrees C until respective static load to failure testing at 30 and 180days. Failure modes were categorized by SEM and tested with Fisher's exact test. Within respective mechanical testing methods the probability of failure curve distributions being significantly different were analyzed by the Wald chi-square statistic. RESULTS: The characteristic fracture toughness at 30- and 180-day storage was 0.82 and 0.87MPam(1/2), while the Weibull Modulus (m) for the failure distributions, was 4.60 and 4.56, respectively. No significant difference was demonstrated in the failure distributions between these groups (p=0.45). The characteristic tensile strength (muTBS(o)) at 30- and 180-day storage was 52.53 and 14.71MPa with an m of 3.04 and 1.56, respectively. Failure distributions for muTBS groups were significantly different (p<0.001). K(IvM) failure modes, regardless of storage time, were within the adhesive joint with 30-day debonds primarily through the top region of the hybrid layer (THL) and after 180-days involving the bottom of the hybrid layer (BHL). The 30-day muTBS group demonstrated a propensity to debond in dentin or resin composite substrates but after 180-days storage debonds again involved the BHL. SIGNIFICANCE: The weak links in the dentin-adhesive resin-resin composite bonded joint may be the interphase regions between the THL and the adhesive resin and the BHL and dentin.
Asunto(s)
Resinas Compuestas/química , Recubrimiento Dental Adhesivo , Recubrimientos Dentinarios/química , Dentina/ultraestructura , Adhesividad , Distribución de Chi-Cuadrado , Cloraminas , Intervalos de Confianza , Restauración Dental Permanente , Análisis del Estrés Dental/instrumentación , Desinfectantes , Elasticidad , Humanos , Funciones de Verosimilitud , Ensayo de Materiales , Microscopía Electrónica de Rastreo , Cementos de Resina/química , Estadística como Asunto , Estrés Mecánico , Propiedades de Superficie , Análisis de Supervivencia , Resistencia a la Tracción , Factores de Tiempo , Compuestos de TosiloRESUMEN
OBJECTIVE: To test the elastic wall concept utilizing adhesive resins of varying stiffness in a low- and high-C-factor cavity design after short- and long-term water storage. METHODS: A flat and box-shaped cavity was restored on occlusal dentin with a resin composite using a filled and unfilled adhesive resin from which microtensile specimens with a 0.5mm(2) cross-sectional area were formed. After storage for 30- and 150-days the microtensile bond strength (muTBS) was determined in a Zwick materials testing machine and the subsequent debond pathway was examined under scanning electron microscopy. Fisher's exact test was used to determine differences in joint and substrate failure modes and a Weibull regression model with gamma frailties was used to test for differences between failure distributions. Tests for three-way and two-way interactions were also completed for storage time, C-factor and adhesive. All tests were at 95% confidence levels. RESULTS: The characteristic strength (TBS degrees ) for the Optibond FL adhesive applied on a flat cavity was 47.57 and 20.90MPa and a box-shaped cavity was 49.26 and 17.49MPa for short- and long-term storage, respectively, while the corresponding TBS degrees for the unfilled Optibond adhesive on the flat cavity design was 36.93 and 32.68MPa and in a box-shaped cavity was 32.84 and 15.46MPa. Combining all groups according to storage time revealed a three-fold increase in the debond pathway including the bottom of the hybrid layer. SIGNIFICANCE: Evidence suggests that the durability of the bonded joint is threatened by hydrolysis and the most susceptible region is the bottom half of the hybrid layer and in low C-factor cavity designs a more flexible adhesive resin liner was more durable.
Asunto(s)
Resinas Compuestas/química , Recubrimiento Dental Adhesivo , Preparación de la Cavidad Dental/clasificación , Recubrimientos Dentinarios/química , Dentina/ultraestructura , Agua/química , Adhesividad , Intervalos de Confianza , Elasticidad , Humanos , Inmersión , Ensayo de Materiales , Microscopía Electrónica de Rastreo , Análisis de Regresión , Cementos de Resina/química , Estadística como Asunto , Estrés Mecánico , Propiedades de Superficie , Resistencia a la Tracción , Factores de TiempoRESUMEN
PURPOSE: Develop a methodology to correlate microtensile bond strength (microTBS) and leakage within the same site of a specimen and evaluate the predominate debond and leakage pathways. MATERIALS AND METHODS: A Class II (MO) slot cavity was prepared in 37 extracted human molars and restored with indirect resin composite restorations. The restored teeth were thermocycled 300X between 5 and 55 degrees C and then stained with silver. Two specimens per tooth were obtained by diamond saw sectioning for measurement of leakage (microm) and microtensile bond strength (MPa) at the gingival wall and then tested for association with regression analysis. Scanning electron microscopy techniques and energy dispersive spectroscopy (EDS) were used to evaluate the debond and leakage pathways. RESULTS: Association between leakage and bond strength was not significant (t=1.31, p=0.198). All 74 samples debonded within the joint, predominantly at the hybrid layer-adhesive resin region with only one specimen including a small portion of resin cement in the debond pathway. Secondary and backscatter imaging and EDS analysis showed leakage in all samples that involved the hybrid layer. CONCLUSION: This study demonstrates a feasible method for evaluating leakage and bond strength at the joint interface within the same specimen.
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Resinas Compuestas , Recubrimiento Dental Adhesivo , Filtración Dental , Análisis del Estrés Dental/métodos , Recubrimientos Dentinarios , Incrustaciones , Filtración Dental/diagnóstico , Permeabilidad de la Dentina , Humanos , Modelos Lineales , Ensayo de Materiales , Microscopía Electrónica de Rastreo , Diente Molar , Modelos de Riesgos Proporcionales , Análisis de Regresión , Cementos de Resina , Resistencia a la TracciónRESUMEN
OBJECTIVES: This investigation was conducted to determine the tensile bond strength of two dental adhesive using a recently introduced "microtensile" bond strength testing design and to verify the failure mode for each test specimen with scanning electron microscopy (SEM). METHODS: Extracted human molars were mounted in stone and the enamel was removed the occlusal surface perpendicular to the long axis of the tooth. A composite resin crown was formed on this flat dentin surface utilizing each dental adhesive system according to manufacturer's instructions. Twenty-four hours later the bonded test specimens were sectioned perpendicular to the adhesive joint, producing six to seven thin slabs per tooth. These dentin/adhesive/composite resin slabs were sectioned free from the stone block and mounted into custom Plexiglas fixtures for trimming and subsequent tensile bond strength testing at 7 d post-bonding. The bond strength of the two adhesives was statistically compared with the t test. The broken specimens were examined with SEM to determine the fracture location or failure mode. Failures for each adhesive system were categorized as either interfacial (joint or mixed) or substrate (dentin and composite) and evaluated by Fisher's exact test. RESULTS: The tensile bond strength and failure modes of All-Bond 2 (Bisco) and Optibond FL (Kerr) were not significantly different. Sixty per cent (12/20) of fractures involving All Bond 2 occurred at the interface, with seven being entirely maintained within the joint, whereas Optibond FL had 35% (7/20) involving some portion of the interface, two totally within the joint. Cohesive fractures of either dentin or composite accounted for 55% of the total failure modes (21/40). The remaining dentin thickness did not affect the measured tensile bond strength. SIGNIFICANCE: This versatile new method permits multiple measurements from a single tooth or small surface areas within a restoration but careful interpretation of the failure mode is required to prevent inappropriate conclusions about the utility of the test.
Asunto(s)
Recubrimiento Dental Adhesivo , Recubrimientos Dentinarios , Metacrilatos/química , Cementos de Resina/química , Coronas , Fracaso de la Restauración Dental , Dentina/química , Recubrimientos Dentinarios/química , Humanos , Ensayo de Materiales/instrumentación , Ensayo de Materiales/métodos , Microscopía Electrónica de Rastreo , Resistencia a la TracciónRESUMEN
OBJECTIVES: Micromechanical retention from the hybrid layer is generally believed to be the mechanism of adhesion of current generation dentin bonding agents. The purpose of this investigation is to evaluate the interfacial fracture toughness of a commercial dentin bonding agent with and without this hybrid layer. METHODS: Ten extracted molars (AB2) were flattened on the occlusal surface, All-Bond 2 Universal Adhesive System (Bisco) was applied according to manufacturer's directions and a resin composite (Prodigy, Kerr) crown was formed. Another group of ten molars (AB2Cl) was handled identically with the exception of a 1 min gentle scrubbing application of 5.25% sodium hypochlorite after acid etching to remove the acid-exposed collagen. Plane-strain chevron-notch short bar fracture toughness specimens were fabricated from all 20 composite crowns and tested according to ASTM E1304-89. Each group was tested to failure in tensile mode at 0.1 mm min-1 and the maximum load at failure was used to determine plane-strain fracture toughness (KQvM). Weibull parameters were calculated and fracture probability distributions were tested for significant difference at the 95% confidence level. Scanning electron microscopy was employed on broken specimens (18/20) to describe the failure mode. RESULTS: Weibull distributions were not significantly different with characteristic plane-strain fracture toughness from maximum load (KQvM0) of 0.97 MPa m1/2 and 0.81 MPa m1/2 and a Weibull modulus of 4.7 and 3.9, respectively, for AB2 and AB2Cl. All AB2 samples failed within the adhesive joint, while the AB2Cl crack propagated from the interphase of adhesive resin and dentin to 1-2 microns into dentin. SIGNIFICANCE: Under the conditions of this study, the presence of collagen did not contribute to a significantly stronger bonded joint. Interfacial fracture toughness evaluation of the dentin-resin composite bimaterial interface shows promise for future investigations.
Asunto(s)
Recubrimiento Dental Adhesivo , Recubrimientos Dentinarios/química , Metacrilatos/química , Colágeno/química , Resinas Compuestas/química , Recubrimiento Dental Adhesivo/métodos , Dentina , Humanos , Ensayo de Materiales , Microscopía Electrónica de Rastreo , Hipoclorito de Sodio , Propiedades de Superficie , Análisis de Supervivencia , Resistencia a la TracciónRESUMEN
The purpose of this study was (1) to evaluate the effect of a 2-minute exposure of 5% NaOCl following acid conditioning of the dentin on the shear bond strength for two adhesive systems and (2) to examine the ultrastructure of the resindentin interface under SEM. The mesial and distal surfaces of 28 extracted human third molars were ground to expose dentin, then polished with 600-grit SiC. Teeth were randomly assigned to four test groups (n = 14) and received the following treatments: Scotchbond Multi-Purpose (SBMP)--Samples were conditioned with 37% phosphoric acid, rinsed and left moist, SBMP primer and adhesive were applied according to the manufacturer's directions, and Restorative Z-100 composite resin was bonded to the dentin surface. SBMP/NaOCl--The same procedures were followed as for SBMP except the surfaces were treated with 5% NaOCl for 2 minutes, after acid conditioning. All-Bond 2 (AB2)--The same technique was followed as for SBMP, using AB2 according to the manufacturer's recommendations. AB2/NaOCl--The same procedure was followed as for SBMP/NaOCl, using AB2. Specimens were thermocycled in a water bath 300 times between 5 degrees-55 degrees C, then sheared in a Zwick Universal Testing Machine. A one-way ANOVA and Duncan's Multiple Range Test were used for statistical analysis of the data. A 2-minute exposure of dentin to 5% NaOCl following acid conditioning of the dentin had no significant effect on the dentin shear bond strength for Scotchbond Multi-Purpose, but significantly increased the bond strength of All-Bond 2 specimens. The interfacial structure of the dentin to resin bond for two dentin treatments and two adhesive systems was studied morphologically under the scanning electron microscope. Argon ion beam etching and acid demineralization clearly revealed the hybrid layer for the conventional treatment with phosphoric acid and indicated an absence of this resin-impregnated collagen network in those specimens treated with both phosphoric acid and NaOCl.
Asunto(s)
Resinas Compuestas/química , Recubrimiento Dental Adhesivo , Recubrimientos Dentinarios/química , Dentina/efectos de los fármacos , Cementos de Resina , Hipoclorito de Sodio/farmacología , Grabado Ácido Dental , Análisis de Varianza , Colágeno , Recubrimiento Dental Adhesivo/métodos , Dentina/ultraestructura , Humanos , Ensayo de Materiales , Metacrilatos , Microscopía Electrónica de Rastreo , Dióxido de Silicio , Capa de Barro Dentinario , Propiedades de Superficie , CirconioRESUMEN
The importance of eight nucleoside 2'-deoxyribosyltransferase residues for catalysis was investigated by site-directed mutagenesis. Each residue was selected because of its proximity to nucleophile Glu-98 or on its potential contribution to intrinsic protein fluorescence. Mutation of Asp-72, Asp-92, Tyr-7, Trp-12, and Met-125 resulted in over a 90% activity loss whereas mutation of Tyr-157, Trp-64, and Trp-127 produced less than a 80% activity loss. The magnitude of the perturbation on catalysis by mutation, however, was dependent on donor substrate. The kcat values for dIno hydrolysis by these mutants were greater than 25% of that for native enzyme. Although mutant and native enzymes bound substrate analogues with comparable affinities, Km values for dIno hydrolysis varied over a 1000-fold range. The pH dependence of Glu-98 esterification by dCyd suggested that amino acids with pK values of 4.2 and 7.5 were relevant for catalysis. The intrinsic protein fluorescence was attributed primarily to Trp-127 (approximately 80%). Pre-steady-state kinetic parameters for deoxyribosylation of mutant enzymes by dCyd, dThd, and dAdo were determined by monitoring changes in enzyme fluorescence. Collectively, results from mutagenesis suggest that, depending upon substrate, either Asp-92 or Asp-72 functions as the general acid catalyst, and that this enzyme undergoes a change in conformation upon Glu-98 deoxyribosylation.
Asunto(s)
Pentosiltransferasa/química , Pentosiltransferasa/metabolismo , Sitios de Unión , Catálisis , Cinética , Modelos Químicos , Modelos Moleculares , Mutagénesis Sitio-Dirigida , Conformación Proteica , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Espectrometría de FluorescenciaRESUMEN
BACKGROUND: Nucleoside 2-deoxyribosyltransferase plays an important role in the salvage pathway of nucleotide metabolism in certain organisms, catalyzing the cleavage of beta-2'-deoxyribonucleosides and the subsequent transfer of the deoxyribosyl moiety to an acceptor purine or pyrimidine base. The kinetics describe a ping-pong-bi-bi pathway involving the formation of a covalent enzyme-deoxyribose intermediate. The enzyme is produced by a limited number of microorganisms and its functions have been exploited in its use as a biocatalyst to synthesize nucleoside analogs of therapeutic interest. RESULTS: We describe the crystal structure of the enzyme with and without bound ligand. The native structure was solved by the single isomorphous replacement with anomalous scattering method (SIRAS) and refined to 2.5 A resolution resulting in a crystallographic R factor of 16.6%. The enzyme comprises a single domain that belongs to the general class of doubly-wound alpha/beta proteins; it also exhibits a unique nucleoside-binding motif. X-ray analysis of enzyme-purine and enzyme-pyrimidine complexes presented here reveals that the active site lies in a cleft formed by the edge of the beta sheet and two alpha helices and contains side chains from two subunits. CONCLUSIONS: These results indicate residues that may be important in substrate binding and catalysis and thus may serve as a framework for elucidating the mechanism of enzyme activity. In particular, the proposed nucleophile, Glu98, lies in the nucleoside-binding pocket at an appropriate position for nucleophilic attack. A comparison of the enzyme interactions with both a purine and pyrimidine ligand provides some insight into the structural basis for enzyme specificity.
Asunto(s)
Sitios de Unión , Pentosiltransferasa/química , Cristalografía por Rayos X , Lactobacillus/enzimología , Unión Proteica , Estructura Secundaria de Proteína , Purinas/química , Purinas/metabolismo , Pirimidinas/química , Pirimidinas/metabolismo , Especificidad por SustratoRESUMEN
The etched resin technique with unfilled acrylic resin has been successful for Class IV and VI defects on anterior incisor teeth and has proven more useful than alternatives in the immediate treatment of traumatized teeth. The advantages of this approach are ease of operation, excellent retention, conservative tooth preparation, excellent esthetics, lower cost to patient, reduced additional insult to recently traumatized teeth in stabilizing them, stable color, and lack of trauma to opposing teeth (Fig. 10).