Biomineralization in Barnacle Base Plate in Association with Adhesive Cement Protein.

Barnacles strongly attach to various underwater substrates by depositing and curing a proteinaceous cement that forms a permanent adhesive layer. The protein MrCP20 present within the calcareous base plate of the acorn barnacle Megabalanus rosa (M. rosa) was investigated for its role in regulating biomineralization and growth of the barnacle base plate, as well as the influence of the mineral on the protein structure and corresponding functional role. Calcium carbonate (CaCO3) growth on gold surfaces modified by 11-mercaptoundecanoic acid (MUA/Au) with or without the protein was followed using quartz crystal microbalance with dissipation monitoring (QCM-D), and the grown crystal polymorph was identified by Raman spectroscopy. It is found that MrCP20 either in solution or on the surface affects the kinetics of nucleation and growth of crystals and stabilizes the metastable vaterite polymorph of CaCO3. A comparative study of mass uptake calculated by applying the Sauerbrey equation to the QCM-D data and quantitative X-ray photoelectron spectroscopy determined that the final surface density of the crystals as well as the crystallization kinetics are influenced by MrCP20. In addition, polarization modulation infrared reflection-absorption spectroscopy of MrCP20 established that, during crystal growth, the content of ß-sheet structures in MrCP20 increases, in line with the formation of amyloid-like fibrils. The results provide insights into the molecular mechanisms by which MrCP20 regulates the biomineralization of the barnacle base plate, while favoring fibril formation, which is advantageous for other functional roles such as adhesion and cohesion.

The tooth on-a-chip: a microphysiologic model system mimicking the biologic interface of the tooth with biomaterials.

The tooth has a unique configuration with respect to biomaterials that are used for its treatment. Cells inside of the dental pulp interface indirectly with biomaterials via a calcified permeable membrane, formed by the dentin matrix and several thousands of dentinal tubules (∼2 µm in diameter). Although the cytotoxic response of the dental pulp to biomaterials has been extensively studied, there is a shortage of in vitro model systems that mimic the dentin-pulp interface and enable an improved understanding of the morphologic, metabolic and functional influence of biomaterials on live dental pulp cells. To address this shortage, here we developed an organ-on-a-chip model system which integrates cells cultured directly on a dentin wall within a microfluidic device that replicates some of the architecture and dynamics of the dentin-pulp interface. The tooth-on-a-chip is made out of molded polydimethylsiloxane (PDMS) with a design consisting of two chambers separated by a dentin fragment. To characterize pulp cell responses to dental materials on-chip, stem cells from the apical papilla (SCAPs) were cultured in odontogenic medium and seeded onto the dentin surface, and observed using live-cell microscopy. Next, to evaluate the tooth-on-a-chip as a platform for materials testing, standard dental materials used clinically (2-hydroxyethylmethacrylate - HEMA, phosphoric acid - PA, and Adper-Scotchbond - SB) were tested for cytotoxicity, cell morphology, and metabolic activity on-chip, and compared against standardized off-chip controls. All dental materials had cytotoxic effects in both on-chip and off-chip systems in the following order: HEMA > SB > PA (p < 0.05), and cells presented consistently higher metabolic activity on-chip than off-chip (p < 0.05). Furthermore, the tooth-on-a-chip enabled real-time tracking of gelatinolytic activity in a model hybrid layer (HL) formed in the microdevice, which suggests that dental pulp cells may contribute to the proteolytic activity in the HL more than endogenous proteases. In conclusion, the tooth-on-a-chip is a novel platform that replicates near-physiologic conditions of the pulp-dentin interface and enables live-cell imaging to study dental pulp cell response to biomaterials.

Os efeitos das Metaloproteinases da matriz extracelular - MMPS e clorexidina no mecanismo de adesão dentária / The effects of metaloproteinases of the extracellular matrix - MMPs and chlorexidine in the dental adhesion mechanism

Introdução: A adesão da resina composta à dentina ocorre pela formação da camada híbrida. Assim, sua degradação ocasiona a perda da resistência de união na interface resina/dentina, influenciando na longevidade da restauração. Após o condicionamento ácido e aplicação do sistema adesivo na dentina desmineralizada, fibras colágenas não envolvidas por sistema adesivo ficam desprotegidas e suscetíveis ao ataque das metaloproteinases (MMPs). Objetivos: Esta revisão buscou esclarecer o efeito das MMPs na degradação da camada híbrida e os efeitos da clorexidina no processo de adesão. Materiais e métodos: Foi realizada uma revisão da literatura por meio de uma busca bibliográfica nas bases de dados Pubmed/ Medline, Scielo e Google Acadêmico, utilizados estudos publicados nos anos de 2005 a 2018. Foi realizada a busca pelos seguintes descritores: Dentistry, MMPs, Chlorhexidine. Resultados: Estas enzimas, presentes na própria dentina, são reativadas pelo ácido fosfórico ou pelos monômeros ácidos dos adesivos autocondicionantes e iniciam a degradação. A aplicação da clorexidina (CHX) na dentina, após o condicionamento ácido, impede ou retarda a degradação das fibras de colágeno da camada híbrida. Conclusão: Concluiu-se que a ligação adesiva à dentina diminui com o passar dos anos devido à ação das MMPs que degradam o colágeno não infiltrado por monômeros adesivos na parte mais profunda da camada híbrida. Além disso, a clorexidina como inibidor terapêutico em sistemas adesivos convencionais é capaz de inibir as MMPs e assim a ligação adesiva à dentina pode ser mantida estável por um período de tempo mais longo.

Introduction: The adhesion of the composite resin to the dentin occurs by the formation of the hybrid layer. Thus, its degradation causes loss of union resistance on interface resin / dentin interface, directly influencing the longevity of the restoration. After the acid etching and the application of the adhesive system into demineralized dentin, collagen fibers not involved by adhesive system get unprotected and susceptibles to attack by metalloproteinases (MMPs). The enzymes, present in the dentin itself, are rehabilitated by phosphoric acid or by the acids monomers of the self-etching adhesives initiating degradation. The application of chlorhexidine (CHX) in the dentin, after acid conditioning, prevents or slows down the degradation of the collagen fibers of the hybrid layer. This literature review sought to clarify the effect of MMPs on the degradation of the hybrid layer and the effects of chlorhexidine on the adhesion process. It was concluded that the adhesive bonding to dentin decreases with the passage of years due in part to the action of MMPs, which degrade collagen not infiltrated by adhesive monomers in the deepest part of the hybrid layer. In addition, the use of chlorhexidine as a therapeutic inhibitor in conventional adhesive systems is capable of inhibiting the MMPs and thus the adhesive bonding to the dentin can be kept stable for a longer period of time.

PAXX and Xlf interplay revealed by impaired CNS development and immunodeficiency of double KO mice.

The repair of DNA double-stranded breaks (DNAdsb) through non-homologous end joining (NHEJ) is a prerequisite for the proper development of the central nervous system and the adaptive immune system. Yet, mice with Xlf or PAXX loss of function are viable and present with very mild immune phenotypes, although their lymphoid cells are sensitive to ionizing radiation attesting for the role of these factors in NHEJ. In contrast, we show here that mice defective for both Xlf and PAXX are embryonically lethal owing to a massive apoptosis of post-mitotic neurons, a situation reminiscent to XRCC4 or DNA Ligase IV KO conditions. The development of the adaptive immune system in Xlf-/-PAXX-/- E18.5 embryos is severely affected with the block of B- and T-cell maturation at the stage of IgH and TCRß gene rearrangements, respectively. This damaging phenotype highlights the functional nexus between Xlf and PAXX, which is critical for the completion of NHEJ-dependent mechanisms during mouse development.

Morphology of the Dentin-resin Interface yielded by Two-step Etch-and-rinse Adhesives with Different Solvents.

AIM: The study aimed to analyze the morphology of the dentin-resin interface yielded by two-step etch-and-rinse adhesive systems with different solvents and compositions. MATERIALS AND METHODS: A total of 32 dentine disks were prepared and randomly assigned to four groups of one-bottle etch-and-rinse adhesive systems containing different solvents: group I, Adper Scotchbond-IXT™ (ethanol/water); group II, XP-Bond™ (tertiary butanol); group III, Prime and Bond NT® (acetone); and group IV, One Coat bond® (5% water). Adhesive systems were applied onto dentin disks, which were then thermal cycled, divided into two hemi-disks (n = 16), and prepared for field-emission scanning electron microscopy to examine the dentin-resin interdiffusion zone. Microphotographs were scanned and data were processed. Data were compared with analysis of variance multivariant test after Kolmogorov-Smirnov and Shapiro-Wilk tests using Statistic Package for the Social Sciences. RESULTS: The adhesive layer thickness average found was group I: 45.9 ± 13.41 urn, group II: 20.6 ± 16.32 urn, group III: 17.7 ± 11.75 urn, and group IV: 50.7 ± 27.81 urn. Significant differences were found between groups I and IV and groups II and III (p < 0.000). Groups I (3.23 ± 0.53 µm) and II (3.13 ± 0.73 µm) yielded significantly thicker hybrid layers than groups III (2.53 ± 0.50 µm) and IV (1.84 ± 0.27 µm) (p < 0.003). Group III presented a less homogeneous hybrid layer, with some gaps. Tag length average was greater in groups II (111.0 ± 36.92 µm) and IV (128.9 ± 78.38 µm) than in groups I (61.5 ± 18.10 µm) and III (68.6 ± 15.84 µm) (p < 0.008). CONCLUSION: Adhesives systems with different solvents led to significant differences in the dentin-resin interface morphology. Solvents role in adhesives bond strength should be considered together with the other adhesive system components. CLINICAL SIGNIFICANCE: The adhesive containing tertiary butanol, in addition, seems to originate a good-quality hybrid layer and long, entangled tags and also appears to have greater ability to originate microtags, which may indicate higher bond strength.

Enamel shear bond strength of different primers combined with an orthodontic adhesive paste.

AIM: The aim of this study was a comparison of shear bond strength (SBS) on tooth enamel of different primers combined with the adhesive paste Transbond XT. MATERIALS AND METHODS: Forty bovine teeth were used in order to create 40 test blocks. The blocks were divided into four groups of 10 blocks each: group A - sample primer (SP); group B - Opal Seal (OS); group C - Transbond Plus SEP (TSEP); group D - Transbond XT Primer (TXT). After surface preparation and application of the primer, respectively, two stainless steel brackets were fixed on each tooth by using Transbond XT. Accordingly, 80 brackets were debonded (n=20). Shear bond strength and adhesive remnant index (ARI) scores were evaluated. Statistical analyses were performed by using the Student's t-test and Mann-Whitney U test. RESULTS: All tested groups revealed high shear bond strength in a similar size range. There were no significant differences between the groups concerning shear bond strength. The ARI scores of group C showed significantly lower ARI scores (0 and 1) than that of group D. Apart from that there was no statistical difference. CONCLUSION: In combination with the adhesive paste Transbond XT, all tested primers were suitable for fixing orthodontic brackets. The primers could be changed according to the clinical situation.

Streptococcus mutans forms xylitol-resistant biofilm on excess adhesive flash in novel ex-vivo orthodontic bracket model.

INTRODUCTION: During orthodontic bonding procedures, excess adhesive is invariably left on the tooth surface at the interface between the bracket and the enamel junction; it is called excess adhesive flash (EAF). We comparatively evaluated the biofilm formation of Streptococcus mutans on EAF produced by 2 adhesives and examined the therapeutic efficacy of xylitol on S mutans formed on EAF. METHODS: First, we investigated the biofilm formation of S mutans on 3 orthodontic bracket types: stainless steel preadjusted edgewise, ceramic preadjusted edgewise, and stainless steel self-ligating. Subsequently, tooth-colored Transbond XT (3M Unitek, Monrovia, Calif) and green Grengloo (Ormco, Glendora, Calif) adhesives were used for bonding ceramic brackets to extracted teeth. S mutans biofilms on EAF produced by the adhesives were studied using the crystal violet assay and scanning electron microscopy. Surface roughness and surface energy of the EAF were examined. The therapeutic efficacies of different concentrations of xylitol were tested on S mutans biofilms. RESULTS: Significantly higher biofilms were formed on the ceramic preadjusted edgewise brackets (P = 0.003). Transbond XT had significantly higher S mutans biofilms compared with Grengloo surfaces (P = 0.007). There was no significant difference in surface roughness between Transbond XT and Grengloo surfaces (P >0.05). Surface energy of Transbond XT had a considerably smaller contact angle than did Grengloo, suggesting that Transbond XT is a more hydrophilic material. Xylitol at low concentrations had no significant effect on the reduction of S mutans biofilms on orthodontic adhesives (P = 0.016). CONCLUSIONS: Transbond XT orthodontic adhesive resulted in more S mutans biofilm compared with Grengloo adhesive on ceramic brackets. Surface energy seemed to play a more important role than surface roughness for the formation of S mutans biofilm on EAF. Xylitol does not appear to have a therapeutic effect on mature S mutans biofilm.

Self-etching zinc-doped adhesives improve the potential of caries-affected dentin to be functionally remineralized.

The aim of this study was to evaluate if mechanical cycling influences bioactivity at the resin-carious dentin interface after bonding with Zn-doped self-etching adhesives. Caries-affected dentin surfaces were bonded with: Clearfil SE bond (SEB), and 10 wt. % ZnO nanoparticles or 2 wt. % ZnCl2 were added into the SEB primer or bonding components. Bonded interfaces were stored during 24 h and then tested or submitted to mechanical loading. Microtensile bond strength was assessed. Debonded dentin surfaces were studied by field emission scanning electron microscopy. Remineralization of the bonded interfaces was evaluated through nanohardness (Hi) and Young's modulus (Ei), Raman spectroscopy/cluster analysis, and Masson's trichrome staining technique. New precipitation of minerals composed of zinc-base salts and multiple Zn-rich phosphate deposits was observed in samples infiltrated with the Zn-doped adhesives. At the hybrid layer, specimens treated with ZnO incorporated in the primer (SEB·P-ZnO), after load cycling, attained the highest Ei and Hi. Load cycling increased Ei at the bottom of the hybrid layer when both, SEB undoped and SEB with ZnCl2 included in the bonding (SEB·Bd-ZnCl2), were used. ZnO incorporated in the primer promoted an increase in height of the phosphate and carbonate peaks, crystallinity, relative mineral concentration, and lower collagen crosslinking. ZnCl2 included in the bonding attained similar results, but relative mineral concentration decreased, associated to higher crosslinking and restricted collagen maturation. In general, a substantial restoration of the mechanical properties of caries-affected dentin substrata occurred when SEB-Zn doped adhesives were used and load cycled was applied, leading to functional and biochemical remineralization.

Microbial management of anaerobic digestion: exploiting the microbiome-functionality nexus.

Anaerobic reactors are mostly operated based on the monitoring of process parameters and empirical expert knowledge due to the limitations of microbial-based management. This review analyzes the requirements to conduct microbial management in anaerobic digestion, emphasizing the importance of understanding the anaerobic microbiome and the need of establishing microbial indicators of optimal performance. The strategies currently applied to shape the reactor microbiome are explored and we assess critically the different types of management (retrospective, prospective and proactive). We conclude that future research should lead to more useful data or insights to accomplish proactive management, seen as stimulation and anticipation rather than remediation.

Effect of imaging powders on the bond strength of resin cement.

The application and incomplete removal of a computer-aided design/computer-aided manufacture imaging powder may affect the dentin surface prior to bonding a ceramic restoration. The purpose of this study was to compare the effect of imaging powder residue on the shear bond strength of a self-adhesive resin cement to dentin. Mounted human third molars were sectioned coronally with a diamond saw to expose the dentin, which was then prepared with a diamond bur mounted in a custom jig. The dentin surface was sprayed with 3 different imaging powders. The 3 powder groups were then divided into 3 subgroups based on the method of powder removal: no rinse, 1-second rinse, and 10-second rinse. A control group was created that had no application of imaging powder. A self-adhesive resin cement was bonded to the surfaces and loaded to failure in a universal testing machine after 24 hours of storage. Data was analyzed with Kruskal-Wallis and Mann-Whitney nonparametric tests. The bonding to dentin surfaces of the powder groups that were rinsed for 1 or 10 seconds were not significantly different from each other or the nonpowdered control. The type of imaging powder did not significantly affect the bond strength. The nonrinsed powdered dentin surface had a significant reduction in bond strength compared to both the control and the rinsed powdered surfaces.

Effect of cleansing methods on saliva-contaminated zirconia--an evaluation of resin bond durability.

The aims of this study were to investigate 1) the influence of cleansing methods after saliva contamination and 2) aging conditions (thermocycling and water storage) on zirconia shear bond strength (SBS) with a resin cement. One hundred and eighty zirconia specimens were sandblasted with 50 µm aluminum oxide particles, immersed in saliva for one minute (with the exception of the control group, [C]), and divided into groups according to the cleansing method, as follows: water rinse (W); 37% phosphoric acid gel (PA); cleaning paste (ie, Ivoclean®) containing mainly zirconium oxide (IC); and 70% isopropanol (AL). Scanning electron microscopy was done to qualitatively evaluate the zirconia surface after each cleansing method. For the SBS test, resin cement buttons were bonded to the specimens using a dedicated jig. SBS was evaluated according to standard protocols after 24 hours, 5000 thermal cycles (TC), or 150 days of water storage. Statistical analysis was performed using two-way analysis of variance and Tukey test (p<0.05). Data showed a significant effect for the 150 days of water storage, TC, and 24 hours of water storage (150 days < TC < 24 hours). Group comparisons showed that PA < AL and W < IC and C. SBS ranged from 10.4 to 21.9 MPa (24 hours), from 6.4 to 14.8 MPa (TC), and from 2.9 to 7.0 MPa (150 days). Failure analysis revealed a greater percentage of mixed failures for the majority of the specimens and a smaller percentage of adhesive failures at the ceramic-resin cement interface. Our findings suggest that Ivoclean® was able to maintain adequate SBS values after TC and 150 days of storage, comparable to the uncontaminated zirconia.

Degradação química e citotoxicidade in vitro de cimentos resinosos fotopolimerizáveis / Chemical degradation and in vitro cytotoxicity of light-cured resin cements

O objetivo deste estudo foi investigar o efeito degradante da saliva artificial e do ácido lático sobre diferentes cimentos resinosos fotopolimerizáveis e avaliar os efeitos citotóxicos dos extratos da degradação em saliva artificial sobre fibroblastos Balb/c 3T3 de ratos. Os cimentos Variolink II (base), AllCem Veneer e RelyX Veneer foram testados. Para caracterização microestrutural destes materiais, análise termogravimétrica (TGA), cálculo do grau de conversão (GC) e análise da distribuição granulométrica das partículas de carga, foram realizados. O GC foi calculado utilizando-se a espectroscopia infravermelha por transformada de Fourier (FTIR). Dez amostras de cada cimento foram construídas em matriz teflon (0,5 mm x 5 mm) e fotoativadas pelo diodo emissor de luz (LED) Elipar S10 (3M ESPE). Após obtenção dos espectros FTIR iniciais (24h após a fotopolimerização), 5 amostras de cada cimento foram, individualmente, imersas em 10 ml de saliva artificial (pH = 7,0) e outras 5 em 10 ml de solução de ácido lático (pH = 4,0), por 18 dias, a 37°C. Depois, as amostras foram submetidas a novas análises espectroscópicas FTIR, sob as mesmas condições iniciais, e posteriormente, seus espectros foram comparados qualitativamente aos obtidos antes do processo de degradação. A viabilidade celular de fibroblastos 3T3, frente aos possíveis efeitos citotóxicos dos resíduos dos cimentos livres na solução de saliva artificial após 18 dias de degradação, foi testada através da redução do brometo de dimetiltiazol-difeniltetrazólico (MTT). As culturas foram expostas aos extratos salivares por 24h e 72h. Células controle foram expostas à solução de saliva artificial pura. Dados foram submetidos a ANOVA e teste Tukey. Os resultados demonstraram maior conteúdo de partículas de carga significante ao cimento Variolink II (65,86%±0,17; p<0,05) quando comparado aos cimentos RelyX Veneer (62,29%±0,30) e AllCem Veneer (62,15%±37,84); distribuição granulométrica trimodal aos cimentos Variolink II (0,2 ­ 3,3µm) e RelyX Veneer (0,6 ­ 29µm), e monomodal ao AllCem Veneer (1 ­ 4,1µm); maior GC ao cimento AllCem Veneer (71,23%±5,53; p<0,05), enquanto RelyX Veneer (66,00%±6,84) e Variolink II (62,24%±2,45) não diferiram entre si (p=0,1306). A solução de ácido lático contribuiu para maiores alterações sobre o conteúdo inorgânico do cimento Variolink II, porém este material demonstrou maior degradação polimérica após imersão em saliva artificial. O cimento RelyX Veneer degradou tanto em saliva artificial quanto em ácido lático, não havendo diferenças em relação à solução degradante. O cimento AllCem Veneer não revelou degradação química após imersão em ambos os meios testados. Em 24h de exposição, os resíduos do cimento AllCem Veneer garantiram a maior viabilidade celular para fibroblastos 3T3 (90,0±6,3), enquanto os do cimento Variolink II a menor (7,43±0,17). Após 72h de exposição, todos os extratos salivares ofereceram citotoxicidades importantes aos fibroblastos. Concluiu-se que os cimentos resinosos avaliados são passíveis de sofrerem degradação em condições que simulam o ambiente oral. Existiu uma relação direta entre degradação química dos cimentos resinosos e efeitos citotóxicos in vitro a fibroblastos 3T3.

The aim of this study was to evaluate the degradation of different light-activated resin cements after aging in artificial saliva and lactic acid solutions, and the in vitro cytotoxicity of salivary extracts obtained from the resin cements degradation on mouse fibroblast (Balb/c 3T3). The resin cements Variolink II, AllCem Veneer and RelyX Veneer were tested. For microstructure characterization, TGA, degree of conversion (DC) and granulometric distribution of filler particles were employed. The DC was calculated from baseline FTIR spectra obtained from uncured and cured samples of each resin cement. These samples were development on teflon mold (0.5 mm x 5.0 mm) and photoactivated by a LED (Elipar S10; 3M ESPE). After baseline FTIR spectra obtaining, 5 samples of each cement were immersed in 10 ml of artificial saliva (pH = 7.0) and the others 5 samples were immersed in 10 ml of lactic acid solution (pH = 4.0), individually, for 18 days, at 37°C. Then, the samples were submitted to new FTIR analyzes, at the same anterior conditions, and their new spectra were compared with the baselines by qualitative FTIR method. The cell viability of 3T3 fibroblast was evaluated of MTT test. The cells were exposed to salivary extracts for 24 h and 72 h. Control cells were exposed to pure artificial saliva solution. The data were submitted to ANOVA and Tukey's test. The results revealed significant higher mass percentage of filler particles for Variolink II (65.86%±0.17; p<0.05) when compared with RelyX Veneer (62.29%±0.30) and AllCem Veneer (62.15%±37.84); trimodal granulometric distribution for Variolink II (0.2 ­ 3.3 µm) and RelyX Veneer (0.6 ­ 29.0 µm), and monomodal distribution for AllCem Veneer (1.0 ­ 4.1 µm); significant higher baseline DC for AllCem Veneer (71.23%; p<0,05), while RelyX Veneer (66.00%±6.84) and Variolink II (62.24%±2.45) were not different from each other (p=0.1306). The lactic acid solution contributed to higher changes of the inorganic matrix for Variolink II cement. However, this cement revealed more polymeric degradation after aging in artificial saliva when compared with lactic acid solution. RelyX Veneer had chemical degradation in both solutions. AllCem Veneer not showed any chemical degradation after aging in both solutions. Within 24 h of exposure, the chemical wastes of the AllCem Veneer cement guaranteed the higher cell viability to 3T3 fibroblasts (90.0±6.30) and there was no difference with the control group (80.3±8.70). The chemical wastes of the Variolink II cement guaranteed the smaller cell viability at the same time (7.43±0.17). After 72 h of exposure, all salivary extracts, except for control group, caused important cytotoxic effects to 3T3 fibroblasts. Then, the resin cements evaluated in this study can suffer chemical degradation under conditions that simulate the oral environment. There was a direct relationship between chemical degradation and in vitro cytotoxicity.

In vivo biocompatibility versus degree of conversion of resin-reinforced cements in different time periods.

This study focused on test the null hypothesis that there is no difference between the degree of conversion and biocompatibility of different resin reinforced glass ionomer cements (RRGICs). Forty-eight male Wistar rats were used, distributed into four groups (n = 12), as follows: Group C (Control, polyethylene), Group FOB (Fuji Ortho Band), Group UBL (Ultra band Lok), and Group MCG (Multicure Glass), in subcutaneous tissue. The events of edema, necrosis, granulation tissue, multinuclear giant cells, young fibroblasts, and collagen formation were analyzed at 7, 15, and 30 days. The degree of conversion was evaluated by the Fourier method. Biocompatibility and degree of conversion were assessed using the Kruskal-Wallis and Dunn tests, and ANOVA and Tukey's test, respectively (P < 0.05). It was observed that, there was significant difference between Groups FOB and UBL for the presence of young fibroblasts at 15 days (P = 0.034) and between the Control and MCG Groups for the presence of multinucleated giant cells at 30 days (P = 0.009). Monomer conversion increased progressively until day 30, with significant difference between Group FOB and Groups UBL and MCG (P = 0.013) at 15 days. The null hypothesis was partially accepted, Fuji Ortho Band showed a less monomer conversion and a smaller number of young fibroblasts in the time of 15 days.

Qualitative evaluation of scanning electron microscopy methods in a study of the resin cement/dentine adhesive interface.

Sample preparation and imaging techniques for scanning electron microscopy (SEM) of dehydrated dental samples can hinder the structural analyses. This study qualitatively evaluated images obtained with two different protocols of SEM preparation and analysis to assess the dentine adhesive interface. The crown and root dentine of 12 bovine incisors were subjected to cementation with the resin cement RelyX U100 or RelyX ARC/SBMP (n = 6). After storage for 7 days in a moist environment at 37 ± 1°C, the dentine samples were dehydrated in an ascending alcohol series, and three specimens from each group were coated with gold or carbon and examined in a high-vacuum (JEOL JSM-6360LV, 10 kV) or low-vacuum (FEI Quanta 200F, 15-30 kV) microscope. Images were obtained at magnifications between 50 and 2,000×, but with different working distances. The use of high vacuum for carbon and gold coating and SEM visualization led to cracks in the samples. A small number of cracks can be described in the specimens subjected to the low-vacuum technique. The protocol for SEM imaging in low vacuum was considered more appropriate for preservation of the integrity of the evaluated structures.

Fabrication of a biodegradable calcium polyphosphate/polyvinyl-urethane carbonate composite for high load bearing osteosynthesis applications.

The formation of biodegradable implants for use in osteosynthesis has been a major goal of biomaterials research for the past 2-3 decades. Self-reinforced polylactide systems represent the most significant success of this research to date, however, with elastic constants up to 12-15 GPa at best, they fail to provide the initial stiffness required of devices for stabilizing fractures of major load-bearing bones. Our research has investigated the use of calcium polyphosphate (CPP), an inorganic polymer in combination with polyvinyl-urethane carbonate (PVUC) organic polymers for such applications. Initial studies indicated that composite samples formed as interpenetrating phase composites (IPC) exhibited suitable as-made strength and stiffness, however, they displayed a rapid loss of properties when exposed to in vitro aging. An investigation to determine the mechanism of this accelerated in vitro degradation for the IPCs as well as to identify possible design changes to overcome this drawback was undertaken using a model IPC system. It was found that strong interfacial strength and minimal swelling of the PVUC are very important for obtaining and maintaining appropriate mechanical properties in vitro.

Effect of salivary esterase on the integrity and fracture toughness of the dentin-resin interface.

Human Salivary Derived Esterases (HSDE) are part of the salivary group of enzymes which show strong degradative activity toward the breakdown of one of the most common monomers used in dental composites and adhesives, 2,2-[4(2-hydroxy 3-methacryloxypropoxy)-phenyl] propane (Bis-GMA), to form the degradation product 2,2-bis [4 (2,3-hydroxy-propoxy)phenyl] propane (Bis-HPPP). This study was aimed to evaluate the effects of HSDE on the biodegradation and fracture toughness of the adhesive resin-dentin interface. Adhesive resin (Scotchbond Multi Purposes), resin composite (Z250) and mini short-rod specimens, were either not incubated; or incubated in phosphate-buffered saline (PBS) or HSDE media for up to 180 days (37 degrees C, pH 7.0). The amount of Bis-HPPP was analyzed by high performance liquid chromatography and mini-SR specimens were tested for fracture toughness using universal testing machine following 30, 90, or 180-day incubation periods. Significantly higher amounts of Bis-HPPP were produced in HSDE than in PBS incubated specimens (p < 0.05). Non-incubated mini-SR specimens had the higher fracture-toughness values, while specimens incubated for 180-days in HSDE had the lowest fracture toughness (p < 0.05). This study suggests that biodegradation is an on-going clinically relevant process that progressively compromises the integrity of the critical resin restoration-adhesive interface, as well as the resin-composite component with time.

Long-term effect of carbodiimide on dentin matrix and resin-dentin bonds.

OBJECTIVES: To characterize the interaction of 1-Ethyl-3-[3-dimethylaminopropyl] carbodiimide Hydrochloride (EDC) with dentin matrix and its effect on the resin-dentin bond. METHODS: Changes to the stiffness of demineralized dentin fragments treated with EDC/N-hydroxysuccinimide (NHS) in different solutions were evaluated at different time points. The resistance against enzymatic degradation was indirectly evaluated by ultimate tensile strength (UTS) test of demineralized dentin treated or not with EDC/NHS and subjected to collagenase digestion. Short- and long-term evaluations of the strength of resin-dentin interfaces treated with EDC/NHS for 1 h were performed using microtensile bond strength (microTBS) test. All data (MPa) were individually analyzed using ANOVA and Tukey HSD tests (alpha = 0.05). RESULTS: The different exposure times significantly increased the stiffness of dentin (p < 0.0001, control-5.15 and EDC/NHS-29.50), while no differences were observed among the different solutions of EDC/NHS (p = 0.063). Collagenase challenge did not affect the UTS values of EDC/NHS group (6.08) (p > 0.05), while complete degradation was observed for the control group (p = 0.0008, control-20.84 and EDC/NHS-43.15). EDC/NHS treatment did not significantly increase resin-dentin muTBS, but the values remained stable after 12 months water storage (p < 0.05). CONCLUSIONS: Biomimetic use of EDC/NHS to induce exogenous collagen cross-links resulted in increased mechanical properties and stability of dentin matrix and dentin-resin interfaces.

In vitro enzymatic biodegradation of adhesive resin in the hybrid layer.

Penetration of adhesives into the demineralized dentin surface and their subsequent conversion are critically important to longevity of the adhesive resin (AR)-dentin bond. The durability of the resin-dentin bond is investigated by monitoring the change of adhesive concentration within the hybrid layer (HL) of aged specimens using Raman spectroscopy. Absolute molar concentrations of Bis-GMA and HEMA were measured across the HL of resin-dentin specimens 24 h after photopolymerization and after 24-week storage in one of three media: artificial saliva (SAL), SAL containing cholesterol esterase to attack resin (EST), and SAL containing bacterial collagenase to attack collagen (COL). No significant difference among these groups for both Bis-GMA and HEMA molar concentrations at 24-h storage was found; however, concentrations decreased from the AR to the middle of the HL. Concentrations remained unchanged at any resin-dentin position after aging in SAL. In the HL, concentrations significantly decreased with aging in COL and tended to decrease in EST. While showing potential enzymatic biodegradative effects of endogenous matrix metalloproteinases and salivary esterases, this methodology may also prove to be a valuable assessment of new chemistries and future approaches to improve resin-dentin bond performance. (c) 2010 Wiley Periodicals, Inc. J Biomed Mater Res, 2010.

Urethane dimethacrylate: a molecule that may cause confusion in dental research.

In recent years, the elution of monomers from dental materials has been a cause for public concern. Urethane dimethacrylate, commonly abbreviated to UDMA, is one of the monomers that are most often tested with regard to elution from and cytotoxicity of resin-based materials. Although each chemical name represents the chemical type, chemical structure, and molecular weight of a molecule, it does not seem to be the same with UDMA. In the present paper, the different forms of UDMA are presented. These include those used by dental manufacturers to produce composite materials and the different types of urethane dimethacrylate used in studies concerning the elution of monomers from composite materials. High-performance liquid chromatography (HPLC) is usually used to detect the eluted monomers, but it does not appear to be adequate in determining the different forms of UDMA. The combination of HPLC with mass spectrometry is shown to be able to specifically identify the compounds eluted in addition to those compounds used as standards in the various studies. The fact that the same name is given to different molecules causes confusion about the results of studies testing the elusion of monomers from composite materials and their possible toxicity.

Effect of bacterial collagenase on resin-dentin bonds degradation.

The objective of this study is to evaluate the effect of a bacterial collagenase on the degradation of resin-dentin bonds. Human dentin surfaces were bonded with: an etch-&-rinse self-priming adhesive (SB), a two-step self-etching primer/adhesive (SEB), and a 1-step self-etching adhesive (OUB). Composite build-ups were constructed. The bonded teeth were stored (24 h, 3 months, 1 year) in distilled water or in a buffered bacterial collagenase solution. Half of the specimens were stored as intact bonded teeth (Indirect Exposure/IE). The other half were sectioned into beams prior to storage (Direct Exposure/DE). After storage the intact teeth were sectioned into beams and all specimens were tested for microtensile bond strengths (MTBS). ANOVA and multiple comparisons tests were performed. Fractographic analysis was performed by scanning electron microscopy. The inclusion of bacterial collagenase in the storing solution did not lower the MTBS values over those seen in specimens stored in water. SB and SEB bonds strength were equal, and were superior to OUB. After 3 months of DE, SB and OUB bonded specimens showed decreases in MTBS; similar reductions required 1 year for SEB/DE. MTBS did not decrease in IE specimens except for OUB. Resin and collagen dissolution were evident in DE groups after storing.