Effect of conditioning protocols and ultrasonic application of an infiltrant resin in white spot lesions

Abstract The aim of this study was to evaluate different conditioning protocols and sonic/ultrasonic application of an infiltrant resin (IR) in artificial white spot lesions (AWSL). the V/L surfaces of 48 molars were induced to an AWSL and divided in 6 groups, according to the conditioning protocols and application technique: 15% hydrochloric acid (HA) + manual application of the IR; HA + 37% phosphoric acid (PA) + manual application of the IR; HA + ultrasonic application (U) of the IR; HA + sonic application (S) of the IR; PA+HA+S; and PA+HA+U. For the Penetration Depth (PD), the crowns were etched with HA for 120s. The IR Icon® (DMG) was applied according to the manufacturer`s instructions. The crowns were dye penetrated (0.1% red fluorophore rhodamine B isothiocyanate for 12h) and bleached with 30% hydrogen peroxide for 12 h. The discs were immersed in a 50% ethanol solution, containing 100 µM of sodium fluorescein. The PD (in µm) was measured using confocal laser scanning microscopy (20x). The bond strength (BS) was performed by michoshear test (0.5 mm/min). Data were submitted to 2-way ANOVA and Tukey (α=0.05). For BS, the interaction was not significant (p>0.05). For PD, the main factors were significant (application - p<0.001; conditioning technique - p=0.003). The ultrasonic application showed the highest PD values. PA+HA presented higher results than HA. The sonic/ultrasonic applications and the use of phosphoric acid prior to hydrochloric acid improved PD of the infiltrant resin. Conditioning protocols or application techniques did not influence BS values.

Resumo: O objetivo deste estudo foi avaliar diferentes protocolos de condicionamento e a aplicação sônica/ultrassônica de uma resina infiltrante (RI) em lesões de mancha branca produzidas artificialmente (LMBA). As superfícies vestibulares/linguais de 48 molares foram induzidas à formação de LMBA e divididas em 6 grupos, de acordo com os protocolos de condicionamento e técnica de aplicação da resina infiltrante: ácido hidroclorídrico 15% (AH) + aplicação manual da RI; AH + ácido fosfórico 37% (AF) + aplicação manual da RI; AH + aplicação ultrassônica da RI (U); AH + aplicação sônica da RI (S); AF + AH + S; e AF + AH + U. para o grau de penetração (GP), condicionou-se as coroas com AH por 120 s. A RI Icon® (DMG) foi aplicada de acordo com as instruções do fabricante. As coroas foram coradas (rodamina B 0,1% por 12 h) e clareadas com peróxido de hidrogênio 30% por 12 h. os discos foram imersos em solução de etanol 50%, contendo 100 µM de fluoresceína sódica. O GP (em µm) foi mensurado por meio de microscopia confocal a laser (20´). A resistência de união (RU) foi calculada pelo teste de microcisalhamento (0,5 mm/min). Os dados foram submetidos ao teste ANOVA 2 fatores e Tukey (α=0,05). Para RU, a interação não foi significante (p>0,05). Para GP, os fatores principais foram significantes (técnica de aplicação - p<0,001; protocolos de condicionamento - p=0,003). A aplicação U mostrou os maiores valores de GP. AF+AH demonstrou resultados superiores ao grupo AH. As aplicações sônica/ultrassônica e o uso do ácido fosfórico antes do ácido hidroclorídrico aumentaram o GP da resina infiltrante. Os protocolos de condicionamento ou as técnicas de aplicação não influenciaram os valores de RU.

Comparative transcriptomic analysis reveals adriamycin-induced apoptosis via p53 signaling pathway in retinal pigment epithelial cells / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

OBJECTIVE@#This paper applied a transcriptomic approach to investigate the mechanisms of adriamycin (ADR) in treating proliferative vitreoretinopathy (PVR) using ARPE-19 cells.@*METHODS@#The growth inhibitory effects of ADR on ARPE-19 cells were assessed by sulforhodamine B (SRB) assay and propidium iodide (PI) staining using flow cytometry. The differentially expressed genes between ADR-treated ARPE-19 cells and normal ARPE-19 cells and the signaling pathways involved were investigated by microarray analysis. Mitochondrial function was detected by JC-1 staining using flow cytometry and the Bcl-2/Bax protein family. The phosphorylated histone H2AX (γ-H2AX), phosphorylated checkpoint kinase 1 (p-CHK1), and phosphorylated checkpoint kinase 2 (p-CHK2) were assessed to detect DNA damage and repair.@*RESULTS@#ADR could significantly inhibit ARPE-19 cell proliferation and induce caspase-dependent apoptosis in vitro. In total, 4479 differentially expressed genes were found, and gene ontology items and the p53 signaling pathway were enriched. A protein-protein interaction analysis indicated that the TP53 protein molecules regulated by ADR were related to DNA damage and oxidative stress. ADR reduced mitochondrial membrane potential and the Bcl-2/Bax ratio. p53-knockdown restored the activation of c-caspase-3 activity induced by ADR by regulating Bax expression, and it inhibited ADR-induced ARPE-19 cell apoptosis. Finally, the levels of the γ-H2AX, p-CHK1, and p-CHK2 proteins were up-regulated after ADR exposure.@*CONCLUSIONS@#The mechanism of ARPE-19 cell death induced by ADR may be caspase-dependent apoptosis, and it may be regulated by the p53-dependent mitochondrial dysfunction, activating the p53 signaling pathway through DNA damage.

Efeito de adição de rodamina B e fluoresceína sódica a sistemas adesivos não simplificados: aspectos fotofísicos e físico-químicos / Effect of addition of rhodamine B and fluorescein to conventional etch-and-rinse and self-etching adhesive systems: photophysical and physical-chemical aspects

A adição de corantes fluorescentes a adesivos odontológicos possibilita a investigação da distribuição espacial desses materiais na interface dente-restauração, utilizando-se a microscopia confocal de varredura a laser (MCVL). A literatura indica falta de padronização na aplicação de agentes fluorescentes com tal finalidade. Esse estudo sistematizou estratégias para a adição de rodamina B (RB) e fluoresceína sódica (FS) a um sistema adesivo convencional de três passos, Adper Scotchbond Multi-Purpose (MP), e um autocondicionante de dois passos, Clearfil SE Bond (SE), considerados "padrão-ouro" na Odontologia. Os objetivos principais foram (a) determinar a menor faixa de concentrações de RB e FS necessária para produzir imagens satisfatórias da interface dentina-adesivo e (b) avaliar o efeito da adição desses corantes sobre algumas propriedades das resinas. Os adesivos foram marcados com RB ou FS em concentrações decrescentes (0,5, 0,1, 0,02 e 0,004 mg/mL) por meio de um método de dispersão semidireto. O comportamento fotofísico/ fluorescente dos adesivos marcados foi investigado por espectroscopia de fotoluminescência e MCVL. Paralelamente, avaliaram-se os adesivos quanto ao grau de conversão (GC) e ao ângulo de contato (AC). Tanto os resultados de GC como os de AC foram submetidos à análise de variância com dois fatores (adesivo e tratamento) com α = 0,05, seguida de teste post-hoc de Tukey. Os máximos comprimentos de onda de emissão e de excitação da RB e da FS foram influenciados pelo meio polimérico e pela concentração de corante de modo geral. A MCVL preliminar de amostras de adesivo polimerizado, realizada sob condições experimentais padronizadas, mostrou que o comportamento fluorescente da RB em MP e SE foi muito semelhante na mesma concentração de corante, mas o mesmo não pôde ser dito do comportamento da FS, que foi notavelmente inferior no adesivo autocondicionante, SE, na concentração mais alta. Em dentina, os adesivos preparados com RB nas concentrações-alvo de 0,1 e 0,02 mg/mL apresentaram fluorescência ótima; já aqueles preparados com 0,004 mg/mL produziram fraco sinal. Adesivos preparados com FS a 0,5 mg/mL apresentaram ótima fluorescência na interface de adesão, enquanto que concentração menor desse corante não produziu sinal suficiente. Padrões morfológicos aparentemente atípicos foram observados na interface de adesão, quando da associação do adesivo SE com o corante FS. A adição de RB e FS nas quatro concentrações indicadas aos adesivos MP e SE não afetou o GC nem o AC em comparação com os grupos de controle correspondentes. Em suma, a RB mostra-se um corante mais versátil que a FS na avaliação morfológica das interfaces dentina-MP e dentina-SE via MCVL. A menor faixa de concentrações de RB nos adesivos MP e SE, na qual é possível produzir imagens satisfatórias das interfaces, situa-se entre 0,1­0,02 mg/mL. Já o corante FS deve ser adicionado a esses adesivos a pelo menos 0,5 mg/mL para produzir níveis de fluorescência satisfatórios na interface de adesão. A não ocorrência de efeitos deletérios sobre a polimerização e a molhabilidade das resinas estabelece uma margem de segurança para a incorporação desses agentes fluorescentes (em concentração ≤ 0,5 mg/mL) nesses sistemas monoméricos.(AU)

The addition of fluorescent dyes to dental adhesives makes it possible to investigate the spatial distribution of such resin-based materials in the tooth-restoration interface, using confocal laser scanning microscopy (CLSM). Literature indicates a lack of standardization on the application of fluorescent agents for this purpose. This work presents strategies for adding rhodamine B (RB) and fluorescein sodium salt (FS) to a three-step etch-and-rinse adhesive system, Adper Scotchbond Multi-Purpose (MP), and a two-step self-etching one, Clearfil SE Bond (SE), both regarded as "gold standard" in restorative dentistry. The main objectives were (a) to determine the lowest range of RB and FS concentrations required to produce suitable images of the dentin-adhesive interface via CLSM and (b) to investigate potential effects of addition of these dyes on some resin properties. The adhesives were labeled with RB or FS at decreasing concentrations (0.5, 0.1, 0.02 and 0.004 mg/mL) by means of a semi-direct dispersion method. The photophysical/fluorescent behavior of the labeled resins was investigated by photoluminescence spectroscopy and by CLSM. The adhesives were also investigated with regards to the degree of conversion (DC) and contact angle (CA). A two-way ANOVA of "adhesive" and "treatment" was conducted on DC and CA separately, followed by Tukey's test. The maximum emission and excitation wavelengths of RB and FS were influenced by the host polymer and the dye concentration in general. The preliminary CLSM of cured adhesive samples, performed with standardized settings, showed that the fluorescent behavior of RB in MP and SE was very similar in the same dye concentration, unlike the behavior of FS, which was lower in the self-etching adhesive for the highest dye concentration. In dentin, the adhesives prepared with RB at the target concentrations of 0.1 and 0.02 mg/mL presented optimal fluorescence; those with 0.004 mg/mL produced poor signal. Adhesives prepared with FS at 0.5 mg/mL presented optimal fluorescence at the bonding interface, whereas lower concentrations of FS did not produce sufficient signal. Atypical morphological features were observed at the bonding interface, when adhesive SE was used with FS. The addition of RB and FS at the four decreasing concentrations to adhesives MP and SE did not affect DC or CA compared to the corresponding controls. In short, RB is more versatile than FS for the morphological characterization of dentin-MP and dentin-SE interfaces via MCVL. The lowest range of RB concentrations in adhesives MP and SE that can produce suitable images of the bonding interface lies between 0.1­0.02 mg/mL. The dye FS should be added to these adhesives at 0.5 mg/mL at least to produce satisfactory fluorescence levels at the bonding interface. Since negative effects on polymerization and wettability of the resins were not observed, the use of RB and FS (in concentration ≤ 0.5 mg/mL) together with MP and SE should be reliable in terms of resin properties.(AU)

Water interaction and bond strength to dentin of dye-labelled adhesive as a function of the addition of rhodamine B

ABSTRACT Objective This study investigated the effect of the fluorescent dye rhodamine B (RB) for interfacial micromorphology analysis of dental composite restorations on water sorption/solubility (WS/WSL) and microtensile bond strength to dentin (µTBS) of a 3-step total etch and a 2-step self-etch adhesive system. Material and Methods The adhesives Adper Scotchbond Multi-Purpose (MP) and Clearfil SE Bond (SE) were mixed with 0.1 mg/mL of RB. For the WS/WSL tests, cured resin disks (5.0 mm in diameter x 0.8 mm thick) were prepared and assigned into four groups (n=10): MP, MP-RB, SE, and SE-RB. For µTBS assessment, extracted human third molars (n=40) had the flat occlusal dentin prepared and assigned into the same experimental groups (n=10). After the bonding and restoration procedures, specimens were sectioned in rectangular beams, stored in water and tested after seven days or after 12 months. The failure mode of fractured specimens was qualitatively evaluated under optical microscope (x40). Data from WS/WSL and µTBS were assessed by one-way and three-way ANOVA, respectively, and Tukey’s test (α=5%). Results RB increased the WSL of MP and SE. On the other hand, WS of both MP and SE was not affected by the addition of RB. No significance in µTBS between MP and MP-RB for seven days or one year was observed, whereas for SE a decrease in the µTBS means occurred in both storage times. Conclusions RB should be incorporated into non-simplified DBSs with caution, as it can interfere with their physical-mechanical properties, leading to a possible misinterpretation of bonded interface.

Effect of an Alcohol-Based Caries Detector on the Surface Tension of Sodium Hypochlorite Preparations

The purpose of this study was to evaluate the effect of an alcohol-based caries detector (Kurakay) on the surface tension of a conventional sodium hypochlorite (NaOCl) preparation, and a product containing a surface-active agent (Chlor-XTRA). The surface tensions of the following solutions were tested: NaOCl, a mixture of NaOCl and Kurakay 9:1 w/w, Chlor-XTRA, a mixture of Chlor-XTRA and Kurakay 9:1 w/w. Ten measurements per test solution were made at 20 C, using an optical method called the "Pendant drop method", with a commercially available apparatus. The addition of Kurakay reduced the surface tension for NaOCl (p<0.05) whilst no significant difference was detected for Chlor-XTRA (p>0.05). Statistically significant differences between the NaOCl and Chlor-XTRA groups were found (p<0.05). The addition of an alcohol-based caries detector resulted in a reduction of the original surface tension values for NaOCl only. Taking into account the fact that mixtures of NaOCl and Kurakay have been used to assess the penetration of root canal irrigants in vitro, the related changes in surface tension are a possible source of bias.

O objetivo deste estudo foi avaliar o efeito de um detector de cárie com álcool (Kurakay) sobre a tensão superficial de um preparado convencional de hipoclorito de sódio (NaOCl) e um produto contendo um agente surfactante (Chlor-XTRA). Foram testadas as tensões superficiais das seguintes soluções: NaOCl, uma mistura de NaOCl e Kurakay na proporção de 9:1 em peso, Chlor-XTRA e um mistura de Chlor-XTRA e Kurakay na proporção de 9:1 em peso. Dez medidas foram feitas com cada solução, a 20 °C, utilizando um método óptico chamado "Método da gota pendente" (Pendant drop method) usando aparelhos disponíveis. Adição do Kurakay reduziu a tensão superficial do NaOCl (p<0,05), mas não houve diferença significante para Chlor-XTRA (p>0,05). Foram encontradas diferenças estatisticamente significantes entre os grupos NaOCl e Chlor-XTRA (p<0,05). Adição de um detector de cárie com álcool resultou na diminuição do valor original da tensão superficial apenas para NaOCl. Considerando que as misturas de NaOCl e Kurakay tem sido usadas para estudar a penetração in vitro dos irrigantes radiculares, as diferenças acima são uma possível fonte de desvio nos resultados.

Rapid Determination of Chimerism Status Using Dihydrorhodamine Assay in a Patient with X-linked Chronic Granulomatous Disease Following Hematopoietic Stem Cell Transplantation

Chronic granulomatous disease (CGD) is a rare genetic disease, which is caused by defects in the NADPH oxidase complex (gp91phox, p22phox, p40phox, p47phox, and p67phox) of phagocytes. This defect results in impaired production of superoxide anions and other reactive oxygen species (ROS), which are necessary for killing bacterial and fungal microorganisms and leads to recurrent, life-threatening bacterial and fungal infections and granulomatous inflammation. The dihydrorhodamine (DHR) flow cytometry assay is a useful diagnostic tool for CGD that can detect absent or reduced NADPH oxidase activity in stimulated phagocytes. We report a patient with X-linked CGD carrying a novel mutation of the CYBB gene whose chimerism status following hematopoietic stem cell transplantation (HSCT) has been rapidly determined using the DHR assay. The level of DHR activity correlates well with short tandem repeat PCR analysis. Considering the advantages of this simple, rapid, and cost-effective procedure, serial measurement of DHR assay would facilitate the rapid determination of a patient's engraftment status, as a supplementary monitoring tool of chimerism status following HSCT.

Evaluation of adsorption potential of adsorbents: a case of uptake of cationic dyes.

Adsorption potential of a commercial activated carbon (FS300) has been evaluated for the uptake of cationic dyes namely methylene blue (MB) and rhodamine B (RB). Though, there are numerous studies in literature which report the sorption of MB (more than 40 studies) and RB (more than 10), however none of these use a common parameter to report the capacity of the sorbent. A protocol, based on the equilibrium dye concentration has been proposed to measure the sorption potential of a sorbent. The Langmuir model can very well describe the experimental equilibrium data for both dyes (coefficient of correlation > 0.999). MB (Qm = 312.5 mg g(-1)) is more adsorbable than the RB (Qm = 144.9 mg g(-1)). Molecular weight and chemical structure of dye molecules seem to affect the dye uptake. The effect of pH on dye uptake has also been evaluated by varyingpH from 3 to 11. Uptake of MB increases with pH, wherein RB removal decreases with pH. Dyes could not be desorbed either by distilled water (0.06 and 0.11% for MB and RB respectively), or by 0.1 NHCl (0.136 and 3.0% for MB and RB respectively) indicating, chemical adsorption type of adsorbent-adsorbate interactions.