Process development and characterization of recombinant nucleocapsid protein for its application on COVID-19 diagnosis.

COVID-19 pandemic was caused by the severe acute respiratory syndrome coronavirus 2 (Sars-CoV-2). The nucleocapsid (N) protein from Sars-CoV-2 is a highly immunogenic antigen and responsible for genome packing. Serological assays are important tools to detect previous exposure to SARS-CoV-2, complement epidemiological studies, vaccine evaluation and also in COVID-19 surveillance. SARS-CoV-2 N (r2N) protein was produced in Escherichia coli, characterized, and the immunological performance was evaluated by enzyme-linked immunosorbent assay (ELISA) and beads-based array immunoassay. r2N protein oligomers were evidenced when it is associated to nucleic acid. Benzonase treatment reduced host nucleic acid associated to r2N protein, but crosslinking assay still demonstrates the presence of higher-order oligomers. Nevertheless, after RNase treatment the higher-order oligomers reduced, and dimer form increased, suggesting RNA contributes to the oligomer formation. Structural analysis revealed nucleic acid did not interfere with the thermal stability of the recombinant protein. Interestingly, nucleic acid was able to prevent r2N protein aggregation even with increasing temperature while the protein benzonase treated begin aggregation process above 55 °C. In immunological characterization, ELISA performed with 233 serum samples presented a sensitivity of 97.44% (95% Confidence Interval, CI, 91.04%, 99.69%) and a specificity of 98.71% (95% CI, 95.42%, 99.84%) while beads-based array immunoassay carried out with 217 samples showed 100% sensitivity and 98.6% specificity. The results exhibited an excellent immunological performance of r2N protein in serologic assays showing that, even in presence of nucleic acid, it can be used as a component of an immunoassay for the sensitive and specific detection of SARS-CoV-2 antibodies.

Brazilian meningococcal C conjugate vaccine: physicochemical, immunological, and thermal stability characteristics.

High temperature is known to cause some instability in polysaccharide-protein conjugated vaccines and studies under stress conditions may be useful in determining whether short-term accidental exposure to undesired conditions can compromise product quality. In this study, we examined the structural stability of three industrial batches of Brazilian Meningococcal C conjugate bulk (MPCT) incubated at 4, 37, and 55 °C for 5 weeks. The effect of exposure to the storage temperatures was monitored by HPLC-SEC, CZE, CD and NMR techniques. The immunological significance of any physicochemical changes observed in MPCT was determined by SBA and ELISA assays of serum from immunized mice. Fluorescence emission spectra at 4 and 37 °C were similar among all samples and compatible with the native fold of the carrier protein. Fluorescence spectra of MPCT stored at 55 °C decreased in intensity and had a significant red-shift, indicating conformational changes. Far-UV CD spectra revealed a trend toward loss of structural conformation as storage temperature was increased to 55 °C. The NMR data showed modified signal intensity of the aromatic and aliphatic residues, mainly for samples incubated at 55 °C, suggesting a partial loss of tertiary structure. About 50% free saccharide content was found in bulks stored at 55 °C, but no difference was observed in the IgG or SBA titers. The present study showed physicochemical methods alone are insufficient to predict the biological activity of a MPCT conjugate vaccine without extensive validation against immunological data. However, they provide a sensitive means of detecting changes induced in a vaccine exposed to adverse environmental condition.

In vitro cytotoxicity of self-curing acrylic resins of different colors.

OBJECTIVE: The aim of this study was to assess the in vitro cytotoxicity of acrylic resins of different colors over time. METHODS: Specimens were divided into 4 groups (n = 6) according to the color of the acrylic resin (Orto Class, Clássico, Campinas, São Paulo, Brazil): Group 1, clear acrylic resin; Group 2, pink acrylic resin; Group 3, blue acrylic resin; and Group 4, green acrylic resin. All specimens were fabricated according to the mass manipulation technique and submitted to mechanical polishing protocol. The control was performed with an amalgam specimen (C+), a glass specimen (C-) and cell control (CC). Specimens were immersed in Minimum Eagle's Medium (MEM) and incubated for 24 h at 37ºC. The extracts from the experimental material were filtered and mixed with L929 fibroblast. Cytotoxicity was evaluated at four different times, 24, 48, 72 and 168 h. After contact, cells were incubated for 24 h and added to 100 µ of 0.01% neutral red dye. The cells were incubated for 3 h for pigment incorporation and fixed. Cells viability was determined by a spectroscopic (BioTek, Winooski, Vermont, USA) with a 492-nm wavelength λ=492 nm). RESULTS: There were no statistical differences between the experimental groups and the CC and C- groups. CONCLUSION: Clear, pink, blue and green self-curing acrylic resins fabricated by means of the mass manipulation technique and mechanically polished are not cytotoxic. Neither the pigment added to the self-curing acrylic resin nor the factor of time influenced the cytotoxicity of the material.

In vitro cytotoxicity of self-curing acrylic resins of different colors

OBJECTIVE: The aim of this study was to assess the in vitro cytotoxicity of acrylic resins of different colors over time. METHODS: Specimens were divided into 4 groups (n = 6) according to the color of the acrylic resin (Orto Class, Clássico, Campinas, São Paulo, Brazil): Group 1: clear acrylic resin; group 2: pink acrylic resin; group 3: blue acrylic resin and group 4: green acrylic resin. All specimens were fabricated according to the mass manipulation technique and submitted to mechanical polishing protocol. The control was performed with an amalgam specimen (C+), a glass specimen (C-) and cell control (CC). Specimens were immersed in Minimum Eagle's Medium (MEM) and incubated for 24 h at 37o C. The extracts from the experimental material were filtered and mixed with L929 fibroblast. Cytotoxicity was evaluated at 4 different times, 24, 48, 72 and 168 h. After contact, cells were incubated for 24 h and added to 100 µ of 0.01% neutral red dye. The cells were incubated for 3 h for pigment incorporation and fixed. Cells viability was determined by a spectroscopic (BioTek, Winooski, Vermont, USA) with a 492-nm wavelength λ=492 nm). RESULTS: There were no statistical differences between the experimental groups and the CC and C- groups. CONCLUSION: Clear, pink, blue and green self-curing acrylic resins fabricated by means of the mass manipulation technique and mechanically polished are not cytotoxic. Neither the pigment added to the self-curing acrylic resin nor the factor of time influenced the cytotoxicity of the material. .

OBJETIVO: avaliar, in vitro, a citotoxicidade de resinas acrílicas autopolimerizáveis, de diferentes cores, ao longo do tempo. MÉTODOS: os corpos de prova foram divididos em quatro grupos (n = 3), de acordo com a cor da resina acrílica utilizada (Orto Class, Clássico, São Paulo/SP), sendo: grupo 1, acrílica incolor; grupo 2, acrílica rosa; grupo 3, acrílica azul; e, grupo 4, acrílico verde. Todos os corpos de prova foram confeccionados pela técnica de massa e polidos mecanicamente. Um corpo de prova de amálgama, um de vidro e célula constituíram o controle positivo (C+), controle negativo (C-), e controle de célula (CC), respectivamente. Em seguida, esses foram imersos em meio mínimo essencial de Eagle (MEM) por 24h, quando se removeu o sobrenadante e colocou-os em contato com fibroblastos L929. Avaliou-se a citotoxicidade em quatro períodos: 24, 48, 72 e 168h. Após o contato com o meio, as células foram incubadas por 24h e adicionou-se 100µ do corante vermelho neutro a 0,01%. Posteriormente, as células foram incubadas por 3h, para incorporação do corante, e fixadas. A contagem das células viáveis foi realizada em espectrofotômetro (BioTek, Winooski, EUA), com um comprimento de onda de 492nm (λ = 492nm). RESULTADOS: não houve diferença estatística entre os grupos experimentais e os grupos CC e C-. CONCLUSÇÕES: as resinas acrílicas autopolimerizáveis incolor, rosa, azul e verde, manipuladas pela técnica de massa e polidas mecanicamente não são citotóxicas. O corante utilizado em resinas autopolimerizáveis e tempo não influenciam na citotoxocidade do material. .

Citotoxicidade in vitro de enxaguatório bucal em diferentes períodos de tempo / In vitro cytotoxicity of mouthwash in different time periods

O objetivo deste trabalho foi avaliar a citotoxicidade do enxaguatório bucal Plax Tradicional. Avaliou-se em diferentes tempos: 1, 15, 30, 45, 60 e 120 segundos quanto seu efeito citotóxico em fibroblastos gengivais L929. Utilizou-se 3 grupos controle: positivo (C+) detergente celular Tween 80, negativo (C-) PBS, e controle de célula (CC) onde as células não foram expostas a nenhum material. O ensaio de citotoxicidade foi realizado utilizando cultura celular de fibroblasto de camundongo (L929). Após contato do enxaguatório com as células, as mesmas foram colocadas em contato com o corante vital vermelho neutro utilizando-se a técnica “dye uptake”. Os valores da quantidade de células viáveis foram submetidos à análise da variância (ANOVA) para determinar se havia diferença estatística entre os grupos, e posteriormente, ao teste de Tukey (p<0.05). Os resultados demonstraram citotoxicidade do enxaguatório com diferenças estatísticas para os grupos CC e C- (p<0.05). Em relação ao controle positivo, o Plax foi mais citotóxico nos tempos 45, 60 e 120 segundos. A citotoxicidade foi diretamente proporcional ao tempo de exposição às culturas de células. Dessa forma, pode- -se concluir que o enxaguatório Plax tradicional é altamente citotóxico a fibroblastos gengivais

The aim of this study was to evaluate the cytotoxicity of Traditional Plax mouthwash. It was evaluated at different times: 1, 15, 30, 45, 60, and 120 seconds as their cytotoxic effect on gingival fibroblasts L929. We used three control groups: positive (C +) cell detergent Tween 80 was negative (C-), PBS, and cell control (CC) where the cells were not exposed to any material. The cytotoxicity assay was performed using cell culture of mouse fibroblast (L929). After contacting the mouthwash with the cells, they were placed in contact with the vital dye neutral red using the technique (dye uptake). The values ​of the amount of viable cells were subjected to analysis of variance (ANOVA) to determine whether there were statistical differences between groups, and subsequently Tukey test (p <0.05). The results showed cytotoxicity of mouthwash with statistical differences for the CC and C- (p <0.05). In relation to the positive control, Plax was more cytotoxic in the days 45, 60, and 120 seconds. Cytotoxicity was directly proportional to exposure time to cell cultures. Thus we can conclude that the traditional Plax mouthwash is highly cytotoxic to gingival fibroblasts

Cytotoxicity of alginate for orthodontic use

OBJECTIVE: To evaluate the cytotoxicity of three different alginate impression materials for orthodontic use. METHODS: Three different brands of alginate were divided into three groups, namely, Group JCO (Jeltrate Chromatic Ortho), OP (Orthoprint) and CO (Cavex Orthotrace). Three control groups were also included: Group C+ (positive control), consisting of detergent Tween 80; Group C- (negative control), consisting of PBS, and Group CC (cell control), consisting of cells not exposed to any material. After manipulating the materials according to the respective manufacturer instructions, samples were made with the use of silicon rings. Then the samples were immersed in Eagle's minimum essential medium (MEM) for 2 minutes. The supernatants were then removed and brought into direct contact with L929 fibroblasts. After exposure to the medium, the cells were incubated for 24 hours. Then 100 µl of 0.01% neutral red dye were added. The cells were incubated again for 3 hours so that the dye could be absorbed. After this 3-hour period, the cells were fixed to perform the viable cell count, using a spectrophotometer (BioTek, Winooski, Vermont, USA) at a wavelength of 492 nm. RESULTS: Statistical differences were found when Groups CC and C- were compared with the other experimental groups. Group JCO had the highest cytotoxicity, followed by Groups OP and CO. CONCLUSION: Based on the results obtained in this work, it was concluded that all alginate impression materials are potentially cytotoxic.

OBJETIVO: avaliar a citotoxicidade de três diferentes alginatos de uso ortodôntico. MÉTODOS: foram avaliados três diferentes alginatos divididos em três grupos, denominados grupo JCO (Jeltrate Chromatic Ortho), OP (Orthoprint) e CO (Carrex Orthotrace). Três grupos controle também participaram: controle + (C+), constituído pelo detergente celular Tween 80; controle - (C-) PBS; e controle de célula (CC) onde as células não foram expostas a nenhum material. Após manipulação dos materiais, seguindo as orientações do fabricante, foram confeccionados corpos de prova utilizando-se anéis de silicone. Em seguida, esses foram imersos em meio mínimo essencial de Eagle (MEM) por 2min, onde, então, procedeu-se à remoção do sobrenadante e à colocação em contato com fibroblastos L929. Após contato com o meio, as células foram incubadas por mais 24h onde, então, foi adicionado o corante vermelho neutro a 0,01%. Passado esse período, foram fixadas e, então, realizada contagem de células viáveis em espectrofotômetro (BioTek, Vermont, EUA) em um comprimento de onda de 492nm. RESULTADOS: os resultados demonstraram diferenças estatística entre os grupos CC e C- com os demais. O grupo experimental JCO mostrou-se com maior citotoxicidade, seguido pelos grupso OP e CO. CONCLUSÕES: pode-se concluir, com a realização desse trabalho, que todos os alginatos testados mostraram caráter citotóxico.

Cytotoxicity of separation orthodontic elastics / Citotoxicidade de elásticos ortodônticos de separação

OBJECTIVE: To test the hypothesis that there is no difference in cytotoxicity between separating elastics of different manufacturers. METHODS: The present article compared latex elastics (4.0 mm, 4.4 mm and 4.8 mm) of four different manufacturers. The sample was allocated to seven groups of 9 elastics: Group A (American Orthodontics, green color, modules), Groups M1 and M2 (Morelli, blue color, modules and free in pack respectively), Groups M3 and M4 (Morelli, green color, modules and free in pack respectively), Group U (Uniden, blue color, free in pack) and Group T (Tecnident, blue color, free in pack) regarding their possible cytotoxic effects on oral tissues. Cytotoxicity assays were performed using cell culture medium containing epithelioid-type cells (Hep-2 line) derived from human laryngeal carcinoma and submitted to the methods for evaluating the cytotoxicity by the "dye-uptake" test, at time intervals 24, 48, 72 and 168 h. Data were compared by analysis of variance (ANOVA) and Tukey's test (p < 0.05). RESULTS: Results showed statistically significant difference (p < 0.05) between group U and all the other Groups (A, M1, M2, M3, M 4 and T) at 24 and 48 hours. CONCLUSIONS: Uniden elastics evoked more cell lysis at 24 and 48 h, although, all brands showed biocompatibility from 72 h onwards.

OBJETIVO: o propósito do presente estudo foi testar a hipótese que não existe diferença de citotoxicidade entre elásticos de diferentes marcas. MÉTODOS: foram comparadas entre si 4 marcas de elásticos de separação (4,0mm, 4,4mm e 4,8mm) intrabucais de látex quanto ao possível efeito citotóxico nos tecidos bucais, divididos em 7 grupos de 9 elásticos cada: grupo A (cor verde - modular, American Orthodontics), grupos M1 e M2, (cor azul - modular e a granel, respectivamente, Morelli), grupos M3 e M4 (cor verde - modular e a granel, respectivamente, Morelli), grupo U (cor azul - a granel, Uniden) e grupo T (cor azul - a granel, Tecnident). O ensaio de citotoxicidade foi realizado utilizando-se cultura de células da linhagem HEp-2 (do tipo epitelióide, que tem origem em carcinoma de laringe humana), sendo submetido o material ao teste para células viáveis em vermelho neutro ("dye-uptake"), no tempo de 24, 48, 72 e 168 horas. A análise de variância e comparação múltipla (ANOVA) e o teste de Tukey foram utilizados (p<0,05). RESULTADOS: os resultados evidenciaram diferença estatisticamente significativa dos grupos A, M1, M2, M3, M4 e T com o grupo U nos tempos de 24 e 48h (p<0,05). CONCLUSÃO: pôde-se evidenciar que os elásticos da marca Uniden causaram alta quantidade de lise celular em 24 e 48h, porém, todas as marcas mostraram-se biocompatíveis a partir de 72h.

Avaliação in vitro da citotoxicidade de enxaguatórios bucais Listerine® / In vitro cytotoxicity of Listerine® mouthwash

Objetivo: Avaliar a citotoxicidade de enxaguatórios bucais Listerine em diferentes períodos de tempo. Material e Métodos: Avaliaram-se os enxaguatórios Listerine: Vanilla Mint, Clássico, Cool Citrus, Menta, Tarta Control e Whitening em diferentes tempos: 1, 15, 30, 45, 60 e 120 segundos quanto ao seu efeito citotóxico em fibroblastos gengivais L929. Utilizaram-se 3 grupos controle: positivo (C+) detergente celular Tween 80, negativo (C-) PBS e controle de célula (CC) em que as células não foram expostas a nenhum material. O ensaio de citotoxicidade foi realizado, utilizando-se cultura celular de fibroblasto de camundongo (L929). Após contato do enxaguatório com as células, estas foram colocadas em contato com o corante vital vermelho neutro, utilizando-se a técnica ("dye uptake"). Os valores da quantidade de células viáveis foram submetidos à análise da variância (ANOVA) para determinar se havia diferenças estatísticas entre os grupos e posteriormente ao teste de Tukey (p<0.05). Resultados: Os resultados demonstraram que todos os enxaguatórios apresentaram citotoxicidade em todos os tempos avaliados. Os mais citotóxicos foram o Tarta Control e Whitening, e o que apresentou menos citotoxicidade foi o Clássico. Essa citotoxicidade foi diretamente proporcional ao tempo de exposição às culturas de células. Conclusão: Pode-se concluir com a realização deste trabalho que todos os enxaguatórios avaliados apresentaram citotoxicidade.

Objective: Evaluate the cytotoxicity of Listerine mouthwash in different time periods. Materials and Methods: We evaluated the mouthwash Listerine: Mint Vanilla, Classic, Cool Citrus, Mint, Control and Tarta Whitening at different times: 1, 15, 30, 45, 60 and 120 seconds as their cytotoxic effect on gingival fibroblasts L929. We used three control groups: positive (C +) cell detergent Tween 80 was negative (C-), PBS, and cell control (CC) where the cells were not exposed to any material. The cytotoxicity assay was performed using cell culture of mouse fibroblast (L929). After contacting the mouthwash with the cells, they were placed in contact with the vital dye neutral red used the technique (dye uptake). The values of the amount of viable cells, were subjected to analysis of variance (ANOVA) to determine whether there were statistical differences between groups, and subsequently Tukey test (p <0.05). Results: The results showed that all mouthrinses showed cytotoxicity at all times evaluated. The most cytotoxic were Tarta Control and Whitening, and showed less cytotoxicity than was the Classic. This cytotoxicity was directly proportional to exposure time to cell cultures. Conclusion: It can be concluded with the completion of this work showed that all mouthrinses evaluated cytotoxicity.

Citotoxicidade de soldas elétricas a ponto: estudo in vitro / Cytotoxicity of electric spot welding: an in vitro study

OBJETIVO: o processo de solda envolve íons metálicos capazes de provocar lise celular. Diante disso, o objetivo deste estudo foi testar a hipótese de que existe citotoxicidade entre diferentes tipos de ligas (CrNi, TMA, NiTi) utilizadas em Ortodontia, submetidas à solda elétrica a ponto. MÉTODOS: três tipos de ligas foram avaliados neste estudo. Foram confeccionados 36 corpos de prova, 6 para cada combinação entre os fios, divididos em 6 grupos - grupo AA (aço com aço), grupo AT (aço com TMA), grupo AN (aço com NiTi), grupo TT (TMA com TMA), grupo TN (TMA com NiTi) e grupo NN (NiTi com NiTi) - que foram submetidos à solda a ponto para avaliação quanto ao possível efeito citotóxico nos tecidos bucais. Previamente, os corpos de prova foram limpos com álcool isopropílico e esterilizados em luz ultravioleta. O ensaio de citotoxicidade foi realizado utilizando-se cultura de células (linhagem L929, fibroblastos de camundongos), submetida ao teste para células viáveis em vermelho neutro ("dye-uptake") no tempo de 24h. A análise de variância e comparação múltipla (ANOVA) e teste de Tukey foram utilizados (p< 0,05). RESULTADOS: os resultados demonstraram que não houve diferença estatisticamente significativa entre os grupos experimentais (P> 0,05). Foi observada maior viabilidade celular no grupo TT, seguido dos grupos AT, TN, AA, NA e NN. CONCLUSÃO: pôde-se evidenciar que solda em fios de liga de NiTi causaram maior quantidade de lise celular. Soldas elétricas a ponto demonstraram pequena capacidade de causar lise celular.

OBJECTIVE: The welding process involves metal ions capable of causing cell lysis. In view of this fact, the aim of this study was to test the hypothesis that cytotoxicity is present in different types of alloys (CrNi, TMA, NiTi) commonly used in orthodontic practice when these alloys are subjected to electric spot welding. METHODS: Three types of alloys were evaluated in this study. Thirty-six test specimens were fabricated, 6 for each wire combination, and divided into 6 groups: Group SS (stainless steel), Group ST (steel with TMA), Group SN (steel with NiTi), Group TT (TMA with TMA), Group TN group (TMA with NiTi) and Group NN (NiTi with NiTi). All groups were subjected to spot welding and assessed in terms of their potential cytotoxicity to oral tissues. The specimens were first cleaned with isopropyl alcohol and sterilized with ultraviolet light (UV). A cytotoxicity assay was performed using cultured cells (strain L929, mouse fibroblast cells), which were tested for viable cells in neutral red dye-uptake over 24 hours. Analysis of variance and multiple comparison (ANOVA), as well as Tukey test were employed (p<0.05). RESULTS: The results showed no statistically significant difference between experimental groups (P>0.05). Cell viability was higher in the TT group, followed by groups ST, TN, SS, NS and NN. CONCLUSIONS: It became evident that the welding of NiTi alloy wires caused a greater amount of cell lysis. Electric spot welding was found to cause little cell lysis.

Avaliação in vitro da Citotoxicidade de Enxaguatórios Bucais com e sem Álcool / In vitro assessment of the cytotoxicity of mouthwashes with and without alcohol

O objetivo do presente trabalho foi verificar a hipótese de que enxaguatórios bucais sem álcool apresentam menor citotoxicidade quando comparado aos com álcool. Avaliou-se os enxaguatórios Plax: Convencional (com álcool) e Sem álcool em diferentes tempos: 1, 15, 30, 45, 60 e 120 segundos quanto seu efeito citotóxico em fibroblastos gengivais L929. Utilizou-se 3 grupos controle: positivo (C+) detergente celular Tween 80, negativo (C-) PBS, e controle de célula (CC) onde as células não foram expostas a nenhum material. O ensaio de citotoxicidade foi realizado utilizando cultura celular de fibroblasto de camundongo (L929). Após contato do enxaguatório com as células, as mesmas foram colocadas em contato com o corante vital vermelho neutro utilizado-se a técnica ("dye uptake"). Os valores da quantidade de células viáveis, foram submetidos à análise da variância (ANOVA) para determinar se havia diferenças estatísticas entre os grupos, e posteriormente ao teste de Tukey (p<0.05). Os resultados demonstraram maior citotoxicidade dos enxaguatórios com álcool. Os enxaguatórios sem álcool apresentaram maior quantidade de células viáveis no entanto ainda assim apresentaram diferenças estatísticas com os grupos controle de células e controle negativo (p<0.05). A citotoxicidade foi diretamente proporcional ao tempo de exposição às culturas de células. Dessa forma pode-se concluir que a hipótese foi confirmada em partes, haja vista que os enxaguatórios sem álcool apresentaram menor citotoxicidade que os com álcool, mas ainda apresentaram citotoxicidade quando comparado aos grupos controle de célula e negativo.

Cytotoxicity of dental alginates / Citotoxicidad de alginatos dentales

Alginate, or irreversible hydrocolloid, is one of the most accepted impression materials used in dentistry. However, some substances existing in these materials can be toxic. The aim of this study was to assess the cytotoxicity of alginates for dental applications. Fourteen different alginates were assessed: Jeltrate, Jeltrate Plus, Jeltrate Chromatic, Alga Gel, Printer Gel, Ava Gel, New Print, Kromopan 100, Tropicalgin, Cavex Orthotrace, Hydrogum, Orthoprint, Cavex Color Change, and Qualitygel. Three control groups were also used in this study: positive control group (C+) consisting of cell detergent Tween 80, negative control group (C-) consisting of PBS, and cell control group (CC) consisting of non-exposed cells. After manipulating the materials according to the manufacturer’s instructions, samples were made by using silicone rings. Next, the samples were immersed into Eagle’s minimum essential medium (MEM) for 2 minutes followed by removal of supernatants and contact with L929 fibroblasts. After contact with the medium, the cells were incubated for further 24 hours in which 100µl of 0.01 por ciento neutral red stain were added. Cells were incubated again for 3 hours so that the stain could be absorbed. After this period, the cells were fixed and viable cell counting was performed by using a spectrophotometer (BioTek, Winooski, Vermont, USA) at wavelength of 492 nm. The results demonstrated statistical differences between CC and C- groups in relation to other ones (p<0.05). No statistical differences were observed between Jeltrate Plus and Hydrogum groups, between Jeltrate and Jeltrate Chromatic, Printer Gel, Tropicalgin, and Qualitygel groups, and between Jeltrate Chromatic and Alga Gel, Ava Gel, New Print, Kromopan 100, Cavex Orthotrace, Hydrogum, Orhtoprint, and Cavex Color Change groups. One can conclude, based on the results of this study, that all alginate materials were found to be cytotoxic.

El alginato o hidrocoloide irreversible, es uno de los materiales de impresión más aceptados y utilizados en odontología. Sin embargo, algunas substancias existentes en estos materiales pueden ser tóxicas. El objetivo de este estudio fue evaluar la citotoxicidad de los alginatos para aplicaciones dentales. Fueron evaluados 14 alginatos diferentes: Jeltrate, Jeltrate Plus, Jeltrate Chromatic, Alga Gel, Printer Gel, Ava Gel, New Print, Kromopan 100, Tropicalgin, Cavex Orthotrace, Hydrogum, Orthoprint, Cavex Color Change y Qualitygel. También se utilizaron tres grupos de control también se utilizaron en este estudio: grupo control positivo (C+) que consiste en células de detergente Tween 80, el grupo de control negativo (C-) que consiste en PBS, y el grupo de células de control (CC) que consiste de las células no expuestas. Después de la manipulación de los materiales de acuerdo a las instrucciones del fabricante, las muestras fueron hechas mediante el uso de anillos de silicona. A continuación, las muestras se sumergieron en medio mínimo esencial de Eagle (MEM) durante 2 minutos, seguido de la eliminación de los sobrenadantes y el contacto con los fibroblastos L929. En caso de contacto con el medio, las células fueron incubadas durante 24 horas más en 100ml de tinción roja neutra al 0,01 por ciento. Las células se incubaron nuevamente durante 3 horas para que la tinción pueda ser absorbida. Después de este período, las células fueron fijadas y el recuento de células viables se realizó mediante un espectrofotómetro (BioTek, Winooski, Vermont, EE.UU.) a la longitud de onda de 492 nm. Los resultados demostraron diferencias estadísticamente significativas entre los grupos de CC y C- en relación con los demás (p<0,05). No se observaron diferencias estadísticas entre los grupos Jeltrate Plus y Hydrogum, entre Jeltrate y los grupos Jeltrate Chromatic, Printer Gel, Tropicalgin y Qualitygel, y entre Jeltrate Chromatic y los grupos Alga Gel, Ava Gel, New Print, ...

Citotoxicidade de luvas de procedimentos em látexSupermax® / Cytotoxic of the procedures gloves in latex Supermax®

O objetivo foi avaliar a citotoxicidade de luvas de procedimento em látex. Foram avaliados três diferentestipos de luvas de procedimento divididos em três grupos assim denominados: 1 (Supermax®),2 (Supermax -Powder Free®) e 3 (Supermax-Microtexturizado®). Utilizou-se 3 grupos controle: positivo(C+) detergente celular Tween 80, negativo (C-) PBS, e controle de célula (CC) cujas não foramexpostas a nenhum material. Após confecção dos corpos de prova, os mesmos foram imersos em meiomínimo essencial Eagle (MEM) por 24 hrs. Passado esse período procedeu-se a remoção do sobrenadantee colocação em contato com os fibroblastos L929. Após contato com o meio, as células foramincubadas por mais 15, 30 e 60 minutos, então foram adicionados 100μl do corante vermelho neutro a0,01%. Uma vez coradas, as mesmas foram fixadas com formoaldeido e então realizada contagem decélulas viáveis em espectrofotômetro (BioTek, Winooski, Vermont, USA). Os valores da quantidade decélulas viáveis, foram submetidos à análise de variância (ANOVA) para determinar se havia diferençasestatísticas entre os grupos, e posteriormente ao teste de Tukey (p<0.05). Nos tempos de 15, 30 e 60minutos os grupos 1 e 3 foram os que demonstraram maior quantidade de células viáveis respectivamente.Com 1 hora de contato todos os grupos demonstraram alta toxicidade celular quando comparadoaos C- e CC. Todas as luvas foram citotóxicas nos tempos avaliados. A toxicidade aumentou como aumento de contato das células.

The objective was to evaluate the cytotoxicity of procedure latex gloves. Three different types ofgloves procedure divided into three groups so called: 1 (Supermax®), 2 (Supermax -- Powder Free®)and 3 (Supermax-Microtexturizado®). We used 3 control groups: positive (C +) cell detergent Tween80, negative (C-) PBS, and control of cell (CC) whose cells were not exposed to any material. Aftermaking the bodies of evidence, they were immersed in Eagle minimum essential medium (MEM) for24 hrs. After this period there has been the removal of supernatant and placed in contact with thefibroblasts L929. After contact with the medium, the cells were incubated for another 15, 30 and 60minutes, which were then added 100 the dye neutral red to 0.01%. Once stained, they were fixedwith formoaldeido and then performed counting of viable cells in spectrophotometer (BioTek, Winooski,Vermont, USA). The values of the quantity of viable cells were subjected to analysis of variance(ANOVA) to determine whether there were statistical differences between groups, and then the Tukeytest (p <0.05). In times of 15, 30 and 60 minutes the groups were 1 and 3 showed that the greatestamount of viable cells respectively. 1 hour of contact with all groups showed high cellular toxicity whencompared to the C-and CC. All gloves were cytotoxic in the time evaluated. The toxicity increased withthe increase of cell contact.

Cytotoxicity of orthodontic elastics: in vitro investigation with on L929 mouse fibroblasts

Aim: To test the hypothesis that there is no difference in the cytotoxicity among natural latex elastics of different manufacturers using a L929 cell line culture. Methods: Different latex intra oral elastics (I.D. = 5/16", 4.5 oz.) were tested. The sample was divided into 7 groups of 15 elastic seach: Group AO (American Orthodontics), Group GAC (GAC International), Group TP (TP Orthodontics), Group AD (Aditek), Group AB (Abzil), Group MO (Morelli) and Group UN (Uniden).Cytotoxicity assays were performed by using cell culture medium containing L-929 line cells(mouse fibroblast). The cytotoxicity was evaluated by using the “dye-uptake” test, which wasemployed at two different moments (1 and 24 h). Data were compared by ANOVA and Tukey’s test(P < 0.05). Results: The results showed a significant difference (P < 0.05) between all groups and the group CC (cell control) at 1 and 24 h. Groups AD, AB, MO and UN were noticeably more cytotoxic than the groups AO, GAC and TP at 1 h. After 24 h, a significant decrease in cell viability was observed in all groups. Conclusions: Intraoral elastics from American Orthodontics, GACand TP Orthodontics trademarks induced less cell lysis than Aditek, Abzil, Morelli and Uniden trademarks.

Citotoxicity of orthodontic mini-implants / Citotoxidade de mini-implantes ortodônticos

OBJECTIVE: Based in the premise that Ti-6Al-4V orthodontic mini-implants can release metal ions into the body fluids, this research is aimed assess the cytotoxic effect of orthodontic mini-implant on L929 fibroblast cells. MATERIAL AND METHODS: Eighteen orthodontic mini-implants made of Ti-6Al-4V alloy were divided into 6 groups: 1 (golden colour, SIN), 2 (silver colour, SIN), 3 (Neodent™), 4 (INP™), 5 (Mondeal™), and 6 (Titanium Fix™). The mini-implants were immersed into Eagle’s minimum essential medium for 24 hours, where supernatant removal and contact with L929 fibroblasts were performed. Cytotoxicity was evaluated in four different periods of time: 24, 48, 72, and 168 hours. After being in contact with the mini-implants immersed, the cells were incubated for further 24 hours and then 100 ml of 0.01% neutral-red staining solution were added. After this period of time, they were fixed and a spectrophotometer was used for counting the viable cells. RESULTS: After the 24 hours period, statistical differences were found by comparing groups 1 and 2 to groups 3,4,5, and C+ (p < 0.05). After the 48 hours period, groups 1 and 2 were shown to be statistically different in relation to groups 3, 4, and C+. After the 72 hours period, statistical differences were found only in group 1 compared to groups 4, 5, 6, CC, and C+ (p < 0.05). After 7 days, no statistical differences were found between the mini-implants. CONCLUSION: Although mini-implants are made of the same alloy, there are differences in their cytotoxicity because of the different concentrations of chemical elements used for manufacturing them.

OBJETIVO: Baseando-se na premissa de que mini-implantes ortodônticos podem liberar íons metálicos nos fluidos corporais, pesquisou-se o efeito citotóxico de mini-implantes ortodônticos em fibroblastos L929. MATERIAL E MÉTODO: Dezoito mini-implantes ortodônticos confeccionados em liga Ti-6Al-4V foram divididos em seis grupos: 1 (dourados, SIN), 2 (prateados SIN), 3 (Neodent), 4 (INP), 5 (Mondeal) e 6 (Titanium Fix). Os mini-implantes foram imersos em meio mínimo essencial Eagle por 24 horas, onde efetuou-se remoção do supernadante e contato com fibroblastos L929. A citotoxicidade foi avaliada em 4 diferentes tempos: 24, 48, 72 e 168 horas. Após contato com os mini-implantes imersos, as células foram incubadas por mais 24 horas; então, 100 ml de solução corante neutra-vermelha foram adicionados. Após, foram fixadas e um espectrofotômetro foi usado para contar as células viáveis. RESULTADOS: Após o período de 23 horas, compararam-se os grupos 1 e 2 aos grupos 3, 4, 5 e C+. Após 72 horas, diferenças estatísticas foram encontradas somente no grupo 1, comparado aos grupos 4, 5, 6, CC e C+ (p < 0,05). Após 7 dias, não foram encontradas diferenças estatisticamente significantes entre os diversos mini-implantes.

Cytotoxicity of orthodontic separating elastics.

BACKGROUND: Separating elastics may be cytotoxic to the interdental gingival tissues. Both latex and non-latex separating elastics are widely used and both types should be biocompatible. OBJECTIVE: To determine if latex and non-latex orthodontic separating elastics are cytotoxic. METHODS: The cytotoxicity of natural latex (Groups A, D and O) and non-latex (Group M) orthodontic separating elastics were determined by incubating 15 elastics of each type in Eagle's essential medium (MEM), removing the supernatant after 24, 48, 72 and 168 hours and adding it to cultures of L-929 mouse fibroblasts in growth medium (MEM plus glutamine, garamicine, fungizone, sodium bicarbonate, buffered saline and foetal calf serum). To verify the cell response in extreme situations, three additional groups were included: Group CC (cell control), consisting of L-929 cells not exposed to supernatants from the maintenance medium with the elastics; Group C+ (positive control), consisting of Tween 80; Group C- (negative control), consisting of phosphate buffered saline solution. The positive and negative controls were incubated in MEM maintenance medium for 24, 48, 72 and 168 hours and the extracted elutes were added to L-929 line cells incubated in the growth medium. The viability of the cells was determined with neutral red (dye-uptake method) at 24, 48, 72 and 168 hours. The data were analysed with the analysis of variance (ANOVA) and Tukey's multiple comparison test. The significance level was p < or = 0.05. RESULTS: The elastics in Groups A, D and O induced greater cell lysis at 72 hours compared to the other experimental times. There were statistically significant differences between the cytotoxicity of the elastics in Groups A, D and O in relation to Group CC for experimental times of 24, 48, 72 and 168 hours (p > 0.05). There was not, however, a statistically significant difference between Groups D and CC at 24 hours. CONCLUSION: The latex and non-latex orthodontic separating elastics tested were considered to be biocompatible.

Evaluation of cytotoxicity of accessories used for traction of impacted teeth

Aim: To test the hypothesis that gold-coated orthodontic accessories used for canine traction are less cytotoxic than those made of stainless steel. Methods: Six different orthodontic accessories were evaluated, three of them made from stainless steel (1 – bracket, 2 – button, 3 – mesh pad) and three made from a gold-coated alloy (4 – small mesh pad, 5 button, 6 – big mesh pad). Three control groups were also analyzed: Positive control (C+), consisting of Tween 80 cell detergent;Negative control (C-), consisting of PBS; and Cell control (CC), consisting of cells not exposed to any material. Dye-uptake technique, in which neutral red dye is incorporated into viable cells, was used to assess the cytotoxicity of the accessories. Viable cell counting was performed using a spectrophotometer. Data were analyzed statistically by A NOVA and Tukey’s test. Results:Statistically significant differences (P< 0.05) were found between Groups 1-3 and Groups 4-6. However, no differences were found between Groups 1-3 and Groups C- and CC, and neither between Groups 4-6 and Group C+. Conclusions: The tested hypothesis was not confirmed since gold-coated orthodontic accessories were found to be more cytotoxic than those made of stainless steel.

In vitro cytotoxicity of latex orthodonticelastics / Citotoxicidad de los elásticos ortodònticos de látex

Natural latex does not fall into the category of materials known to be entirely inoffensive. The objective of the present in vitro study is to test the hypothesis that there is no difference in the cytotoxicity between natural latex elastics of different colours. The present article compared different latex intra-oral elastics (5/16 = 7.9 mm). The sample was divided into four groups according to their manufacturer: Group N (Natural latex elastic, Morelli), Group R (Red colour elastic, Morelli) Group Y (Yellow colour elastic, Morelli) and Group G (Green colour elastic, Morelli). Cytotoxicity assays were performed by using cell culture medium containing L-929 line cells (mouse fibroblast). The cytotoxicity was evaluated by using the “dyeuptake” test, which was employed at two different moments (1 and 24 h). Data were compared by analysis of variance (ANOVA) and Tukey’s test (p<0.05). The results showed a significant difference (p<0.05) between the groups N, R, Y, G and the negative cytotoxicity control at 1 and 24 h (p<0.05), it did not have presented significant difference between the groups N, R, Y, G tested (p>0.05) at 1 and 24 h. Morelli intra-oral elastics were found to be highly cytotoxic, regardless of their colour and immersion time.

El látex natural no entra en la categoría de materiales que se sabe del todo inofensivo. El objetivo del presente estudio in vitro es poner a prueba la hipótesis de que no hay diferencia en la citotoxicidad entre elásticos de látex natural de diferentes colores. El presente artículo compara diferentes elásticos intraorales de látex (5/16 =7,9 mm). La muestra se dividió en cuatro grupos según su fabricante: Grupo N (elástico látex, Morelli), Grupo I (elástico de color rojo, Morelli) Grupo Y (elástico de color amarillo, Morelli) y el Grupo G (elástico color verde, Morelli). Pruebas de citotoxicidad se realizaron mediante el uso de medio de cultivo celular que contiene líneas celulares L-929 (fibroblastos de ratón). La citotoxicidad se evaluó mediante el test “dye-uptake”, que se empleó en dos momentos diferentes (1 y 24 h). Los datos se compararon mediante análisis de varianza (ANOVA) y test de Tukey (p<0,05). Los resultados mostraron una diferencia significativa (p<0,05) entre los grupos N, R, Y, G y la negativa citotoxicidad del control en 1 y 24 h (p<0,05), no han presentado diferencias significativas entre los grupos N, R , Y, G probado (p>0,05) en 1 y 24 h. Elásticos intraorales Morelli resultaron ser altamente citotóxicos, independientemente de su color y tiempo de inmersión.

Antiviral activity of the green marine alga Ulva fasciata on the replication of human metapneumovirus.

We evaluated the antiviral activity of the marine alga, Ulva fasciata, collected from Rasa beach and Forno beach, Búzios, Rio de Janeiro, Brazil on the replication of human metapneumovirus (HMPV). The algae extracts were prepared using three different methodologies to compare the activity of different groups of chemical composites obtained through these different methodologies. Four out of the six extracts inhibited nearly 100% of viral replication. The results demonstrated that the majority of the extracts (five out of six) possess virucidal activity and therefore have the ability to interact with the extracellular viral particles and prevent the infection. On the other hand, only two extracts (from Forno beach, obtained by maceration and maceration of the decoction) were able to interact with cell receptors, hindering the viral entry. Finally, only the extract of algae collected at Forno beach, obtained by maceration presented intracellular activity. To our knowledge, this is a pioneer study on antiviral activity of marine algae against HMPV. It is also the first on antiviral activity against HMPV ever done in Brazil. The study also shows the effect of different environment factors and different chemical procedures used to obtain the extract on its biological properties.

Antiviral activity of the green marine alga Ulva fasciata on the replication of human metapneumovirus / Atividade antiviral da alga verde marinha Ulva fasciata na replicação do metapneumovírus humano

We evaluated the antiviral activity of the marine alga, Ulva fasciata, collected from Rasa beach and Forno beach, Búzios, Rio de Janeiro, Brazil on the replication of human metapneumovirus (HMPV). The algae extracts were prepared using three different methodologies to compare the activity of different groups of chemical composites obtained through these different methodologies. Four out of the six extracts inhibited nearly 100 percent of viral replication. The results demonstrated that the majority of the extracts (five out of six) possess virucidal activity and therefore have the ability to interact with the extracellular viral particles and prevent the infection. On the other hand, only two extracts (from Forno beach, obtained by maceration and maceration of the decoction) were able to interact with cell receptors, hindering the viral entry. Finally, only the extract of algae collected at Forno beach, obtained by maceration presented intracellular activity. To our knowledge, this is a pioneer study on antiviral activity of marine algae against HMPV. It is also the first on antiviral activity against HMPV ever done in Brazil. The study also shows the effect of different environment factors and different chemical procedures used to obtain the extract on its biological properties.

Neste artigo, foi avaliada a atividade antiviral da alga marinha Ulva fasciata, coletada nas Praias do Forno e Rasa, em Búzios, Rio de Janeiro, Brasil, sobre a replicação do metapneumovírus humano (HMPV). Os extratos desta alga foram preparados utilizando três diferentes metodologias, visando a comparação da atividade de diferentes grupos de compostos químicos que são obtidos dependendo da metodologia empregada. Quatro, do total de seis extratos foram capazes de inibir praticamente 100 por cento da replicação viral. Os resultados demonstram também que a maioria dos extratos (cinco, dos seis), possui atividade virucida e, portanto, possuem a habilidade de interagir com a partícula viral extracelularmente impedindo a infecção. Por outro lado, apenas dois extratos (coletado da Praia do Forno e, preparado através de maceração e maceração do decocto) foram capazes de se ligar a receptores celulares, impossibilitando assim a entrada das partículas virais nas células. Finalmente, apenas o extrato que foi preparado por maceração da alga coletada na Praia do Forno, demonstrou atividade intracelular. Até onde sabemos, este é um estudo pioneiro sobre a atividade antiviral de algas marinhas sobre o HMPV. É também o primeiro estudo sobre atividade antiviral sobre HMPV realizado no Brasil. O estudo também mostra o efeito de diferentes condições ambientais e procedimentos químicos utilizados na preparação do extrato sobre suas propriedades biológicas.

Cytotoxicity of latex and non-latex orthodontic elastomeric ligatures on L929 mouse fibroblasts.

This study investigated the cytotoxicity exists between latex and non-latex Orthodontic elastomeric ligatures. Six elastomeric ligatures (1 latex, 2 latex-free and 3 polyurethane) from different manufacturers were divided into 6 groups of 15 elastics each: A (Latex-free, American Orthodontics), M (Polyurethane, Morelli), G (Polyurethane,GAC International), Te (Polyurethane, Tecnident), TP (Natural latex,TP Orthodontics) and U (Latex-free,3M Unitek). The cytotoxicity assay was performed using cell cultures (L929 mouse fibroblast cell line), which were subjected to the cell viability test with neutral red ("dye-uptake") at 1, 2, 3, 7 and 28 days. Data were analyzed statistically by ANOVA and Tukey's test (α=0.05). No statistically significant differences (p>0.05) were observed between Groups M and Te in all experimental periods, except at 2 days. No significant differences (p>0.05) in cell viability were found either among Groups A, G, TP and U or between Groups M and Te at 24 h or among Groups CC, A, G, TP and U at 2 and 28 days. It may be concluded that latex-free elastomeric ligatures from American Orthodontics and Unitek trademarks induced less cell lysis compared to latex and polyurethane ligatures.