Actividad antimicrobiana in vitro del extracto de hojas de Bixa orellana L. contra bacterias anaerobias asociadas a la vaginosis bacteriana y Lactobacillus spp / In vitro antimicrobial activity of Bixa orellana L. leaves extract against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp.

Objetivos . Describir la actividad antimicrobiana in vitro del extracto metanólico de las hojas de Bixa orellana L. contra las bacterias anaerobias asociadas a la vaginosis bacteriana y Lactobacillus spp. Materiales y métodos . Se incluyeron en el estudio ocho cepas de referencia ATCC; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula y Lactobacillus crispatus, y 22 aislamientos clínicos; once aislados de Gardnerella vaginalis y once aislados de Lactobacillus. La susceptibilidad antimicrobiana se determinó mediante el método de difusión en agar. La concentración mínima inhibitoria (CMI) y la concentración bactericida mínima (CBM) fueron determinadas utilizando el método de dilución en agar y un método de dilución modificado, respectivamente. Resultados . Todas las cepas de referencia ATCC tuvieron un alto nivel de susceptibilidad al extracto, con excepción de P. vibia, V. parvula y L. crispatus. Interesantemente, los aislamientos clínicos de G. vaginalis y la cepa ATCC de G. vaginalis fueron los más susceptibles al extracto dados los bajos valores de CMI (1,0 - 2,0 mg/mL) y CBM (1,0 - 4,0 mg/mL), mientras que, los aislamientos clínicos de Lactobacillus spp. y la cepa ATCC de L. crispatus fueron los menos susceptibles debido a los altos valores de CMI (32,0 mg/mL) y CBM (≥ 32,0 mg/mL). Conclusiones . Los experimentos in vitro sugieren que el extracto posee propiedades antibacterianas selectivas dada su alta actividad contra bacterias anaerobias asociadas a vaginosis bacteriana y baja actividad contra especies de Lactobacillus.

Objective. To describe the in vitro antimicrobial activity of the methanolic extract of Bixa orellana L. leaves against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp. Materials and methods. Eight ATCC reference strains; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula, and Lactobacillus crispatus, and twenty-two clinical isolates; eleven Gardnerella vaginalis and eleven Lactobacillus strains, were included in the study. The antimicrobial susceptibility was determined by the agar diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by using agar dilution and a modified dilution plating method, respectively. Results. All ATCC reference strains showed high levels of susceptibility to the extract, except P. vibia, V. parvula and L. crispatus. Interestingly, all G. vaginalis clinical isolates and the G. vaginalis ATTC strain were the most susceptible to the extract, given their low MIC (1.0 - 2.0 mg/mL) and MBC (1.0 - 4.0 mg/mL) values, whereas, the Lactobacillus spp. clinical isolates and the L. crispatus ATCC strain were the least susceptible bacteria given their high MIC (32.0 mg/mL) and MBC (≥ 32.0 mg/mL) values. Conclusions. In vitro experiments suggest that the extract possesses selective antimicrobial properties given its high activity against bacterial vaginosis-associated anaerobic bacteria and low activity against Lactobacillus species.

Evaluation of different protocols for culturing mesenchymal stem cells derived from murine bone marrow

ABSTRACT Introduction: Culturing bone marrow mesenchymal stem cells (BM-MSCs) is a key point in different fields of research, including tissue engineering and regenerative medicine and studies of the bone marrow microenvironment. However, isolating and expanding murine BM-MSCs in vitro has challenged researchers due to the low purity and yield of obtained cells. In this study, we aimed to evaluate five different protocols to culture murine BM-MSCs in vitro. Methods: All protocols were based on the adhesion capacity of BM-MSCs to the tissue culture plastic surface and varied in the types of plate, culture media, serum, additional supplementation and initial cell density. Flow cytometry analysis was used to investigate lineage purity after expansion. Results: The expression of CD45 and CD11b was detected in the cultures generated according to all protocols, indicating low purity with the presence of hematopoietic cells and macrophages. The cellular growth rate and morphology varied between the cultures performed according to each protocol. Cells cultured according to protocol 5 (8 × 107cells/plate, Roswell Park Memorial Institute (RPMI) culture medium during first passage and then Iscove's Modified Delbecco's Medium (IMDM) culture medium, both supplemented with 9% fetal bovine serum, 9% horse serum, 12μM L-glutamine) presented the best performance, with a satisfactory growth rate and spindle-shape morphology. Conclusion: Our results point out that the purity and satisfactory growth rate of murine BM-MSC cultures are not easily achieved and additional approaches must be tested for a proper cell expansion.

Avaliação da composição química de um óleo de copaíba (Copaifera spp) e seu efeito em células-tronco mesenquimais / Evaluation of the chemical composition of a copaiba oil (Copaifera spp) and its effect on mesenchymal stem cells

A oleorresina obtida de copaíferas é amplamente utilizada na medicina tradicional brasileira. Este estudo avaliou a composição química por Cromatografia Gasosa (CG) e o efeito da oleorresina de Copaifera officinalis em células-tronco. Para isso as células foram tratadas com a oleorresina nas concentrações de 0,5, 20, 110, 140, 170 ou 200 µg/ml por 24h. A avaliação por CG identificou os sesquiterpenos ß-cariofileno, trans-α-bergamoteno e óxido de cariofileno II como os compostos majoritários da oleorresina. As menores concentrações de oleorresina utilizadas apresentaram resultados semelhantes ao grupo controle e as maiores concentrações diminuíram significativamente a viabilidade celular e apresentaram maior citotoxicidade. Como conclusão, os principais componentes encontrados na oleorresina de copaíba foram os sesquiterpenos e as baixas concentrações testadas não foram citotóxicas. O aumento das concentrações de oleorresina de copaíba promoveu diminuição da viabilidade celular e aumento dos efeitos citotóxicos nas células-tronco. Embora a oleorresina de copaíba tenha uso etnofarmacológico na cicatrização, este estudo demonstrou efeito citotóxico em células-tronco, as quais estão relacionadas ao processo de regeneração corpóreo. Portanto, deve-se ter cuidado com a dosagem de oleorresina a ser utilizada, uma vez que este estudo in vitro mostrou citotoxicidade e um impacto negativo na viabilidade das células-tronco nas mais altas concentrações testadas. [au]

Oleoresinobtained from Copaifera trees is extensively used in Brazilian traditional medicine. This study hasevaluated the chemical composition and effect of Copaifera officinalisoleoresin on stem cells. The oleoresin was analyzed by Gas Chromatography (GC) and the cells were treated with the oleoresin at concentrations of 0.5, 20, 110, 140, 170 or 200 µg/ml for 24h for cellular tests. GC identified the sesquiterpenes beta-caryophyllene, trans-alpha-bergamotene, and caryophyllene oxide II as the main compounds in oleoresin. The cell viability and cytotoxicity assays showed the lowest concentrations of oleoresin used presented similar results to the control group and the higher concentrations tested significantly decreased cell viability and increased cytotoxicity. In a conclusion, the main components found in copaiba oleoresin were sesquiterpenes and the low tested concentrations were not cytotoxic. The increased concentrations of copaiba oleoresin promoted a decrease in cell viability and an increase of cytotoxicity in the stem cells. Although copaiba oleoresin has ethnopharmacology use in healing, this study showed toxicity in stem cells, which are related to the corporeal regeneration process. Therefore, caution must be taken with the dosage of the oleoresin to be used since this in vitro study showed cytotoxicity and a negative impact on stem cell viability at the higher tested concentrations. [au]

Atividade fotoprotetora e antioxidante de compostos fenólicos: uma revisão sistemática de testes in vitro / Photoprotective and antioxidant activity of phenolic compounds: a systematic review of in vitro tests / Actividad fotoprotectora y antioxidante de compuestos fenólicos: una revisión sistemática de ensayos in vitro

RESUMO Introdução: Os compostos fenólicos, devido a sua estrutura química, possuem a capacidade de absorver a energia ultravioleta e reduzir a formação de radicais livres. Objetivo: Avaliar a atividade fotoprotetora e antioxidante de compostos fenólicos a partir da observação de resultados in vitro e verificar a importância do uso de modelos biológicos nessa perspectiva. Metodologia: Foi realizada uma pesquisa de artigos publicados, na base de dados Pubmed, entre 2010 e 2020, que atendessem aos objetivos deste trabalho, 44 artigos foram selecionados. Resultados: Os métodos instrumentais utilizados para avaliação da atividade fotoprotetora apresentaram boa correlação in vivo e mostram-se rápidos e eficazes na determinação do fator de proteção solar. Além desses, têm-se aplicado métodos biológicos para a avaliação de aspectos que não são mensurados por métodos físico-químicos, relacionado aos danos ao DNA, decorrentes da exposição solar. Para a avaliação da atividade antioxidante, o método do radical DPPH foi empregado em 92,6 % dos estudos analisados e foi observado que os antioxidantes podem incrementar a proteção solar e, ainda, auxiliar na estabilidade de filtros solares sintéticos. Conclusão: Os compostos fenólicos, especialmente aqueles com propriedades antioxidantes, podem ser utilizados como agentes fotoprotetores em formulações tópicas para reduzir os danos à pele induzidos pela radiação UV.

SUMMARY Introduction: Phenolic compounds, due to their chemical structure, can absorb ultraviolet energy and reduce the formation of free radicals. Aim: To evaluate the photoprotective and antioxidant activity of phenolic compounds from the observation of in vitro results and to verify the importance of the use of biological models in this perspective. Methodology: A search for articles published in the Pubmed database was carried out between 2010 and 2020, which met the objectives of this work, 44 articles were selected. Results: According to the literature, the instrumental methods used to assess independent photoprotective activity, good correlation in vivo, and demonstrating rapid and effective determination of the sun protection factor. In addition to these, biological methods have been provided for the evaluation of aspects not measured by physical-chemical methods, related to DNA damage, resulting from sun exposure. For the evaluation of antioxidant activity, the DPPH radical method was registered in 92.6 % of published studies and it was observed that antioxidants can increase sun protection and also help in the stability of synthetic sunscreens. Conclusion: Phenolic compounds, especially with antioxidant properties, can be used as photoprotective agents in topical formulations to reduce skin damage induced by UV radiation.

RESUMEN Introducción: Los compuestos fenólicos, por su estructura química, tienen la capacidad de absorber la energía ultravioleta y reducir la formación de radicales libres. Objetivo: Evaluar la actividad fotoprotectora y antioxidante de compuestos fenólicos a partir de la observación de resultados in vitro y comprobar la importancia del uso de modelos biológicos en esta perspectiva. Metodología: Se realizó una búsqueda de artículos publicados en la base de datos Pubmed entre 2010 y 2020, que cumplieron con los objetivos de este trabajo, se seleccionaron 44 artículos. Resultados: Los métodos instrumentales utilizados para evaluar la actividad fotoprotectora mostraron una buena correlación in vivo y demostraron ser rápidos y eficientes en la determinación del factor de protección solar. Además de estos, se aplicaron métodos biológicos para evaluar aspectos no medidos por métodos físico-químicos, relacionados con el daño en el ADN por exposición solar. Para la evaluación de la actividad antioxidante se utilizó el método radical DPPH en el 92,6% de los estudios analizados y se observó que los antioxidantes pueden aumentar la protección solar y también ayudar en la estabilidad de los protectores solares sintéticos. Conclusión: Los compuestos fenólicos, especialmente aquellos con propiedades antioxidantes, pueden utilizarse como agentes fotoprotectores en formulaciones tópicas para reducir el daño cutáneo inducido por la radiación UV.

Parámetros para la evaluación visual e instrumental del color dental en estudios in-vitro. Revisión de la literatura / Parameters for visual and instrumental assessment of tooth colour in in-vitro studies. Literature review

Objetivo: establecer los parámetros para la evaluación visual e instrumental del color dental en estudios in-vitro a partir de la literatura científca publicada entre 2015 y 2021. Métodos: se realizó la búsqueda en las bases de datos: PubMed, Web of Science, Science Direct, Scopus, Scielo y Lilacs; también en el motor de búsqueda Google Académico y las bibliotecas de las editoriales Wiley y Springer. Las palabras clave utilizadas fueron tooth, color, in-vitro, color perception, shade matching, thresholds, appearance, surrounding, "CIELAB" y "CIEDE2000". Teniendo en cuenta los criterios de elegibilidad, se seleccionaron los estudios de acuerdo al título, resumen y texto completo. Resultados: la búsqueda arrojó un total de 37 publicaciones que se agruparon en tres tópicos: 1. toma de color visual: condiciones ambientales, observadores y nivelación; 2. toma de color instrumental: instrumentos; y 3. procesamiento de datos: cálculo de la diferencia de color y umbrales de perceptibilidad (PT) y aceptabilidad (AT). Conclusiones: los aspectos más importantes en la evaluación visual son la iluminación, el ambiente para registro (sitio, entorno y fondo alrededor de la muestra), las condiciones geométricas de visualización, los observadores y el uso de guías. En la evaluación instrumental es relevante elegir el aparato apropiado de acuerdo con su precisión y reproducibilidad, como los espectroradiómetros y los espectrofotómetros de uso clínico. Se presenta el procesamiento de datos para establecer las variaciones de cada coordenada, las diferencias de color (ΔE): CIELAB y CIEDE2000, los umbrales y los lineamientos.

Objective: To establish the parameters for the visual and instrumental evaluation of tooth color in in-vitro studies based on the scientifc literature published between 2015 and 2021. Methods: The search was carried out in the databases of PubMed, Web of Science, Science Direct, Scopus, Scielo, Lilacs; search engine Google Scholar and publishers' library of Wiley and Scielo, using the keywords "tooth", "color", "in vitro", "color perception", "shade matching", "thresholds", "appearance", "surrounding", "CIELAB", and "CIEDE2000". The literature was selected according to the title, abstract and full text taking into the eligibility criteria. Results: It yielded a total of 37 publications, which were grouped into three topics: 1. visual color acquisition: environmental conditions for color acquisition, observers and levelling. 2. instrumental color sampling: instruments. 3. Data processing: Calculation of color diference and perception thresholds (PT) and acceptability thresholds (AT). Conclusions: The most important aspects in the visual assessment are lighting, the environment for color registration (site, environment and background around the sample), the geometric conditions of visualization, the observers and the use of guides. Regarding the instrumental assessment of color, the appropriate devices must be chosen according to its precision and reproducibility, being the spectrophotometers and spectroradiometers the most precise ones. It is presented how the data processing is carried out to establish the variations of each coordinate, the color diferences (ΔE): CIELAB and CIEDE2000, thresholds and guidelines.

Estudo comparativo para avaliação da eficácia cicatrizante de formulações tópicas contendo Triticum aestivum L. (sinônimo Triticum vulgare) em modelo de pele humana nativa / Comparative study to evaluate the wound healing efficacy of topical formulations containing Triticum aestivum L. (Triticum vulgare) in a native human skin model

Introdução: extratos vegetais e ativos derivados de plantas tem sido desenvolvidos com o objetivo de melhorar e potencializar o processo de cicatrização cutânea, dentre eles, o Triticum aestivum L. (sinônimo Triticum vulgare). Objetivo: avaliar o efeito do extrato de grão inteiro (EGTA-PR) e extrato aquoso (EATA-FI) de Triticum aestivum L. na cicatrização cutânea em pele humana ex vivo. Métodos: fragmentos de pele obtidos de cirurgia plástica eletiva foram submetidos a lesões teciduais e tratados com os extratos durante oito dias para avaliação histológica da reepitelização e marcação proteica do fator de crescimento epidérmico (EGF). Resultados: EGTA-PR e EATA-FI aceleraram o processo de reepitelização em cultura de pele humana submetida a lesão tecidual. Adicionalmente, foi observado um aumento da marcação proteica de EGF após o tratamento com EGTA-PR. Conclusão: EGTA-PR apresentou um melhor desempenho na reepitelização quando comparado ao EATA-FI, pois apresentou uma maior marcação proteica para EGF em cultura de pele humana. Da mesma forma, os resultados histológicos mostraram que a redensificação dérmica obtida com o EGTA-PR foi visualmente superior à observada com EATA-FI. Os resultados obtidos são promissores e corroboram as diversas ações biológicas já reportadas na literatura para extrato de Triticum aestivum L. nas etapas da cicatrização tecidual.

Introduction: Plant extracts and actives derived from plants were developed to improve and enhance the skin healing process including Triticum aestivum L. (Triticum vulgare). Purpose: To evaluate the effect of whole grain extract (EGTA-PR) and aqueous extract (EATA-FI) of Triticum aestivum L., on ex vivo skin healing. Methods: Skin fragments obtained from elective plastic surgery were subjected to tissue damage and treated with extracts for eight days for histological evaluation of re-epithelialization and immunofluorescence for epidermal growth factor (EGF). Results: EGTA-PR and EATA-FI accelerated the re-epithelialization process in human skin culture submitted to tissue injury. Additionally, we observed increased EGF protein labeling after treatment with EGTA-PR. Conclusion: EGTA-PR showed a better performance in re-epithelialization when compared to EATA-FI, as it presented a higher protein labeling for EGF in human skin culture. Likewise, the histological results showed that the dermal redensification obtained with EGTA-PR was visually superior to that observed with EATA-FI. The results obtained are promising and corroborate the several biological actions already reported in the literature for Triticum aestivum L. extract in tissue healing stages

Estrategias para la evaluación de extractos de polifenoles en modelos in vitro de cáncer de vías digestivas / Strategies for Evaluation of Polyphenol Extracts on in vitro Models of Digestive Tract Cancer / Estratégias para a avaliação de extratos de polifenóis em modelos in vitro de câncer do sistema digestivo

Introducción: Los polifenoles son compuestos que se encuentran naturalmente en alimentos como frutas, verduras, té, vino y chocolates, a los que se les atribuyen beneficios a la salud humana por su capacidad antioxidante. El cáncer de las vías digestivas se encuentra entre la tercera y quinta causas de muerte para la población, por lo que se ha incrementado el interés por realizar los estudios dirigidos a encontrar compuestos polifenólicos que ayuden en su prevención o tratamiento. Objetivo: Identificar las estrategias disponibles para la evaluación de polifenoles en células de cáncer de vías digestivas. Metodología: Búsqueda de literatura en bases de datos como Ovid, Pubmed, Science Direct y Elsevier Journal. Se seleccionaron artículos en los cuales se reporta el efecto biológico de los polifenoles sobre líneas celulares de cáncer de vías digestivas publicados entre 2012 y 2022. Resultados: Varios estudios han reportado el uso de un buen número de líneas celulares como modelos in vitropara estudios de polifenoles en cáncer y han resaltado las líneas AGS y HT-29, además de técnicas para la caracterización de los polifenoles, como el ensayo 2,2-Difenil-I-Picril Hidrazilo (DPPH). Sin embargo, para evaluar el efecto biológico se identifican diversas pruebas que deben analizarse antes de su implementación. Conclusiones: En la literatura se identifica que existen varias alternativas y estrategias para la evaluación de extrac-tos vegetales en cultivos in vitro de cáncer de vías digestivas; no obstante, antes de pasar al diseño experimental, deben tenerse en cuenta una serie de consideraciones para garantizar la utilidad de los resultados.

Introduction: Polyphenols are compounds naturally found in foods such as fruits, vegetables, tea, wine and chocolates, and it was attributed with benefits to human health due to their antioxidant capacity. Cancer of the digestive tract is between the third and fifth cause of death for the population, increasing the interest in carrying out studies aimed at finding polyphenolic compounds that help in their prevention or treatment. Objective: Identify the available strategies for the evaluation of polyphenols in digestive tract cancer cells. Method: A literature search was performed in databases such Ovid, Pubmed, Science Direct and Elsevier Journal and selected articles reporting the biological effect of polyphenols on digestive tract cancer cell lines, published between 2012 and 2022. Results: Currently studies report the use of a good number of cell lines as in vitro models for poly-phenol studies in cancer highlighting the AGS and HT-29 lines, in addition to techniques for the characterization of polyphenols such as the α, α-diphenyl-ß-picrylhydrazyl DPPH assay, however, to evaluate the biological effect various tests are identified that should be analyzed before implemen-tation. Conclusions: The literature identifies that there are many alternatives and strategies for the evaluation of plant extracts in in vitro cultures of digestive tract cancer, however, before moving on to the experimental design, a number of considerations should be taken into account to ensure the usability of the results

Introdução: Os polifenóis são compostos encontrados naturalmente em alimentos como frutas, legumes, chá, vinho e chocolates, aos quais são atribuídos benefícios para a saúde humana devido à sua capacidade antioxidante. O câncer do sistema digestivo está entre a terceira e a quinta principais causas de morte na população, o que levou a um interesse crescente em estudos destinados a encon-trar compostos polifenólicos que ajudem a prevenir ou tratar esse tipo de câncer. Objetivo: Identificar as estratégias disponíveis para a avaliação dos polifenóis nas células cancerosas do sistema digestivo. Metodologia: Pesquisa bibliográfica em bases de dados como Ovid, Pubmed, Science Direct e Elsevier Journal. Foram selecionados artigos que relatam o efeito biológico dos polifenóis em linhas celulares de câncer do sistema digestivo, publicados entre 2012 e 2022. Resultados: Vários estudos relataram a utilização de várias linhas celulares como modelos in vitro para estudos de polifenóis no câncer destacando as linhas AGS e HT-29, bem como técnicas para a ca-racterização de polifenóis, como o ensaio 2,2-Difenil-I-Picril Hidrazil (DPPH). No entanto, para avaliar o efeito biológico, são identificados vários testes que devem ser analisados antes da sua aplicação. Conclusões: A literatura identifica que existem várias alternativas e estratégias para a avaliação de extratos de plantas em culturas in vitro de câncer do sistema digestivo; no entanto, antes de passar à concepção experimental, é necessário ter em conta uma série de considerações para garantir a uti-lidade dos resultados

Effect of implant crestal position on primary stability before and after loading: an in vitro study / Efeito do posicionamento crestal do implante na estabilidade primária antes e após carga: um estudo in vitro

Introduction: Primary stability is one of the goals of modern implant dentistry and if achieved, reduces treatment time for prosthetic rehabilitation and the number of interventions made in patients mouth. Several companies state as protocol for connical conection implants, a subcrestally positioning. Objective: This in vitro study aimed to evaluate the effect of placing a conical connection implant equicrestally and subcrestally on static and loading condition in two types of bone density. Material and method: A total of 200 bone cylinders were extracted from femur of pigs, standardized by means of x-rays and computerized microtomography scan (microCT) and separated in low and high density specimens. The implants were placed on the center of the bone cylinders and were evaluated before and after loading by means of microCT and histomorphometry. Result: The results showed that placing the evaluated implant subcrestally provided better primary stability and performance on static and loading situations on low and high density bone. Conclusion: Placing implant subcrestally improve primary stability outcomes under loading and static situations.

Introdução: A estabilidade primária é um dos objetivos da implantodontia moderna e, caso atingida, reduz o tempo de tratamento para a reabilitação protéticas e o número de intervenções realizadas. Diversas empresas preconizam a posição subcrestal no uso de implantes com conexão cônica interna. Objetivo: Este estudo in vitro avaliou o efeito do posicionamento de implantes de conexão conica interna sub e equicrestal sob condições estáticas e em função, considerando dois tipos de densidades ósseas. Material e método: um total de 200 espécimes de osso extraído do femur de suínos e padronizados por meio de radiografias e microtomografias computadorizadas foram separados em densidade alta e baixa. Implantes foram instalados no centro dos especimes e for a avaliados por meio de microCT e histomorfometria. Resultado: Os resultados demonstraram que a colocação de implante subcrestalmente promoveu melhor estabilidade primária e performance em todas as situações, irrespectivamente à densidade óssea. Conclusão: A colocação de implantes subcrestalmente melhora a estabilidade primária em todas as situações, sendo indicada quando da utilização de conexões cônicas internas.

A review of alternative methods to the use of animals in safety evaluation of cosmetics

ABSTRACT Alternative methods to the use of animals in research have been a global trend, mainly after the publication of the 3R's principle (Replacement, Reduction, and Refinement), proposed by Russel and Burch. In the cosmetic sector, safety and efficacy assessments using animals have generated controversial debates. For this reason, in vitro research techniques are widely used to assess acute toxicity; corrosivity and irritation; skin sensitization; dermal and percutaneous absorption; repeated dose toxicity; reproductive toxicity; mutagenicity and genotoxicity; carcinogenicity; toxicokinetic studies; photo-induced toxicity; and human data. Although there are many methodologies described, validated, and widely used in the cosmetic area, the evaluation of the safety of cosmetic ingredients and products is still an expanding field. It needs global collaboration among regulatory agencies, universities, and industry, to meet several unmet needs in the fields of sensitization, carcinogenicity, systemic action, among other issues involving safety of users of cosmetic products. This review article will cover the currently most relevant in vitro models regarding irritation, corrosion, sensitization, mutagenicity, genotoxicity, and phototoxicity, to help to choose the most appropriate test for evaluating the safety and toxicity of cosmetic ingredients and products.

Establishment and functional identification of in vitro culture system of small intestine organoid in normal mice / 中华肾脏病杂志

Objective:To establish an in vitro culture system of small intestinal organoid in normal mice and perform functional identification, and to provide an in vitro research tool for material transport in the intestine under chronic kidney disease. Methods:The small intestinal crypts of C57BL/6J mice were isolated, extracted and cultured in an in vitro three dimension culture system. The formation of small intestinal organoid was observed with inverted microscope. The tissue structure of the small intestinal organoid was observed by hematoxylin and eosin staining. The cellular composition of the small intestinal organoid was identified by immunofluorescence. The expression of substance absorption-related transporters in the small intestinal organoid was detected by real time fluorescence quantitative PCR. Results:The small intestinal crypts were successfully extracted. The organoids of small intestine and different intestinal segments were successfully constructed. The cultured organoids had vigorous proliferation ability and maintained proliferation ability after passing through generations. Immunofluorescence results showed that the small intestinal organoids expressed mucin2, chromogranin A, oflm4 and lysozyme, which were different types of intestinal cell biomarkers. The PCR results showed that small intestinal organoids expressed calcium, phosphate and sodium absorption-related transporters, and the mRNA expression levels of major transporters for sodium and phosphate absorption in different intestinal segments-like organs were consistent with those in vivo, which was consistent with the characteristics of small intestinal segmental absorption. Conclusions:The successful construction of small intestine and different intestinal segments organoids, and the first observation of the expression of substance absorption-related transporters in such organoid, provide a stable and convenient in vitro research tool for the development of intestinal substance transport in chronic kidney disease.

Alimentos con potencial efecto inmunomodulador y antiviral a propósito de la pandemia COVID-19 / Potential Immunomodulatory and Antiviral Food Effect in The Context of COVID-19 Pandemic

Resumen Antecedentes: los tratamientos con base en medicamentos contra la COVID-19 no han sido aprobados hasta la actualidad. La forma más efectiva de enfrentar este problema de salud pública es la prevención con una adecuada alimentación, medidas de higiene y protección. Los alimentos han sido históricamente utilizados por la población para mejorar su nutrición y complementar el tratamiento o prevención de enfermedades. Se conocen los diversos compuestos bioactivos de algunos alimentos, que en estudios experimentales demostraron su acción antiviral e inmunomoduladora. Objetivo: identificar los compuestos bioactivos o preparados de alimentos con potencial efecto inmunomodulador, inmunoestimulante y antiviral contra el coronavirus. Materiales y métodos: se realizó una búsqueda en Google Scholar, Scopus y en la Biblioteca Virtual de Salud en Bases de datos de Medicina Tradicional, Complementaria e Integrativa utilizando los términos food, immunomodulatory, immunostimulatory y antiviral en cuatro búsquedas sucesivas. Resultados: se obtuvieron 93 artículos y se identificó mayor evidencia sobre el efecto antiviral e inmunológico contra el coronavirus en nueve alimentos: Allium sativum, Cinnamomum zeylanicum, Citrus sinensis, Zingiber officinale, Vitis vinífera, Allium cepa, Curcuma longa, Punica granatum y Sambucus nigra. Los cuatro primeros mostraron actividad contra el SARS-CoV-2. Conclusiones: se evidenció el efecto inmunológico y antiviral contra el coronavirus de nueve alimentos; sin embargo, son estudios in silico e in vitro, por ello se requiere mayor investigación preclínica y clínica que lo confirmen.

Abstract Background: Up to date drug-based treatments for COVID-19 have not been approved. The most effective way to face this public health problem is prevention with adequate nutrition, hygiene, and protection measures. Food has historically been used by people to improve their nutrition and complement the treatment or prevention of diseases. The various bioactive compounds of some foods are known, which in experimental studies demonstrated their antiviral and immunomodulatory action. Objective: To identify bioactive compounds of foods or food preparations with potential immunomodulatory, immunostimulatory, and antiviral effects against coronavirus. Material and Methods: A search was carried out in Google Scholar, Scopus, and Virtual Health Library on Traditional Complementary and Integrative Medicine using the terms food, immunomodulatory, immunostimulatory, and antiviral in four successive searches. Results: 93 articles were obtained, and further evidence of antiviral and immunological effect against coronavirus was identified on nine foods: Allium sativum, Cinnamomum zeylanicum, Citrus sinensis, Zingiber officinale, Vitis vinifera, Allium cepa, Curcuma longa, Punica granatum, and Sambucus nigra. The first four of them showed activity against SARS-CoV-2. Conclusions: The immunological and antiviral effect against coronavirus of nine foods was evidenced; however, they are in silico and in vitro studies, therefore it is required further preclinical and clinical research to confirm this.

Techniques for removing root canal posts: an vitro study / Técnicas para remoção de pinos intrarradiculares: um estudo in vitro

Objetivo: O objetivo deste estudo foi responder qual é a melhor técnica para remoção de pinos de fibra de vidro e pinos metálicos fundidos considerando tempo, preservação da estrutura dentária e custos. Métodos: Foi realizado um estudo in vitro, randomizado e cego. Sessenta dentes tratados endodonticamente foram randomizados em dois grupos de acordo com o tipo de pino. Uma segunda randomização foi realizada para cada tipo de técnica de remoção (ultrassom, broca ou combinada; n=10). One-way ANOVA foi usado para comparar o tempo de remoção do pino, teste t pareado comparou a quantidade de estrutura dentária removida e Kruskal Wallis seguido pelo teste post hoc de Mann-Whitney foram usados para análise de custo (P = 0,05). Resultados: Não houve diferença no tempo de remoção do pino considerando o tipo ou técnica do pino (P=0,630). A perda média de estrutura dentária na região cervical foi superior a 30% quando apenas uma ponta diamantada foi usada para remover pino de fibra de vidro (P<0,00001) e pino metálico fundido (P=0,008). Conclusão: De acordo com os resultados deste estudo, podemos concluir que a seleção da técnica de remoção dependerá da habilidade do operador, e sempre ocorrerá alguma perda de estrutura dentária, embora seja esperada uma perda maior quando apenas pontas diamantadas são usadas para este fim.(AU)

Objective: The aim of this study was to answer which is the best technique to removing glass fiber post and cast metal post considering time, preservation of tooth structure, and costs. Methods: An in vitro, randomized, blinded study was conducted. Sixty endodontically treated teeth were randomized into two groups according to post type. A second randomization was performed for each type of removal technique (ultrasound, drill, or combined; n=10). One-way ANOVA was used to compare the time to post removal, paired t-test compared the amount of tooth structure removed and Kruskal Wallis followed by post hoc Mann-Whitney test were used for cost analysis (P=0.05). Results: There was no difference in post removal time considering post type or technique (P=0.630). The average loss of tooth structure in the cervical region was greater than 30% when only a diamond bur was used to remove fiber glass post (P<0.00001) and cast metal post (P=0.008). Conclusion: According to the results of this study, we can conclude that the selection of the removal technique will depend on the skill of the operator, and some loss of tooth structure will always occur, although greater loss is expected when only diamond burs are used for this purpose.(AU)

Direct pedicle screw insertion pullout strength / Resistência ao arrancamento do parafuso pedicular de inserção direta

ABSTRACT Objective: Study the in vitro pullout strength of SpineGuard/Zavation Dynamic Surgical Guidance Z-Direct Screw (DSG Screw), a screw pedicle designed to be inserted using a direct insertion technique. Methods: DSG Screws of 5.5 mm and 6.5 mm were introduced into polyurethane blocks with a density of 10 PCF (0,16 g/cm3). According to the experimental group, screws were inserted without pilot hole, with pilot without tapping, undertapping and line-to-line tapping. Screw pullout tests were performed using a universal test machine after screw insertion into polyurethane blocks. Results: Screws inserted directly into the polyurethane blocks without pilot hole and tapping showed a statistically higher pullout strength. Insertion of the screw without tapping or with undertapping increases the pullout screw strength compared to line-to-line tapping. Conclusion: DSG Screw showed the highest pullout strength after its insertion without pilot hole and tapping. Level of Evidence V, Expert Opinion.

RESUMO Objetivo: Estudar a resistência ao arrancamento in vitro do parafuso de inserção direta da SpineGuard/Zavation (parafuso DSG), um parafuso pedicular projetado para ser inserido usando a técnica de inserção direta. Métodos: Parafusos DSG de 5,5 mm e 6,5 mm foram introduzidos em blocos de poliuretano com densidade de 10 PCF (0,16 g/cm3). De acordo com o grupo experimental, os parafusos foram inseridos sem orifício piloto, com orifício e sem macheamento e macheamento diâmetro inferior com mesma geometria. Os testes de resistência dos parafusos foram realizados usando uma máquina de teste universal após a inserção dos parafusos nos blocos de poliuretano. Resultados: Os parafusos inseridos diretamente nos blocos de poliuretano sem orifício piloto e sem macheamento apresentaram uma resistência de arrancamento com significância estatística maior. A inserção do parafuso sem macheamento ou com macheamento com diâmetro inferior apresenta maior resistência ao arrancamento em comparação com o macheamento do mesmo diâmetro. Conclusão: O parafuso DSG apresentou a maior resistência ao arrancamento após sua inserção sem orifício piloto e sem macheamento. Nível de Evidência V, Opinião do Especialista.

Efecto del Camphenol Plus sobre el músculo liso vascular de vena porta en ratas Wistar / Effect of Camphenol Plus on portal vein vascular smooth muscle in Wistar rats

RESUMEN Introducción: uno de los antisépticos comúnmente empleado en Estomatología desde el pasado siglo y que mantiene su uso hasta la actualidad, lo constituye el Camphenol Plus. Son escasos los reportes científicos de su efecto sobre el endotelio y la dinámica contráctil del músculo liso vascular, en especial de tejidos venosos como la vena porta hepática. Objetivo: determinar el efecto del Camphenol Plus sobre el músculo liso vascular de la vena porta. Métodos: se realizó una investigación experimental preclínica, con la utilización de 21 venas porta obtenidas de ratas Wistar. Las preparaciones realizadas se colocaron en baño de órganos, se registró la tensión desarrollada por el músculo liso vascular tras la adición de diez microlitros de Camphenol Plus, en diferentes concentraciones y durante diferentes intervalos de tiempo. Resultados: el Camphenol Plus, tras la preactivación del musculo liso vascular de la vena porta, indujo vasorelajación, la que se incrementó durante todo el tiempo de estudio y según el incremento de las concentraciones del medicamento. Existieron diferencias significativas entre los valores de tensión promedios registrados en los diferentes intervalos de tiempo con los de la tensión espontánea basal y la tensión base inicial. Conclusiones: el Camphenol Plus, indujo "in vitro", relajación de la musculatura lisa de la vena porta a través de un acoplamiento excitación-contracción de tipo farmacomecánico.

ABSTRACT Introduction: Camphenol Plus is one of the antiseptics commonly used in Dentistry since the last century and still in use today. There are few scientific reports of its effect on the endothelium and contractile dynamics of vascular smooth muscle, especially in venous tissues such as the hepatic portal vein. Objective: to determine the effect of Camphenol Plus on the vascular smooth muscle of the portal vein. Methods: a preclinical experimental investigation was carried out using 21 portal veins obtained from Wistar rats. The preparations were placed in an organ bath and the tension developed by the vascular smooth muscle was recorded after the addition of ten microliter of Camphenol Plus, at different concentrations and during different time intervals. Results: Camphenol Plus, after the preactivation of the vascular smooth muscle of the portal vein, induced relaxation, which increased throughout the study time and according to the increase in drug concentrations. There were significant differences between the average tension values recorded in the different time intervals with those of the basal spontaneous tension and the initial baseline tension. Conclusions: Camphenol Plus induced "in vitro" relaxation of portal venous smooth muscles through a pharmacomechanical excitation-contraction coupling.

Alternative method for the evaluation of monovalent inactivated foot and mouth disease virus vaccine

Foot and mouth disease is a highly contagious viral disease of cloven-hoofed animals that has a significant economic impact on livestock. A recent outbreak was detected and recorded as exotic strain of foot and mouth disease virus SAT2 (Serotype SAT2, topotype VII, Lib-12 lineage). The emergency vaccine was produced and assessed in vivo and large number of vaccine batches were urgently needed. The present work was aimed to provide a rapid evaluation of inactivated foot and mouth disease SAT2 oily vaccine to exclude the unsatisfactory batches during emergency circumstances and to reduce time, effort and cost. The extraction of foot and mouth disease antigen content from oily adjuvanted vaccine was carried out using isopropyl myristate and benzyl alcohol methods. The extracted viral antigen was identified by foot and mouse disease serotyping ELISA and 146S content was quantified using sucrose density gradient analysis. Evaluations were carried out instantly and at 2h, 6h and 24h. The results indicated the efficiency of benzyl alcohol to breakdown the oil emulsion either MONTANIDE™ ISA 206 VG or MONTANIDE™ ISA 50 V2, while the isopropyl myristate was efficient for MONTANIDE™ ISA 50 V2 only. The identification and quantification of 146S for extracted antigen using benzyl alcohol indicated significant stable records at different time intervals for the vaccine batches, while the extraction using isopropyl myristate indicated unstable records at different time intervals. It was concluded that the evaluation of monovalent foot and mouse disease vaccine could be conducted in vitro, using serotyping ELISA and quantification of 146S for the extracted antigen, either using benzyl alcohol or isopropyl myristate (MONTANIDE™ ISA50 V2 only), with the consideration that 146S content should not less than 4 μg/mL(AU)

La fiebre aftosa es una enfermedad viral altamente contagiosa de los animales de pezuña hendida que tiene un impacto económico significativo en el ganado. Se detectó un brote reciente que se registró como causado por una cepa exótica del virus de la fiebre aftosa (serotipo SAT2, topotipo VII, linaje Lib-12). La vacuna de emergencia se elaboró y evaluó in vivo, existiendo una urgente necesidad de contar con un gran número de lotes de la misma. El presente trabajo tuvo como objetivo proporcionar una evaluación rápida de la vacuna oleosa inactivada (SAT2) contra la fiebre aftosa, para excluir los lotes insatisfactorios durante circunstancias de emergencia, reduciendo tiempo, esfuerzo y costo. La extracción del contenido de antígeno de fiebre aftosa, de la vacuna oleosa adyuvada, se llevó a cabo utilizando miristato de isopropilo y alcohol bencílico. El antígeno viral extraído se identificó utilizando un ELISA de serotipificación y se cuantificó el contenido de 146S mediante análisis de gradiente de densidad de sacarosa. Las evaluaciones se realizaron de forma instantánea y a las 2h, 6h y 24h. Los resultados indicaron la eficacia del alcohol bencílico para separar la emulsión de aceite para MONTANIDE ™ ISA 206 VG o MONTANIDE™ ISA 50 V2, mientras que el miristato de isopropilo fue eficaz para MONTANIDE™ ISA 50 V2 únicamente(AU)

Change in the activity of glucosylceramide synthase in human hepatoma cell line with hepatitis B virus infection in vitro / 临床肝胆病杂志

ObjectiveTo investigate the change in the activity of glucosylceramide synthase, the key enzyme in glycosphingolipid metabolism and synthesis, in Huh7 cells infected by hepatitis B virus (HBV) in vitro. MethodsBlood samples were collected from nine previously untreated patients with acute hepatitis B who attended Department of Infectious Diseases, The First Hospital of Lanzhou University, from June to August, 2019, and the blood samples collected from seven healthy individuals who underwent physical examination were established as control. Huh7 cells were inoculated with the high-copy HBV particles (＞9.9×107 IU/ml) in the serum of patients with HBV infection (infection group), and Huh7 cells co-cultured with the serum of healthy individuals were established as control group. The expression levels of HBsAg and HBV DNA in the cytoplasm of HBV-infected Huh7 cells were measured, and the correlation between GCS activity and virus was analyzed. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups, and a Pearson correlation analysis was performed. ResultsCompared with the control group, the infection group had a significant reduction in the number of cells, an increase in cell volume, and cell membrane fragmentation. The infection group had a significant increase in the expression of HBsAg in cytoplasm at 4 hours, 8 hours, 2 days, and 5 days after infection (P＜0.05); the expression level of HBV DNA tended to increase significantly from 4 hours after infection to 8 hours, 2 days, and 5 days after infection (16.67±11.55 IU/ml vs 112.01±25.94 IU/ml/328.01±10350 IU/ml/101.60±49.84 IU/ml, P＜0.001), with the highest level at 2 days after infection. During HBV infection, the activity of GCS gradually increased with the increase in viral replication from 4 hours after infection (126.21±9.59 IU/ml) and reached a peak at 2 days after infection (226.53±36.27 IU/ml), with a significant difference between the infection group and the control group at 2 days after infection (226.53±36.27 IU/ml vs 136.50±1544 IU/ml, t=3.956, P=0.016 7). The activity of GCS was positively correlated with HBV DNA level (r=0.576 8, P=0047 1). ConclusionHuh7 cells are successfully infected with the high-copy HBV particles in the serum of patients with HBV infection, which mimics the characteristics of HBV infection in vitro to a certain degree. The activity of GCS may be associated with HBV infection, suggesting that glycosphingolipid synthesis and metabolism may be closely associated with HBV.

Biological and mechanical characterization of commercial and experimental periodontal surgical dressings

Abstract The objective of this study was to evaluate the biocompatibility and mechanical properties of two commercially available and one experimental periodontal dressing materials. The cytotoxicity of Periobond ® , Barricaid ® and one experimental periodontal dressing based on Exothane ® 8 monomer was tested on 3T3/NIH mouse fibroblast. Genotoxicity was assessed by micronuclei formation, and cell alterations were analyzed using light microscopy. Both biological assays were performed using the eluate obtained from specimens after 24, 72, or 168 hours of incubation. Mechanical characterization was assessed through the ultimate tensile strength and the water sorption and solubility tests. The significance level of α = 0.05 was used for all statistical analyses. All the materials promoted a cell viability lower than 60% in all evaluated times. In general, the cell viability was significantly reduced after 72 and 168h of specimens' incubation. Considering the factor material, there were not statistical differences in the cell viability (p = 0.156). The genotoxicity was not statistically significant among the groups in the different periods of time (p > 0.05). Differences in the ultimate tensile strength values were not statistically significant different among the groups (p = 0.125). Periobond ® showed the higher water sorption values (p < 0.001). Regarding solubility, there were no statistical differences between the groups (p = 0.098). All the periodontal dressing materials evaluated in this study exerted a cytotoxic effect against mouse fibroblasts, and their toxicity became more evident over time. Among the materials evaluated, the experimental light-cure type has shown overall similar properties to the commercial references.

Estudio comparativo in vitro de la filtración apical con tres técnicas de obturación / Estudo comparativo in vitro de filtração apical com três técnicas de obturação / A comparative in vitro study of apical leakage with three obturation techniques

Resumen Objetivo: Comparar la microfiltración apical de dientes obturados con tres diferentes técnicas: condensación lateral, condensación vertical y sistema obtura II in vitro. Métodos: Se utilizaron 75 dientes uniradiculares, divididos en 5 grupos, Grupo 1: condensación lateral, Grupo 2: condensación vertical, Grupo 3: sistema obtura II, Grupo 4: control positivo, Grupo 5: control negativo. Las raíces fueron sometidas a filtración apical por difusión indirecta con tinta china, diafanizadas y observadas con un microscopio Vasconcellos. Se realizaron pruebas estadístico descriptivo con el programa GraphPad prism 8. Resultados: El grupo 1 presentó mayor filtración (0,6603±0,5063) con una diferencia estadísticamente significativa en comparación al grupo 3 (p<0.05), sin embargo, no hubo extrusión de la gutapercha a diferencia de los otros dos grupos. Conclusiones: La técnica de sistema Obtura II presentó un mejor sellado apical, lo que condujo a una disminución en la filtración apical, demostrando ser más efectiva en comparación a las otras técnicas utilizada en el estudio.

Resumo Objetivo: Comparar a microfiltração apical de dentes obturados com três diferentes técnicas: condensação lateral, condensação vertical e sistema obtura II. Métodos: foram utilizados 75 dentes unirradiculares, divididos em 5 grupos, Grupo 1: condensação lateral, Grupo 2: condensação vertical, Grupo 3: sistema obtura II, Grupo 4: controle positivo, Grupo 5: controle negativo. As raízes foram submetidas à filtração apical por difusão indireta com tinta da Índia, diafanizadas e observadas ao microscópio Vasconcellos. Os testes estatísticos descritivos foram realizados com o programa GraphPad prism 8. Resultados: O Grupo 1 apresentou maior filtração (0,6603 ± 0,5063) com diferença estatisticamente significante em relação ao Grupo 3 (p <0,05), porém não houve extrusão de guta-percha ao contrário dos outros grupos onde havia. Conclusões: A técnica do sistema Obtura II apresentou melhor selamento apical, o que levou à diminuição da filtração apical, mostrando-se mais eficaz em relação às outras técnicas utilizadas no estudo.

Abstract Objective: To compare the in vitro apical microleakage of teeth sealed with three techniques: lateral condensation, vertical condensation, and Obtura II. Methods: A total of 75 single-root teeth were divided into five groups: Group 1 - lateral condensation, Group 2 - vertical condensation, Group 3 - Obtura II system, Group 4 - positive control, Group 5 - negative control. The roots were subjected to apical leakage through indirect diffusion with Chinese ink, diaphonized, and observed under a Vasconcellos microscope. Descriptive statistical tests were conducted with GraphPad Prism 8. Results: Group 1 showed greater microleakage (0.6603 ± 0.5063) with a statistically significant difference compared to group 3 (p < 0.05). However, there was no gutta-percha extrusion, unlike in the other groups. Conclusion: The Obtura II technique showed better sealing, which led to decreased apical microleakage, thus proving to be more effective than the other techniques studied.

Evaluation of physical and biological properties of conventional glass ionomer cements / Avaliação das propriedades físicas e biológicas de cimentos de ionômero de vidro convencionais

Objetivo: avaliar as propriedades físicas e biológicas dos cimentos de ionômero de vidro convencionais (CIVs). Metodologia: foram avaliados os seguintes CIVs: Fuji IX (GC Europe, Bélgica), Ketac Molar (3M ESPE, Estados Unidos), Maxxion R (FGM, Brasil) e Vitro Molar (Nova DFL, Brasil). O tempo de presa, a alteração dimensional, a radiopacidade, a sorção e a solubilidade em água foram avaliados para todos os materiais. A resistência à compressão foi analisada em intervalos de 1h, 24h, 7 dias e 28 dias; e liberação de íons fluoreto em 3h, 24h e 72h. A viabilidade celular foi avaliada após 24 e 48 horas com células de fibroblastos. A análise estatística foi realizada por meio do software SigmaPlot 12 (Systat Inc, San Jose, CA, EUA), com nível de significância estabelecido em α = 0,05. Resultado: apenas o Fuji IX teve um tempo de presa dentro da faixa recomendada pela Especificação Padrão da ADA 96 (2012), não superior a 6 minutos. Vitro Molar e Maxxion R apresentaram radiopacidade que não estava de acordo com a especificação ADA 96 (2012). Maxxion R e Vitro Molar mostraram uma alteração dimensional estatisticamente semelhante. Quanto às propriedades mecânicas, o Fuji IX foi o único CIV que apresentou aumento da resistência à compressão durante o período de avaliação de 28 dias. O Ketac Molar apresentou a maior viabilidade celular, enquanto o Maxxion R apresentou citotoxicidade severa e o maior valor cumulativo de liberação de flúor. Conclusão: Fuji IX e Ketac Molar apresentaram as propriedades físicas e biológicas mais adequadas entre os CIVs avaliados.(AU)

Objective: to evaluate the physical and biological properties of conventional glass ionomer cements (GICs). Methodology: the following GICs were evaluated: Fuji IX (GC Europe, Belgium), Ketac Molar (3M ESPE, United States), Maxxion R (FGM, Brazil) and Vitro Molar (Nova DFL, Brazil). Setting time, dimensional change, radiopacity, water solubility and water absorption were evaluated for all materials. Compressive strength was analyzed after intervals of 1h, 24h, 7 days and 28 days; and release of fluoride ions at 3 am, 24 am and 72 am. Cell viability was assessed after 24 and 48 hours with fibroblast cells. Statistical analysis was performed using SigmaPlot 12 software (Systat Inc, San Jose, CA, USA), with a significance level set at α = 0.05. Result: only the Fuji IX had an adjustment time within the range recommended by the Standard Specification of ADA 96 (2012) of not exceeding 6 minutes. Vitro Molar and Maxxion R had radiopacity that was not in accordance with the ADA 96 (2012) specification. Maxxion R and Vitro Molar showed a statistically similar dimensional change. As for mechanical properties, Fuji IX was the only GIC that showed an increase in compressive strength during the evaluation period of 28 days. Ketac Molar showed the highest cell viability, while Maxxion R showed severe cytotoxicity and the highest cumulative fluoride release value. Conclusion: Fuji IX and Ketac Molar showed the most appropriate physical and biological properties among the evaluated GICs.(AU)

Descubrimiento de fármacos basado en imagenología de células vivas / Drug discovery based on live cell imaging

RESUMEN El diseño eficiente de compuestos aprovechando las características estructurales de las moléculas y la búsqueda eficiente de dianas terapéuticas, ha proporcionado herramientas efectivas en la investigación de nuevos tratamientos cuando esta se enfoca en mecanismos celulares de la enfermedad. Los cambios fenotípicos producidos por la interacción in vitro entre molécula-diana, pueden controlarse cuantitativamente mediante imagenología de células vivas. Para garantizar una interacción adecuada, es necesario considerar diferentes elementos cruciales: 1. Las características estructurales y la dinámica molecular del compuesto a evaluar. 2. La relevancia del blanco para la fisiopatología de interés. Sin embargo, el desconocimiento del panorama general en el descubrimiento de fármacos, desde problemáticas estructurales y celulares, ha enlentecido la búsqueda de nuevos tratamientos. Esta revisión descriptiva de tema presenta algunos aspectos estructurales importantes para la caracterización de compuestos como candidatos terapéuticos, y aproximaciones experimentales para desarrollo de sistemas celulares. Los tópicos discutidos se enfocan en la monitorización por imagenología de células vivas y así mismo proporcionamos ejemplos relevantes. La monitorización de efectos fenotípicos producidos por interacciones entre candidato químico y blanco terapéutico en un sistema celular puede favorecer la búsqueda eficiente de moléculas potencialmente terapéuticas.

SUMMARY The efficient compounds' design taking advantage of the molecule's structural characteristics and efficient search for therapeutic targets has provided effective tools for the research of new treatments when this is focused on disease cellular mechanisms. Phenotypic changes produced by in vitro interaction between molecules and targets can be monitored quantitatively by live cell imaging. To guarantee adequate interaction, it is necessary to consider different crucial elements: 1. Structural characteristics and molecular dynamics of the evaluated compound. 2. Target relevance for the concern physiopathology. However, overview's ignorance of the drug discovery, from structural and cellular problems, has slowed the new treatments research. This literature review presents some important structural aspects for compounds' characterization as therapeutic candidates and experimental approaches for cellular systems development. Subjects discussed are focused on live cell imaging and we also provide relevant examples. Phenotypic monitoring of interactions' produced effects between the chemical candidate and therapeutic target in a cellular system can favor the efficient search of potentially therapeutic molecules.