RESUMO
The TRPV1 receptor, which is known to contribute significantly to pain perception, has recently been identified as a useful tool for predicting eye stinging potential in cosmetics. In this study, HEK-293 cells with high TRPV1 expression were utilized to evaluate calcium influx related to receptor activation triggered by chemicals and cosmetic formulations. The cells were exposed to increasing concentrations of substances to cause or not some aggression to the eye, and TRPV1 activity was assessed by measuring intracellular FURA-2 AM fluorescence signal. To confirm TRPV1 channel activation, capsazepine, a capsaicin antagonist, was employed in addition to using capsaicin as a positive control. The study's results indicate that this novel model can identify compounds known to cause some aggression to the eye, such as stinging, considering a cut-off value of 60% of Ca2+ influx exposed to the lowest evaluated concentration (0.00032%). When applied to the cosmetic baby formulation, although the presented model exhibited higher sensitivity by classifying as stinging formulations that had previously undergone clinical testing and were deemed non-stinging, the assay could serve as a valuable in vitro tool for predicting human eye stinging sensation and can be used as a tier 1 in an integrated testing strategy.
Assuntos
Cálcio , Cosméticos , Canais de Cátion TRPV , Humanos , Cosméticos/toxicidade , Células HEK293 , Canais de Cátion TRPV/metabolismo , Cálcio/metabolismo , Olho/efeitos dos fármacos , Capsaicina/análogos & derivados , Capsaicina/farmacologia , Alternativas aos Testes com AnimaisRESUMO
In vitro models of the dental pulp microenvironment have been proposed for the assessment of biomaterials, to minimise animal use in operative dentistry. In this study, a scaffold/3-D dental pulp cell culture interface was created in a microchip, under simulated dental pulp pressure, to evaluate the cell-homing potential of a chitosan (CH) scaffold functionalised with calcium aluminate (the 'CHAlCa scaffold'). This microphysiological platform was cultured at a pressure of 15 cm H2O for up to 14 days; cell viability, migration and odontoblastic differentiation were then assessed. The CHAlCa scaffold exhibited intense chemotactic potential, causing cells to migrate from the 3-D culture to its surface, followed by infiltration into the macroporous structure of the scaffold. By contrast, the cells in the presence of the non-functionalised chitosan scaffold showed low cell migration and no cell infiltration. CHAlCa scaffold bioactivity was confirmed in dentin sialophosphoprotein-positive migrating cells, and odontoblastic markers were upregulated in 3-D culture. Finally, in situ mineralised matrix deposition by the cells was confirmed in an Alizarin Red-based assay, in which the CHAlCa and CH scaffolds were adapted to fit within dentin discs. More intense deposition of matrix was observed with the CHAlCa scaffold, as compared to the CH scaffold. In summary, we present an in vitro platform that provides a simple and reproducible model for selecting and developing innovative biomaterials through the assessment of their cell-homing potential. By using this platform, it was shown that the combination of calcium aluminate and chitosan has potential as an inductive biomaterial that can mediate dentin tissue regeneration during cell-homing therapies.
Assuntos
Compostos de Alumínio , Compostos de Cálcio , Quitosana , Animais , Alicerces Teciduais/química , Polpa Dentária , Materiais Biocompatíveis/química , Diferenciação Celular , Células Cultivadas , Engenharia TecidualRESUMO
The current study was designed to check the anthelmintic activities of some local plants. Seeds of Amomum (A.) subulatum and Vitex (V.) negundo in different solvents were subjected to in vitro (adult motility assay; AMA and egg hatch assay; EHA) and in vivo (faecal egg count reduction test; FECRT) anthelmintic activity testing protocols using Haemonchus (H.) contortus as an experimental model. The results of AMA, EHA, and FECRT were statistically analysed through linear regression and Duncan multiple range test. In AMA test, at 50 mg mL-1 concentration, the percent mortality of H. contortus was higher in A. subulatum than V. negundo, whereas, in EHA test, A. subulatum was proven better ovicidal (LC50=14.2 µg mL-1) than V. negundo (LC50= 65.7405 µg mL-1). The FECRT also indicated the better efficacy of A. subulatum than V. negundo against natural infection of gastrointestinal (GI) parasites. The crude powder of plants used in this study showed 29.6% to 57.7% anthelmintic. The reduction rate was found higher for A. subulatum (3 g kg-1) as compared to V. negundo (7 g kg-1). Reagrding efficacy analysis of solvents used for plants extract, ethyl acetate and chloroform were found better in increasing ovicidal activity in adult worms (in vitro testing), whereas, the crude aqueous methanol was found better than the crude powders in in vivo testing. It will be beneficial to document the indigenous knowledge to standard scientific procedures for their validation. This study will help to motivate the farmers to make a better choice of cultivation of the indigenous plants because of their varying efficacies as an alternative preventive approach against the GI parasitic infections.
O presente estudo foi desenhado para verificar as propriedades anti-helmínticas de algumas plantas locais. Sementes de Amomum (A.) subulatum e Vitex (V.) negundo em diferentes solventes foram submetidas à análise de atividade anti-helmíntica in vitro (ensaio de motilidade de adultos; AMA e teste de eclosão de ovos; EHA) e in vivo (teste de redução da contagem de ovos nas fezes; TRCOF), usando o Haemonchus (H.) contortus como modelo experimental no protocolo de teste. Os resultados dos testes AMA, EHA e TRCOF foram analisados estatisticamente por meio de regressão linear e teste de Duncan. No teste AMA, na concentração de 50 mg mL-1, o percentual de mortalidade de H. contortus foi maior com o uso de A. subulatum do que com V. negundo, enquanto, no teste EHA, A. subulatum apresentou maior ação ovicida (LC50=14,2 µg mL- 1) do que V. negundo (LC50= 65,7405 µg mL-1). O TRCOF também indicou a melhor eficácia do uso de A. subulatum do que de V. negundo contra a infecção natural de parasitas gastrointestinais (GI). O extrato bruto seco das plantas utilizadas neste estudo apresentou 29,6% a 57,7% de atividade anti-helmíntica. A taxa de redução observada com o uso de A. subulatum (3 g kg-1) foi maior que com o uso de V. negundo (7 g kg-1). Em relação à análise da eficácia dos solventes utilizados para o extrato de plantas, o acetato de etila e o clorofórmio apresentaram maior ação ovicida em vermes adultos (testes in vitro), enquanto o extrato bruto metanólico aquoso apresentou maior eficácia do que os extratos brutos secos em testes in vivo. Consideramos vantajoso documentar o conhecimento indígena relativos aos procedimentos científicos padronizados, para sua validação. Este estudo irá servir de motivação para que os agricultores façam escolhas melhores referentes ao cultivo das plantas indígenas devido às suas diferentes eficácias comprovadas, servindo como alternativa para a abordagem preventiva contra as infecções parasitárias GI.
Assuntos
Doenças Parasitárias , Plantas Medicinais , Vitex/parasitologia , Amomum/parasitologia , Anti-HelmínticosRESUMO
Microparticulate systems such as microparticles, microspheres, microcapsules or any particle in a micrometer scale (usually of 1-1000 µm) are widely used as drug delivery systems, because they offer higher therapeutic and diagnostic performance compared to conventional drug delivery forms. These systems can be manufactured with many raw materials, especially polymers, most of which have been effective in improving the physicochemical properties and biological activities of active compounds. This review will focus on the in vivo and in vitro application in the last decade (2012 to 2022) of different active pharmaceutical ingredients microencapsulated in polymeric or lipid matrices, the main formulation factors (excipients and techniques) and mostly their biological activities, with the aim of introducing and discussing the potential applicability of microparticulate systems in the pharmaceutical field.
Assuntos
Sistemas de Liberação de Medicamentos , Polímeros , Composição de Medicamentos/métodos , Sistemas de Liberação de Medicamentos/métodos , Polímeros/química , Excipientes , Cápsulas , Microesferas , Tamanho da PartículaRESUMO
Phthalocyanine (Pc) dyes are photoactive molecules that can absorb and emit light in the visible spectrum, especially in the red region of the spectrum, with great potential for biological scopes. For this target, it is important to guarantee a high Pc solubility, and the use of suitable pyridinium units on their structure can be a good strategy to use effective photosensitizers (PSs) for photodynamic therapy (PDT) against cancer cells. Zn(II) phthalocyanines (ZnPcs) conjugated with thiopyridinium units (1-3) were evaluated as PS drugs against B16F10 melanoma cells, and their photophysical, photochemical, and in vitro photobiological properties were determined. The photodynamic efficiency of the tetra- and octa-cationic ZnPcs 1-3 was studied and compared at 1, 2, 5, 10, and 20 µM. The different number of charge units, and the presence/absence of a-F atoms on the Pc structure, contributes for their PDT efficacy. The 3-(4',5'-dimethylthiazol-2'-yl)-2,5-diphenyl tetrazolium bromide (MTT) assays on B16F10 melanoma cells show a moderate to high capacity to be photoinactivated by ZnPcs 1-3 (ZnPc 1 > ZnPc 2 > ZnPc 3). The best PDT conditions were found at a Pc concentration of 20 µM, under red light (λ = 660 ± 20 nm) at an irradiance of 4.5 mW/cm2 for 667 s (light dose of 3 J/cm2). In these conditions, it is noteworthy that the cationic ZnPc 1 shows a promising photoinactivation ratio, reaching the detection limit of the MTT method. Moreover, these results are comparable to the better ones in the literature.
RESUMO
In breast cancer treatment, chemotherapy resistance is a major problem where many receptive tumors rebound and develop resistance. When provided in combination, cancer drugs are most successful, thus reducing the risk of developing resistant cancer cells. However, the evaluation of combination therapies has increased rapidly in recent years. Consequently, by repurposing old treatments, the discovery of additional medicines that may interact synergistically with chemotherapy is considered a current medical aim through discovering a new cancer medication or therapeutic strategy. The purpose of this research is to increase the anti-cancer activity of carboplatin (CP) by increasing the apoptotic effect of breast cancer cells (MCF-7) during in vitro experiments in combination with oxytetracycline. Our results showed a high synergistic effect between oxytetracycline and carboplatin, MCF-7 representative cell treated with carboplatin with/without different concentrations of oxytetracycline (5% and 10% of IC50). Oxytetracycline, which potentiated the action of carboplatin and/or had notable activity was reported as a single agent. This research demonstrated the synergistic relationship between oxytetracycline and carboplatin in viability assays. Surprisingly, our findings suggest that inhibiting treatment strategies can extend carboplatin's therapeutic window, potentially allowing for cancer therapy.(AU)
No tratamento do câncer de pulmão a resistência à quimioterapia é o maior problema no qual muitos tumores receptivos apresentam um rebote e desenvolvem a resistência. Quando oferecidas em combinações, as drogas anticancerígenas apresentam maior taxa de sucesso, reduzindo assim o risco de desenvolvimento de células cancerígenas resistentes. Contudo, a avaliação das terapias de combinação tem crescido muito rapidamente. Consequentemente, por reaproveitamento de tratamentos antigos, a descoberta de novos medicamentos adicionais que podem interagir sinergicamente com a quimioterapia que é considerada como auxílio médico na corrente busca à descoberta de novas medicações anticancerígenas ou estratégias terapêuticas. O propósito da presente pesquisa é aumentar a atividade anticancerígena da Carboplatina (CP) pelo incremento do efeito apoptótico de células de câncer pulmonar (MCF-7) em experimentos in vitro pela combinação com oxitetraciclina. Os resultados obtidos confirmaram elevado efeito sinérgico entre oxitetraciclina e Carboplatina em células MCF-7 representativas tratadas com Carboplatina, com e sem diferentes concentrações de oxitetraciclina (5% e 10% de IC50). A oxitetracilina que potencializou a ação da Carboplatina e/ou teve uma notável atividade relatada como um agente isolado. A pesquisa demonstrou a relação sinérgica entre oxitetraxiclina e Carboplatina nos ensaios de viabilidade. Surpreendentemente, os resultados obtidos sugeriram que as estratégias de tratamento inibidor podem aplicar um janela terapêutica da Carboplatina com potencial para a terapia do câncer.(AU)
Assuntos
Oxitetraciclina/farmacologia , Neoplasias da Mama/tratamento farmacológico , Carboplatina/farmacologia , Técnicas In Vitro/veterinária , Sinergismo Farmacológico , Células MCF-7/efeitos dos fármacosRESUMO
This study aimed to isolate and select in vitro bacteria with probiotic potential for the Amazon ornamental fish Nannostomus beckfordi. For isolate, twelve fish underwent surgery procedure to remove their intestinal tract, macerate and then inoculate in the plate petri containing de Man Rugosa Sharped Agar (MRS). After bacterial growth (48 hours at 35ºC), selected strains were inoculated in MRS broth and submitted to resistance test with NaCl (0.5, 1.0, 1.5, 2.0, 2.5 and 3.0%), pH (4, 5, 6, 8 and 9) and bile salts (5% w/v). Inhibition test against pathogenic bacteria Aeromonas hydrophila, Pseudomonas aeroginosa, Streptococcus agalactiae and Aeromonas Jandaei was also performed. Within the isolated strains group (23 strains), only six (S1, S2, S3, S4, S5 and S6) showed probiotic potential. Strains S1 and S6 showed the greater resistance for NaCl (0.5% and 1%) and pH (5 and 6), but only S1 obtained better results to resist the bile salts. Even against pathogenic bacteria, the S1 showed the best results with inhibition halos greater than 9 mm. In the end, this bacterial strain (S1) was identified as Enterococcus faecium 11037CHB. Thus, this is the first report regarding isolated autochthonous bacterium E. faecium with probiotic potential of N. beckfordi.
O objetivo deste estudo foi isolar e selecionar in vitro bactérias com potencial probiótico do peixe ornamental Amazônico Nannostomus beckfordi. Para o isolamento, retirou-se o intestino de 12 espécimes, que foram macerados, homogeneizados e semeados em placa de petri contento Ágar Man Rogosa e Sharpe (MRS). Posteriormente ao crescimento bacteriano (48 horas a 35ºC), as cepas selecionadas foram mantidas em caldo MRS e submetidas a testes de resistência a NaCl (0,5, 1,0, 1,5, 2,0 e 2,5 e 3,0%), pH (4, 5, 6, 8 e 9) e sais biliares (5% p/v). O antagonismo foi realizado frente as bactérias patogênicas Aeromonas hydrophila, Pseudomonas aeroginosa, Streptococcus agalactiae e Aeromonas jandaei. Das cepas isoladas (23 cepas), apenas seis (C1, C2, C3, C4, C5 e C6) apresentaram potencial probiótico. As cepas C1 e C6 tiveram maior resistência (p<0,05) para o NaCl (0,5 e 1%) e pH (5 a 6), na presença de sais biliares somente a C1 teve a melhor resistência de crescimento. Para o antagonismo frente as bactérias patogênicas, a C1 apresentou halo de inibição maior que 9 mm. Sendo esta cepa bactéria (C1) identificada como Enterococcus faecium 11037 CHB. Portanto, este é o primeiro relato do isolamento da bactéria autóctone E. faecium em N. beckfordi com potencial probiótico.
Assuntos
Animais , Probióticos/isolamento & purificação , Characidae/microbiologia , Enterococcus faecium/isolamento & purificaçãoRESUMO
Chagas disease is a neglected tropical disease caused by the protozoan parasite Trypanosoma cruzi. There is an urgent need for safe, effective, and accessible new treatments since the currently approved drugs have serious limitations. Drug development for Chagas disease has historically been hampered by the complexity of the disease, critical knowledge gaps, and lack of coordinated R&D efforts. This review covers some of the translational challenges associated with the progression of new chemical entities from preclinical to clinical phases of development, and discusses how recent technological advances might allow the research community to answer key questions relevant to the disease and to overcome hurdles in R&D for Chagas disease.
RESUMO
Abstract Background: Soybean milk by-product (SMBP) is a potential alternative feed ingredient in swine diets due to its high protein content. However, information on energy and nutritional values of SMBP used as swine feed ingredient is limited. Objective: To estimate energy values and protein digestibility of SMBP in pigs based on in vitro assays. Methods: Four SMBP samples were obtained from 3 soybean milk-producing facilities. In vitro total tract disappearance (IVTTD) and in vitro ileal disappearance (IVID) of dry matter (DM) in the SMBP samples were determined. In vitro ileal disappearance of crude protein was determined by analyzing crude protein content in undigested residues after determining IVID of DM. Digestible and metabolizable energy of SMBP were estimated using gross energy, IVTTD of DM, and prediction equations. Results: Sample 4 had greater IVTTD of DM than that of sample 3 (97.7 vs. 94.4%, p<0.05), whereas IVID of DM in sample 4 was lower compared with sample 1 (53.5 vs. 65.0%, p<0.05). In vitro ileal disappearance of crude protein in sample 2 was greater than that in sample 1 and 3 (92.6 vs. 90.6 and 90.1%; p<0.05). The estimated metabolizable energy of SMBP ranged from 4,311 to 4,619 kcal/kg as-is basis and the value of sample 3 was the least (p<0.05) among SMBP samples. Conclusion: Energy values and protein digestibility should be determined before using SMBP in swine diets.
Resumen Antecedentes: El subproducto de la leche de soja (SMBP) es un ingrediente alimenticio alternativo con uso potencial en dietas porcinas dado su alto contenido de proteína. Sin embargo, la información sobre sus valores energéticos y nutricionales para alimentación de cerdos es muy limitada. Objetivo: Estimar los valores de energía y la digestibilidad de la proteína del SMBP en cerdos con base en ensayos in vitro. Métodos: Se obtuvieron cuatro muestras de SMBP de tres empresas productoras de leche de soja. Se determinaron la desaparición de tracto total in vitro (IVTTD) y la desaparición ileal in vitro (IVID) de la materia seca (DM) en las muestras de SMBP. La desaparición ileal in vitro de proteína cruda se determinó analizando el contenido de proteína cruda en residuos no digeridos después de determinar la IVID de la DM. La energía digestible y metabolizable de SMBP se estimó utilizando la energía bruta, IVTTD de la DM y ecuaciones de predicción. Resultados: La muestra 4 tuvo una mayor IVTTD de la DM que la muestra 3 (97,7 vs. 94,4%, p<0,05), mientras que la IVID de la DM en la muestra 4 fue menor en comparación con la muestra 1 (53,5 vs. 65,0%, p<0,05). La desaparición ileal in vitro de la proteína cruda en la muestra 2 fue mayor que la de las muestras 1 y 3 (92,6 vs. 90,6 y 90,1%; p<0,05). La energía metabolizable estimada de SMBP varió de 4.311 a 4.619 kcal/kg (en base húmeda) y el valor de la muestra 3 fue el menor (p<0.05) entre las muestras de SMBP. Conclusión: Los valores de energía y la digestibilidad de la proteína deben determinarse antes de usar el SMBP en dietas porcinas.
Resumo Antecedentes: O subproduto do leite de soja (SMBP) é um potencial ingrediente alternativo na dieta de suínos, considerando seu alto teor de proteínas. No entanto, as informações sobre os valores energéticos e nutricionais do SMBP usado como ingrediente alimentar para suínos são limitadas. Objetivo: Estimar valores energéticos e digestibilidade protéica do SMBP em suínos com base em ensaios in vitro. Métodos: Foram obtidas quatro amostras de SMBP de três instalações produtores de leite de soja. Foram determinados o desaparecimento total do trato in vitro (IVTTD) e o desaparecimento ileal in vitro (IVID) da matéria seca (DM) nas amostras de SMBP. O desaparecimento ileal in vitro da proteína bruta foi determinado pela análise do conteúdo de proteína bruta em resíduos não digeridos após a determinação da IVID do DM. A energia digerível e metabolizável do SMBP foi estimada usando energia bruta, IVTTD do DM e equações de predição. Resultados: a amostra 4 apresentou maior IVTTD de DM do que a amostra 3 (97,7 vs. 94,4%, p<0,05) enquanto a IVID do DM na amostra 4 foi menor em comparação com a amostra 1 (53,5 vs. 65,0%, p<0,05). O desaparecimento ileal in vitro da proteína bruta na amostra 2 foi superior ao da amostra 1 e 3 (92,6 vs. 90,6 e 90,1%; p<0,05). A energia metabolizável estimada do SMBP variou de 4.311 a 4.619 kcal/kg no estado em que se encontra e o valor da amostra 3 foi o menor (p<0,05) entre as amostras do SMBP. Conclusão: os valores energéticos e a digestibilidade das proteínas devem ser determinados antes do uso do SMBP nas dietas suínas.
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Las afecciones causadas por el Virus de la Hepatitis B (VHB) constituyen una de las grandes problemáticas de salud pública a nivel mundial. En la actualidad la principal estrategia de prevención es una vacuna obtenida mediante la tecnología de ADN recombinante a partir de la expresión del gen viral que codifica el antígeno de superficie de la hepatitis B (HBsAg). En Argentina, esta vacuna está incorporada en el Calendario Nacional de Vacunación desde el año 2000, y es deber de la Autoridad Reguladora Nacional (ARN) verificar la calidad, seguridad y eficacia de cada lote liberado al mercado. Considerando que la determinación del contenido antigénico en vacunas representa uno de los ensayos esenciales de control de calidad y que, para ello, es necesario contar con metodologías estandarizadas que permitan obtener resultados reproducibles y confiables, en el Laboratorio de Inmunobiológicos del Instituto Nacional de Medicamentos se estandarizó un método para la identificación y cuantificación del HBsAg en vacunas contra la Hepatitis B empleando un kit comercial de ELISA cuyo uso clínico está previsto para la detección de HBsAg en suero o plasma humano. Para ello, se analizaron dos muestras: una preparación de antígeno HBsAg purificado y una vacuna contra la Hepatitis B ADN recombinante comercial y se evaluaron los parámetros de especificidad, exactitud, repetibilidad, precisión intermedia y linealidad. En todos los casos, se cumplieron con los criterios de aceptación establecidos para cada parámetro, por lo cual se concluye que la metodología analítica es adecuada para la identificación y cuantificación del HBsAg.