In Vitro Cytotoxicity Evaluation of Three Root-End Filling Materials in Human Periodontal Ligament Fibroblasts

Abstract The aim of this study was to evaluate in vitro the cytotoxicity on human periodontal ligament fibroblasts of three root-end filling materials: MTA Angelus(r), EndoSequence Root Repair Material Putty(r) and Super EBA(r). A primary culture of human periodontal ligament fibroblasts was previously obtained in order to evaluate the cytotoxicity of the three extracts from the root-end filling materials after 2 and 7 days of setting. Serial dilutions of these extracts (1:1, 1:2, 1:4 and 1:8) were evaluated at 1, 3 and 7 days using the methyl-thiazol-tetrazolium (MTT) colorimetric assay. Cell viability was evaluated as percentage of the negative control group, which represented 100% cell viability. Statistical analyses were done with t-test, ANOVA and Kruskal-Wallis test at a significance level of 5%. It was found that the main difference among root-end filling materials was in the higher dilutions (p<0.05), but there was a similar behavior in lower dilutions (p>0.05). Cell viability of MTA Angelus(r) was superior for 2-day setting (p<0.05), compared with the other two root-end fillings. There were no statistically significant differences between 7-day set MTA Angelus(r) and EndoSequence Root Repair Material Putty(r). Super EBA(r) showed the lowest percentage of cell viability at higher dilutions (p<0.05). Therefore, MTA Angelus(r) and EndoSequence Root Repair Material Putty(r) were less cytotoxic in the highest dilution (1:1) compared with Super EBA(r).

Resumo O objetivo deste trabalho foi avaliar in vitro a citotoxicidade em fibroblastos do ligamento periodontal humano de três cimentos de retrobturação: MTA Angelus(r), EndoSequence Root Repair Material Putty(r) e Super EBA(r). Uma cultura de fibroblastos primários do ligamento periodontal humano foi obtida anteriormente a fim de avaliar a citotoxicidade dos três extratos dos cimentos de retrobturação após 2 e 7 dias de endurecimento. As diluições em série destes extratos (1:1, 1:2, 1:4 e 1:8) foram avaliados em 1, 3 e 7 dias empregando o ensaio colorimétrico metil-tiazol-tetrazólio (MTT). A viabilidade celular foi calculada em base da porcentagem do grupo de controle negativo, que representou 100% de viabilidade de células. As análises estatísticas foram realizadas com o teste t, ANOVA e teste de Kruskal-Wallis a um nível de significância de 5%. Verificou-se que a principal diferença entre os cimentos de retrobturação estava nas diluições mais elevadas (p<0,05) e houve um comportamento semelhante nas diluições mais baixas (p>0,05). A viabilidade celular dos fibroblastos do ligamento periodontal humano foi superior para MTA Angelus(r) de 2 dias de endurecimento (p<0,05), em comparação com os outros materiais de retrobturação. Não houve diferença significante entre MTA Angelus(r) e EndoSequence Root Repair Material Putty(r) de 7 dias de endurecimento. Super EBA(r) mostrou a menor percentagem da viabilidade celular nas diluições mais altas (p<0,05). Portanto, os cimentos de retrobturação MTA Angelus(r) e EndoSequence Root Repair Material Putty(r) foram menos citotóxicos na diluição mais alta (1:1) em comparação com Super EBA(r).

Actividad citotóxica de physalis peruviana (aguaymanto) en cultivos celulares de adenocarcinoma colorectal, próstata y leucemia mieloide crónica / Cytotoxic effect of physalis peruviana in cell culture of colorectal and prostate cancer and chronic myeloid leukemia

INTRODUCCIÓN: Las plantas han sido empleadas como drogas por siglos. Sin embargo, se ha investigado poco sobre su gran potencial como fuentes de nuevos agentes terapéuticos. El objetivo del presente trabajo fue evaluar la actividad citotóxica de los extractos etanólicos de tallos y hojas de Physalis peruviana, sobre las líneas celulares HT-29, PC-3, K-562 y VERO. MATERIALES Y MÉTODOS: Las líneas celulares HT-29, PC-3, K-562 y VERO, fueron expuestas a cuatro concentraciones de extractos etanólicos de tallos y hojas de Physalis peruviana. Asimismo, a diferentes concentraciones de cisplatino y 5-Fluorouracilo (5-FU), que se usaron como controles positivos. Se hallaron los porcentajes de crecimiento en 48 horas, luego se determinó la concentración inhibitoria 50 (IC50) mediante análisis de regresión lineal y el índice de selectividad de cada muestra. RESULTADOS: Los extractos etanólicos de tallos y hojas de Physalis peruviana mostraron actividad citotóxica: Los CI50 en μg/mL en hojas y tallos fueron, 0.35 (r=-0.95 p<0.025) y 0.37 (r=-0,90 p<0.05) para HT-29; 0.87 (r=-0.98 p<0.01) y 1.01 (r=-0.95 p<0.025) para PC-3; 0.02 (r=-0.98 p<0.01) y 0.03 (r=-0.98 p<0.01) para K-562; 4.9 (r=-0.95 p<0.025) y 6.2 (r=-0.98 p<0.01) para VERO. Los CI50 para los antineoplásicos fueron: para el cisplatino: 4.2 (r=-0.96 p<0.025), 10.3 (r=-0.97 p<0.025), 0.15 (r=-0.98 p=0.01), y 1.1 (r=-0.98 p=0.01). Parael 5-FU: 2.3 (r=-0.97 p<0.025), 17.9 (r=-0.95 p<0.025), 0.15 (r=-0.98 p=0.01), y 1.1 (r=-0.94 p=0.05) para HT-29, PC-3, K562 y VERO, respectivamente. Los índices de selectividad de los extractos de tallos y hojas, estuvieron entre 5.6 y 245 para las líneas celulares tumorales evaluadas; por el contrario, el cisplatino y el 5-FU, solo alcanzaron valores entre 0.11 y 7.3...

INTRODUCTION: The plants have been used as drugs for centuries. However, limited research has been done on its great potential as sources of new therapeutic agents. The purpose of this study was to evaluate Physalis peruviana cytotoxic activity on cell lines HT-29, PC-3, K-562 and VERO. Materials and Methods: The HT-29 cell lines, PC-3, K-562 and VERO, were exposed to four concentrations of P. peruviana ethanolic leave and stem extracts, also at different concentrations of cisplatin and 5-fluorouracil (5-FU), which were used as positive controls. We found rates of growth within 48 hours, then we determined the inhibitory concentration 50 (IC50) using linear regression analysis and the index of selectivity of each sample. RESULTS: The P. peruviana ethanolic leave and stem extracts showed cytotoxic activity. The IC50 in μg/mL in leaves and stems were, 0.35 (r =-0.95 p <0.025) and 0.37 (r =- 0.90 p <0.05 ) for HT-29; 0.87 (r =-0.98 p <0.01) and 1.01 (r =-0.95 p <0.025) for PC-3; 0.02 (r =-0.98p <0.01) and 0.03 (r =-0.98 p <0.01) for K-562; 4.9 (r =-0.95 p <0.025) and 6.2 (r =-0.98 p <0.01) for VERO. The IC50 for antineoplastic were: for cisplatin: 4.2 (r =-0.96 p <0.025),10.3 (r =-0.97 p <0.025), 0.15 (r =-0.98 p = 0.01) and 1.1 (r =- 0.98 p = 0.01); for 5-FU: 2.3 (r =-0.97 p <0.025), 17.9 (r =-0.95 p <0.025), 0.15 (r =-0.98 p = 0.01) and 1.1 (r =-0.94 p = 0.05) for HT-29, PC-3, K562 and VERO respectively. The leaves and stems extracts selectivity index were between 5.6 and 245 for tumor cell lines evaluated, by contrast, cisplatin and 5-FU, only showed values between 0.11 and 7.3. CONCLUSIONS: The P. peruviana leaves and steams ethanolic extracts were more cytotoxic than cisplatin and 5 FU, on the lines HT-29, PC-3 and K562. Furthermore the P. peruviana cytotoxic effects were less than cisplatin and 5-FU for VERO control cells lines.

Actividad citotóxica del extracto etanólico de gnaphalium spicatum keto keto en cultivos de líneas celulares tumorales humanas / Cytotoxic activity of an ethanolic extract of gnaphalium spicatum keto keto on human tumor cell lines

Objetivos. Evaluar la actividad citotóxica de extractos etanólicos de raíces, tallos, hojas y flores de Gnaphalium spicatum sobre algunas líneas celulares tumorales humanas. Materiales y métodos. Las líneas celulares HT-29, H-460, MCF-7, M-14, PC-3, DU-145, K-562, y 3T3, fueron expuestas a cuatro concentraciones de extractos etanólicos de raíces, tallos, hojas y flores de Gnaphalim spicatum, asimismo, a diferentes concentraciones de cisplatino, que se usó como control positivo. Se halló los porcentajes de crecimiento en 48 horas. Luego se determinó la concentración inhibitoria 50 (CI50) mediante análisis de regresión lineal, el índice de selectividad de cada muestra y finalmente, la relación dosis-respuesta entre las concentraciones de los extractos y cisplatino, con los porcentajes de crecimiento. Resultados. El extracto etanólico de las raíces de Gnaphalium spicatum mostró mayor actividad citotóxica en la líneas celulares MCF-7 yK-562. Los CI50 en ¦Ìg/mL fueron de 98 (r= -0,98 p menor que 0,01) y 46 (r= -0,97 p menor que 0,01), respectivamente. Asimismo, su citotoxicidad en la l¨ªnea celular 3T3 fue de 215 (r= 0,97 p menor que 0,01). Los CI50 del cisplatino en ¦Ìg/mL fueron de 2 (r=-0,96 p menor que 0,01), 7,7 (r=-0,98 p menor que 0,01) y 3 (r=-0,97p menor que 0,01), para MCF-7, K-562 y 3T3, respectivamente. Los índices de selectividad del extracto de raíces y cisplatino fueron 2,2 y 0,3 para MCF-7, y 4,7 y 1,5 para K-562, respectivamente. Los extractos de tallos, hojas y flores obtuvieron CI50 mayor que a 0,250 mg/mL en todas la líneas celulares evaluadas. Conclusiones. Los extractos etanólicos de tallos, hojas y flores de Gnaphalium spicatum no mostraron actividadcitotóxica en este bioensayo. Los extractos de raíces mostraron citotoxicidad en las todas las líneas celulares tumorales, excepto en M-14. Además fueron menos citotóxicos en relación al cisplatino en la línea celular 3T3.

Objectives. To evaluate the cytotoxic activity ethanolic extracts of roots, stems, leaves and flowers of Gnaphalium spicatum in somehuman tumor cell lines. Material and methods. The following cell lines: HT-29, H-460, MCF-7, M-14, PC-3, DU-145, K-562, and 3T3, were exposed to four different concentrations of ethanolic extracts from roots, stems, leaves and flowers of Gnaphalium spicatum, and also to different concentrations of cisplatin, which was used as a positive control. Percentage growth was assessed after 48 hours. The minimalinhibitory concentration for 50 per cent of the cells (IC50) was determined using linear regression analysis; also, the selectivity index for each sample and the dose-response relationship between the extract concentrations and cisplatin, as well as growth percentages were determined. Results. The ethanolic extract of Gnaphalium spicatum roots showed its highest cytotoxic activity in MCF-7 and K-562 cell lines. IC50 values, expressed in ¦Ìg/mL, were 98 (r=-0.98; p <0.01) and 46 (r= -0.97; p minor that 0.01), respectively. Cytotoxicity against the 3T3 cellline was 215 (r= 0.97; p minor that 0.01). IC50 values for cisplatin were 2 (r= -0.96 p minor that 0.01), 7.7 (r= -0.98; p minor that 0.01), and 3 (r= -0.97; p minor that 0.01), for theMCF7, K562 and 3T3 cell lines, respectively. The selectivity index of the ethanolic extract from Gnaphalium spicatum roots and cisplatinwere 2.2 and 0.03 for the MCF-7 cell line, and 4,7 and 1.5 for the K-562 cell line, respectively. Extracts from stems, leaves and flowers of Gnapahlium spicatum acheived IC50 levels major that 0.250 mg/mL in every cell lines tested. Conclusions. The ethanolic extracts of stems, leaves and flowers of Gnaphalium spicatum did not show cytotoxic activity in this bioassay. The extract from roots showed cytotoxicity in all tumor cell lines, except in M-14. Furthermore, it was less cytotoxic than cisplatin in 3T3 cell line.

Double-blind randomized placebo-controlled study of bixa orellana in patients with lower urinary tract symptoms associated to benign prostatic hyperplasia

OBJECTIVE: To determine the efficacy of Bixa Orellana (BO) in patients with benign prostatic hyperplasia (BPH) presenting moderate lower urinary tract symptoms (LUTS). MATERIALS AND METHODS: It is a prospective double-blind randomized placebo-controlled study. One thousand four hundred and seventy eight patients presenting moderate LUTS associated to BPH were interviewed, from whom we selected 136 to fulfill the criteria of inclusion and exclusion. Assignation was performed at random in blocks of four to receive B0 at a dose of 250 mg 3 times a day or placebo (Pbo) for 12 months, 68 patients were assigned to each group. From the patients in the study we obtained data of demographic, epidemiologic, symptom score, uroflowmetry and post void residual urine variables. RESULTS: Basically both groups were compared clinically, demographically and biochemically. Throughout the study variations of symptom score, mean delta symptom score during each visit and the final average delta were similar for both groups (BO - 0.79 ± 1.87 and Pbo - 1.07 ± 1.49) (p = 0.33). Similarly variations of Qmax mean, Qmax average delta and final average delta were similar (BO 0.44 ± 1.07 and Pbo 0.47 ± 1.32) (p = 0.88). Variations of post void residual urine mean, post void residual urine average delta in each visit and the final average delta were similar for both groups (BO 4.24 ± 11.69 and Pbo 9.01 ± 18.66) (p = 0.07). No differences were found in the answers of clinically significant improvement assessed with relative risk and risk differences, even though the proportion of adverse effects was similar for both groups. CONCLUSION: Patients with BPH that present moderate LUTS did not show any benefit receiving BO when compared to placebo.

Actividad cicatrizante del látex de Jatropha curcas (Angiospermae: Euforbiaceae) / Cicatrization effect of Jatropha curcas latex (Angiospermae: Euforbiaceae)

The wound healing effects of Jathopha curcas latex upon surgical wound produced in Balb/c mice skin, were studied with a modification of the Hoowes-Sooy-Harvey method. The effects of topical treatment using single 50 ul doses of latex at different dilutions (10 per cent to 100 per cent) was compared with a multiple dose treatment (four 25 ul/dose q12h, latex 5 per cent to 100 per cent). The single dose treatment with 10 per cent, 50 per cent or 100 per cent latex and the multiple dose treatment with dilutions between 5 per cent and 10 per cent, have a healing effect but only on males. The multiple dose treatment with 50 per cent or pure undiluted latex produced caustic lesions to treated skin

Citotoxicidad inducida por veneno de serpientes peruanas sobre fibroblastos de ratón / Cytotoxity induced by Peruvian snake venom on fibroblasts of mice

The cytotoxic effect of venoms from six crotalinae Peruvian snakes (Bathrops atrox; B. brazili; B. pictus; B. barnetti; Lachesis m. muta y Crotalus durissus terrificus) was studied in an in vitro system of BALB/c 3T3 fibroblasts grown in Dulbecco modified minimal essential medium at 37-C in a humidified atmosphere of 5% CO2 - 95% air. The viability of the cells was evaluated 24 hours after the treatment with the different venoms, using the method of exclusion of trypan blue. The six venoms produced cytotoxic effects at 24 hours on the 3T3 fibroblasts. The venom from B atrox was the most potent (DE50=162 ng/ml) and that from B. barnetti the least (DE50=7182ng/ml)

Evolución de la inmunidad pasiva y respuesta a la vacunación contra el sarampión / Follow up of the pasive immunity and response to antimesles vaccination

Treinta y cuatro niños recién nacidos fueron seguidos hasta los 10 meses de edad. Evaluaciones clínicas y antropométricas se llevaron a cabo al nacimiento y a los 3,6 y 9 meses. Una pequeña muestra de sangre fue obtenida durante estas evaluaciones con la finalidad de tederminar el título de las inmunoglobulinas G (igG) anti-sarampión usando la técnica de ELISA modificada en nuestro laboratorio. A los 9 meses de edad todos los niños fueron vacunados contra contra sarampión y el título de igG anti-sarampión fue determinado nuevamente a los 10 meses de edad. El estado nutricional de acuerdo a las medidas antropométricas (usando los standards de crecimiento del NCHS) estuvieron generalmente dentro del rango normal para todos los niños durante el estudio. Treinta y tres niños seroconvirtieron despúes de la vacunación contra el sarampión y un niño presentó valores de seroconversión dudosos. La relación Hijo/Madre de igG antisarampión al nacimiento fue de 1.40 para todos los niños, siendo 1.70 para los varones y 1.15 para las mujeres. Se encontró que la pérdida de anticuerpos anti-sarampión fue en función de los títulos iniciales (al nacimiento), de modo que a la edad de 9 meses, los tres grupos de niños (con títulos inicales bajo, intermedio y alto) presentaron títulos promedio idénticos. Se propone la hipótesis de que el estado nutricional o el incremento en la frecuencia de la exposición de los niños al agente infecciosossería el responsable de la pérdida temprana de los anticuerpos anti-sarampión provenientes de la madre