ABSTRACT
Objetivo: Desarrollar una nueva técnica para la transferencia de cianometacrilato de muy baja densidad sobre las superficies internas y externas del corazón. Material y métodos: Se combinó el cianometacrilato curado por luz ultravioleta de muy baja densidad con colorante comercial para demostrar su capacidad de transferencia sobre las superficies externas e internas de un modelo de corazón in vitro. Se mezclaron 0,5 ml de cianometacrilato con 0,2 ml tinta, y el material se inyectó sobre la superficie del corazón en aire seco, lo que permite su fijación durante 2-3 seg. A continuación, toda la preparación se sumergió en solución salina y se agitó vigorosamente para eliminar el compuesto no ligado. Se realizó un experimento similar sin cianometacrilato y con cianometacrilato de alta viscosidad (3000 cps). Se comprobó visualmente después del lavado que una cantidad significativa del compuesto se liga a las superficies del corazón en comparación con la tinta sola. Luego, se investigó la transferencia efectiva del compuesto a las superficies internas y externas del corazón después de haber sumergido el tejido en solución salina. Entre diversas técnicas, la de pincelado fue la más efectiva para la transferencia orientada del compuesto. Mediante esta técnica fue muy fácil transferir el compuesto sobre las superficies endocárdica y epicárdica. También otras áreas específicas, como la orejuela auricular izquierda, las superficies internas del ventrículo izquierdo en el origen del músculo papilar y el ápex ventricular izquierdo, fueron investigadas exitosamente. La fuente de luz ultravioleta fue un dispositivo en forma de lapicera basado en un diodo emisor de luz. Luego del tratamiento con luz ultravioleta, se observó cierta precipitación del compuesto en algunas zonas. El experimento se repitió con tres muestras para determinar el resultado. Conclusiones: La transferencia de cianometacrilato de muy baja viscosidad curado por luz ultravioleta es potencialmente útil para el estudio de las superficies internas y externas del corazón. Esta técnica podría servir para la transferencia de factor de crecimiento.
Background: Cyan methacrylate is a compound with remarkable adhesion properties used in different specific areas, including medicine, where it can be applied in some entities and is also subject of research in the field of cardiology Objective: The aim of this study was to develop a novel technique for the transfer of very-low viscosity cyan methacrylate on the inner and outer surfaces of the heart. Methods: Very-low viscosity ultraviolet curable cyan methacrylate was mixed with commercially available dye to demonstrate its ability of transfer on the inner and outer surfaces of an in-vitro heart model. Cyan methacrylate (0.5 ml) was mixed with 0.2 ml of ink, and the material was injected on the surface of the heart in dry air, allowing its fixation for 2-3 seconds. Subsequently, the whole preparation was immersed in saline, and was vigorously shaken to remove the unbound compound. A similar experi-ment was performed without cyan methacrylate, and with high viscosity cyan methacrylate (3000 cps). A significant amount of the compound was visually found to attach to the surfaces of the heart compared with ink alone. Then, after soaking the tissue in saline solution, effective transfer of the compound was assessed on the inner and the outer surfaces of the heart. Results: Among various techniques, the paintbrush technique was the most effective one for the targeted transfer of the com-pound. With this specific areas, such as instead of specific areas, as technique, it was easier to transfer the compound on the epicardial and endocardial surfaces. Also, other specific areas, as the left atrial appendage, the left ventricular inner surfaces at the origin of the papillary muscle and the left ventricular apex were successfully investigated. The ultraviolet light source was a pen-shaped device based on a light emission diode. Some compound precipitation was observed in some areas following ultraviolet treatment. The experiment was repeated with three different samples to determine the results. Conclusions: Very-low viscosity ultraviolet curable cyan methacrylate transfer is potentially useful to study the inner and outer surfaces of the heart. This technique could be useful for growth factor transfer.
ABSTRACT
Objective To study the radiation sensitive enhancement ratio (SER) of NS398 on esophageal cancer stem cells and adherent tumor cells and analyze the radioresistance related protein expressions.Methods ECA109 esophageal cancer stem cells were cultured in serum-free medium.Expression levels of cell surface maker CD44 + and CD271 + were analyzed by flow cytometry.MTT assay was used to detect cell proliferation after the treatments with NS398 and irradiation(0,4 and 8 Gy).The sensitization effects of NS398 on the parental cells and its spheroid were evaluated by clone formation assay.Western blot assay was performed to determine protein expressions.Results Serum-free medium was successfully applied to isolate the cancer stem cells with spherical properties.CD271 + in the spheroid cells was notable higher than that in the parent cells (t =3.81,P ＜ 0.05).After irradiation,the proliferation rate of parental cells was higher than that in spheroid cells.After the combination treatment of NS398 and irradiation,SF2 value of parental cells was lower than spheroid cells(t =2.91,P ＜ 0.05)and the SER of NS398 on parental cells was greater than spheroid cells.The expressions of Bmi-1,c-Myc,β-catenin and Cyclin D1 in spheroid cells were higher than those in parental cells (t =8.09,7.90,7.50,7.15,P＜0.05).Cyclin D1 expression levels under both cell situations increased after 4 Gy irradiation (t =9.74,6.67,P ＜0.05).Compared to the 4 Gy irradiation alone group,the β-catenin and Cyclin D1 expression levels in both parental cells (t =10.15,12.12,P ＜ 0.05) and spheroid cells (t =3.23,7.45,P ＜ 0.05) decreased in the combination group.Conclusions Esophageal cancer stem cells with high level of CD271 can be isolated with serum-free medium and it is radioresistant where β-catenin and its downstream proteins may be involved.