RESUMEN
Casiopeinas are a group of copper-based antineoplastic molecules designed as a less toxic and more therapeutic alternative to cisplatin or Doxorubicin; however, there is scarce evidence about their toxic effects on the whole heart and cardiomyocytes. Given this, rat hearts were perfused with Casiopeinas or Doxorubicin and the effects on mechanical performance, energetics, and mitochondrial function were measured. As well, the effects of Casiopeinas-triggered cell death were explored in isolated cardiomyocytes. Casiopeinas III-Ea, II-gly, and III-ia induced a progressive and sustained inhibition of heart contractile function that was dose- and time-dependent with an IC50 of 1.3 ± 0.2, 5.5 ± 0.5, and 10 ± 0.7 µM, correspondingly. Myocardial oxygen consumption was not modified at their respective IC50, although ATP levels were significantly reduced, indicating energy impairment. Isolated mitochondria from Casiopeinas-treated hearts showed a significant loss of membrane potential and reduction of mitochondrial Ca2+ retention capacity. Interestingly, Cyclosporine A inhibited Casiopeinas-induced mitochondrial Ca2+ release, which suggests the involvement of the mitochondrial permeability transition pore opening. In addition, Casiopeinas reduced the viability of cardiomyocytes and stimulated the activation of caspases 3, 7, and 9, demonstrating a cell death mitochondrial-dependent mechanism. Finally, the early perfusion of Cyclosporine A in isolated hearts decreased Casiopeinas-induced dysfunction with reduction of their toxic effect. Our results suggest that heart cardiotoxicity of Casiopeinas is similar to that of Doxorubicin, involving heart mitochondrial dysfunction, loss of membrane potential, changes in energetic metabolites, and apoptosis triggered by mitochondrial permeability.
Asunto(s)
Antineoplásicos/efectos adversos , Cardiotoxicidad/etiología , Mitocondrias Cardíacas/efectos de los fármacos , Compuestos Organometálicos/efectos adversos , Animales , Antineoplásicos/administración & dosificación , Apoptosis/efectos de los fármacos , Cardiotoxicidad/metabolismo , Cardiotoxicidad/patología , Complejos de Coordinación/efectos adversos , Complejos de Coordinación/química , Cobre/efectos adversos , Cobre/química , Masculino , Mitocondrias Cardíacas/metabolismo , Compuestos Organometálicos/administración & dosificación , Ratas , Ratas WistarRESUMEN
Surface modification in nanostructured mesoporous silica particles (MSNs) can significantly increase the uptake in myocardial cells. Herein, MSNs particles were synthesized and chemically functionalized to further assess their biocompatibility in rat myocardial cell line H9c2. The surface modification resulted in particles with an enhanced cellular internallization (3-fold increase) with respect to pristine particles. Apoptosis events were not evident at all, while necrosis incidence was significant only at a higher doses (>500µg/mL). In particular, the percentage of necrotic cells decrease in a statistically significant manner for the functionalized particles at lower doses than 100µg/mL. This study concludes that the proposed surface functionalization of MSNs particles does not compromise their viability on H9c2 cells, and therefore they could potentially be used for biomedical purposes. Fourier-transform infrared, Raman, TGA/DSC, N2 adsorption-desorption, and TEM techniques were used to characterize the as-prepared materials. Confocal microscopy and flow cytometry analyses were carried out to measure the histograms of cell complexity and the half maximal inhibitory concentration, respectively. Reactive oxygen species generation was accessed using assays with MitoSOX and Amplex Red fluoroprobes.
Asunto(s)
Miocitos Cardíacos , Adsorción , Animales , Nanopartículas , Nanoestructuras , Ratas , Dióxido de SilicioRESUMEN
Given the well-known physical properties of graphene oxide (GO), numerous applications for this novel nanomaterial have been recently envisioned to improve the performance of biomedical devices. However, the toxicological assessment of GO, which strongly depends on the used material and the studied cell line, is a fundamental task that needs to be performed prior to its use in biomedical applications. Therefore, the toxicological characterization of GO is still ongoing. This study contributes to this, aiming to synthesize and characterize GO particles and thus investigate their toxic effects in myocardial cells. Herein, GO particles were produced from graphite using the Tour method and subsequent mild reduction was carried out to obtain low-reduced GO (LRGO) particles. A qualitative analysis of the viability, cellular uptake, and internalization of particles was carried out using GO (~54% content of oxygen) and LRGO (~37% content of oxygen) and graphite. GO and LRGO reduce the viability of cardiac cells at IC50 of 652.1±1.2 and 129.4±1.2µg/mL, respectively. This shows that LRGO particles produce a five-fold increase in cytotoxicity when compared to GO. The cell uptake pattern of GO and LRGO particles demonstrated that cardiac cells retain a similar complexity to control cells. Morphological alterations examined with electron microscopy showed that internalization by GO and LRGO-treated cells (100µg/mL) occurred affecting the cell structure. These results suggest that the viability of H9c2 cells can be associated with the surface chemistry of GO and LRGO, as defined by the amount of oxygen functionalities, the number of graphitic domains, and the size of particles. High angle annular dark-field scanning transmission electron microscopy, dynamic light-scattering, Fourier-transform infrared, Raman, and X-ray photoelectron spectroscopies were used to characterize the as-prepared materials.
Asunto(s)
Endocitosis/efectos de los fármacos , Grafito/toxicidad , Miocitos Cardíacos/citología , Nanoestructuras/toxicidad , Animales , Muerte Celular/efectos de los fármacos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Citometría de Flujo , Concentración 50 Inhibidora , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/ultraestructura , Nanoestructuras/ultraestructura , Oxidación-Reducción , Espectroscopía de Fotoelectrones , Ratas , Espectroscopía Infrarroja por Transformada de Fourier , Espectrometría RamanRESUMEN
AIMS: Ongoing inflammation and endothelial dysfunction occurs within the local microenvironment of heart failure, creating an appropriate scenario for successful use and delivery of nanovectors. This study sought to investigate whether cardiovascular cells associate, internalize, and traffic a nanoplatform called mesoporous silicon vector (MSV), and determine its intravenous accumulation in cardiac tissue in a murine model of heart failure. METHODS AND RESULTS: In vitro cellular uptake and intracellular trafficking of MSVs was examined by scanning electron microscopy, confocal microscopy, time-lapse microscopy, and flow cytometry in cardiac myocytes, fibroblasts, smooth muscle cells, and endothelial cells. The MSVs were internalized within the first hours, and trafficked to perinuclear regions in all the cell lines. Cytotoxicity was investigated by annexin V and cell cycle assays. No significant evidence of toxicity was found. In vivo intravenous cardiac accumulation of MSVs was examined by high content fluorescence and confocal microscopy, with results showing increased accumulation of particles in failing hearts compared with normal hearts. Similar to observations in vitro, MSVs were able to associate, internalize, and traffic to the perinuclear region of cardiomyocytes in vivo. CONCLUSIONS: Results show that MSVs associate, internalize, and traffic in cardiovascular cells without any significant toxicity. Furthermore, MSVs accumulate in failing myocardium after intravenous administration, reaching intracellular regions of the cardiomyocytes. These findings represent a novel avenue to develop nanotechnology-based therapeutics and diagnostics in heart failure.
Asunto(s)
Insuficiencia Cardíaca/diagnóstico , Insuficiencia Cardíaca/terapia , Corazón/fisiología , Corazón/fisiopatología , Miocitos Cardíacos/fisiología , Nanoestructuras/uso terapéutico , Animales , Materiales Biocompatibles , Modelos Animales de Enfermedad , Insuficiencia Cardíaca/fisiopatología , Humanos , Inyecciones Intravenosas , Masculino , Ratones , Ratones Endogámicos C57BL , Miocardio , Polímeros , SilicioRESUMEN
Stress-induced cardiomyopathy, triggered by acute catecholamine discharge, is a syndrome characterized by transient, apical ballooning linked to acute heart failure and ventricular arrhythmias. Rats receiving an acute isoproterenol (ISO) overdose (OV) suffer cardiac apex ischemia-reperfusion damage and arrhythmia, and then undergo cardiac remodeling and dysfunction. Nevertheless, the subcellular mechanisms underlying cardiac dysfunction after acute damage subsides are not thoroughly understood. To address this question, Wistar rats received a single ISO injection (67 mg/kg). We found in vivo moderate systolic and diastolic dysfunction at 2 wk post-ISO-OV; however, systolic dysfunction recovered after 4 wk, while diastolic dysfunction worsened. At 2 wk post-ISO-OV, cardiac function was assessed ex vivo, while mitochondrial oxidative metabolism and stress were assessed in vitro, and Ca(2+) handling in ventricular myocytes. These were complemented with sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA), phospholamban (PLB), and RyR2 expression studies. Ex vivo, basal mechanical performance index (MPI) and oxygen consumption rate (MVO2) were unchanged. Nevertheless, upon increase of metabolic demand, by ß-adrenergic stimulation (1-100 nM ISO), the MPI versus MVO2 relation decreased and shifted to the right, suggesting MPI and mitochondrial energy production uncoupling. Mitochondria showed decreased oxidative metabolism, membrane fragility, and enhanced oxidative stress. Myocytes presented systolic and diastolic Ca(2+) mishandling, and blunted response to ISO (100 nM), and all these without apparent changes in SERCA, PLB, or RyR2 expression. We suggest that post-ISO-OV mitochondrial dysfunction may underlie decreased cardiac contractility, mainly by depletion of ATP needed for myofilaments and Ca(2+) transport by SERCA, while exacerbated oxidative stress may enhance diastolic RyR2 activity.
Asunto(s)
Señalización del Calcio , Cardiomiopatías/metabolismo , Daño por Reperfusión Miocárdica/metabolismo , Estrés Oxidativo , Agonistas Adrenérgicos/toxicidad , Animales , Proteínas de Unión al Calcio/genética , Proteínas de Unión al Calcio/metabolismo , Cardiomiopatías/etiología , Cardiomiopatías/fisiopatología , Células Cultivadas , Ventrículos Cardíacos/citología , Ventrículos Cardíacos/metabolismo , Ventrículos Cardíacos/fisiopatología , Isoproterenol/toxicidad , Ratones , Mitocondrias Cardíacas/metabolismo , Contracción Miocárdica , Daño por Reperfusión Miocárdica/fisiopatología , Miocitos Cardíacos/efectos de los fármacos , Miocitos Cardíacos/metabolismo , Consumo de Oxígeno , Ratas , Ratas Wistar , Canal Liberador de Calcio Receptor de Rianodina/genética , Canal Liberador de Calcio Receptor de Rianodina/metabolismo , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico/genética , ATPasas Transportadoras de Calcio del Retículo Sarcoplásmico/metabolismoRESUMEN
BACKGROUND: It is generally accepted that an excessive production of reactive oxygen species plays an important role in acute renal failure secondary to ischemia and reperfusion. tert-Butylhydroquinone (tBHQ) is a well-known antioxidant. In this study, we evaluated whether tBHQ pretreatment prevented renal damage induced by ischemia and reperfusion (I/R). METHODS: Four groups of rats were studied: (a) control-sham (CT), (b) tBHQ-sham (tBHQ), (c) I/R and (d) tBHQ + I/R. Intraperitoneal (i.p.) injections of tBHQ (50 mg/kg) were given to the tBHQ and tBHQ + I/R groups and 3% ethanol/isotonic saline solution to the CT and I/R groups. Animals were killed 24 hours after I/R. RESULTS: tBHQ attenuated I/R-induced renal dysfunction, structural damage, oxidative/nitrosative stress, glutathione depletion and the decrease in several antioxidant enzymes. CONCLUSION: The renoprotective effect of tBHQ on I/R injury was associated with the attenuation in oxidative/nitrosative stress and the preservation of antioxidant enzymes.