ABSTRACT
Introducción: La Troponina I (TnI) plasmática es el biomarcador "Gold" estándar utilizado en diagnóstico de Infarto Agudo al Miocardio (IAM), indicando necrosis cardíaca. Las microvesículas extracelulares (MVEC), participan en comunicación celular, por lo que estudiar su distribución entregaría información respecto del evento isquémico, antesala del infarto. Objetivo: Estudiar las MVECs plasmáticas en pacientes con Síndrome Coronario Agudo (SCA) y compararlas con los niveles de TnI. Métodos: Plasma de 22 pacientes controles se recolectó 0-2hrs post-ingreso a urgencia. Plasma de 45 pacientes SCA se recolectó 0-2, 6-8 y 10-14hrs post ingreso, junto con la toma de muestra para estudio de TnI. Las MVECs plasmáticas fueron enriquecidas mediante kit comercial. La determinación de la concentración y tamaño MVECs se realizó por NTA (Nanoparticles Tracking Assay) usando el equipo Nanosight. Resultados: La concentración promedio de MVECs 0-2 hrs post ingreso fue 7,2 veces superior en plasma de pacientes con SCA vs controles y la moda del tamaño disminuyó en pacientes con SCA. La TnI no mostró diferencias significativas en 0-2 hrs post ingreso en el grupo estudiado. La concentración de las MVEC disminuyó significativamente después de 10-14 hrs post ingreso, mientras que la concentración promedio TnI se mantuvo invariable demostrando el aumento de MVECs previo al incremento de TnI. Conclusión. El aumento de MVECs previo al incremento de la TnI en pacientes infartados, sugiere que las MVECs aumentan en la fase previa del IAM, como respuesta al daño tisular. Actualmente, estudiamos el contenido molecular de las MVECs, para establecer un método diagnóstico del Síndrome Coronario Agudo basado en MVECs.
Background: Troponin I (TnI) is the gold standard used to establish the diagnosis of myocardial infarction (AMI), indicating the presence of myocardial necrosis. Extracellular micro vesicles are involved in cellular communication. Their distribution may provide information relating to the development of AMI in patients with acute coronary syndromes (ACS) Aim: to study plasma levels of ECMV compared to those of TnI in patients with ACS. Methods: The plasma levels of TnI and ECMV from 22 control patients coming to the emergency units was compared to plasma from 45 patients with ACS. Levels of both parameters were determined 0-2, 6-8 and 10-14 hours post admission. ECMVs were enriched by means of a commercial kit. Concentration and size of ECMV was determined by NTA (Nanoparticles tracking assay) using the Nanosight equipment. Results: Plasma concentration of ECMV was 7.2 times higher than that of TnI 0-2 hrs post admission. The mode of ECMV size was lower in patients with ACS. Concentration of ECMV had decreased significantly 10-14 hrs post admission, whereas the TnI levees remained stable. Conclusion: The increase in ECMV earlier than TnI in AMI suggests that ECMV are elevated in the pre-AMI phase, as a response to early tissue damage. A study of cellular content of ECMV, being carried out, may lead to develop a method for the early diagnosis of AMI in patients with ACS.
Subject(s)
Humans , Male , Female , Adult , Middle Aged , Aged , Extracellular Vesicles/physiology , Myocardial Infarction/blood , Myocardial Infarction/metabolism , Troponin I/blood , Acute Coronary Syndrome/blood , Acute Coronary Syndrome/metabolism , Analysis of Variance , Biomarkers/blood , Cell Tracking/methods , Exosomes/physiology , NanoparticlesABSTRACT
PURPOSE: We aimed to investigate the effectiveness of ferritin as a contrast agent and a potential reporter gene for tracking tumor cells or macrophages in mouse cancer models. MATERIALS AND METHODS: Adenoviral human ferritin heavy chain (Ad-hFTH) was administrated to orthotopic glioma models and subcutaneous colon cancer mouse models using U87MG and HCT116 cells, respectively. Brain MR images were acquired before and daily for up to 6 days after the intracranial injection of Ad-hFTH. In the HCT116 tumor model, MR examinations were performed before and at 6, 24, and 48 h after intratumoral injection of Ad-hFTH, as well as before and every two days after intravenous injection of ferritin-labeled macrophages. The contrast effect of ferritin in vitro was measured by MR imaging of cell pellets. MRI examinations using a 7T MR scanner comprised a T1-weighted (T1w) spin-echo sequence, T2-weighted (T2w) relaxation enhancement sequence, and T2*-weighted (T2*w) fast low angle shot sequence. RESULTS: Cell pellet imaging of Ad-hFTH in vitro showed a strong negatively enhanced contrast in T2w and T2*w images, presenting with darker signal intensity in high concentrations of Fe. T2w images of glioma and subcutaneous HCT116 tumor models showed a dark signal intensity around or within the Ad-hFTH tumor, which was distinct with time and apparent in T2*w images. After injection of ferritin-labeled macrophages, negative contrast enhancement was identified within the tumor. CONCLUSION: Ferritin could be a good candidate as an endogenous MR contrast agent and a potential reporter gene that is capable of maintaining cell labeling stability and cellular safety.
Subject(s)
Animals , Female , Humans , Male , Mice , Brain Neoplasms/diagnostic imaging , Cell Line, Tumor , Cell Tracking/methods , Colonic Neoplasms/diagnostic imaging , Contrast Media/administration & dosage , Disease Models, Animal , Ferritins/administration & dosage , Genes, Reporter , Glioma/diagnostic imaging , Injections, Intravenous , Macrophages , Magnetic Resonance Imaging/methods , Neoplasm Transplantation , Skin Neoplasms/diagnostic imaging , Time FactorsABSTRACT
En los últimos años ha surgido un gran interés en conocer la biodistribución de las células madre en el organismo después que son infundidas o inyectadas directamente en una parte del cuerpo. Para esto se han usado diferentes procederes, entre ellos, el marcaje de las células con diferentes fluorocromos, tales como la proteína con fluorescencia verde y la proteína con fluorescencia roja; o bien se les hace una transfección con plasmidos que codifican proteínas fluorescentes o se emplean sondas moleculares fluorescentes para la identificación de cromosomas. Recientemente se han introducido técnicas imagenológicas de avanzada no invasivas, entre las que tenemos la resonancia magnético nuclear, así como procederes basados en el marcaje con radionúclidos para la obtención de imágenes detectadas por tomografía por emisión de positrones (PET, del inglés positron emission tomography), o por tomografía computarizada por emisión de fotón único (SPECT, del inglés single- photon emission computed tomography).
In past years there was an increasing interest by to know about the biodistribution of stem cells in organism after its perfusion or direct injection in a part of the body. Thus, we used different procedures including the cell labeling with distinct fluorochromes, such as the green fluorescence protein and the red fluorescence protein or a plasmid transfection codifying the fluorescent proteins of fluorescent molecular stents to identify the chromosomes. Recently non-invasive leading imaging techniques have been introduced including nuclear magnetic resonance. As well as procedures based on radionuclide labeling to obtain the positron emission tomography (PET) images or by photon-emission computed tomography (SPECT).