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Evaluación de la citotoxicidad y genotoxicidad en células V79 y HaCaT asociado a la exposición a nanopartículas de carbón de La Loma Cesar

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datacite.rightshttp://purl.org/coar/access_right/c_16ec
dc.contributor.advisorLeón Mejía, Grethel
dc.contributor.advisorAcosta Hoyos, Antonio José
dc.contributor.authorMiranda Guevara, Alvaro de Jesús
dc.date.accessioned2024-04-25T22:40:47Z
dc.date.available2024-04-25T22:40:47Z
dc.date.issued2024
dc.description.abstractEl carbón es un mineral que en la actualidad representa una fuente de energía clave en la economía mundial. Durante décadas ha sido utilizado para la generación de electricidad, calefacción y como materia prima en la industria. La extracción de este mineral involucra la remoción de capas de tierra y roca para acceder a las capas de carbón debajo de la superficie, proceso que deja una huella ambiental significativa, y que ha desencadenado preocupaciones cruciales relacionadas con la salud, especialmente a través de la generación del polvo proveniente de las actividades de minería. En este sentido se considera que la composición de las partículas de carbón, el tamaño y forma, juegan un papel fundamental en las afecciones respiratorias de las poblaciones humanas. El objetivo principal de este estudio fue analizar los efectos citotóxicos y genotóxicos in vitro de nanopartículas de carbón en células V79 y HaCaT. Mediante el método de separación en medio ácido se aislaron las nanopartículas. Posteriormente estas nanopartículas fueron usadas para exponer células V79 y HaCaT a diferentes concentraciones. A través del ensayo con rezasurina y sulforodamina se determinaron los efectos de estas partículas en la viabilidad celular, y se seleccionaron las concentraciones 50, 150 y 300 μg/mL para realizar los ensayos de genotoxicidad, ensayo cometa y micronúcleos. La microscopía de fuerza atómica proporcionó una visión detallada de la topografía de las nanopartículas, destacando su propensión a la aglomeración. Mediante SEM-EDS se evidenció la forma y diversidad química de estas nanopartículas, constituidas principalmente por elementos como carbono (C), oxígeno (O), hierro (Fe), calcio (Ca) y mediante cromatografía de gases acoplada a espectrometría de masas (GC/MS), se determinaron los hidrocarburos aromáticos policíclicos (HAP) presentes en las nanopartículas como fluoranteno, naftaleno, antraceno, 7H-benzo[c]fluoreno, fenantreno, pireno, benzo[a]antraceno, criseno y algunos derivados alquílicos. La evaluación de la genotoxicidad mediante marcadores como el ensayo cometa, la formación de micronúcleos y la inmunomarcación empleando anticuerpos anti-Gamma H2AX, mostró un efecto dosis-respuesta evidenciando la capacidad de las nanopartículas de carbón para inducir inestabilidad genética y muerte celular. El uso de la técnica de temperatura melting y PCR en tiempo real permitió evidenciar una posible alteración en la estructura y estabilidad del ADN debido a la interacción físico-química de este con las nanopartículas de carbón. En conclusión, este estudio destaca la relación entre las características específicas de las nanopartículas de carbón para llegar a comprender el entendimiento de sus interacciones a nivel celular, molecular y sentar las bases para dilucidar los mecanismos relacionados con el desarrollo de diferentes enfermedades respiratorias.spa
dc.description.abstractEl coal is a mineral that currently represents a key energy source in the global economy. For decades, it has been used for electricity generation, heating, and as a raw material in industry. The extraction of this mineral involves the removal of layers of soil and rock to access the coal seams beneath the surface, a process that leaves a significant environmental footprint and has triggered crucial health concerns, especially through the generation of dust from mining activities. In this regard, it is considered that the composition, size, and shape of coal particles play a fundamental role in respiratory conditions in human populations. The main objective of this study was to analyze the in vitro cytotoxic and genotoxic effects of coal nanoparticles on V79 and HaCaT cells. Nanoparticles were isolated using the acid medium separation method. Subsequently, these nanoparticles were used to expose V79 and HaCaT cells to different concentrations. The effects of these particles on cell viability were determined through resazurin and sulforhodamine assays, and concentrations of 50, 150, and 300 μg/mL were selected for genotoxicity assays, comet assay, and micronuclei assay. Atomic force microscopy provided a detailed view of the topography of the nanoparticles, highlighting their tendency to agglomerate. SEM-EDS revealed the shape and chemical diversity of these nanoparticles, primarily composed of elements such as carbon (C), oxygen (O), iron (Fe), calcium (Ca), and through gas chromatography coupled with mass spectrometry (GC/MS), polycyclic aromatic hydrocarbons (PAHs) present in the nanoparticles were determined, such as fluoranthene, naphthalene, anthracene, 7Hbenzo[c]fluorene, phenanthrene, pyrene, benzo[a]anthracene, chrysene, and some alkyl derivatives. The evaluation of genotoxicity using markers such as the comet assay, micronucleus formation, and immunostaining employing antibodies against Gamma H2AX showed a dose-response effect, demonstrating the ability of coal nanoparticles to induce genetic instability and cell death. The use of melting temperature technique and real-time PCR allowed for the evidence of a possible alteration in the structure and stability of DNA due to the physicochemical interaction with coal nanoparticles. In conclusion, this study highlights the relationship between the specific characteristics of coal nanoparticles to understand their interactions at the cellular and molecular levels and lay the groundwork for elucidating mechanisms related to the development of various respiratory diseases.eng
dc.format.mimetypepdf
dc.identifier.urihttps://hdl.handle.net/20.500.12442/14552
dc.language.isospa
dc.publisherEdiciones Universidad Simón Bolívarspa
dc.publisherFacultad de Ciencias Básicas y Biomédicasspa
dc.rights.accessrightsinfo:eu-repo/semantics/restrictedAccess
dc.subjectNanopartículasspa
dc.subjectCarbónspa
dc.subjectCitotoxicidadspa
dc.subjectGenotoxicidadspa
dc.subjectDaño oxidativospa
dc.subjectHAPspa
dc.subjectMetalesspa
dc.subjectMuerte celularspa
dc.subjectNanoparticleseng
dc.subjectCoaleng
dc.subjectCytotoxicityeng
dc.subjectGenotoxicityeng
dc.subjectOxidative damageeng
dc.subjectPAHseng
dc.subjectMetalseng
dc.subjectCell deatheng
dc.titleEvaluación de la citotoxicidad y genotoxicidad en células V79 y HaCaT asociado a la exposición a nanopartículas de carbón de La Loma Cesarspa
dc.type.driverinfo:eu-repo/semantics/doctoralThesis
dc.type.spaTesis de doctorado
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