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Nanotoxicologia: métodos para avaliação do perfil toxicológico de sistemas nanoparticulares

Authors: Costa, Inês Henriques da Silva e;

Nanotoxicologia: métodos para avaliação do perfil toxicológico de sistemas nanoparticulares

Abstract

A nanotecnologia é uma ciência promissora em diversas áreas, incluindo a indústria farmacêutica, na qual o seu principal objetivo é o desenvolvimento de novas terapias. As nanopartículas lipídicas sólidas (SLN) foram desenvolvidas com o objetivo de ultrapassar os limites dos sistemas coloidais tradicionais como lipossomas, emulsões e as nanopartículas poliméricas. São partículas com tamanhos inferiores ao mícron, entre 50-1000nm, constituídas por lípidos fisiológicos, biodegradáveis, e biocompatíveis, que se mantêm sólidos à temperatura corporal e ambiente. Neste projeto é abordada a nanotecnologia associada a estas partículas, como os excipientes utilizados, os métodos de produção, as suas principais aplicações e a sua toxicidade. Apesar destas nanopartículas apresentarem todos os pré-requisitos para serem considerados sistemas de transporte seguros, sendo esperada baixa ou até nenhuma toxicidade, a sua utilização médica requer mais evidência sobre a sua segurança. A maioria dos dados disponíveis na literatura sobre o comportamento destas partículas in vitro parece confirma-la, mas alguns resultados discordantes também já foram apresentados. O stress oxidativo é um dos danos mais largamente associados às nanopartículas, mas nas SLN este efeito não tem tido amplamente estudado. Os poucos estudos realizados revelam resultados contraditórios, o que nos incentivou a averiguar a possível indução de stress oxidativo por uma série de cSLN em HepG2, uma linha celular do carcinoma hepatocelular. Averiguámos a geração de espécies reativas de oxigénio (ROS) e a alteração na atividade dos sistemas antioxidantes enzimáticos após exposição às cSLN. O ensaio de DCFH-DA revelou um enorme aumento da presença ROS, a atividade da superóxido dismutase aumentou e a atividade da glutationa redutase diminuiu drasticamente (início de apoptose), o que nos indica que estas nanopartículas causam, realmente, stress oxidativo nesta linha celular. Em adição, investigámos se tal toxicidade é causada pelo lípido catiónico usado nas formulações em causa (brometo de cetiltrimetilamônio, CTAB). Os resultados direcionam-no como uma razão para tal toxicidade, mas não a única. Contudo, os nossos resultados foram contraditórios e mais investigação deve ser levada a cabo. Nanotechonology is a promising science in several fields, including the pharmaceutical industry, in which its main goal is to develop new therapeutics. Solid lipid nanoparticles (SLN) were developed to overcome the limits of traditional colloidal carriers such as liposomes, emulsions and polymeric nanoparticles. They are submicron particles with size in the range of 50-1000 nm and are composed of physiological, biodegradable and biocompatible lipids that remain solid at body and room temperature. In this thesis it is described the nanotechnology associated to SLN, materials used as excipients, methods of production, their main applications and their toxicology. Although these particles present all pre-requisites to be considered safe drug carries, being expected to present low or even none toxicity, their use in medicine requires more evidence of their safety. The available data on the in vitro behavior of these systems seem to confirm it, but some contradictory results have also been reported. Oxidative stress is one of the most reported damage associated to nanoparticles, but in SLN it has not been widely studied. The few performed studies have revealed some contradictory result, which led us to study the possibility of oxidative stress induction by a set of cSLN in HepG2, a human hepatocellular carcinoma cell line. We ascertained the generation of reactive oxygen species (ROS) and the alteration on antioxidant enzymes’ activity after exposure to the cSLN. DCFH-DA assay revealed great increase of ROS presence, the activity of superoxide dismutase was increased and the activity of gluthatione redutase was largely decreased (apoptosis initiation), indicating that these nanoparticles caused, indeed, oxidative stress in this cell line. In addition, we investigated if such toxicity is caused by the cationic lipid used in the formulations (cetyl trimethylammonium bromide, CTAB), our main suspect. The results direct it as one, but not the only, reason of such toxicity. Nevertheless, our results were contradictory and more investigations must be carried out.

Country
Portugal
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Keywords

Nanotoxicidade, Oxidative stress, Solid lipid nanoparticles, Nanopartículas lipídicas sólidas, Stress oxidativo, Nanotoxicology

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This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
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popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
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influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
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impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
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