GUSTAVO HENRIQUE COSTAVARCA

GUSTAVO HENRIQUE COSTAVARCA

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An updated review of macro, micro, and nanostructured hydrogels for biomedical and pharmaceutical applications
2020 - LIMA, CAROLINE S.A. de; BALOGH, TATIANA S.; VARCA, JUSTINE P.R.O.; VARCA, GUSTAVO H.C.; LUGAO, ADEMAR B.; CAMACHO-CRUZ, LUIS A.; BUCIO, EMILIO; KADLUBOWSKI, SLAWOMIR S.
Hydrogels are materials with wide applications in several fields, including the biomedical and pharmaceutical industries. Their properties such as the capacity of absorbing great amounts of aqueous solutions without losing shape and mechanical properties, as well as loading drugs of different nature, including hydrophobic ones and biomolecules, give an idea of their versatility and promising demand. As they have been explored in a great number of studies for years, many routes of synthesis have been developed, especially for chemical/permanent hydrogels. In the same way, stimuli-responsive hydrogels, also known as intelligent materials, have been explored too, enhancing the regulation of properties such as targeting and drug release. By controlling the particle size, hydrogel on the micro- and nanoscale have been studied likewise and have increased, even more, the possibilities for applications of the so-called XXI century materials. In this paper, we aimed to produce an overview of the recent studies concerning methods of synthesis, biomedical, and pharmaceutical applications of macro-, micro, and nanogels.
Simultaneous intramolecular crosslinking and sterilization of papain nanoparticles by gamma radiation
2020 - FAZOLIN, GABRIELA N.; VARCA, GUSTAVO H.C.; FREITAS, LUCAS F. de; ROKITA, BOZENA; KADLUBOWSKI, SLAWOMIR; LUGAO, ADEMAR B.
Papain-based nanoparticles were recently developed using radiation technologies and proven effective to generate nanosized crosslinked papain particles with preserved enzymatic activity. The applications of such nanostructured systems are expected to be similar to native papain with considerable biopharmaceutical advantages and concern drug loading among other biotechnological applications. The nature of such crosslinks and the possibility to provide simultaneous sterilization have been hypothesized but remain not totally clarified. This manuscript advances the discussion on the radiation-induced synthesis of protein nanoparticles by approaching the nature of the crosslinking and the possible contribution of bityrosine linkages and disulfide bridges to the overall nanoparticle assembly as well as the feasibility of the simultaneous sterilization process under the preestablished conditions of processing. Papain nanoparticles were synthesized and characterized according to size, proteolytic activity, bityrosine, cysteine content and molecular weight by SDS-PAGE upon sonication at 40 kHz. Bacterial identification and the sterility tests were performed in accordance with ISO 11737 prior to and after inoculating 106 CFU of Corynebacterium xerosis. Our experiments evidenced the crosslinking of rather intra- than intermolecular nature and a contribution of cysteine bridges and bityrosine linkages to the stabilization and formation of the papain nanoparticle assembly. The technique was effective to promote simultaneous crosslinking and sterilization at the established conditions of processing and may be validated in accordance with the ISO 11137.
The effects of radiation and experimental conditions over papain nanoparticle formation
2020 - FAZOLIN, GABRIELA N.; VARCA, GUSTAVO H.C.; KADLUBOWSKI, SLAWOMIR; SOWINSKI, SEBASTIAN; LUGAO, ADEMAR B.
Papain is a natural enzyme extracted from the fruit of Carica papaya Linnaeus, successfully applied in the pharmaceutical area as a drug carrier and debridement agent for wounds. In recent studies papain nanoparticles were synthesized and crosslinked with the use of ionizing radiation in the search for biopharmaceutical advantages as well as the development of bioactive nanocarriers. This study addresses the effects of buffer molarity and irradiation conditions on papain nanoparticles formation. Nanoparticles were synthesized on ice bath using ethanol (20%, v/v) as a cosolvent and crosslinked by gamma radiation using a 60Co source. Experimental variables included the synthesis in deionized water and in 1, 10, 25 and 50mM phosphate buffer, under different temperatures of −20 °C, 0 °C and 20 °C before and throughout the irradiation period, and using radiation dose rates of 0.8, 2, 5 and 10 kGy h−1 to reach the dose of 10 kGy. Proteolytic activity was quantified using Nα- benzoyl-DL-arginine-p-nitroanilide hydrochloride. Nanoparticle size and crosslinking by means of bityrosine were evaluated by dynamic light scattering and fluorescence measurements, respectively. Buffer molarity and radiation dose rate were identified to influence bityrosine formation and proteolytic activity without impacting nanoparticle size. Variations in temperature impacted bityrosine formation exclusively. Optimized conditions for papain nanoparticle synthesis were achieved using 50mM phosphate buffer at the dose rate of 5 kGy h−1 and temperature of 0 °C throughout the process.
The effect of radiation dose rate over the formation of protein-based nanoparticles for nanosized delivery of chemo and radiotherapeutics
2018 - VARCA, G.H.C.; FAZOLIN, G.N.; FERREIRA, A.H.; OLIVEIRA, J.P.R. de; MARQUES, F.; LUGAO, A.B.
Recent studies demonstrated the development of papain and bovine serum albumin nanoparticles using gamma radiation (10 kGy) in presence of 20-30% (v/v) ethanol. With the purpose of producing stable and well defined nanocarriers, this work aims to determine the influence of different dose rates over protein nanoparticle formation. For this purpose, papain and BSA nanoparticles were synthetized in phosphate buffer (50 mM, pH 7.2) and ethanol (20-30%, v/v) using a radiation dose of 10 kGy and dose rate of 0.8, 2, 5 and 10 kGy.h-1. After irradiation, samples were evaluated by dynamic light scattering, fluorescence and proteolytic activity to verify the size, secondary structure and monitoring of the enzymatic activity, respectively. For papain nanoparticles it was observed that the dose rate did not influence the particle size formation, however crosslinking evidenced by bityrosine showed that samples irradiated at 0.8 and 5 kGy.h-1 presented higher bityrosine levels. On the other hand, BSA nanoparticles presented different results if compared to papain NPs. Different dose rates caused different and non-linear size increase for each condition, following the order: 5 > 10 > 0.8 > 2 kGy.h-1. However, in terms of crosslinking formation, a linear increase was registered, as at 0.8 kGy.h-1 the smallest signal was achieved, whereas at 10 kGy.h- 1 the highest signal was recorded. In conclusion, BSA nanoparticles were more sensitive to different radiation dose rates than nanopapain. Optimized results in terms of size increase and higher bityrosine levels were observed for the samples irradiated at 5 kGy.h-1, in which nanoparticle formation will occur faster if compared to the synthesis carried out under distinct conditions. As final applications of the system concert their use for the delivery of chemo or radiotherapeutics, the loading of paclitaxel, a well-known chemotherapeutic agent, and radiolabeling with tecntetium- 99m, a radioisotope suitable for biomedical applications, have also been performed with high efficiency, thus demonstrating a proof of concept of such systems.
Influência do cosolvente na formação de nanopartículas de papaína induzidas por radiação
2018 - FAZOLIN, G.N.; VARCA, G.H.C.; LUGAO, A.B.
Biomateriais à base de proteínas são de grande interesse biomédico e vem mostrando relevância nas linhas de pesquisas atuais por serem uma alternativa no carreamento de fármacos, além de apresentar efeitos colaterais reduzidos ou inexistentes. A papaína, enzima proteolítica utilizada neste trabalho, é uma alternativa no estudo de sistemas nano estruturados por se mostrar promissora no carreamento de fármacos. A técnica utilizada para síntese desta nanopartícula utiliza o etanol como cosolvente. Além de promover desolvatação na proteína, o etanol tem a função de proteger a enzima durante o processo de radiação – utilizado para reticulação e esterilização. Observou-se anteriormente que o etanol auxilia no aumento do tamanho da nanopartícula, contudo, não se sabe qual a influência deste cosolvente durante o processo. Sendo assim, o objetivo principal deste trabalho é estudar a influência do etanol nas nanopartículas de papaína. A síntese será realizada na presença (20%) e ausência do etanol, tampão fosfato 50 Mm pH 7,4 e radiação ionizante a 10 kGy durante 2 horas (5 kGy/hora). As amostras foram avaliadas através da técnica de espalhamento dinâmico de luz, fluorescência e bioatividade a fim de verificar o tamanho, a estrutura secundária e acompanhamento da atividade enzimática, respectivamente. Após a irradiação, as amostras foram filtradas, congeladas e liofilizadas por 48 horas. As amostras foram resuspendidas em duas condições: água deionizada e na presença de etanol. Observou-se quando resuspendidas em água, as amostras voltam ao tamanho nativo (± 4 nm) enquanto na presença de etanol este tamanho é mantido como no controle (± 7 nm). As análises de fluorescência demonstraram que a reticulação por radiação continuou existente e a bioatividade decaiu 30% em ambos os casos, quando comparada com a bioatividade inicial. Conclui-se que o etanol contribui para a formação do tamanho da nanopartícula durante a síntese porem quando evaporado perde-se a solvatação e mantem o aumento de tamanho somente promovido pela radiação.
Radiation-synthesized protein-based drug carriers: Size-controlled BSA nanoparticles
2016 - QUEIROZ, R.G.; VARCA, G.H.C.; KADLUBOWSKI, S.; ULANSKI, P.; LUGAO, A.B.
Irradiation as an alternative route for protein crosslinking: Cosolvent free BSA nanoparticles
2016 - VARCA, GUSTAVO H.C.; QUEIROZ, RODRIGO G.; LUGAO, ADEMAR B.
Radiation synthesized protein-based nanoparticles: A technique overview
2014 - VARCA, GUSTAVO H.C.; PEROSSI, GABRIELA G.; GRASSELLI, MARIANO; LUGAO, ADEMAR B.
Synthesis and characterization of hydrogels composed by different types of poly(vinyl alcohol)
2010 - ALCANTARA, MARA T.S.; VARCA, GUSTAVO H.C.; AMARAL, RENATA H.; MATTOS, THAMIRES C.B.; RIELLA, HUMBERTO G.; LUGAO, ADEMAR B.
Polymer hydrogels as optimized delivery systems
2013 - BATISTA, JORGE G.S.; VARCA, GUSTAVO H.C.; FERRAZ, CAROLINE C.; GARRIDO, GABRIELA P.; DINIZ, BRUNA M.; CARVALHO, VINICIUS S.; LUGAO, ADEMAR B.