ALEX ALVES RODRIGUES
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Artigo IPEN-doc 30589 Development of spheroids in co-culture of prostate tumor with human fibroblasts, using the hanging-drop technique with plate inversion for analysis of gamma irradiation by 60Co2024 - SILVA, T.M.; RODRIGUES, A.A.; SILVA, G.D.; SANTOS, E.C.; LIMA, P.F.; PRUDENTE, S.R.; PAZ, M.M.; VIEIRA, D.P.Artigo IPEN-doc 30354 Radioactive gold nanoparticles coated with BSA2024 - BARBEZAN, ANGELICA B.; ROSERO, WILMMER A.A.; VIEIRA, DANIEL P.; RIGO, MARIA E.Z.; SILVA, GIOVANA D. da; RODRIGUES, ALEX A.; ALMEIDA, LUIS F. de; SILVA, FABIO F.A. da; RIVERA, ANDY G.; SILVA, NATANAEL G. da; BERNARDES, EMERSON S.; ZEITUNI, CARLOS A.; ROSTELATO, MARIA E.C.M.Background: Nanotechnology has revolutionized medicine, especially in oncological treatments. Gold nanoparticles (AuNPs) stand out as an innovative alternative due to their biocompatibility, potential for surface modification, and effectiveness in radiotherapeutic techniques. Given that prostate cancer ranks as one of the leading malignancies among men, there's a pressing need to investigate new therapeutic approaches. Methods: AuNPs coated with bovine serum albumin (BSA) were synthesized and their cytotoxicity was assessed against prostate tumor cell lines (LNCaP and PC-3), healthy prostate cells (RWPE-1), and endothelial control cells (HUVEC) using the MTS/PMS assay. For in vivo studies, BALB/C Nude mice were employed to gauge the therapeutic efficacy, biodistribution, and hematological implications post-treatment with BSA-coated AuNPs. Results: The BSA-coated AuNPs exhibited cytotoxic potential against PC-3 and LNCaP lines, while interactions with RWPE-1 and HUVEC remain subjects for further scrutiny. Within animal models, a diverse therapeutic response was observed, with certain instances indicating complete tumor regression. Biodistribution data emphasized the nanoparticles' affinity towards particular organs, and the majority of hematological indicators aligned with normative standards. Conclusions: BSA-coated AuNPs manifest substantial promise as therapeutic tools in treating prostate cancer. The present research not only accentuates the nanoparticles' efficacy but also stresses the imperative of optimization to ascertain both selectivity and safety. Such findings illuminate a promising trajectory for avant-garde therapeutic modalities, holding substantial implications for public health advancements.Dissertação IPEN-doc 30204 Desenvolvimento e implementação de um sistema piloto otimizado para a gestão de biotérios de roedores2023 - RODRIGUES, ALEX A.A utilização de animais em procedimentos biomédicos é uma prática amplamente difundida, envolvendo mais de 115 milhões de animais anualmente. Estes animais, obrigatoriamente, devem viver dentro de instalações adequadas denominados de biotérios, que, por sua vez, deve oferecer bem-estar aos animais seguindo, o conceito dos 3R’s. Além disto, os biotérios devem seguir normas e leis que visam a saúde, o bem-estar, a segurança e o correto funcionamento dos mesmos, contudo, cada biotério tem suas especificidades e particularidades. Devido a alta demanda de trabalho e normas a serem seguidas, a gestão de um biotério deve ser otimizada e contar com auxílio de ferramentas tecnológicas, contudo, no Brasil este tipo de gestão ainda não existe. O atual modelo de gerenciamento no Brasil, que muitas vezes depende de métodos rudimentares, impacta negativamente a eficiência na administração de recursos humanos, de materiais e, inclusive, dos animais. Deste modo, o objetivo do presente projeto foi criar um sistema piloto automatizado, customizável e inovador para otimizar a gestão de biotérios e atender às principais demandas. Para a criação da primeira versão do programa, foram utilizadas as ferramentas da Microsoft Power Apps, Sharepoint e Power Automate respectivamente. Os resultados demonstram que foi possível criar um programa simples e acessível, que atende às principais demandas do biotério Nanci do Nascimento, no IPEN. Este programa foi testado diversas vezes e com diferentes colaboradores e funcionou perfeitamente, conforme o esperado. Com base nestes resultados, concluímos que conseguimos criar um programa de computador que funciona como uma ferramenta tecnológica e inovadora para otimizar a gestão de biotérios de modo customizável, assim, atendendo às particularidades de cada instituição.Resumo IPEN-doc 30292 Production of a double-network hydrogel using sodium alginate and nano-structured cellulose to 3D cell cultures2023 - SILVA, GIOVANA D. da; SAKATA, SOLANGE K.; ASSIS, JOAO V.A. de; SANTOS, ESTHER C. dos; PRUDENTE, SULEYNA R.; RODRIGUES, ALEX A.; FALCAO, PATRICIA L.; VIEIRA, DANIEL P.Introduction and objective 2D cell cultures have limitations regarding on tissue representativity. 3D cell cultures can use hydrogels of alginate with cellulose with adequate viscoelasticity properties for cell growth, being from plant sources, abundant and low cost. This work consisted of producing a biocompatible gel from plant sources for threedimensional cultures, promoting polymeric matrices for cells, helping in cell interactions and nutrient transport, providing mechanical support, self-assembly capacity, biodegradation, ability to reticulation, stability control and mechanical resistance. Methodology Transformation of microcrystalline cellulose into nanofibers was achieved freezing aqueous suspensions on presence of 4M NaOH to proper dissociation of fibers. To obtain suitable dispersion, sodium citrate was added to prevent aggregation. Suspensions were analyzed by Scanning Electron Microscope (SEM), Fourier Transform Infrared Spectroscopy (FTIR), Zeta Potential. For cell viability analysis, murine fibroblastic cell lines (NIH/3T3) were plated (2.5 x 105 cells per well) 24-well plate embedded in gel (100µL). Results and discussion The analysis of cellulose suspensions through SEM, showed a significant change in the size and shape of the structures after hydrolysis, indicating the obtention of structures on a nanometric scale. For the analysis of cellulose aggregation, the zeta potential values indicated that after the addition of sodium citrate, greater dispersion was obtained between the cellulose structures, enabling resistance to the structure in a uniform way. FTIR analysis showed changes in the covalent bonds of the products. Cell viability assay showed structures containing fibroblast cells, alginate and cellulose with 1 cycle of freezing with citrate showed an intact gel structure, with cell aggregates indicating possible cell growth, while the one with only alginate showed dead cells and showed that the hydrogel did not induce cellular toxicity. These results suggest that the hydrolysis of microcrystalline cellulose can lead to obtaining cellulose nanofibers with potential for applications in tissue engineering. Conclusions Hydrogels, they have potential for applications in tissue engineering, since they have mechanical resistance and cell viability. In addition, hydrogels from exclusively vegetable sources, since these are in large quantity, low cost and environmental impact, given that the alginate comes from brown algae found in several coastal regions and the cellulose can be extracted from renewable sources or various vegetable waste from agroindustry.Resumo IPEN-doc 30279 Development of human fibroblast spheroids with hanging-drop inverted plates2023 - RODRIGUES, ALEX A.; SAMPAIO, MARLOS C.; SANTOS, ESTHER C. dos; PRUDENTE, SULEYNA R.; LIMA, MAYELLE M.P.; SILVA, GIOVANA D. da; MATHOR, MONICA B.; VIEIRA, DANIEL P.Introduction and objective: 2D cultures have limitations in cell growth. 3D cultures, on the other hand, have become a valuable and powerful tool for biomedical research in recent decades. Due to their resemblance to living systems and cellular interactions, this type of culture can be developed using various methodologies, including nanoparticles, hydrogels, and layers of agarose, among others. Considering the need for testing and validating new molecules and effective therapies for treating various diseases, the objective of this study is to standardize a 3D human fibroblast culture model. Methodology: HF002-J, human fibroblast cells, were cultured at 37ºC in a humid atmosphere containing 5% CO2, maintained in RPMI 1640 medium supplemented with 10% fetal bovine serum and 1% antibiotics. When reaching 60- 70% confluence, the cells were detached using a 0.05% trypsin solution. Spheroids were prepared using the hangingdrop technique adapted from [1] 440 µL of medium containing cell variations ranging from 2 × 103 to 6 × 104 cells per well of a 96-well plate were deposited, generating a positive meniscus. The plate was inverted and incubated as described. Results and discussion: The present study aimed to evaluate the development of cellular spheroids after 4 days of culture using different cell preparations. Our results demonstrated that the preparations used produced compact spheroids, characterized by homogeneous sizes in the range of 500 to 1000 μm. When analysing the images obtained by wide-field fluorescence microscopy, we observed that the proportions of unviable cells labelled with fluorophores varied significantly according to the initial number of cells used in the preparations. Notably, increasing the initial number of cells resulted in a proportional increase in the number of non-viable cells present in the formed spheroids. These results suggest that the initial cell density can affect the development and viability of the formed spheroids. It is possible that too high cell density led to greater competition for nutrients and space, resulting in greater cell mortality and less viable spheroids. Conclusions: Based on the results obtained, it was possible to develop an initial prototype of spheroids from human fibroblast cells that can resemble tissues in vivo due to their cellular interactions, thus providing a new tool for the study of drugs and treatments.Resumo IPEN-doc 30278 High-throughput production of tumor spheroids (melanoma and colon carcinoma) using simple plate treatment and automated fluorescence microscopy analysis2023 - PRUDENTE, SULEYNA R.; ASSIS, JOAO V.A. de; SANTOS, ESTHER C. dos; SILVA, GIOVANA D. da; RODRIGUES, ALEX A.; ROCHA, LEONARDO W.P. de S.; FALCAO, PATRCIA L.; VIEIRA, DANIEL P.Introduction and objective: Cancer is currently one of the leading causes of death in the world. The objective of this work is the formation of viable spheroids from cells of melanoma (SKMEL-37) and colon adenocarcinoma (HT29-MTX) cell lines and their evaluation regarding cell viability to enable the use of threedimensional cell culture as an alternative to the use of experimental animal models. Methodology: Cells were maintained in RPMI 1640 medium and kept in an incubator at 37°C, 5% CO₂ with controlled humidity. Upon reaching 60-70% confluence, cells were washed with phosphate buffered saline (PBS) and detached using trypsin solution. Afterwards, they were seeded in 24-well plates pre-treated with PluronicⓇ F-127 (0.5g/mL in 2-propanol) and turned back in incubator for 72 hours. Then, the formed spheroids were stained with Hoechst 33342 and SYTOX® Green solution, incubated for 60 minutes and images were acquired automatically in a HTS equipment (INCell 2500 HS, Cytiva). Results and discussion: Properly cohesive spheroids were obtained for both lineages, 20-30 per well. After 72h, only a small fraction of cells (about 5%) were considered unviable by SYTOX® staining. Principal Component Analysis (PCA) using 13 variables, and further Principal Component Regression (PCR) showed that nuclei mean and maximum intensities (Hoechst), and nuclei volume are the most relevant variables, corelated to number of plated cells. Days in culture appeared to not correlate with other variables. Conclusions: It was concluded that the methodology for the production of spheroids for melanoma and colon adenocarcinoma cell lines presented is simple, fast and cheap, in which, in 72 hours, the spheroids form freely, without restriction of shape and size and presenting low cell death, being also compatible with the high throughput screening technique (HTS). Nuclei volume and intensity can be used in future analysis to assess cell global viability in spheroids.Resumo IPEN-doc 30247 Desenvolvimento de esferóides de melanoma humano como modelo alternativo ao uso de animais de laboratório2023 - RODRIGUES, ALEX A.; SAMPAIO, MARLOS C.; SILVA, GIOVANA D. da; SANTOS, ESTHER C. dos; PRUDENTE, SULEYNA R.; LIMA, MAYELLE M.P.; VIEIRA, DANIEL P.Resumo IPEN-doc 29185 Análise histórica das atividades do biotério Nanci do Nascimento (IPEN)2022 - RODRIGUES, ALEX A.; CORTEZ, MARLOS; MASCARENHAS, NEIDE F.; REIS, ISMARIA S.; LIMA, MAYELLE M.P.; GODOI, ELIANA L.; VIEIRA, DANIEL P.