DANIEL PEREZ VIEIRA

Resumo

Scientific Researcher at IPEN since 2010, but my academic experience comes since 1998. Research fields comprising biological effects of ionising radiation on tumoral and non-tumoral in vitro models, pre-clinical tests of radiopharmaceuticals, development of methods of genotoxicity assessment, gene therapy, effects of radiomodifiers on tumor models and organotypic 3D cell culture. Experience on protozoology and immunology research fields. - Post-Graduation Professor at Universidade de São Paulo - Nuclear Technology Programs - Visiting Professor at Instituto Israelita de Ensino e Pesquisa Albert Einstein - Radiopharmacy (post-graduation course) (Text obtained from the Currículo Lattes on October 8th 2021)


Pesquisador responsável pelo Lab. de Radiobiologia do Centro de Biotecnologia - CEBIO (IPEN-CNEN/SP). Head of Lab. of Radiobiology - CEBIO (IPEN/CNEN-SP). (Texto extraído do Currículo Lattes em 08 out. 2021)

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Agora exibindo 1 - 10 de 96
  • Artigo IPEN-doc 30388
    Synergizing immune balance
    2024 - TEIXEIRA, BRUNA H.; GONCALVES, KARINA de O.; VIEIRA, DANIEL P.; COURROL, LILIA C.
    The multifaceted health benefits of curcumin (Curcuma longa), attributed to its antioxidant, antitumor, and anti-inflammatory activities, have drawn significant scientific attention. Curcumin shows promise as a potential modulator of macrophage polarization, offering a natural strategy for managing inflammation and promoting tissue repair. However, a limiting factor for this beneficial molecule is its limited bioavailability due to its low solubility in water. This study aimed to quantify the effect of curcumin gold nanoparticle (CurAuNP)-mediated ultrasound irradiation on THP-1-derived macrophages as potential therapeutic targets. The photoreduction method was applied to synthesize the gold nanoparticles with curcumin as a ligand (CurAu). The effect of adding polyethylene glycol in the synthesis process was studied (CurAuPEG). CurAuNP characterization included UV/Vis, Zeta potential, transmission electron microscopy, and FTIR. The amount of singlet oxygen released by curcumin and CurAuNPs was quantified by observing 1.3-diphenylisobenzofuran quenching upon ultrasound irradiation (1 MHz and 1 W/cm2). The results indicated that ultrasound therapy for 4 min with CurAuNPs significantly enhanced singlet oxygen generation and reduced macrophage viability compared to curcumin alone. The increased sonoluminescence and curcumin delivery facilitated by CurAuNPs led to greater curcumin activation. Consequently, CurAuNPs could offer promising therapeutic options for modulating macrophage polarization in pro-inflammatory and anti-inflammatory stages.
  • Artigo IPEN-doc 30364
    Evaluation of low-kV energy X-ray radiation effect on breast cancer cells incubated with gold nanoparticles capped with aminolevulinic acid, methyl aminolevulinate, and gamma-aminobutyric acid
    2024 - LOPES, ISABELA S.; SANTOS, NOEMY R.; SILVA, GIOVANA D. da; SANTOS, LEONARDO C. dos; POTIENS, MARIA da P.A.; VIEIRA, DANIEL P.; COURROL, LILIA C.
    Gold nanoparticles (AuNPs) have unique properties, including size-dependent optical and electronic characteristics, biocompatibility, and ease of functionalization, making them attractive for various diagnostic and therapeutic applications. The goal of this work was to verify the potential application of gold nanoparticles capped with aminolevulinic acid (ALA), methyl aminolevulinate (MALA), and gamma-aminobutyric acid (GABA) in lowenergy X-ray diagnostics and therapy of breast cancer. ALA and MALA induce the accumulation of protoporphyrin IX (PpIX) in cancer cells. PpIX, in turn, can be excited by Cherenkov radiation, producing reactive oxygen species after energy or electron transfer from the triplet excited state of PpIX to molecular oxygen, which induces cell death by apoptosis or necrosis. The results indicated that ALA and MALA nanoparticles promoted reduced cell viability by approximately 20 % with X-ray irradiation at an energy level of 35 kV for 5 min. Breast cancer cells possess GABA receptors, allowing for targeted effects by GABANPs, which can enhance contrast and improve diagnostic potential. GABAAuNPs also demonstrated decreased cell viability in similar to 10 % following X-ray irradiation, making them a promising material for breast cancer diagnosis and treatment.
  • Artigo IPEN-doc 30354
    Radioactive gold nanoparticles coated with BSA
    2024 - 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.
  • Resumo IPEN-doc 30297
    Sodium alginate and nanocellulose hydrogel as scaffold to in vitro 3D prostate cancer irradiated model
    2023 - SILVA, G.D.; SAKATA, S.K.; ASSIS, J.V.A.; VIEIRA, D.P.
    Introdução: Recently, traditional cell culture systems structured in 2 dimensions using monolayers of cells in culture media are being replaced by 3D structures, in which cells can be organized in spheroids. To obtain these structures, hydrogels can be used as permeable to gas and nutrients scaffolds, also providing physical support to cells. This work aimed to produce a double network hydrogel containing sodium alginate (SA) and nanocellulose (NC), obtained by irradiation of microcrystalline cellulose, and its ability to maintain in culture of human prostate adenocarcinoma. Objetivos: To analyze whether SA+NC gels can keep viable 3D LNCap (prostatic carcinoma) in vitro, with and without exposure to radiation (0 and 2Gy gamma). Métodos: 0.75g of microcrystalline cellulose (Sigma-Aldrich, 435236) dispersed in water were irradiated (300 kGy) (25 mm column height) in an electron beam source (Dynamitron® Job 188 ,RDI- Radiation Dynamics Inc.). The precipitated powder was washed in water by centrifugation. Nanocellulose pellet was added to a sodium alginate (2.5%) dissolved in PBS. LNCaP cells were maintained in RPMI 1640 medium in monolayers in culture flasks and controlled atmosphere (37º, 5% CO2). 24-well plates were used, pre-treated with Pluronic® F-127 solution (0.5g/mL in 2-propanol). The hydrophobic portions of Pluronic molecules were directed towards the center of the well, thus preventing cell adhesion to the culture plastic. In each well 1x105 cells were added, forming clusters of cells after 3 days. Clusters were removed and added to the hydrogel seeded in 96-well plates. Crosslinking was achieved using 100 μL of 2mM CaCl2 solution on top of the gels. After gelation, the saline solution was removed and the wells received 100μL of culture medium and were submitted to gamma irradiation with doses of 0 and 2 Gy (GammaCell, Canada), and further kept in incubator for 24h. Medium was replaced by fresh medium with Hoescht 33342 (10mg/mL) and SYTOX? Green (5mM) and kept in an incubator for 30 minutes. Plates were imaged in an INCell Analyzer 2500HS and images were obtained to determine the dead cell count. Resultados: Visual evidence of spheroids enclosed in gels showed increased cell viability in SA+NC comparing to SA gels only. No visual differences were observed in irradiated (2Gy) spheroids. Conclusão: SA+NC gels can sustain cell viability and cause no changes in cell radioresistance, being a suitable model to in vitro studies.
  • Resumo IPEN-doc 30292
    Production of a double-network hydrogel using sodium alginate and nano-structured cellulose to 3D cell cultures
    2023 - 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 plates
    2023 - 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 analysis
    2023 - 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 30277
    Comparison of image deconvolution methods to improve image quality of 3D cell cultures acquired using HCS/HTS equipment
    2023 - ASSIS, JOAO V.A. de; VIEIRA, DANIEL P.
    Introduction and objective: Accurate imaging analysis of 3D in vitro cell cultures became critical to evaluation of morphological and some physiological aspects of cells in these aggregates. Fluorescence microscopy protocols allow analysis of a plethora of molecular markers, DNA content and integrity and assessment of morphological changes upon different conditions. Light diffraction by plastic (or glass) substrate, and through sample and mounting/culture media distorts events, and thus images must require deconvolution methods to proper visualization and analysis. The work used 2 different algorithms and 3 iteration numbers to deconvolve images of spheroids acquired in large scale using HTS/HCS equipment. Methodology: Murine fibroblasts (NIH/3T3) were seeded on non-adhesive 96-well plates in RPMI 1640 medium. After 72 hours in culture, formed spheroids received pan-nuclear (Hoescht 33342) and nucleus of dead cells (SYTOX® Green) staining and imaged using HCS/HTS equipment (INCell 2500 HS, Cytiva). PSF files were generated using fluorescent beads in same cultured medium. Using DeconvolutionLab2 [1], a Fiji [2] plugin, spheroid images were deconvolved using the Tikhonov-Miller or Richardson-Lucy algorithms, with 10, 20 or 30 iterations. Orthogonal views of deconvolved stacks were analysed to find best results. Results and discussion: Both algorithms rendered similar results, with good reproducibility and resolution of light distortions. No real advantages were perceived in 30-iterations experiments, although 20 iterations rendered best images, regarding shape of nuclei (must be as spherical as possible) and drastic reduction of light refraction between slices. Conclusions: The work found that both methods were very easy to apply to stacks, with similar results. 20 iterations must be the better option to deconvolve spheroid images, uniting efficiency and saving computational resources. Richardson-Lucy can be the method of choice, as produced slightly better results.
  • Resumo IPEN-doc 30274
    Antitumoral efficacy of gold nanoparticles with polyphenols in breast cancer and metastatic cells in vitro
    2023 - SPADREZANO, ISADORA; RODRIGUES, MURILO A.V.; VIEIRA, DANIEL P.; LUGAO, ADEMAR B.
    Introduction and objective: Gold nanoparticles (AuNPs) present beneficial properties in cancer diagnosis and therapy, as they can be coated and biofunctionalized with bioactive molecules through surface modification using relatively non‐toxic reagents, enabling the reduction of metal ions, stabilization, and selective detection of cellular receptors [1]. The reduction of gold by phytochemicals to form nanoparticles represents promising methods in Green Nanotechnology. This study aims to compare the binding capacity and evaluate the antiproliferative potential of gold nanoparticles coated with tannic acid (TA‐AuNPs) against human breast tumor cell lines. Cytotoxicity was determined using the MTS method. Methodology: The synthesis of TA‐AuNPs was established through the chemical reduction, and the purification method was based on centrifugation, where the centrifuged sample (P1) was resuspended after removing and storing the supernatant (S1), thus generating three samples, including the non‐centrifuged (NC). The confirmation of TA‐AuNPs formation was achieved by UV‐Visible absorption spectroscopy, and the size determination of TA‐AuNPs was carried out using dynamic light scattering and Transmission Electron Microscopy (TEM) techniques. The Zeta potential was used to determine the stability of TA‐AuNPs. Human breast adenocarcinoma cell lines (MCF‐7 and MDA‐MB‐231) were used, and the cytotoxicity determination was performed using the MTS assay. Results and discussion: The synthesis of TA‐AuNPs showed a change in color, indicating the formation of spherical AuNPs. UV‐Visible analysis was performed for preliminary characterization, where the absorption band at a wavelength of 529 nm, correlated with the localized surface plasmon resonance band, indicates the presence of approximately 20 nm AuNPs in all samples. To evaluate the size of TA‐AuNPs, the samples were analyzed using the dynamic light scattering method. The hydrodynamic diameters measured indicate that centrifugation induces greater aggregation of the AuNPs. However, based on the polydispersity values ranging from 0.087 ‐ 0.186, the AuNPs showed uniform sizes. The Zeta potential was an effective indicator used as a criterion to classify and quantify stability, with the samples exhibiting magnitudes above 40 mV, indicating high stability. TEM images confirmed the presence of TA‐AuNPs in the samples and provided information about their dimensions, around approximately 20 nm with spherical morphology. The cytotoxicity assessment for validating TA‐AuNPs demonstrated that the coating plays a crucial role in the degree of internalization, and a higher quantity of specific receptors resulted in a greater internalization of TA‐ AuNPs. It was observed that there was a higher percentage of cell viability in MCF‐7 cells compared to MDA‐MB‐ 231 cells. Conclusions: The synthesis was efficient in obtaining TA‐AuNPs. The TA‐AuNPs showed a narrow size distribution, predominantly spherical morphology, and low PDI, indicating significant potential for biomedical applications in the healthcare field. The results satisfactorily and efficiently demonstrated that TA‐AuNPs are effective against breast cancer cells (MCF‐7) and exhibit higher efficiency in targeting metastatic breast cancer cells (MDA‐MB‐ 231), highlighting their significant potential for various medical applications in the field of nanomedicine.
  • Resumo IPEN-doc 30247
    Desenvolvimento de esferóides de melanoma humano como modelo alternativo ao uso de animais de laboratório
    2023 - RODRIGUES, ALEX A.; SAMPAIO, MARLOS C.; SILVA, GIOVANA D. da; SANTOS, ESTHER C. dos; PRUDENTE, SULEYNA R.; LIMA, MAYELLE M.P.; VIEIRA, DANIEL P.