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Item Evaluation of Photodynamic Therapy Using AuNPs@Ce6 in 3D Cultures of Triple-Negative Breast Cancer(MDPI) Ventura, Aveline; Gonçalves, Giulia Capizzani; Soares, Cristina Pacheco; Sant’anna, Luciana Barros; Marmo, Vitor Luca Moura; Sibelino, Sônia Khouri; Raniero, Leandro JoséConventional cancer treatments have limited efficacy for aggressive subtypes such as triple-negative breast cancer (TNBC), which points to the importance of new therapeu- tic strategies. Functionalized nanoparticles in conjunction with photodynamic therapy (PDT) represent a promising alternative. Additionally, 3D cell culture emerges as a more effective model, as it better replicates the structural and functional characteristics of the tumor microenvironment. In this study, 3D microtumors of TNBC were cultivated and treated with PDT using gold nanoparticles functionalized with Chlorin e6 (AuNPs@Ce6). Cell viability was assessed using the MTT colorimetric assay, combined with histological analysis using hematoxylin-eosin staining. The MTT assay and histological evaluation of the 3D spheroids demonstrated that PDT with AuNPs@Ce6 effectively reduced cell viability and induced necrotic morphological changes, while maintaining biocompatibility with the non-irradiated control group. These findings reinforce the potential of this ap- proach for further investigation in TNBC models and underscore the value of 3D cultures as physiologically relevant and ethical alternatives to animal testing.Item Curcumin-coated iron oxide nanoparticles for photodynamic therapy of breast cancer(Springer Nature Link) Ferreira, Virginia Rezende; Ventura, Aveline; Candido, Marcela Aparecida; Ferreira-Strixino, Juliana; Raniero, Leandro JoséBreast cancer is the deadliest cancer among women and its treatment using traditional methods leads the patient to experience adverse effects. However, photodynamic therapy (PDT) is a non-invasive therapy modality that works through a photosensi- tizing agent, which treating activated by a suitable light source, releases reactive oxygen species capable of treating cancer. Furthermore, recent research indicates that combining PDT and nanoparticles can enhance therapeutic effects. In this way, the synthesis of IONPs (iron oxide nanoparticles) was carried out, and their subsequent coating was done with curcumin (IONPs@curcumin) so that they could act as therapeutic agents against breast cancer. Curcumin solubility tests were carried out to achieve the best results, with ethanol as a solvent, in different concentrations of ethanolic curcumin solution, with the optimal outcome observed at a concentration of 1 mM. Subsequently, the stability analysis was conducted by adjusting the pH of the medium, revealing that at pH 10, the IONPs@curcumin exhibited the best stability and dispersion conditions. Then, cytotoxicity tests of IONPs@curcumin were carried out on the MDA-MB-468 triple-negative breast cancer cell line, under experimental conditions without irradiation and subjected to PDT. The results revealed a viability greater than 70%, as it did not exhibit cytotoxicity for cells in the dark. After 1 h of incubation, the PDT associated with IONPs@curcumin showed 32% of cell viability at a concentration of 30 mg/mL.Item Nano-Enabled Colorimetric Assay for the Detection of Paracoccidioides lutzii: Advancing Diagnostics with Nanotechnology(MDPI) Comparato Filho, Olavo de Osti; Cândido, Marcela Aparecida; Ventura, Aveline; Morais, Flavia Villaça; Raniero, Leandro JoséDeforestation is a common occurrence driven by agricultural expansion, urbanization, and infrastructure development. These activities often lead to increased human interaction with ecosys- tems, potentially exposing individuals to Paracoccidioides spores (P. brasiliensis and P. lutzii) found in the soil, resulting in Paracoccidioidomycosis (PCM). This fungal infection is endemic to specific regions in Latin America, such as Brazil, Colombia, Venezuela, and Argentina. Diagnosis typically involves a combination of clinical assessment, imaging techniques, and laboratory examinations. P. lutzii lacks the glycoprotein Gp43, a key antigenic protein utilized in serological tests for PCM diagnosis. In this study, a colorimetric test employing gold nanoparticles (AuNPs) and label-free methodology was employed for P. lutzii detection. The effectiveness of the label-free colorimetric test was assessed using a total of 100 samples. This detection was achieved through the amplification of the gp43 gene and the use of a specific probe (5′CAGGGGTGCG3′) in conjunction with AuNPs. The receiver operating characteristic curve was employed to assess the test, revealing that the method can accurately detect P. lutzii with a sensitivity of 100% and a specificity of 100%. The findings indicate a substantial impact on remote endemic regions attributable to the implementation of cost-effective diagnostic methodologies.