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Item Aplicação de bionanossensor para diagnóstico da superexpressão do EGFR in vitro(2023-03-07) Raniero, Leandro José; Soares, Cristina Pacheco; Campos, Andrea Porto Carreiro; Vieira, Paula Fonseca Antunes; São José dos CamposO Receptor do Fator de Crescimento Epidérmico (EGFR) é codificado pelo gene EGFR. A superexpressão do receptor EGFR pode estar ligada a malignidades com pior prognóstico e suas vias de sinalização EGFR são ativadas pela proteína EGF, regulando as ações celulares. Como resultado, o receptor EGFR está sendo estudado para uma variedade de diagnósticos de tumores, estimulando o desenvolvimento de abordagens inovadoras para aumentar a qualidade e a eficiência dos exames diagnósticos. Os nanomateriais podem reconhecer células cancerígenas por direcionamento específico de vias moleculares, ressaltando a importância da nanomedicina. Neste estudo, bionanossensores foram sintetizados para detecção da EGFR, pelo processo de funcionalização da proteína EGF e Clorina e6 (Ce6) em nanopartículas de ouro. Essas nanopartículas utilizam o direcionamento ativo para entrega da proteína EGF ao receptor EGFR, e a Ce6 serve como uma molécula marcadora fluorescente. Bionanossensores foram examinados in vitro em linhagens celulares MDA-MB-468 e M059J. A distribuição, absorção, internalização e intensidade de fluorescência dos bionanossensores in vitro foi determinada por Microscopia Confocal e Citometria de Fluxo. Os resultados mostram que ambas as linhagens celulares analisadas acumulam bionanossensores, no entanto, a MDA-MB- 468 possui maior intensidade devido às suas propriedades de superexpressão do receptor EGFR. Os achados apontam para propriedades ideais para a concepção de um diagnóstico inovador, cuja acurácia se dará pela detecção de células superexpressas do receptor de EGFR em estágios precoces do câncer.Item Specific nanomarkers fluorescence: in vitro analysis for EGFR overexpressed cells in triple-negative breast cancer and malignant glioblastoma(Elsevier) Vieira, Paula Fonseca Antunes; Jesus, Viviane Paula dos Santos; Cândido, Marcela Aparecida; Pacheco-Soares, Cristina; Castilho, Maiara Lima; Raniero, Leandro JoséBackground: Epidermal Growth Factor Receptor (EGFR receptor) is encoded by the EGFR gene. EGFR receptor signaling pathways are activated by EGF protein, regulating cell actions. Overexpression of EGFR receptor may be linked to malignancies with a poor prognosis. As a result, EGFR receptor is being studied for a variety of tumor diagnostics, spurring the development of innovative approaches to increase quality and efficiency. Nanomaterials can recognize cancer cells by specifically targeting of molecular pathways, underscoring the importance of nanomedicine. In this study, we synthesized EGFR-specific nanomarkers by functionalizing EGF protein and Chlorin e6 in gold nanoparticles. These nanoparticles use active targeting to deliver EGF protein to EGFR receptor, and Chlorin e6 serves as a fluorescent marker molecule Methods: Nanomarkers were examined in vitro in MDA-MB-468 and M059J cell lines. Confocal microscopy and flow cytometry were used to examine the distribution, uptake, internalization, and fluorescence intensity of nanomarkers in vitro Results: The results show that both lines examined accumulate nanomarkers. However, MDA-MB-468 had the highest intensity due to its EGFR receptor overexpression properties Conclusion: The findings point to ideal properties for detecting EGFR receptor overexpressed cells.Item Chlorin e6-EGF conjugated gold nanoparticles as a nanomedicine based therapeutic agent for triple negative breast cancer(Elsevier) Castilho, Maiara Lima; Jesus, Viviane Paula dos Santos; Vieira, Paula Fonseca Antunes; Hewitt, Kevin Cecil; Raniero, Leandro JoséTo develop a treatment modality for triple-negative breast cancer, we investigated the efficacy of a bifunctional theranostic nanoprobes (BN) during Photodynamic Therapy (PDT) on human breast carcinoma and normal human cells. The BN is a 21 nm gold nanoparticles functionalized with Chlorin e6 (Ce6) and Epidermal Growth Factor (EGF). Attachment to gold nanoparticle stabilizes Ce6 while EGF acts as a cancer cell targeting agent. Fluorescence Spectroscopy and Confocal Fluorescence Microscopy revealed a gradual uptake of nanoprobes into cancer cells at an average rate of 63 BN/min. Cell viability assays showed that 0.2 μg/mL BN concentration was highly cytotoxic to cancer cells (86 %), but not normal cells. At this concentration, 58 % cancer cells were necrotic and 38 % apoptotic, while the reactive oxygen species (ROS) was 9-fold higher in cancer cells compared to normal. Overall, results suggest that BN mediated PDT can achieve targeted cancer cell death with high efficiency.