Preliminary evaluation of zeolite-based platforms as potential dual drug delivery systems against microbial infections in the tumor microenvironment

Several zeolite-based delivery systems (ZDS) built with faujasite structure were prepared containing silver (Ag+) and 5-Fluorouracil (5-FU) as antimicrobial and antineoplastic agents, respectively. The idea behind this drug combination is an answer to the increasing evidence of colonization of tumor microenvironments by pathogenic microorganisms and their active role in tumor growth. Two ZDS with a fixed load of 5-FU and different silver loads, Ag7(5-FU).info:eu-repo/semantics/publishedVersio

Machine learning-assisted optimization of drug combinations in zeolite-based delivery systems for melanoma therapy

Two independent artificial neural network (ANN) models were used to determine the optimal drug combination of zeolite-based delivery systems (ZDS) for cancer therapy. The systems were based on the NaY zeolite using silver (Ag+) and 5-fluorouracil (5-FU) as antimicrobial and antineoplastic agents. Different ZDS samples were prepared, and their characterization indicates the successful incorporation of both pharmacologically active species without any relevant changes to the zeolite structure. Silver acts as a counterion of the negative framework, and 5-FU retains its molecular integrity. The data from the A375 cell viability assays, involving ZDS samples (solid phase), 5-FU, and Ag+ aqueous solutions (liquid phase), were used to train two independent machine learning (ML) models. Both models exhibited a high level of accuracy in predicting the experimental cell viability results, allowing the development of a novel protocol for virtual cell viability assays. The findings suggest that the incorporation of both Ag and 5-FU into the zeolite structure significantly potentiates their anticancer activity when compared to that of the liquid phase. Additionally, two optimal AgY/5-FU@Y ratios were proposed to achieve the best cell viability outcomes. The ZDS also exhibited significant efficacy against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus); the predicted combination ratio is also effective against S. aureus, underscoring the potential of this approach as a therapeutic option for cancer-associated bacterial infections.FCT - Fundação para a Ciência e a Tecnologia (UIDB/04469/2020).A.R.B. and V.I. express their gratitude to the Portuguese Foundation for Science and Technology (FCT) for providing funding through the Ph.D. Grants SFRH/BD/141058/2018 and UI/BD/152219/2021, respectively. This research work has received financial support from national funds provided by FCT/MCTES (PIDDAC) under the projects UID/QUI/0686/2020 (CQ-UM), UIDB/04469/2020 (CEB), and UIDP/50026/2020 (ICVS). Additionally, the projects of BioTecNorte (operation NORTE-01-0145-FEDER-000004 and NORTE-01-0145-FEDER-000055) are supported by the Northern Portugal Regional Operational Program (NORTE 2020) under the Portugal 2020 Partnership Agreement, cofunded by the European Regional Development Fund (ERDF). This work was also supported by the “Contrato Programa” UIDB/04050/2020 funded by national funds through the FCT I.P. The authors also thank Patrícia R. Correia for their contribution to cell viability studies

Nanoestruturas à base de silício como sistemas de libertação de fármacos para aplicações biológicas e médicas

Tese de doutoramento em Ciências da SaúdeRecentemente, a nanomedicina tem oferecido vantagens promissoras para os desafios colocados pelas abordagens convencionais utilizadas na terapia e diagnóstico do cancro. Um design cuidadoso pode facilitar o desenvolvimento de nanopartículas versáteis, com múltiplos atributos terapêuticos e/ou diagnósticos numa única estrutura. Entre os nanomateriais disponíveis, as nanopartículas à base de silício apresentam vários benefícios, como evidenciado nas diversas aplicações biomédicas documentadas na literatura. Assim, este projeto teve como objetivo desenvolver nanopartículas à base de silício com especial destaque para o seu potencial no campo da terapia do cancro. Resultados recentes sugerem que bactérias intratumorais podem desempenhar um papel na resistência às drogas anticancerígenas frequentemente utilizadas, contribuindo para a complexidade do tratamento. Por isso, a exploração das propriedades das nanopartículas culminou na criação de diferentes nanossistemas com dupla ação anticancerígena e antibacteriana. Mais especificamente, este trabalho de doutoramento incluiu o uso de materiais baseados em silício como zeólitos, nanopartículas de silício poroso e sílica mesoporosa. O estudo começou com o desenvolvimento de zeólitos com corantes fluorescentes, como potenciais agentes de bioimagem, económicos e estáveis, em células de cancro da mama, que revelaram propriedades luminescentes multicoloridas. Além disso, explorou-se a incorporação de 5-fluorouracilo e prata na estrutura de zeólitos faujasite para aplicações anticancerígenas e antibacterianas. Foram usadas ferramentas de aprendizagem automática para determinar combinações ótimas de sistemas de zeólitos, posteriormente validadas por resultados experimentais, num modelo de cancro de pele. O projeto também envolveu o desenvolvimento de um nanosistema duplo com propriedades antibacterianas e antitumorais, usando nanopartículas de silício poroso para a entrega simultânea de doxorrubicina e prata. Finalmente, explorou-se o uso de nanopartículas de sílica mesoporosa em combinação com outros materiais para criar uma plataforma híbrida e teranóstica, com propriedades magnéticas. O design foi desenvolvido para obter propriedades anticancerígenas e antibacterianas, através da combinação de doxorrubicina e ciprofloxacina, respetivamente. Em resumo, os nanossistemas à base de silício desenvolvidos neste trabalho, representam uma contribuição valiosa para uma melhor compreensão das suas potenciais aplicações biomédicas.In recent years, nanomedicine has been offering promising solutions to the challenges posed by conventional approaches used in cancer diagnosis and therapy. Achieving an optimal balance between the nanoparticle's advantages and disadvantages is an ongoing challenge. The creation of nanoparticles containing multiple therapeutic and/or diagnostic attributes within a single structure requires a careful design. Among available nanomaterials, silicon-based nanoparticles provide several advantages, evident by the extensive research and diverse biomedical applications documented in the literature. Thus, this dissertation aimed to develop silicon-based nanoparticles tailored for biomedical applications, with a particular focus on their potential in the field of cancer therapy. Emerging evidence suggests that intratumor bacteria have a role in the resistance to commonly used anticancer drugs, contributing to the treatment's complexity. Thus, the nanoparticle properties were explored to create dual-action systems with both anticancer and antibacterial activities. More specifically, this work included the use of materials based on silicon such as zeolites, porous silicon, and mesoporous materials. The study of zeolites started with the development of fluorescent dye-containing zeolites as potential cost-effective and stable bioimaging agents in breast cancer cells, revealing multicolor luminescent properties. Additionally, the incorporation of 5-fluorouracil and silver into a faujasite zeolite structure was explored for anticancer and antibacterial applications. Machine learning tools were used to determine optimal zeolite system combinations, further validated by experimental data in a skin cancer cell model. The project also involved the development of a dual nanocarrier with antibacterial and anticancer properties using porous silicon nanoparticles for targeted delivery of doxorubicin and silver. Finally, the use of mesoporous silica nanoparticles was explored in combination with other materials to yield a hybrid theranostic platform with magnetic properties. The nanoparticles were tailored to display dual anticancer and antibacterial properties, by the combination of doxorubicin with ciprofloxacin, respectively. In summary, the silicon-based nanosystems developed within this thesis present a valuable contribution to unveiling their potential for applications in the biomedical field.I would like to thank the European Union, Portuguese Government, and Portuguese Foundation for Science and Technology for the funding (SFRH/BD/141058/2018 and COVID/BD/152997/2022) and the opportunity to work on my PhD thesis, in the scope of the Portugal 2020 and Norte 2020 programs. I would like to express my appreciation to the Luso-American Development Foundation (FLAD) for the FLAD Grant R&D@PhD 2021

On-site and rapid optical assay to test biological samples

Dissertação de mestrado em Técnicas de Caracterização e Análise QuímicaCurrently there is a need for diagnosis processes, which uses urine samples and offer quick, easy, and accurate results, since most of the traditional systems require time, material, facilities, and specialized personnel, which can affect the efficiency of the therapy chosen for the patient. In this context, this work reports two different strategies to improve the stability and latency of the results provided by a device specifically developed for the analysis of urine samples on diapers and early medical condition assessment. The main goal was to obtain stable, rapid, and easily interpretable results, by employing colorimetric detection, without the need of electricity to analyze several urine biomarkers. The device design already comprises components to provide the conditions for the results stability, such as a self-locking system and a cover layer. These same conditions were tested, and the outcomes suggest that the device has great potential to keep valid results for long periods. The encapsulation of the dye molecules, normally used in the bioassays, into hosts like zeolites nanostructures, which were used to enhance color stability, was also studied in order to have a chemical approach to this issue. These obtained dye nanomaterials were characterized by employing several techniques (Room Temperature Fourier Transform Infrared (FTIR), N2 adsorption isotherms, scanning electron microscopy (SEM/EDX), thermogravimetric analysis (TGA) and powder X-ray diffraction (XRD)) and, the results show that the encapsulation of the dyes was successfully achieved, preserving both the guest and host structures. Regarding the colorimetric results, a long-time stability of the dye colors was achieved due to their encapsulation into the zeolite nanostructures. The choice of the type of paper for the μPAD and the zeolite nanostructures properties influences the colorimetric response. The results obtained in this work were promising for the studied assays, and the strategy could also be extended to other biomarkers.Atualmente, existe uma necessidade de processos de diagnóstico que utilizem amostras de urina e ofereçam resultados rápidos, fáceis e precisos, uma vez que a maioria dos métodos tradicionais requerem tempo, material, instalações e pessoas especializadas, o que pode afetar a eficiência da terapia escolhida para o paciente. Neste contexto, este trabalho explora duas estratégias diferentes para melhorar a estabilidade e latência dos resultados obtidos por um dispositivo especificamente desenvolvido para a análise de urina em fraldas e avaliação médica prévia (monitorização da saúde). O objetivo principal foi obter resultados estáveis, rápidos e fáceis de interpretar, ao utilizar a deteção colorimétrica, sem necessidade de eletricidade para analisar vários biomarcadores da urina. O design do dispositivo já contém componentes para fornecer as condições para a estabilidade dos resultados, tal como um sistema de fecho automático e uma camada de cobertura. Estas mesmas condições foram testadas e os resultados sugerem que o dispositivo tem grande potencial para manter a validade dos resultados por longos períodos de tempo. O encapsulamento de moléculas de corantes, normalmente utilizadas em ensaios biológicos, em hospedeiros como zeólitos, no sentido de aumentar a estabilidade das cores das moléculas, foi também estudado com o objetivo de explorar uma abordagem química ao problema. Os nanomateriais obtidos foram caracterizados por várias técnicas (Espetroscopia de infravermelhos (FTIR), isotérmicas de adsorção de N2, microscopia eletrónica de varrimento (SEM/EDX), análise termogravimétrica (TGA) e difração de raio-X (XRD)) e os resultados confirmam que o encapsulamento das moléculas, com a preservação das estruturas dos corantes e dos respetivos hospedeiros. Em relação aos resultados colorimétricos, foi alcançada uma estabilidade das cores dos corantes por um longo período de tempo devido ao seu encapsulamento nas estruturas dos zeólitos. A escolha do tipo de papel para os μPADs e as propriedades dos zeólitos demostraram ter influência na resposta colorimétrica. Os resultados obtidos neste trabalho são promissores para os ensaios estudados, e o método pode também ser estendido a outros biomarcadores

A doçaria conventual vila-condense

A “Doçaria Conventual Vila-Condense” é a compilação de todos os dados, observações e aprendizagens obtidas durante a condução de um projeto que se centrou na compreensão da confeção de doces conventuais em Vila do Conde, no papel e caraterísticas das doceiras e na função desempenhada pelos estabelecimentos que os comercializam. Ao longo desta memória são abordadas a história desta tradição na cidade, a possibilidade do seu poder identitário, as considerações mais teóricas e práticas relativas à confeção dos doces, o trajeto percorrido desde a recolha de materiais bibliográficos até à conclusão do filme “Meias-luas”, e todas as vantagens, particularidades e dificuldades associadas à escolha de um meio visual para a apresentação de um projeto do foro académico, sem descurar os aspetos artísticos, criativos e estéticos indispensáveis à produção de um filme.Doçaria Conventual Vila-Condense” is the collection of all the data, observations and learnings obtained during the course of a project focused on the comprehension of the making of Vila do Conde’s convent sweets, on the role played by the sweet’s confectioners, and on the function displayed by the establishments which sell them. Throughout this memoire we will deal with the history of the tradition, with the possibility of an identity trait, with the more theorical and practical considerations regarding the making of the sweets, with the path that had to be trailed from the gathering of bibliographical information to the conclusion of the film “Meias-luas”, and with all the advantages, particularities and difficulties connected to the choosing of a visual medium of presentation of an academic project, without neglecting all of the artistic, creative and aesthetic features which are detachable from the production of a film

Recycling of natural and waste materials as supports for green silver nanoparticles as efficient catalysts in photodegradation of organic pollutants

The use of a sustainable technology to synthesize green silver nanoparticles (AgNP) from renewable resources is very appealing. Here, a green protocol for the production AgNP/composites is introduced, via in situ reduction of Ag+ ions on sustainable materials (clays and waste materials) using eucalyptus leaves extract (ELE) as a reducing and stabilizing agent, without the addition of any harmful or expensive organic solvent, surfactant or dangerous compounds. The synthesized AgNP/composites were evaluated as photocatalysts to degrade Indigo Carmine (IC) dye as a model pollutant under visible light. Kaolin and pistachio shells showed to be promising supports for green AgNP, with decolorization efficiency of 85.3(±1.8)% and 48.3(±4.1)%, respectively, after 120min of irradiation. The best catalytic results were obtained with kaolin as support and so the influence of different synthesis parameters of AgNP/kaolin nanocomposite was evaluated. The catalytic results indicated that increasing the amount of AgNO3 and ELE in the nanocomposites, increased their photocatalytic performance. Nanocomposites synthesized at room temperature showed smaller amount of AgNP and higher decolorization efficiency, compared to those biosynthesized at 50 °C. The evaluation of active species responsible for the photodegradation of IC showed that O2- and H+ radicals are species involved in the reaction. The AgNP/kaolin showed also to be efficient to degrade both atrazine and sulfamethoxazole after 120min of irradiation.info:eu-repo/semantics/publishedVersio

Doxorubicin delivery performance of superparamagnetic carbon multi-core shell nanoparticles: pH dependence, stability and kinetic insight

In the past decade, magnetic nanoparticles (MNPs) have been among the most attractive nanomaterials used in different fields, such as environmental and biomedical applications. The possibility of designing nanoparticles with different functionalities allows for advancing the biomedical applications of these materials. Additionally, the magnetic characteristics of the nanoparticles enable the use of magnetic fields to drive the nanoparticles to the desired sites of delivery. In this context, the development of new MNPs in new approaches for drug delivery systems (DDSs) for cancer treatment has increased. However, the synthesis of nanoparticles with high colloidal stability triggered drug delivery, and good biocompatibility remains a challenge. Herein, multi-core shell MNPs functionalized with Pluronic ® F-127 were prepared and thoroughly characterized as drug carriers for doxorubicin delivery. The functionalized nanoparticles have an average size of 17.71 ± 4.2 nm, high water colloidal stability, and superparamagnetic behavior. In addition, the nanoparticles were able to load 936 μg of DOX per mg of functionalized nanomaterial. Drug release studies at different pH values evidenced a pH-triggered DOX release effect. An increase of 62% in cumulative drug release was observed at pH simulating tumor endosome/lysosome microenvironments (pH 4.5) compared to physiological conditions (pH 7.4). In addition, an innovative dynamic drug delivery study was performed as a function of pH. The results from this test confirmed the pH-induced doxorubicin release capability of carbon multi-core shell MNPs. The validity of traditional kinetic models to fit dynamic pH-dependent drug release was also studied for predictive purposes.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and to the ERDF for financial support by funds FCT/MCTES to CIMO (UIBD/00690/2020) and RTChip4Theranostics (NORTE-01-0145-FEDER-029394). This work is a result of these projects. This research was funded by RTChip4Theranostics – Real time Liver-on-a-chip platform with integrated micro(bio)sensors for preclinical validation of graphene-based magnetic nanocarriers towards cancer theranostics, with the reference NORTE-01-0145-FEDER-029394, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (ERDF); and CIMO (UIDB/00690/2020) through ERDF under the Program PT2020. Adriano S. Silva thanks his doctoral Grant with reference SFRH/BD/151346/2021 financed by the Portuguese Foundation for Science and Technology (FCT), with funds from NORTE2020, under the MIT Portugal Program.info:eu-repo/semantics/publishedVersio

Modification of microfluidic paper-based devices with dye nanomaterials obtained by encapsulation of compounds in Y and ZSM5 zeolites

Zeolite nanostructures were used as hosts for dyes normally used in some bioassays (pH and protein), and applied on microfluidic paper-based analytical devices (PADs). The obtained dye nanomaterials were characterized by several techniques (structural (FTIR and XRD), surface (SEM/EDX), textural (N2 adsorption) analyses and dye loading determination by TGA analysis), to confirm the stability of both host and guest. After their deposition on the papers surface, the color change was studied by analyzing the RGB values, taking into account the lifetime and the results stability of the PADs. These results confirmed that both the lifetime and the color change of the modified devices were stable over one week. It was also demonstrated that the choice of the paper for the PAD as well as the zeolite nanostructures properties influences the colorimetric response. This strategy by encapsulating dyes into zeolites solve current challenges in colorimetric sensors in where long-term stability of the colorimetric results is an issue. The results obtained were promising for the studied assays, and the strategy could also be extended to other bioassays.A.R.B. thanks for the ERASMUS+ program to her grant. This work has been developed under the scope of the projects: BioTecNorte (NORTE-01-0145-FEDER-000004), AIProcMat@N2020 (NORTE-01-0145-FEDER-000006), projects supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). This work also has been funded by ERDF through COMPETE2020–Programa Operacional Competitividade e Internacionalização (POCI), Project POCI-01-0145-FEDER-006984, Associate Laboratory LSRE-LCM and by national funds through FCT, Fundacão para a Ciência e a Tecnologia for project PTDC/AAGTEC/5269/2014 and Centre of Chemistry (UID/QUI/00686/2013 and UID/QUI/0686/2016). The support from the Oslofjordfondet project, «Touchsensor for enklere og raskere urinprøvetaking og analyse (no. 234972)» and NSFC project no. 61650410655 is also acknowledged.info:eu-repo/semantics/publishedVersio

Surface functionalization of zeolite-based drug delivery systems enhances their antitumoral activity in vivo

Supplementary data to this article can be found online at https://doi. org/10.1016/j.msec.2020.111721.Zeolites have attractive features making them suitable carriers for drug delivery systems (DDS). As such, we loaded the anticancer drug 5-fluorouracil (5-FU), into two different zeolite structures, faujasite (NaY) and Linde Type L (LTL), to obtain different DDS. The prepared DDS were tested in vitro using breast cancer, colorectal carcinoma, and melanoma cell lines and in vivo using the chick embryo chorioallantoic membrane model (CAM). Both assays showed the best results for the Hs578T breast cancer cells, with a higher potentiation for 5-FU encapsulated in the zeolite LTL. To unveil the endocytic mechanisms involved in the internalization of the zeolite nanoparticles, endocytosis was inhibited pharmacologically in breast cancer and epithelial mammary human cells. The results suggest that a caveolin-mediated process was responsible for the internalized zeolite nanoparticles. Aiming to boost the DDS efficacy, the disc-shaped zeolite LTL outer surface was functionalized using amino (NH2) or carboxylic acid (COOH) groups and coated with poly-L-lysine (PLL). Positively functionalized surface LTL nanoparticles revealed to be non-toxic to human cells and, importantly, their internalization was faster and led to a higher tumor reduction in vivo. Overall, our results provide further insights into the mechanisms of interaction between zeolite-based DDS and cancer cells, and pave the way for future studies aiming to improve DDS anticancer activity.NV and ARB thank FCT (Foundation for Science and Technology) for their Ph.D. grants (SFRH/BD/97797/2013 and SFRH/BD/141058/2018, respectively). This work has been funded by ICVS Scientific Mi croscopy Platform, member of the national infrastructure PPBI - Portu guese Platform of Bioimaging (PPBI-POCI-01-0145-FEDER-022122); by National funds, through the Foundation for Science and Technology (FCT) - project UIDB/50026/2020 and UIDP/50026/2020; and by the projects NORTE-01-0145-FEDER-000013 and NORTE-01-0145-FEDER 000023, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). This work was also supported by FCT, under the scope of the projects: PTDC/ AAGTEC/5269/2014 and Centre of Chemistry (UID/QUI/00686/2013 and UID/QUI/0686/2016).info:eu-repo/semantics/publishedVersio