By Type: Journal Article

15 years on siRNA delivery: Beyond the State-of-the-Art on inorganic nanoparticles for RNAi therapeutics, Conde, João, Ambrosone Alfredo, Hernandez Yulán, Tian Furong, McCully Mark, Berry Catherine C., Baptista Pedro V., and T Claudia , NANO TODAY, Volume In Press, (2015) Abstract

RNAi has always captivated scientists due to its tremendous power to modulate the phenotype of living organisms. This natural and powerful biological mechanism can now be harnessed to down-regulate specific gene expression in diseased cells; opening up endless opportunities. Since most of the conventional siRNA delivery methods are limited by a narrow therapeutic index and significant side and off-target effects, we are now in the dawn of a new age in gene therapy driven by nanotechnology vehicles for RNAi therapeutics. Here, we outlook the "do's and dont's" of the inorganic RNAi nanomaterials developed in the last 15 years and the different strategies employed are compared and scrutinized, offering important suggestions for the next 15.

RNAi-based glyconanoparticles trigger apoptotic pathways for in vitro and in vivo enhanced cancer-cell killing, Conde, Joao, Tian Furong, Hernandez Yulan, Bao Chenchen, Baptista Pedro V., Cui Daxiang, Stoeger Tobias, and de la Fuente Jesus M. , Nanoscale, Volume 7, Number 19, p.9083-9091, (2015) Abstract

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Characterization of antiproliferative potential and biological targets of a copper compound containing 4'-phenyl terpyridine, Mendo, Ana Soraia, Figueiredo Sara, Roma-Rodrigues Catarina, Videira Paula A., Ma Zhen, Diniz Mário, Larguinho Miguel, Costa Pedro, Lima João C., Pombeiro Armando J. L., Baptista Pedro V., and Fernandes Alexandra R. , JBIC Journal of Biological Inorganic Chemistry, Volume 2, Issue 6, p.935-948, (2015) AbstractWebsite

Several copper complexes have been assessed as anti-tumor agents against cancer cells. In this work, a copper compound [Cu(H2O){OS(CH3)2}L](NO3)2 incorporating the ligand 4'-phenyl-terpyridine antiproliferative activity against human colorectal, hepatocellular carcinomas and breast adenocarcinoma cell lines was determined, demonstrating high cytotoxicity. The compound is able to induce apoptosis and a slight delay in cancer cell cycle progression, probably by its interaction with DNA and induction of double-strand pDNA cleavage, which is enhanced by oxidative mechanisms. Moreover, proteomic studies indicate that the compound induces alterations in proteins involved in cytoskeleton maintenance, cell cycle progression and apoptosis, corroborating its antiproliferative potential.

Significance of the balance between intracellular glutathione and polyethylene glycol for successful release of small interfering RNA from gold nanoparticles, McCully, Mark, Hernandez Yulan, Conde Joao, Baptista Pedro V., de la Fuente Jesus M., Hursthouse Andrew, Stirling David, and Berry Catherine C. , Nano Research, Volume 8, Number 10, p.3281-3292, (2015) Abstract

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POxylated Polyurea Dendrimers: Smart Core-Shell Vectors with IC50 Lowering Capacity, Restani, Rita B., Conde Joao, Pires Rita F., Martins Pedro, Fernandes Alexandra R., Baptista Pedro V., and Bonifacio Vasco D. B. , MACROMOLECULAR BIOSCIENCE, Volume 15, Issue 8, p.1045–1051, (2015) AbstractWebsite

The design and preparation of highly efficient drug delivery platforms using green methodologies is at the forefront of nanotherapeutics research. POxylated polyurea dendrimers are efficiently synthesized using a supercritical-assisted polymerization in carbon dioxide. These fluorescent, pH-responsive and water-soluble core-shell smart nanocarriers show low toxicity in terms of cell viability and absence of glutathione depletion, two of the major side effect limitations of current vectors. The materials are also found to act as good transfection agents, through a mechanism involving an endosomal pathway, being able to reduce 100-fold the IC50 of paclitaxel.

Mobile based gold nanoprobe TB diagnostics for point-of-need., Veigas, B., Fortunato E., and Baptista P. V. , Methods in molecular biology (Clifton, N.J.), Volume 1256, p.41-56, (2015) Abstract

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Heterocyclic Anticancer Compounds: Recent Advances and the Paradigm Shift towards the Use of Nanomedicine’s Tool Box, Martins, Pedro, Jesus João, Santos Sofia, Raposo Luis R., Roma-Rodrigues Catarina, Baptista Pedro Viana, and Fernandes Alexandra , Molecules, Issue 20, p.16852-16891, (2015) AbstractWebsite

The majority of heterocycle compounds and typically common heterocycle fragments present in most pharmaceuticals currently marketed, alongside with their intrinsic versatility and unique physicochemical properties, have poised them as true cornerstones of medicinal chemistry. Apart from the already marketed drugs, there are many other being investigated for their promising activity against several malignancies. In particular, anticancer research has been capitalizing on the intrinsic versatility and dynamic core scaffold of these compounds. Nevertheless, as for any other promising anticancer drugs, heterocyclic compounds do not come without shortcomings. In this review, we provide for a concise overview of heterocyclic active compounds and families and their main applications in medicine. We shall focus on those suitable for cancer therapy while simultaneously addressing main biochemical modes of action, biological targets, structure-activity relationships as well as intrinsic limitation issues in the use of these compounds. Finally, considering the advent of nanotechnology for effective selective targeting of drugs, we shall discuss fundamental aspects and considerations on nanovectorization of such compounds that may improve pharmacokinetic/pharmacodynamic properties of heterocycles.

Gold nanoprobe-based non-crosslinking hybridization for molecular diagnostics, Larguinho, Miguel, Canto Rafaela, Cordeiro Milton, Pedrosa Pedro, Fortuna Andreia, Vinhas Raquel, and Baptista Pedro V. , Expert Review of Molecular Diagnostics, Volume 15, Number 10, p.1355-1368, (2015) Abstract

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Gold Nanotheranostics: Proof-of-Concept or Clinical Tool?, Pedrosa, Pedro, Vinhas Raquel, Fernandes Alexandra, and Baptista Pedro V. , Nanomaterials, Volume 5, Issue 4, p.1853-1879, (2015) AbstractWebsite

Nanoparticles have been making their way in biomedical applications and personalized medicine, allowing for the coupling of diagnostics and therapeutics into a single nanomaterial—nanotheranostics. Gold nanoparticles, in particular, have unique features that make them excellent nanomaterials for theranostics, enabling the integration of targeting, imaging and therapeutics in a single platform, with proven applicability in the management of heterogeneous diseases, such as cancer. In this review, we focus on gold nanoparticle-based theranostics at the lab bench, through pre-clinical and clinical stages. With few products facing clinical trials, much remains to be done to effectively assess the real benefits of nanotheranostics at the clinical level. Hence, we also discuss the efforts currently being made to translate nanotheranostics into the market, as well as their commercial impact.

The significance of the balance between intracellular glutathione and polyethylene glycol (PEG) for successful siRNA release from gold nanoparticles, McCully, Mark, Hernandez Yulan, Conde João, Baptista Pedro V., de la Fuente Jesus M., Hursthouse Andrew, and St David , Nano Research, (2015) AbstractWebsite

The therapeutic promise of small interfering RNAs (siRNAs) for specific gene silencing is dependent on the
successful delivery of functional siRNAs to the cell cytoplasm. Their conjugation to an established delivery
platform, such as gold nanoparticles, offers a huge potential for treating diseases and advancing our
understanding of cellular processes. The success or failure is dependent on both the uptake of the nanoparticlesinto the cells and subsequent intracellular release of the functional siRNA. In this paper, utilising gold nanoparticle siRNA-mediated delivery against C-MYC, we aimed to determine if we could achieve knockdown in a cancer cell line with low levels of intracellular glutathione, and determine the influence, if any, of polyethylene glycol (PEG) ligand density on knockdown, with a view to determine the optimal nanoparticle
design to achieve C-MYC knockdown. We demonstrate that, regardless of the PEG density, knockdown in cells with relatively low glutathione levels can be achieved, and also the possible effect of steric hindrance in terms of PEG on the availability of the siRNA for cleavage in the intracellular environment. Gold nanoparticle uptake was demonstrated via transmission electron microscopy and mass spectroscopy, whilst knockdown was determined at the protein and physiological levels (cells in S-phase) by in-cell westerns and BrdU incorporation, respectively.

Gold nanoparticle-based theranostics: disease diagnostics and treatment using a single nanomaterial, Vinhas, Raquel, Cordeiro Milton, Carlos Fábio, Mendo Soraia, Fernandes Alexandra, Figueiredo Sara, and Baptista Pedro , Journal of Nanobiosensors in Disease Diagnosis, Issue 4, p.11-23, (2015) AbstractWebsite

Nanotheranostics takes advantage of nanotechnology-based systems in order to diagnose and treat a specific disease. This approach is particularly relevant for personalized medicine, allowing the detection of a disease at an early stage, to direct a suitable therapy toward the target tissue based on the molecular profile of the altered phenotype, subsequently facilitating disease monitoring and following treatment. A tailored strategy also enables to reduce the off-target effects associated with universal treatments and improve the safety profile of a given treatment. The unique optical properties of gold nanoparticles, their ease of surface modification, and high surface-to-volume ratio have made them central players in this area. By combining imaging, targeting, and therapeutic agents in a single vehicle, these nanoconjugates are (ought to be) an important tool in the clinics. In this review, the multifunctionality of gold nanoparticles as theranostics agents will be highlighted, as well as the requirements before the translation of these nanoplatforms into routine clinical practice.

DNA adduct identification using gold-aptamer nanoprobes, Larguinho, Miguel, Santos Sofia, Almeida Joao, and Baptista Pedro V. , Iet Nanobiotechnology, Volume 9, Number 2, p.95-101, (2015) Abstract

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Field Effect Sensors for Nucleic Acid Detection: Recent Advances and Future Perspectives, Veigas, Bruno, Fortunato Elvira, and Baptista Pedro V. , Sensors, Volume 15, Number 5, p.10380-10398, (2015) Abstract

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3h Gold nanobeacons: a potential nanotheranostics platform, Baptista, Pedro Viana , Nanomedicine, Volume 9, Number 15, p.2247-2250, (2014) Abstract

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Insights into the mechanisms underlying the antiproliferative potential of a Co(II) coordination compound bearing 1,10-phenanthroline-5,6-dione: DNA and protein interaction studies, Luis, Daniel V., Silva Joana, Tomaz Ana Isabel, de Almeida Rodrigo F. M., Larguinho Miguel, Baptista Pedro V., Martins Luisa M. D. R. S., Silva Telma F. S., Borralho Pedro M., Rodrigues Cecilia M. P., Rodrigues Antonio S., Pombeiro Armando J. L., and Fernandes Alexandra R. , Journal of Biological Inorganic Chemistry, Volume 19, Number 6, p.787-803, (2014) Abstract

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Gold-Nanobeacons for gene therapy: evaluation of genotoxicity, cell toxicity and proteome profiling analysis, Conde, João, Larguinho Miguel, Cordeiro Ana, Raposo Luis R., Costa Pedro M., Santos Susana, Diniz Mário, Fernandes Alexandra R., and Baptista Pedro Viana , Nanotoxicology, Volume 8, Issue 5, p.521-532, (2014) AbstractWebsite

Antisense therapy is a powerful tool for post-transcriptional gene silencing suitable for down-regulating target genes associated to disease. Gold nanoparticles have been described as effective intracellular delivery vehicles for antisense oligonucleotides providing increased protection against nucleases and targeting capability via simple surface modification. We constructed an antisense gold-nanobeacon consisting of a stem-looped oligonucleotide double-labelled with 3′-Cy3 and 5′-Thiol-C6 and tested for the effective blocking of gene expression in colorectal cancer cells. Due to the beacon conformation, gene silencing was directly detected as fluorescence increases with hybridisation to target, which can be used to assess the level of silencing. Moreover, this system was extensively evaluated for the genotoxic, cytotoxic and proteomic effects of gold-nanobeacon exposure to cancer cells. The exposure was evaluated by two-dimensional protein electrophoresis followed by mass spectrometry to perform a proteomic profile and 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (MTT) assay, glutathione-S-transferase assay, micronucleus test and comet assay to assess the genotoxicity. This integrated toxicology evaluation showed that the proposed nanotheranostics strategy does not exhibit significant toxicity, which is extremely relevant when translating into in vivo systems.

Low cost, safe, disposable, rapid and self-sustainable paper-based platform for diagnostic testing - Lab-on-Paper, Costa, Mafalda, Veigas Bruno, Jacob Jorge, Santos David, Gomes Jacinto, Baptista Pedro V., Martins Rodrigo, Inácio João, and Fortunato Elvira , NANOTECHNOLOGY, Volume 9, Issue 25, p.094006, (2014) AbstractWebsite

There is a strong interest in the use of biopolymers in the electronic and biomedical industries, mainly towards low-cost applications. The possibility of developing entirely new kinds of products based on cellulose is of current interest, in order to enhance and to add new functionalities to conventional paper-based products. We present our results towards the development of paper-based microfluidics for molecular diagnostic testing. Paper properties were evaluated and compared to nitrocellulose, the most commonly used material in lateral flow and other rapid tests. Focusing on the use of paper as a substrate for microfluidic applications, through an eco-friendly wax-printing technology, we present three main and distinct colorimetric approaches: (i) enzymatic reactions (glucose detection); (ii) immunoassays (antibodies anti-Leishmania detection); (iii) nucleic acid sequence identification (Mycobacterium tuberculosis complex detection). Colorimetric glucose quantification was achieved through enzymatic reactions performed within specific zones of the paper-based device. The colouration achieved increased with growing glucose concentration and was highly homogeneous, covering all the surface of the paper reaction zones in a 3D sensor format. These devices showed a major advantage when compared to the 2D lateral flow glucose sensors, where some carryover of the coloured products usually occurs. The detection of anti-Leishmania antibodies in canine sera was conceptually achieved using a paper-based 96-well enzyme-linked immunosorbent assay format. However, optimization is still needed for this test, regarding the efficiency of the immobilization of antigens on the cellulose fibres. The detection of Mycobacterium tuberculosis nucleic acids integrated with a non-cross-linking gold nanoprobe detection scheme was also achieved in a wax-printed 384-well paper-based microplate, by the hybridization with a species-specific probe. The obtained results with the above-mentioned proof-of-concept sensors are thus promising towards the future development of simple and cost-effective paper-based diagnostic devices.

Antibody-drug gold nanoantennas with Raman spectroscopic fingerprints for in vivo tumour theranostics, Conde, Joao, Bao Chenchen, Cui Daxiang, Baptista Pedro V., and Tian Furong , Journal of Controlled Release, Volume 183, p.87-93, (2014) Abstract

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Star-shaped magnetite@gold nanoparticles for protein magnetic separation and SERS detection, Quaresma, Pedro, Osorio Ines, Doria Goncalo, Carvalho Patricia A., Pereira Andre, Langer Judith, Araujo Joao Pedro, Pastoriza-Santos Isabel, Liz-Marzan Luis M., Franco Ricardo, Baptista Pedro V., and Pereira Eulalia , Rsc Advances, Volume 4, Number 8, p.3659-3667, (2014) AbstractWebsite

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Revisiting 30 years of biofunctionalization and surface chemistry of inorganic nanoparticles for nanomedicine, Conde, Joao, Dias Jorge T., Grazu Valeria, Moros Maria, Baptista Pedro V., and de la Fuente Jesus M. , Frontiers in Chemistry, Volume 2, (2014) Abstract

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Experimental optimization of a passive planar rhombic micromixer with obstacles for effective mixing in a short channel length, Bernacka-Wojcik, Iwona, Ribeiro Susana, Wojcik Pawel Jerzy, Alves Pedro Urbano, Busani Tito, Fortunato Elvira, Baptista Pedro Viana, Covas Jose Antonio, Aguas Hugo, Hilliou Loic, and Martins Rodrigo , Rsc Advances, Volume 4, Number 99, p.56013-56025, (2014) Abstract

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Histopathological findings on Carassius auratus hepatopancreas upon exposure to acrylamide: correlation with genotoxicity and metabolic alterations., Larguinho, Miguel, Costa Pedro M., Sousa Gonçalo, Costa Maria H., Diniz Mário S., and Baptista Pedro V. , Journal of Applied Toxicology , Volume 34, Issue 12, (2014)
A promising road with challenges: where are gold nanoparticles in translational research?, Bao, Chenchen, Conde Joao, Polo Ester, del Pino Pablo, Moros Maria, Baptista Pedro V., Grazu Valeria, Cui Daxiang, and de la Fuente Jesus M. , Nanomedicine, Volume 9, Number 15, p.2353-2370, (2014) Abstract

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Anti-cancer precision theranostics: a focus on multifunctional gold nanoparticles, Cabral, Rita M., and Baptista Pedro V. , Expert Review of Molecular Diagnostics, Volume 14, Number 8, p.1041-1052, (2014) Abstract

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Dual-color control of nucleotide polymerization sensed by a fluorescence actuator, Reimão-Pinto, M. M., Cordeiro A., Almeida C., Pinheiro A. V., Moro A., Lima J. C., and Baptista P. V. , Photochem Photobiol Sci, Volume 13, Number 5, p.751-6, (2014) AbstractWebsite

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