By Type: Journal Article

Martins, Pedro, Daniela Rosa, Alexandra R. Fernandes, and Pedro V. Baptista. "Nanoparticle Drug Delivery Systems: Recent Patents and Applications in Nanomedicine." Recent Patents on Nanomedicine 3 (2014): 105-118. AbstractWebsite

Traditional methods of drug delivery present several drawbacks, mainly due to off-target effects that may originate severe side and toxic effect to healthy tissues. Parallel to the development of novel more effective drugs, particular effort has been dedicated to develop and optimize drug delivery vehicles capable of specifically targeting the required tissue/organ and to deliver the cargo only where and when it is needed. New drug delivery systems based on nanoscale devices showing new and improved pharmacokinetic and pharmacodynamics properties like enhanced bioavailability, high drug loading or systemic stability have surged in the past decade as promising solutions to the required therapeutic efficacy. Amongst these nanoscale vectors, nanoparticles for drug delivery, such as polymeric, lipidbased, ceramic or metallic nanoparticles, have been at the forefront of pharmaceutical development. The interest in nanomedicine for treatment and diagnosis is clearly reflected on the increasing number of publications and issued patents every year. Here, we provide a broad overview of novel nanoparticle based drug delivery systems, ranging from polymeric systems to metal nanoparticles, while simultaneously listing the most relevant related patents.

Baptista, Pedro Viana. "Gold nanobeacons: A potential nanotheranostics platform." Nanomedicine 9 (2014): 2247-50.Website
Restani, Rita B., João Conde, Pedro V. Baptista, Maria Teresa Cidade, Ana M. Bragança, Jorge Morgado, Ilídio J. Correia, Ana Aguiar-Ricardo, and Vasco D. B. Bonifacio. "Polyurea dendrimer for efficient cytosolic siRNA delivery." RSC ADVANCES 4 (2014): 54872. AbstractWebsite

The design of small interfering RNA (siRNA) delivery materials showing efficacy in vivo is at the forefront of nanotherapeutics research. Polyurea (PURE-type) dendrimers are ‘smart’ biocompatible 3D polymers that unveil a dynamic and elegant back-folding mechanism involving hydrogen bonding between primary amines at the surface and tertiary amines and ureas at the core. Similarly, to a biological proton pump, they are able to automatically and reversibly transform their conformation in response to pH stimulus. Here, we show that PURE-G4 is a useful gene silencing platform showing no cellular toxicity. As a proof of concept we investigated the PURE-G4-siRNA dendriplex, which was shown to be an attractive platform with high transfection efficacy. The simplicity associated with the complexation of siRNA with polyurea dendrimers makes them a powerful tool for efficient cytosolic siRNA delivery.

Baptista, P. V. "Nanodiagnostics: leaving the research lab to enter the clinics?" Diagnosis (Berl) 1 (2014): 305-309. AbstractWebsite


Larguinho, Miguel, Ana Cordeiro, Mario S. Diniz, Pedro M. Costa, and Pedro V. Baptista. "Metabolic and histopathological alterations in the marine bivalve Mytilus galloprovincialis induced by chronic exposure to acrylamide." Environmental Research 135 (2014): 55-62. Abstract


Baptista, Pedro V. "Nanodiagnostics: leaving the research lab to enter the clinics?" Diagnosis 1 (2014): 305-309. AbstractWebsite

Nanotechnology has provided a plethora of valuable tools that can be applied for the detection of biomolecules and analytes relevant for diagnosis purposes – nanodiagnostics. This surging new field of molecular diagnostics has been revolutionizing laboratory procedures and providing new ways to assess disease biomarkers with increased sensitivity. While most of the reported nanodiagnostics systems are proof-of-concepts that demonstrate their efficacy in the lab, several nanodiagnostics platforms have already matured to a level that open the way for effective translation to the clinics. Nanodiagnostics platforms (e.g., gold nanoparticles containing systems) have been remarkably useful for the development of molecular diagnosis strategies for DNA/RNA detection and characterization, including systems suitable for point-of-care. How near are nanodiagnostics to go from the bench to the bedside?

Larguinho, Miguel, Daniela Correia, Mario S. Diniz, and Pedro V. Baptista. "Evidence of one-way flow bioaccumulation of gold nanoparticles across two trophic levels." Journal of Nanoparticle Research 16 (2014). Abstract


Veigas, Bruno, Alexandra R. Fernandes, and Pedro V. Baptista. "AuNPs for identification of molecular signatures of resistance." Frontiers in Microbiology 5 (2014). Abstract


Carlos, Fábio Ferreira, Orfeu Flores, Gonçalo Doria, and Pedro Viana Baptista. "Characterization of genomic SNP via colorimetric detection using a single gold nanoprobe." Analytical Biochemistry 465 (2014): 1-5. AbstractWebsite

Identification of specific nucleic acid sequences mediated by gold nanoparticles derivatized thiol-modified oligonucleotides (Au-nanoprobes) has been proven to be a useful tool in molecular diagnostics. Here, we demonstrate that, on optimization, detection may be simplified via the use of a single Au-nanoprobe to detect a single nucleotide polymorphism (SNP) in homo- or heterozygote condition. We validated this non-cross-linking approach through the analysis of 20 clinical samples using a single specific Au-nanoprobe for an SNP in the FTO (fat mass and obesity-associated) gene against direct DNA sequencing. Sensitivity, specificity, and limit of detection (LOD) were determined, and statistical differences were calculated by one-way analysis of variance (ANOVA) and a post hoc Tukey's test to ascertain whether there were any differences between Au-nanoprobe genotyped groups. For the first time, we show that the use of a single Au-nanoprobe can detect SNP for each genetic status (wild type, heterozygous, or mutant) with high degrees of sensitivity (87.50%) and specificity (91.67%).

Roma-Rodrigues, Catarina, Alexandra R. Fernandes, and Pedro Viana Baptista. "Exosome in Tumour Microenvironment: Overview of the Crosstalk between Normal and Cancer Cells." Biomed Research International (2014). Abstract


Pedrosa, Pedro, Bruno Veigas, Diana Machado, Isabel Couto, Miguel Viveiros, and Pedro V. Baptista. "Gold nanoprobes for multi loci assessment of multi-drug resistant tuberculosis." Tuberculosis 94 (2014): 332-337. AbstractWebsite

Tuberculosis, still one of the leading human infectious diseases, reported 8.7 million new cases in 2011 alone. Also, the increasing rate of multidrug-resistant tuberculosis (MDRTB) and its treatment difficulties pose a serious public health threat especially in developing countries. Resistance to isoniazid and rifampicin, first line antibiotics, is commonly associated with point mutations in katG, inhA and rpoB genes of Mycobacterium tuberculosis complex (MTBC). Therefore, the development of cheap, fast and simple molecular methods to assess susceptibility profiles would have a huge impact in the capacity of early diagnosis and treatment of MDRTB.

Gold nanoparticles functionalized with thiol-modified oligonucleotides (Au-nanoprobes) have shown the potential to provide a rapid and sensitive detection method for MTBC and single base mutations associated with antibiotic resistance, namely the characterization of the three most relevant codons in rpoB gene associated to rifampicin resistance. Here we extend the Au-nanoprobe approach towards discriminating specific mutations within inhA and rpoB genes in PCR amplified DNA from isolates. Using a multiplex PCR reaction for these two genes, it is possible to assess both loci in parallel, and extend the potential of the Au-nanoprobe method to MDRTB molecular characterization with special application in the most frequent Portuguese genotypes.

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


Luis, Daniel V., Joana Silva, Ana Isabel Tomaz, Rodrigo F. M. de Almeida, Miguel Larguinho, Pedro V. Baptista, Luisa M. D. R. S. Martins, Telma F. S. Silva, Pedro M. Borralho, Cecilia M. P. Rodrigues, Antonio S. Rodrigues, Armando J. L. Pombeiro, and Alexandra R. Fernandes. "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." Journal of Biological Inorganic Chemistry 19 (2014): 787-803. Abstract


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


Larguinho, Miguel, José L. Capelo, and Pedro V. Baptista. "Fast Nucleotide identification using gold nanoparticle-based surface assisted laser desorption/ionisation." Talanta 105 (2013): 417-421.
Conde, João, João Rosa, Jesús M. de la Fuente, and Pedro V. Baptista. "Gold-nanobeacons for simultaneous gene specific silencing and intracellular tracking of the silencing events." Biomaterials 34 (2013): 2516-2523.
Conde, J., J. M. de la Fuente, and P. V. Baptista. "Nanomaterials for reversion of multidrug resistance in cancer: a new hope for an old idea?" Front Pharmacol 4 (2013): 134. AbstractWebsite


Conde, João, Jesus M. de la Fuente, and Pedro V. Baptista. "Nanomaterials for reversion of multidrug resistance in cancer: a new hope for an old idea?" Front. Pharmacol. (2013).
Bernacka-Wojcik, Iwona, Paulo Lopes, Ana Catarina Vaz, Bruno Veigas, Pawel Jerzy Wojcik, Pedro Simões, David Barata, Elvira Fortunato, Pedro V. Baptista, Hugo Águas, and Rodrigo Martins. "Bio-microfluidic platform for gold nanoprobe based DNA detection—application to Mycobacterium tuberculosis." Biosens Bioelectron 48 (2013): 87-93. AbstractWebsite

We have projected and fabricated a microfluidic platform for DNA sensing that makes use of an optical colorimetric detection method based on gold nanoparticles. The platform was fabricated using replica moulding technology in PDMS patterned by high-aspect-ratio SU-8 moulds. Biochips of various geometries were tested and evaluated in order to find out the most efficient architecture, and the rational for design, microfabrication and detection performance is presented. The best biochip configuration has been successfully applied to the DNA detection of Mycobacterium tuberculosis using only 3 µl on DNA solution (i.e. 90 ng of target DNA), therefore a 20-fold reduction of reagents volume is obtained when compared with the actual state of the art.

Conde, João, João Rosa, and Pedro V. Baptista. "Gold-Nanobeacons as a Theranostic System for the Detection and Inhibition of Specific Genes." Nature Protocol Exchange (2013). AbstractWebsite

This protocol describes the synthesis and detailed calibration of a gold nanoparticle-based nanobeacon (Au-nanobeacon) as an innovative theranostic approach for detection and inhibition of sequence-specific DNA and RNA for in vitro and ex vivo applications. Under hairpin configuration, proximity to gold nanoparticles leads to fluorescence quenching; hybridization to a complementary target restores fluorescence emission due to the gold nanobeacons’ conformational reorganization that causes the fluorophore and the AuNP to part from each other. This concept can easily be extended and adapted to assist the in vitro evaluation of silencing potential of a given sequence to be later used for ex vivo gene silencing and RNAi approaches, with the ability to monitor real-time gene delivery action. The time range for the entire protocol is ~8 days, including synthesis, functionalization and calibration of Au-nanobeacons, RNAi and gene silencing assays.

Conde, Joao, Furong Tian, Yulan Hernandez, Chenchen Bao, Daxiang Cui, Klaus-Peter Janssen, M. Ricardo Ibarra, Pedro V. Baptista, Tobias Stoeger, and Jesus M. de la Fuente. "In vivo tumor targeting via nanoparticle-mediated therapeutic siRNA coupled to inflammatory response in lung cancer mouse models." Biomaterials 34 (2013): 7744-7753. Abstract



Under laser radiation, cells labeled with gold nanoparticles (AuNPs) are believed to suffer thermal damage due to the transfer of the absorbed light from the AuNPs to the cells. This process, which involves complex mechanisms such as the rapid electron–phonon decay in the AuNPs, followed by phonon–phonon relaxation, culminates in the localized heating of both the AuNPs and the cells, setting the rational for the use of these nanostructures, under laser light, in cancer photothermal therapy (PTT). Here, we discuss the chemical and biological aspects of this promising new therapeutic approach, including the advantages over conventional cancer therapies and the challenges that scientists still need to overcome to progress toward translation research

Branquinho, R., J. V. Pinto, T. Busani, P. Barquinha, L. Pereira, P. V. Baptista, R. Martins, and E. Fortunato. "Plastic Compatible Sputtered Ta2O5 Sensitive Layer for Oxide Semiconductor TFT Sensors." J. Display Technol 9 (2013): 723-728.
Cordeiro, M., L. Giestas, J. C. Lima, and Pedro Viana Baptista. "Coupling an universal primer to SBE combined spectral codification strategy for single nucleotide polymorphism analysis." J Biotechnol 168 (2013): 90-94. AbstractWebsite

We previously reported a strategy that combines Förster resonance energy transfer (FRET) based spectral codification with a single base extension (SBE) reaction for single nucleotide sequence discrimination in solution. This strategy is capable of unequivocally detect the allele variants present in solution. To extend the use of this tool to any locus of interest, it would be required the development of an universal approach capable of combining a sequence specific SBE primer to an universal sequence labeled and optimized for spectral codification.
Here, we extend this concept to a general strategy by means of a labeled universal oligonucleotide primer (donor), a sequence specific primer that allows for incorporation of the complementary acceptor labeled ddNTP, which allows discrimination the allele variant in the sample via the unambiguous FRET signature of the donor/acceptor pair.

Veigas, Bruno, Pedro Pedrosa, Isabel Couto, Miguel Viveiros, and Pedro V. Baptista. "Isothermal DNA amplification coupled to Au-nanoprobes for detection of mutations associated to Rifampicin resistance in Mycobacterium tuberculosis." Journal of Nanobiotechnology (2013).