Synergistic Antibacterial Effects of Amoxicillin and Gold Nanoparticles: A Therapeutic Option to Combat Antibiotic Resistance.

Compacted Au@16-mph-16/DNA-AMOX (NSi) nanosystems were prepared from amoxicillin (AMOX) and precursor Au@16-mph-16 gold nanoparticles (Ni) using a Deoxyribonucleic acid (DNA) biopolymer as a glue. The synthesized nanocarrier was tested on different bacterial strains of Escherichia coli, Staphylococcus aureus, and Streptococcus pneumoniae to evaluate its effectiveness as an antibiotic as well as its internalization. Synthesis of the nanosystems required previous structural and thermodynamic studies using circular dichroism (CD) and UV-visible techniques to guarantee optimal complex formation and maximal DNA compaction, characteristics which facilitate the correct uptake of the nanocarrier. Two nanocomplexes with different compositions and structures, denoted NS1 and NS2, were prepared, the first involving external Au@16-mph-16 binding and the second partial intercalation. The Ni and NSi nanosystems obtained were characterized via transmission electron microscopy (TEM), zeta potential, and dynamic light scattering (DLS) techniques to measure their charge, aggregation state and hydrodynamic size, and to verify their presence inside the bacteria. From these studies, it was concluded that the zeta potential values for gold nanoparticles, NS1, and NS2 nanosystems were 67.8, -36.7, and -45.1 mV. Moreover, the particle size distribution of the Au@16-mph-16 gold nanoparticles and NS2 nanoformulation was found to be 2.6 nm and 69.0 nm, respectively. However, for NS1 nanoformulation, a bimodal size distribution of 44 nm (95.5%) and 205 nm (4.5%) was found. Minimal inhibitory concentration (MIC) values were determined for the bacteria studied using a microdilution plates assay. The effect on Escherichia coli bacteria was notable, with MIC values of 17 µM for both the NS1 and NS2 nanosystems. The Staphylococcus aureus chart shows a greater inhibition effect of NS2 and NP2 in non-diluted wells, and clearly reveals a great effect on Streptococcus pneumoniae, reaching MIC values of 0.53 µM in more diluted wells. These results are in good agreement with TEM internalization studies of bacteria that reveal significant internalization and damage in Streptococcus pneumoniae. In all the treatments carried out, the antibiotic capacity of gold nanosystems as enhancers of amoxicillin was demonstrated, causing both the precursors and the nanosystems to act very quickly, and thus favoring microbial death with a small amount of antibiotic. Therefore, these gold nanosystems may constitute an effective therapy to combat resistance to antibiotics, in addition to avoiding the secondary effects derived from the administration of high doses of antibiotics.

New Avenues of Research for Nanoparticle Drug Delivery Systems.

Knowledge of the different elements that determine the optimal method for drug loading and delivery nanosystems using nanoparticles of different natures is experiencing a remarkable boom in many scientific fields, especially in medicine, chemistry, biology, materials science and molecular biotechnology [...].

Gold Nanosystems Covered with Doxorubicin/DNA Complexes: A Therapeutic Target for Prostate and Liver Cancer.

Different gold nanosystems covered with DNA and doxorubicin (Doxo) were designed and synthesized for cancer therapy, starting from Au@16-Ph-16 cationic nanoparticles and DNA-Doxo complexes prepared under saturation conditions. For the preparation of stable, biocompatible, and small-sized compacted Au@16-Ph-16/DNA-Doxo nanotransporters, the conditions for the DNA-Doxo compaction process induced by gold nanoparticles were first explored using fluorescence spectroscopy, circular dichroism and atomic force microscopy techniques. The reverse process, which is fundamental for Doxo liberation at the site of action, was found to occur at higher CAu@16-Ph-16 concentrations using these techniques. Zeta potential, dynamic light scattering and UV-visible spectroscopy reveal that the prepared compacted nanosystems are stable, highly charged and of adequate size for the effective delivery of Doxo to the cell. This fact is verified by in vitro biocompatibility and internalization studies using two prostate cancer-derived cell lines (LNCaP and DU145) and one hepatocellular carcinoma-derived cell line (SNU-387), as well as a non-tumor prostate (PNT2) cell line and a non-hepatocarcinoma hepatoblastoma cell line (Hep-G2) model used as a control in liver cells. However, the most outstanding results of this work are derived from the use of the CI+NI combined treatments which present strong action in cancer-derived cell lines, while a protective effect is observed in non-tumor cell lines. Hence, novel therapeutic targets based on gold nanoparticles denote high selectivity compared to conventional treatment based on free Doxo at the same concentration. The results obtained show the viability of both the proposed methodology for internalization of compacted nanocomplexes inside the cell and the effectiveness of the possible treatment and minimization of side effects in prostate and liver cancer.

Use of Nanoparticles to Prevent Resistance to Antibiotics-Synthesis and Characterization of Gold Nanosystems Based on Tetracycline.

Antimicrobial resistance (AMR) is a serious public health problem worldwide which, according to the World Health Organization (WHO), requires research into new and more effective drugs. In this work, both gold nanoparticles covered with 16-3-16 cationic gemini surfactant (Au@16-3-16) and DNA/tetracycline (DNA/TC) intercalated complexes were prepared to effectively transport tetracycline (TC). Synthesis of the Au@16-3-16 precursor was carried out by using trihydrated gold, adding sodium borohydride as a reducing agent and the gemini surfactant 16-3-16 as stabilizing agent. Circular dichroism and atomic force microscopy techniques were then used to ascertain the optimal R range of the relationship between the concentrations of Au@16-3-16 and the DNA/TC complex (R = CAu@16-3-16/CDNA) that allow the obtainment of stable and compact nanosystems, these characteristics being fundamental for their use as antibiotic transporters. Stability studies over time were carried out for distinct selected Au@16-3-16 and Au@16-3-16/DNA-TC nanoformulations using the ultraviolet−visible spectrophotometry technique, checking their stability for at least one month. In addition, in order to know the charge and size distribution of the nanocomplexes, DLS and zeta potential measurements were performed in the solution. The results showed that the characterized nanosystems were highly charged, stable and of a reduced size (<100 nm) that allows them to cross bacterial membranes effectively (>1 µm). Once the different physicochemical characteristics of the gold nanosystems were measured, Au@16-3-16 and Au@16-3-16/DNA-TC were tested on Escherichia coli and Staphylococcus aureus to study their antibacterial properties and internalization capacity in microbes. Differences in the interaction of the precursors and the compacted nanosystems generated were observed in Gram-positive and Gram-negative bacteria, possibly due to membrane damage or electrostatic interaction with internalization by endocytosis. In the internalization experiments, depending on the treatment application time, the greatest bacterial destruction was observed for all nanoformulations explored at 18 h of incubation. Importantly, the results obtained demonstrate that both new nanosystems based on TC and Au@16-3-16 precursors have optimal antimicrobial properties and would be beneficial for use in patients, avoiding possible side effects.

Lysozyme-AuNPs Interactions: Determination of Binding Free Energy.

Investigation and optimization of lysozyme (Lys) adsorption onto gold nanoparticles, AuNPs, were carried out. The purpose of this study is to determine the magnitude of the AuNPs-lysozyme interaction in aqueous media by simple spectrophotometric means, and to obtain the free energy of binding of the system for the first time. In order to explore the possibilities of gold nanoparticles for sensing lysozyme in aqueous media, the stability of the samples and the influence of the gold and nanoparticle concentrations in the detection limit were studied. ζ potential measurements and the shift of the surface plasmon band showed a state of saturation with an average number of 55 Lys per gold nanoparticle. Lysozyme-AuNPs interactions induce aggregation of citrate-stabilized AuNPs at low concentrations by neutering the negative charges of citrate anions; from those aggregation data, the magnitude of the interactions has been measured by using Benesi-Hildebrand plots. However, at higher protein concentrations aggregation has been found to decrease. Although the nanocluster morphology remains unchanged in the presence of Lys, slight conformational changes of the protein occur. The influence of the size of the nanoclusters was also investigated for 5, 10, and 20 nm AuNPs, and 10 nm AuNPs was found the most appropriate.

Colorimetric, Naked-Eye Detection of Lysozyme in Human Urine with Gold Nanoparticles.

The stabilizing effect of lysozymes to salt addition over a gold colloid are exploited in order to detect lysozymes in human urine samples. The present research is aimed at the development of a fast, naked-eye detection test for urinary lysozymuria, in which direct comparison with a colorimetric reference, allows for the immediate determination of positive/negative cases. CIEL*a*b* parameters were obtained from sample absorbance measurements, and their color difference with respect to a fixed reference point was measured by calculating the ΔE76 parameter, which is a measure of how well the colors can be distinguished by an untrained observer. Results show that a simple and quick test can reliably, in less than 15 min, give a positive colorimetric response in the naked eye for concentrations of a urinary lysozyme over 57.2 µg/mL. This concentration is well within the limits of that observed for leukemia-associated lysozymurias, among other disorders.

Biocompatible DNA/5-Fluorouracil-Gemini Surfactant-Functionalized Gold Nanoparticles as Promising Vectors in Lung Cancer Therapy.

The design and preparation of novel nanocarriers to transport cancer drugs for chemotherapy purposes is an important line of research in the medical field. A new 5-fluorouracil (5-Fu) transporter was designed based on the use of two new biocompatible gold nanosystems: (i) a gold nanoparticle precursor, Au@16-Ph-16, stabilized with the positively charged gemini surfactant 16-Ph-16, and (ii) the compacted nanocomplexes formed by the precursor and DNA/5-Fu complexes, Au@16-Ph-16/DNA-5-Fu. The physicochemical properties of the obtained nanosystems were studied by using UV-visible spectroscopy, TEM, dynamic light scattering, and zeta potential techniques. Method tuning also requires the use of circular dichroism, atomic force microscopy, and fluorescence spectroscopy techniques for the prior selection of the optimal relative Au@16-Ph-16 and DNA concentrations (R = CAu@16-Ph-16/CDNA), biopolymer compaction/decompaction, and 5-Fu release from the DNA/5-Fu complex. TEM experiments revealed the effective internalization of the both precursor and Au@16-Ph-16/DNA-5-Fu-compacted nanosystems into the cells. Moreover, cytotoxicity assays and internalization experiments using TEM and confocal microscopy showed that the new strategy for 5-Fu administration enhanced efficacy, biocompatibility and selectivity against lung cancer cells. The differential uptake among different formulations is discussed in terms of the physicochemical properties of the nanosystems.

Encased Gold Nanoparticle Synthesis as a Probe for Oleuropein Self-Assembled Structure Formation.

Stable oleuropein-coated gold nanoparticles in aqueous media were synthesized for the first time. Oleuropein (OLE) concentration in the reaction medium was found to greatly influence the outcome and stability of the resulting nanocolloid, with a marked decrease in particle size being found for the more concentrated oleuropein solutions. The protection mechanisms involved in the stabilized nanosystems were analyzed. Oleuropein self-assembled structures were found to be formed at a concentration threshold of [OLE] > 5 × 10-5 M, and observed through the use of CryoSEM imaging. Those structures were responsible for both the increased stability and the decrease in size observed at the more concentrated solutions.

Author Correction: Design of highly stabilized nanocomposite inks based on biodegradable polymer-matrix and gold nanoparticles for Inkjet Printing.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Citrate and Polyvinylpyrrolidone Stabilized Silver Nanoparticles as Selective Colorimetric Sensor for Aluminum (III) Ions in Real Water Samples.

The use of silver nanoparticles stabilized with citrate and polyvinylpyrrolidone as a sensor for aluminum ions determination is proposed in this paper. These non-functionalized and specific nanoparticles provide a highly selective and sensitive detection system for aluminum in acidic solutions. The synthesized nanoparticles were characterized by transmission electron microscopy. Surface plasmon band deconvolution analysis was applied to study the interaction between silver nanoparticles and aluminum ions in solution. The interaction band in the UV-visible region was used as an analytical signal for quantitation purposes. The proposed detection system offers an effective AND wide linearity range (0.1-103 nM), specificity for Al(III) in THE presence of other metallic ions in solution, as well as high sensitivity (limit of detection = 40.5 nM). The proposed silver-nanoparticles-based sensor WAS successfully used for detecting Al(III) in real water samples.

Design of highly stabilized nanocomposite inks based on biodegradable polymer-matrix and gold nanoparticles for Inkjet Printing.

Nowadays there is a worldwide growing interest in the Inkjet Printing technology owing to its potentially high levels of geometrical complexity, personalization and resolution. There is also social concern about usage, disposal and accumulation of plastic materials. In this work, it is shown that sugar-based biodegradable polyurethane polymers exhibit outstanding properties as polymer-matrix for gold nanoparticles composites. These materials could reach exceptional stabilization levels, and demonstrated potential as novel robust inks for Inkjet based Printing. Furthermore, a physical comparison among different polymers is discussed based on stability and printability experiments to search for the best ink candidate. The University of Seville logo was printed by employing those inks, and the presence of gold was confirmed by ToF-SIMS. This approach has the potential to open new routes and applications for fabrication of enhanced biomedical nanometallic-sensors using stabilized AuNP.

Chitosan as a capping agent: Insights on the stabilization of gold nanoparticles.

A theoretical method based on Dissipative Particle Dynamics (DPD) was applied for the study of the interaction between coarse-grained models of chitosan and gold nanoparticles. These models were used to understand the role of chitosan in the stabilization of gold nanoparticles (5 and 7 nm). In addition, this stabilization was also supported by synthesized and characterized chitosan-gold nanoparticles comparing their stabilities with typical citrate-gold nanoparticles. Finally, the role of chitosan as a capping agent through the steric and electrosteric mechanisms in aqueous media with different pH values was also evaluated.

Exploring Factors for the Design of Nanoparticles as Drug Delivery Vectors.

To achieve optimal results when employing nanoparticles in biomedical fields, choosing the right type of nanoparticle and determining the correct procedure for drug loading are key factors. Each type of nanoparticle presents a determined set of characteristics that are, in some cases, unique. In general, their surface charge, geometry or hydrophilic character may be limiting factors, depending on what their intended application is. Once synthesized, additional factors, such as their interaction with biological systems and liberation mechanisms into the target cells, also need to be taken into account. Multiple advantages arise from the use of nanoparticles, such as the capability to solubilize hydrophobic compounds and an increased bioavailability. Those advantages justify the extensive and delicate study that should be undertaken in order to use them as drug delivery agents. One of the most important factors for the design of a drug delivery system with nanoparticles is achieving a high drug-to-nanoparticle ratio. In this Minireview, all of these key factors, both physicochemical and biological, are described, and special emphasis is placed on loading methods employed to introduce drugs into nanoparticles.

Decorating a single giant DNA with gold nanoparticles.

We decorated a single giant DNA (1.66 × 105 base pairs) with gold nanoparticles through the simple procedure of mild warming, without denaturation of the DNA molecule. Single-molecule observation with fluorescence microscopy revealed that individual decorated DNA molecules stay in the bulk solution by avoiding aggregation and precipitation, and exhibit translational and conformational fluctuation, i.e., Brownian motion. An analysis of the intra-chain fluctuation of single DNA molecules revealed that the apparent spring constant and damping coefficient of a DNA chain increased by ca. 13- and 5-fold, respectively, upon decoration with gold nanoparticles. Observation by transmission electron microscopy revealed that gold nanoparticles were stably attached to the DNA skeleton. UV-visible measurements revealed the absence of any detectable change in surface plasmon resonance, suggesting that the gold nanoparticles assemble without the formation of a densely packed aggregate. CD measurements showed that the secondary structure of decorated DNA is still essentially the B-form.

Understanding and improving aggregated gold nanoparticle/dsDNA interactions by molecular spectroscopy and deconvolution methods.

It is well known that single-stranded DNA (ssDNA) is easily able to adsorb on citrate-capped, non-functionalized gold nanoparticles (AuNPs). However, the affinity of double-stranded DNA (dsDNA) for them is much more limited. The present work demonstrates that long dsDNA suffers from a bending conformational change when anionic nanoparticles are present in solution. A striking decrease in the persistence length of the double helix in the absence of salt is observed through dynamic light scattering (DLS), viscometric, and atomic force microscopy (AFM) methods. Long dsDNA is therefore shown to be able to interact with anionic gold nanoparticles. To date, only ssDNA detection has been described by making use of interparticle cross-linking aggregation mechanisms; however, the data shown in this work allow for the development of new methods for detecting dsDNA in solution by using aggregated AuNPs as a starting point. The aggregation state is induced by the controlled addition of an inert electrolyte. A deconvolution procedure of the experimental plasmon shows how individual bands corresponding to aggregated nanoclusters diminish as the DNA concentration increases in the presence of 0.075 M NaCl.

Covalent and Non-Covalent DNA-Gold-Nanoparticle Interactions: New Avenues of Research.

The interactions of DNA, whether long, hundred base pair chains or short-chained oligonucleotides, with ligands play a key role in the field of structural biology. Its biological activity not only depends on the thermodynamic properties of DNA-ligand complexes, but can and often is conditioned by the formation kinetics of those complexes. On the other hand, gold nanoparticles have long been known to present excellent biocompatibility with biomolecules and are themselves remarkable for their structural, electronic, magnetic, optical and catalytic properties, radically different from those of their counterpart bulk materials, and which make them an important asset in multiple applications. Therefore, thermodynamic and kinetic studies of the interactions of DNA with nanoparticles acting as small ligands are key for a better understanding of those interactions to allow for their control and modulation and for the opening of new venues of research in nanomedicine, analytic and biologic fields. The interactions of gold nanoparticles with both DNA polymers and their smaller subunits; special focus is placed on those interactions taking place with nonfunctionalized gold nanoparticles are reviewed in the present work.

Fluorescent imino and secondary amino chitosans as potential sensing biomaterials.

A variety of fluorescent imino and secondary amino chitosans were synthesized under very mild conditions by reaction of the biopolymer amino functions with aromatic aldehydes in an acidified methanolic suspension. Simultaneous reactions of several aldehydes with chitosan were successfully carried out, and kinetic studies showed that 1-pyrenecarboxaldehyde reacts the fastest among them. An unprecedented study on the evaluation of the degree of N-substitution (DS, ranging from 31.7% to 12.0%) for the chitosan Schiff bases by using solid state CPMAS (13)C NMR is performed. A linear correlation between the DS obtained for the secondary amino chitosans by (1)H NMR (55.3-10.2%) and those obtained by CPMAS (13)C NMR (34.4-13.8%) has allowed us to calculate an empirical correlation factor that could be applied on chitosan-based aromatic systems. The new chiral-labelled chitosan derivatives exhibit a stable fluorescent behaviour, which was used to explore solvent sensoring applications.

Nonfunctionalized Gold Nanoparticles: Synthetic Routes and Synthesis Condition Dependence.

Since Faraday first described gold sol synthesis, synthetic routes to nanoparticles, as well as their applications, have experienced a huge growth. Variations in synthesis conditions such as pH, temperature, reduction, and the stabilizing agent used will determine the morphology, size, monodispersity, and stability of nanoparticles obtained, allowing for modulation of their physical and chemical properties. Although many studies have been made about the synthesis and characterization of individual nanosystems of interest, to our knowledge the common, general traits that all those synthesis share have not been previously compiled. In this review, we aim to offer a global vision of some of the most relevant synthetic procedures reported up to date, with a special focus on nonfunctionalized gold nanoparticle synthetic routes in aqueous media, and to display a broad overview of the influence that synthesis conditions have on the shape, stability, and reactivity of nanoparticle systems.

Interaction of gold nanoparticles mediated by captopril and S-nitrosocaptopril: the effect of manganese ions in mild acid medium.

We report herein results regarding reactivity and assembly of citrate-capped gold nanoparticles (AuNPs) mediated by captopril (cap) and S-nitrosocaptopril (NOcap), two angiotensin converting enzyme inhibitors and antihypertensive agents. The results were compared with that of cysteine (Cys), a thiol-containing amino acid found in plasma. The interparticle interactions were characterized by monitoring the evolution of the surface plasmon resonance band using the spectrophotometric method. The original gold nanoparticles were efficiently modified by small amounts of Mn(+2) ions, which are adsorbed onto the surface of 15.4 nm citrate-capped gold nanoparticles, giving rise to manganese-gold nanoparticles (Mn-AuNPs) that, in mild acid medium, have proved to be highly sensitive and a rapid colorimetric detection method for thiols. Depending on the concentration of the Mn(+2) ions the aggregation of AuNPs can be rapidly induced. The kinetics of the assembly process has been studied. Good first-order kinetics has been observed, with the exception of captopril-mediated nanoparticle aggregation at low concentration of either cap or acid. The rate of Cys-mediated assembly of gold nanoparticles in aqueous 10 mM acetic acid is more than 20-times faster than pure AuNPs and concentrations of Cys as low as 34 nM can be detected in less than 40 min under conditions of stable Mn-AuNPs. Similar effects were observed with cap or NOcap. The assembly-disassembly reversibility is shown with cap and NOcap and depends highly on pH.

Aprendizaje significativo del alumnado de física aplicada del grado en farmacia: evaluación basada en el empleo de cuestionarios / No disponible

Objetivos: Se han detectado claras deficiencias en el proceso de enseñanza-aprendizaje en las prácticas de la asignatura de «Física Aplicada a Ciencias de la Salud» del primer curso del Grado en Farmacia. Por tanto, el objetivo de este trabajo es desarrollar un sencillo programa de evaluación que permita extraer información inmediata sobre la práctica docente, permitiendo modificar la secuencia de actividades y la metodología. Material y Métodos: Se emplearon idénticos cuestionarios iniciales y finales a la práctica docente que fueron diseñados en base a problemas y/o preguntas clave, trabajadas en clase. Los resultados fueron clasificados en distintos modelos y representados, de acuerdo con su complejidad, en escaleras de aprendizaje. Resultados: La clasificación de los resultados de los cuestionarios inicial y final en modelos de pensamiento mostró una evolución positiva del aprendizaje del alumnado. En concreto, el 57% de los alumnos evolucionó hacia esquemas de pensamientos más complejos en el caso de la práctica del Calor Específico, mientras que un 60% de los alumnos lo hicieron en el caso de la Práctica de Arquímedes. Conclusiones: Profesores y alumnos coinciden en que el proceso de enseñanza-aprendizaje resulta mucho más completo y atractivo cuando se fomenta la participación del alumno y la reflexión. Un objetivo en un futuro próximo es tratar de repetir esta experiencia en diferentes grupos de alumnos para comparar resultados, analizar con más profundidad las dificultades en la asimilación de conceptos y diseñar nuevas estrategias docentes

Aim: It have been detected clearly deficiencies in the teaching-learning process for the practice subject of «Physics Applied Health Sciences» which is taught in the first course of the Degree in Pharmacy. Therefore, the motivation of this work is the development of a simple evaluation program that permits to gain prompt information about the teaching process which allows the modification of the activities sequence and methodology. Materials and Methods: Identical initial and final questionnaires were employed in the teaching practice based on problems and/or key questions worked in class. The results were classified in different models and represented, according to their complexity, in stairs of learning. Results: The classification of the questionnaires results in model thinking denoted a positive evolution of the student learning. In particular, in the case of the Determination of the Specific Heat around a 57 % of the students evolved into more complex thoughts schemes, while a 60% of the students did the same for the Archimedes practice. Conclusion: Teachers and students concur that the teaching-learning process is much more complete and attractive when student participation and reflection was encouraged. A goal in the near future could be try to repeat this experience with students of different groups to compare results, analyze in depth the difficulties in the assimilation of concepts and the design of new teaching strategies