Holographic Recording of Unslanted Volume Transmission Gratings in Acrylamide/Propargyl Acrylate Hydrogel Layers: Towards Nucleic Acids Biosensing.

The role of volume hydrogel holographic gratings as optical transducers in sensor devices for point-of-care applications is increasing due to their ability to be functionalized for achieving enhanced selectivity. The first step in the development of these transducers is the optimization of the holographic recording process. The optimization aims at achieving gratings with reproducible diffraction efficiency, which remains stable after reiterative washings, typically required when working with analytes of a biological nature or several step tests. The recording process of volume phase transmission gratings within Acrylamide/Propargyl Acrylate hydrogel layers reported in this work was successfully performed, and the obtained diffraction gratings were optically characterized. Unslanted volume transmission gratings were recorded in the hydrogel layers diffraction efficiencies; up to 80% were achieved. Additionally, the recorded gratings demonstrated stability in water after multiple washing steps. The hydrogels, after functionalization with oligonucleotide probes, yields a specific hybridization response, recognizing the complementary strand as demonstrated by fluorescence. Analyte-sensitive hydrogel layers with holographic structures are a promising candidate for the next generation of in vitro diagnostic tests.

Surface Micro-Patterned Biofunctionalized Hydrogel for Direct Nucleic Acid Hybridization Detection.

The present research is focused on the development of a biofunctionalized hydrogel with a surface diffractive micropattern as a label-free biosensing platform. The biosensors described in this paper were fabricated with a holographic recording of polyethylene terephthalate (PET) surface micro-structures, which were then transferred into a hydrogel material. Acrylamide-based hydrogels were obtained with free radical polymerization, and propargyl acrylate was added as a comonomer, which allowed for covalent immobilization of thiolated oligonucleotide probes into the hydrogel network, via thiol-yne photoclick chemistry. The comonomer was shown to significantly contribute to the immobilization of the probes based on fluorescence imaging. Two different immobilization approaches were demonstrated: during or after hydrogel synthesis. The second approach showed better loading capacity of the bioreceptor groups. Diffraction efficiency measurements of hydrogel gratings at 532 nm showed a selective response reaching a limit of detection in the complementary DNA strand of 2.47 µM. The label-free biosensor as designed could significantly contribute to direct and accurate analysis in medical diagnosis as it is cheap, easy to fabricate, and works without the need for further reagents.

Processing of Holographic Hydrogels in Liquid Media: A Study by High-Performance Liquid Chromatography and Diffraction Efficiency.

The storage of time-stable holographic gratings in hydrogel matrices when the material is immersed in aqueous media is a real challenge at present. The optimization of the storage stages of the holograms must be properly investigated to identify the most suitable development processes. For this reason, this work is focused on the study of the optimization of the washing stages of the hydrogels based on acrylamide and N,N'-methylenebis(acrylamide) once unslanted transmission holograms have been stored. High-performance liquid chromatography and UV-visible measurements have been employed in our system to analyze the composition of the washing solutions. PBST and DMSO:H2O are used as solvents in the washing stages. The diffraction efficiencies are measured during the washing stages and after the storing of the holograms during several days in PBST. Maximum diffraction efficiencies of 38 and 27.6% are reached when PBST and DMSO:H2O are employed, respectively, for the washing process. Holograms show temporal stability after being stored immersed in PBST at 4 °C for 4 days.

DNA -based hydrogels for high-performance optical biosensing application.

Analyte-sensitive DNA-based hydrogels find multiple applications in the field of biosensors due to their adaptable nature. Here, the design of DNA-based hydrogel and its application as sensing platform for the detection of a specific target sequence are presented. DNA-functionalized hydrogel structures were formed via a free radical co-polymerization process. A simple one-step probe immobilization procedure is reported: DNA probe molecules are added to the photoactive polymer mixture, dispensed onto a solid support, or a mold, and covalently attached while the hydrogel is formed through UV light exposure. Such hydrogels can be synthesized with desired recognition ability through the selection of a certain nucleotide sequence. Here we show the application of DNA-based hydrogel to detect the target with high performance in fluorescence microarray format and, additionally, to fabricate holographic surface relief gratings for label-free sensing assays.

Hydrogel-based holographic sensors and biosensors: past, present, and future.

Hydrogel-based holographic sensors consist of a holographic pattern in a responsive hydrogel that diffracts light at different wavelengths depending on the dimensions and refractive index changes in the material. The material composition of hydrogels can be designed to be specifically responsive to different stimuli, and thus the diffraction pattern can correlate with the amount of analyte. According to this general principle, different approaches have been implemented to achieve label-free optical sensors and biosensors, with advantages such as easy fabrication or naked-eye detection. A review on the different approaches, sensing materials, measurement principles, and detection setups, and future perspectives is offered.

Label-free detection of C-Reactive protein using bioresponsive hydrogel-based surface relief diffraction gratings.

Responsive hydrogel-based surface relief gratings have demonstrated great performances as transducers in optical sensing. However, novel and smart designs of hydrogels are needed for the appropriate detection of analytes and biomolecules since the existing materials are very limited to specific molecules. In this work, a biosensing system based on surface relief gratings made of bioresponsive hydrogels has been developed. In particular, the hydrogel contains phosphocholine moieties to specifically recognize C-Reactive protein (CRP). The CRP-Sensing hydrogel capacity to selectively detect CRP was fully demonstrated. Using Direct Laser Interference Patterning, micro-gratings were created on CRP-Sensing hydrogel substrates and applied for the label-free sensing of CRP using a simple laser-based homemade optical setup. Limits of detection (LOD) and quantification (LOQ) in human serum dilutions of 1.07 and 8.92 mg L-1, respectively, were reached. These results demonstrate that the biosensing system allows the selective label-free detection of CRP within concentration ranges around those related to risks of cardiovascular diseases and sepsis. Besides, amplification strategies have been carried out improving the sensitivity, widening the linear range, and reaching better LOD and LOQ (0.30 mg L-1 and 4.36 mg L-1). Finally, all the approaches were tested for the quantification of CRP in certified human serum with recoveries of around 100%.

Microwave-assisted functionalization of carbon nanohorns with oligothiophene units with SERS activity.

Carbon nanohorns have been functionalized with oligothiophene units via the 1,3-dipolar cycloaddition reaction under microwave irradiation and solvent-free conditions. A dramatic Raman enhancement was found for one of the synthesized derivatives. Experimental and in silico studies helped to understand the enhancement, attributed to the modification of electromagnetic fields upon functionalization at the tip of the nanostructures.

Bactericidal Effect of 5-Mercapto-2-nitrobenzoic Acid-Coated Silver Nanoclusters against Multidrug-Resistant Neisseria gonorrhoeae.

Neisseria gonorrhoeae is among the most multidrug-resistant bacteria in circulation today, and new treatments are urgently needed. In this work, we demonstrate the ability of 5-mercapto-2-nitrobenzoic acid-coated silver nanoclusters (MNBA-AgNCs) to kill strains of Neisseria gonorrhoeae. Using an in vitro bactericidal assay, MNBA-AgNCs had been found to show significantly higher anti-gonococcal bioactivity than the antibiotics ceftriaxone and azithromycin and silver nitrate. These nanoclusters were effective against both planktonic bacteria and a gonococcal infection of human cell cultures in vitro. Treatment of human cells in vitro with MNBA-AgNCs did not induce significant release of lactate dehydrogenase, suggesting minimal cytotoxicity to eukaryotic cells. Our results suggest that MNBA-AgNCs hold great potential for topical treatment of localized gonorrhoeae.

Graphene Oxide-Upconversion Nanoparticle Based Portable Sensors for Assessing Nutritional Deficiencies in Crops.

The development of innovative technologies to rapidly detect biomarkers associated with nutritional deficiencies in crops is highly relevant to agriculture and thus could impact the future of food security. Zinc (Zn) is an important micronutrient in plants, and deficiency leads to poor health, quality, and yield of crops. We have developed portable sensors, based on graphene oxide and upconversion nanoparticles, which could be used in the early detection of Zn deficiency in crops, sensing mRNAs encoding members of the ZIP-transporter family in crops. ZIPs are membrane transport proteins, some of which are up-regulated at the early stages of Zn deficiency, and they are part of the biological mechanism by which crops respond to nutritional deficiency. The principle of these sensors is based on the intensity of the optical output resulting from the interaction of oligonucleotide-coated upconversion nanoparticles and graphene oxide in the absence or presence of a specific oligonucleotide target. The sensors can reliably detect mRNAs in RNA extracts from plants using a smartphone camera. Our work introduces the development of accurate and highly sensitive sensors for use in the field to determine crop nutrient status and ultimately facilitate economically important nutrient management decisions.

Production of ready-to-use few-layer graphene in aqueous suspensions.

Graphene has promising physical and chemical properties such as high strength and flexibility, coupled with high electrical and thermal conductivities. It is therefore being incorporated into polymer-based composites for use in electronics and photonics applications. A main constraint related to the graphene development is that, being of a strongly hydrophobic nature, almost all dispersions (usually required for its handling and processing toward the desired application) are prepared in poisonous organic solvents such as N-methyl pyrrolidone or N,N-dimethyl formamide. Here, we describe how to prepare exfoliated graphite using a ball mill. The graphene produced is three to four layers thick and ∼500 nm in diameter on average, as measured by electron microscopy and Raman spectroscopy; can be stored in the form of light solid; and is easily dispersed in aqueous media. Our methodology consists of four main steps: (i) the mechanochemical intercalation of organic molecules (melamine) into graphite, followed by suspension in water; (ii) the washing of suspended graphene to eliminate most of the melamine; (iii) the isolation of stable graphene sheets; and (iv) freeze-drying to obtain graphene powder. This process takes 6-7 or 9-10 d for aqueous suspensions and dry powders, respectively. The product has well-defined properties and can be used for many science and technology applications, including toxicology impact assessment and the production of innovative medical devices.

Targeted killing of prostate cancer cells using antibody-drug conjugated carbon nanohorns.

The ability of carbon nanohorns (CNHs) to cross biological barriers makes them potential carriers for delivery purposes. In this work, we report the design of a new selective antibody-drug nanosystem based on CNHs for the treatment of prostate cancer (PCa). In particular, cisplatin in a prodrug form and the monoclonal antibody (Ab) D2B, selective for PSMA+ cancer cells, have been attached to CNHs due to the current application of this antigen in PCa therapy. The hybrids Ab-CNHs, cisplatin-CNHs and functionalised-CNHs have also been synthesized to be used as control systems. The efficacy and specificity of the D2B-cisplatin-CNH conjugate to selectively target and kill PSMA+ prostate cancer cells have been demonstrated in comparison with other derivatives. The developed strategy to functionalise CNHs is fascinating because it can allow the fine tuning of both drug and Ab molecules attached to the nanostructure in order to modulate the activity of the nanosystem. Finally, the herein described methodology can be used for the incorporation of almost any drugs or Abs in the platforms in order to create new targeted drugs for the treatment of different diseases.

Triazine-Carbon Nanotubes: New Platforms for the Design of Flavin Receptors.

The synthesis of functionalised carbon nanotubes as receptors for riboflavin (RBF) is reported. Carbon nanotubes, both single-walled and multi-walled, have been functionalised with 1,3,5-triazines and p-tolyl chains by aryl radical addition under microwave irradiation and the derivatives have been fully characterised by using a range of techniques. The interactions between riboflavin and the hybrids were analysed by using fluorescence and UV/Vis spectroscopic techniques. The results show that the attached functional groups minimise the π-π stacking interactions between riboflavin and the nanotube walls. Comparison of p-tolyl groups with the triazine groups shows that the latter have stronger interactions with riboflavin because of the presence of hydrogen bonds. Moreover, the triazine derivatives follow the Stern-Volmer relationship and show a high association constant with riboflavin. In this way, artificial receptors in catalytic processes could be designed through specific control of the interaction between functionalised carbon nanotubes and riboflavin.

Carbon nanohorns as integrative materials for efficient dye-sensitized solar cells.

Different nanocarbons, that is, single-wall carbon nanotubes, graphene, single-wall carbon nanohorns (SWCNHs), and their respective oxidized analogs have been used to fabricate novel doped TiO2 electrodes for DSSCs. Our results indicate that all of the nanocarbons significantly enhance the device characteristics when compared to standard TiO2 electrodes. Overall, our most outstanding finding is that SWCNH derivatives are also a plausible material for developing highly-efficient DSSCs.