Unlocking New Avenues: Solid-State Synthesis of Molecularly Imprinted Polymers.

Molecularly imprinted polymers (MIPs) are established artificial molecular recognition platforms with tailored selectivity towards a target molecule, whose synthesis and functionality are highly influenced by the nature of the solvent employed in their synthesis. Steps towards the "greenification" of molecular imprinting technology (MIT) has already been initiated by the elaboration of green MIT principles; developing MIPs in a solvent-free environment may not only offer an eco-friendly alternative, but could also significantly influence the affinity and expected selectivity of the resulting binding sites. In the current study the first solvent-free mechanochemical synthesis of MIPs via liquid-assisted grinding (LAG) is reported. The successful synthesis of the imprinted polymer was functionally demonstrated by measuring its template rebinding capacity and the selectivity of the molecular recognition process in comparison with the ones obtained by the conventional, non-covalent molecular imprinting process in liquid media. The results demonstrated similar binding capacities towards the template molecule and superior chemoselectivity compared to the solution-based MIP synthesis method. The adoption of green chemistry principles with all their inherent advantages in the synthesis of MIPs may not only be able to alleviate the potential environmental and health concerns associated with their analytical (e.g., selective adsorbents) and biomedical (e.g., drug carriers or reservoirs) applications, but might also offer a conceptual change in molecular imprinting technology.

Topical Treatment for Retinal Degenerative Pathologies: A Systematic Review.

The topical administration of medicines is the preferred route in ocular therapy, at least for the anterior segment of the eye. However, the eye's inherent functional and biological barriers all work against the active pharmaceutical ingredient (API) to efficiently reach the targeted retinal structures. The main objective of this article is to offer a systematic review of the scientific literature in recent years, focusing on the latest developments of topical treatment intended for retinal degenerative diseases. Database search returned 102 clinical studies, focused on topical treatment for age macular degeneration, macular edemas (in diabetic retinopathy, surgery related or in retinal dystrophies) or glaucoma. After the exclusion of low-powered studies and those combining vitreo-retinal surgery, 35 articles remained for analysis. Currently, the topical treatment of retinal degenerative diseases is limited by the difficulty to deliver effective drug concentrations to the posterior eye structures. However, in the case of drug classes like NSAIDs, the presence of certain molecular and metabolic features for specific representatives makes the topical administration currently feasible in several clinical contexts. For other drug classes, either a fine-tuning of the API's pharmacokinetic profile or the use of more advanced formulation strategies, such as rationally designed nanostructured drugs and vehicles, crystalline polymorphs or supramolecular complexes, could bring the much awaited breakthrough for a more predictable and controlled delivery towards the retinal structures and could eventually be employed in the future for the development of more effective ways of delivering drugs to the posterior eye, with the ultimate goal of improving their clinical efficacy.

In Vivo Applications of Molecularly Imprinted Polymers for Drug Delivery: A Pharmaceutical Perspective.

Molecularly imprinted polymers (MIPs) have been proven to be a promising candidate for drug delivery systems (DDS) due to their ability to provide a sustained and controlled drug release, making them useful for treating a wide range of medical conditions. MIP-based DDS offer many advantages, including the administration of a smaller drug doses, due to the higher drug payload or targeted delivery, resulting in fewer side effects, as well as the possibility of attaining high concentrations of the drug in the targeted tissues. Whether designed as drug reservoirs or targeted DDS, MIPs are of great value to drug delivery as conventional drug formulations can be redesigned as DDS to overcome the active pharmaceutical ingredient's (APIs) poor bioavailability, toxic effects, or other shortcomings that previously made them less efficient or unsuitable for therapy. Therefore, MIP design could be a promising alternative to the challenging research and development of new lead compounds. Research on MIPs is primarily conducted from a material science perspective, which often overlooks some of their key pharmaceutical requirements. In this review, we emphasize the specific features that make MIPs suitable for clinical use, from both a material science and a biopharmaceutical perspective.

Analytical Perspectives in the Study of Polyvalent Interactions of Free and Surface-Bound Oligonucleotides and Their Implications in Affinity Biosensing.

The high affinity and/or selectivity of oligonucleotide-mediated binding offers a myriad of therapeutical and analytical applications, whose rational design implies an accurate knowledge of the involved molecular mechanisms, concurring equilibrium processes and key affinity parameters. Oligonucleotide-functionalized gold surfaces or nanostructures are regularly employed analytical platforms for the development of label-free optical or electrochemical biosensors, and recently, novel detection platform designs have been increasingly considering the synergistic effect of polyvalent binding, involving the simultaneous interaction of two or several oligonucleotide strands. Considering the general lack of studies involving ternary single-stranded DNA (ssDNA) interactions, a complementary analytical workflow involving capillary gel electrophoretic (CGE) mobility shift assay, microcalorimetry and computational modeling has been deployed for the characterization of a series of free and surface-bound binary and ternary oligonucleotide interactions. As a proof of concept, the DNA analogue of MicroRNA 21 (miR21), a well-known oncogenic short MicroRNA (miRNA) sequence, has been chosen as a target molecule, simulating limiting-case scenarios involved in dual molecular recognition models exploited in affinity (bio)sensing. Novel data for the characterization of oligonucleotide interacting modules is revealed, offering a fast and complete mapping of the specific or non-specific, often competing, binary and ternary order interactions in dynamic equilibria, occurring between various free and metal surface-bound oligonucleotides.

Off-Resonance Gold Nanobone Films at Liquid Interface for SERS Applications.

Extensive effort and research are currently channeled towards the implementation of SERS (Surface Enhanced Raman Spectroscopy) as a standard analytical tool as it has undisputedly demonstrated a great potential for trace detection of various analytes. Novel and improved substrates are continuously reported in this regard. It is generally believed that plasmonic nanostructures with plasmon resonances close to the excitation wavelength (on-resonance) generate stronger SERS enhancements, but this finding is still under debate. In the current paper, we compared off-resonance gold nanobones (GNBs) with on-resonance GNBs and gold nanorods (GNRs) in both colloidal dispersion and as close-packed films self-assembled at liquid-liquid interface. Rhodamine 6G (R6G) was used as a Raman reporter in order to evaluate SERS performances. A 17-, 18-, and 55-fold increase in the Raman signal was observed for nanostructures (off-resonance GNBs, on-resonance GNBs, and on-resonance GNRs, respectively) assembled at liquid-liquid interface compared to the same nanostructures in colloidal dispersion. SERS performances of off-resonance GNBs were superior to on-resonance nanostructures in both cases. Furthermore, when off-resonance GNBs were assembled at the liquid interface, a relative standard deviation of 4.56% of the recorded signal intensity and a limit of detection (LOD) of 5 × 10-9 M could be obtained for R6G, rendering this substrate suitable for analytical applications.

Analytical techniques for multiplex analysis of protein biomarkers.

INTRODUCTION: The importance of biomarkers for pharmaceutical drug development and clinical diagnostics is more significant than ever in the current shift toward personalized medicine. Biomarkers have taken a central position either as companion markers to support drug development and patient selection, or as indicators aiming to detect the earliest perturbations indicative of disease, minimizing therapeutic intervention or even enabling disease reversal. Protein biomarkers are of particular interest given their central role in biochemical pathways. Hence, capabilities to analyze multiple protein biomarkers in one assay are highly interesting for biomedical research. AREAS COVERED: We here review multiple methods that are suitable for robust, high throughput, standardized, and affordable analysis of protein biomarkers in a multiplex format. We describe innovative developments in immunoassays, the vanguard of methods in clinical laboratories, and mass spectrometry, increasingly implemented for protein biomarker analysis. Moreover, emerging techniques are discussed with potentially improved protein capture, separation, and detection that will further boost multiplex analyses. EXPERT COMMENTARY: The development of clinically applied multiplex protein biomarker assays is essential as multi-protein signatures provide more comprehensive information about biological systems than single biomarkers, leading to improved insights in mechanisms of disease, diagnostics, and the effect of personalized medicine.

Correction to: Affinity capillary electrophoresis for identification of active drug candidates in myotonic dystrophy type 1.

Unfortunately the name of Jean Jacques Vanden Eynde was missing as co-author of this contribution. The correct list of authors is: Ioan O. Neaga, Stephanie Hambye, Ede Bodoki, Claudio Palmieri, Jean Jacques Vanden Eynde, Eugénie Ansseau, Alexandra Belayew, Radu Oprean, Bertrand Blankert.

Topical nanodelivery system of lutein for the prevention of selenite-induced cataract.

Cataracts are responsible for half of the world blindness, surgery being the only viable treatment. Lutein, a naturally occurring carotenoid in the eye, has the potential to reduce cataract progression by protecting the eye from photo-oxidative stress. To restore the eye's natural line of defense against photo-oxidative stress, a formulation was developed using zein and poly(lactic-co-glycolic acid) nanoparticles (NPs) embedded in an optimized bioadhesive thermosensitive gel for the delivery of lutein via topical application. Cataracts were induced in Crl:WI rats via selenite injection at 13 days post-partum, followed by 7 days of treatment with free lutein or lutein-loaded NPs administered orally or topically. Cataract severity was significantly reduced in rats treated with topical applications of lutein-loaded NPs compared to the positive control, while no significant differences were observed in rats treated with other lutein formulations including oral and topically applied free lutein.

Affinity capillary electrophoresis for identification of active drug candidates in myotonic dystrophy type 1.

Myotonic dystrophy type 1 (DM1) is an autosomal dominantly inherited degenerative disease with a slow progression. At the present, there is no commercially available treatment, but sustained effort is currently undertaken for the development of a promising lead compound. In the present paper we report the development of a fast, versatile, and cost-effective affinity capillary electrophoresis (ACE) method for the screening and identification of potential drug candidates targeting pathological ARN probes relevant for DM1. The affinity studies were conducted in physiologically relevant conditions using 50 mM HEPES buffer (pH 7.4) in a fused silica capillary dynamically coated with poly(ethylene oxide), by testing a library of potential ligands against (CUG)50 RNA as target probe with a total run time of 4-5 h/ligand. For the most promising ligands, their affinity parameters were assessed and some results formerly reported on the affinity of pentamidine (PTMD) and neomycin against CUG repeats were confirmed. To the best of the authors' knowledge, the estimated binding stoichiometry for some of the tested compounds (i.e., ~ 121:1 for PTMD against the tested RNA probe) is reported for the first time. Additionally, the potential of a novel pentamidine like compound, namely 1,2-ethane bis-1-amino-4-benzamidine (EBAB) with much lower in vivo toxicity than its parent compound has also been confirmed studying its effect on a live cell model by fluorescence microscopy. Further tests, such as the evaluation of the rescue in the mis-splicing of the involved genes, can be performed to corroborate the potential therapeutic value of EBAB in DM1 treatment. Graphical abstract ᅟ.

Improved Enantioselectivity for Atenolol Employing Pivot Based Molecular Imprinting.

In the last few decades, molecular imprinting technology went through a spectacular evolution becoming a well-established tool for the synthesis of highly selective biomimetic molecular recognition platforms. Nevertheless, there is still room for advancement in the molecular imprinting of highly polar chiral compounds. The aim of the present work was to investigate the favorable kosmotropic effect of a ternary complex involving a polar chiral template (eutomer of atenolol) and a functional monomer, bridged by a central metal ion through well-defined, spatially directional coordinate bonds. The efficiency of the chiral molecular recognition was systematically assessed on polymers obtained both by non-covalent and metal-mediated molecular imprinting. The influence on the chromatographic retention and enantioselectivity of different experimental variables (functional monomers, cross-linkers, chaotropic agents, metal ions, porogenic systems, etc.) were studied on both slurry packed and monolithic HPLC columns. Deliberate changes in the imprinting and rebinding (chromatographic) processes, along with additional thermodynamic studies shed light on the particularities of the molecular recognition mechanism. The best performing polymer in terms of enantioselectivity (α = 1.60) was achieved using 4-vinyl pyridine as functional monomer and secondary ligand for the Co(II)-mediated imprinting of S-atenolol in the presence of EDMA as cross-linker in a porogenic mixture of [BMIM][BF4]:DMF:DMSO = 10:1:5, v/v/v.

Zein Nanoparticles Uptake by Hydroponically Grown Soybean Plants.

In the interest of developing and characterizing a polymeric nanoparticle pesticide delivery vehicle to soybeans, zein nanoparticle (ZNP) uptake by the roots and biodistribution to the leaves of soybean plants was measured. Zein was tagged with fluorescein isothiocyanate (FITC) and made into nanoparticles (135 ± 3 nm diameter. 0.202 ± 0.034 PDI and 81 ± 4 mV zeta-potential at pH 6) using an emulsion-diffusion method. After 10 days of hydroponic exposure, association between particles and roots of plants was found to vary based on bulk nanoparticle concentration. While 0.37 mg NP/mg dry weight were detected in roots immersed in 0.88 mg NP/mL nanoparticle suspension, 0.58 mg NP/mg dry weight associated with roots immersed in a high dose nanoparticle suspension of 1.75 mg NP/mL at 10 days. Nanoparticle root uptake followed second order kinetics. A small amount of increased fluorescence was detected in the hydroponically exposed plant's leaves, suggesting that either small amounts of particles or other fluorescent contaminants of zein were up taken by the roots and biodistributed within the plant. To the authors' knowledge, this is the first study in which the uptake and time-dependent association between polymeric nanoparticles and soybeans are quantified.

Application of capacitively coupled contactless conductivity as an external detector for zone electrophoresis in poly(dimethylsiloxane) chips.

In this work, lab-made PDMS microfluidic chips were matched to a capacitively coupled contactless conductivity detector (C(4) D) having external in-plane electrodes (eDAQ, Australia). The advantages of this type of C(4) D are the choice to reversibly place or remove the microchip onto/from the detector and to freely variate the position of the detection (separation length) on the microchip. The thickness of the bottom layer of the PDMS chip was optimized to achieve sensitive detection during the electrophoretic separation. PDMS chips with 100 µm bottom layer used with the C(4) D platform were tested by CZE of a mixture of seven anions and different types of real samples. Using split-flow pressure sample injection and effective length of 6.5 cm, the numbers of theoretical plates were in the range of 4000-6000 (63,000-93,000/m) and the LODs amounted to 3.66-14.7 µmol/L (0.13-2.26 µg/mL) for the studied anions.

Simultaneous enantiospecific recognition of several ß-blocker enantiomers using molecularly imprinted polymer-based electrochemical sensor.

The development of a chiral electrochemical sensor using an electrogenerated molecularly imprinted polymer (MIP)-based ultrathin film using R(+)-atenolol (ATNL) as a template was reported. The proposed sensor exhibited distinctive enantiospecific oxidation peaks toward the R-antipodes of four ß-blocker representatives and additional oxidation peaks common to both enantiomers of each studied ß-blocker, allowing thus the simultaneous analysis of all of their enantiomers in a single analysis. The specific preconditioning of the polymer by alternative exposure to aqueous and nonaqueous medium was proven to be essential for the chiral recognition ability of the obtained sensor. The rebinding property of the MIP film was studied by using a well-known redox probe, a change in the morphology and diffusive permeability of the thin polymeric layer in the presence of its template being observed. The applicability of the optimized analytical procedure was demonstrated by the analysis of ATNL's enantiomers in the range of 1.88 × 10(-7)-1.88 × 10(-5) mol/L. The developed polymeric interface is the first reported transductor of a chiral electrochemical sensor able to exhibit simultaneous enantiospecificity toward several ß-blocker representatives extensively used in the pharmaceutical and biomedical fields, offering good prospects in the simple, cost-effective, and fast assessment of their enantiomeric ratio and total concentration.

Surface mediated chiral interactions between cyclodextrins and propranolol enantiomers: a SERS and DFT study.

The nanoparticles mediated enantioselective recognition of propranolol enantiomers through native cyclodextrin complexation has been investigated by using surface-enhanced Raman spectroscopy (SERS). The highly efficient chiral recognition mechanism is based on a synergistic interaction between spherical noble metal nanoparticles, propranolol enantiomers and native cyclodextrins (CDs). Amongst the native cyclodextrins, ß-CD has the highest chiral recognition ability for propranolol enantiomers, due to its specific shape and cavity size, thus producing the largest difference between the recorded SERS spectra of the two hosted enantiomers. The molecular interaction mechanism responsible for enantioselectivity was furthermore proven by quantum chemical calculations based on density functional theory (DFT). The theoretical calculations and experimental SER spectra allowed the assignment of functional moieties involved in the chiral recognition mechanism. The most important factors governing the highly efficient chiral probing by SERS are the fundamentally different mechanism of interaction between the R- and S-enantiomers and ß-CD and the strength of interaction between the nanoparticle surface and the two propranolol-CD complexes. The role of metallic nanoparticles in the enantioselective recognition process has been experimentally evaluated by using silver and gold nanoparticles as SERS substrates, given their ability to interact differently with the complexes. The viability of this new method for chiral discrimination has been demonstrated for both substrates and could open new avenues for these kinds of applications.

Recent advances in capillary electrochromatography using molecularly imprinted polymers.

There is an increased and continuous need for developing new methods for the separation and quantification of an increasing number of analytes in the environmental, pharmaceutical, pharmacological, and toxicological sciences. CEC is still withholding its popularity, representing a viable alternative to the more conventional techniques (HPLC, GC) due to the numerous advantages, such as, low sample/reagent volumes, high separation efficiencies, hybrid separation principle, etc. One particular promising direction in CEC is the use of molecularly imprinted polymers (MIPs) as stationary phases. They are usually immobilized in the capillary column as a continuous polymeric monolith or as a thin polymer coating attached to the capillary's inner wall. Another emerging trend is the use of MIPs in the form of nanoparticles as a pseudostationary phase. This review discusses the recent developments (2011-2013) in finding the optimal polymerization mixture and the suitable MIP format that should be employed in CEC separations. The most important applications of MIPs in CEC technique are also highlighted.

Development and validation of NIR-chemometric methods for chemical and pharmaceutical characterization of meloxicam tablets.

CONTEXT: Near-Infrared (NIR) spectroscopy is an important component of a Process Analytical Technology (PAT) toolbox and is a key technology for enabling the rapid analysis of pharmaceutical tablets. OBJECTIVE: The aim of this research work was to develop and validate NIR-chemometric methods not only for the determination of active pharmaceutical ingredients content but also pharmaceutical properties (crushing strength, disintegration time) of meloxicam tablets. MATERIALS AND METHODS: The development of the method for active content assay was performed on samples corresponding to 80%, 90%, 100%, 110% and 120% of meloxicam content and the development of the methods for pharmaceutical characterization was performed on samples prepared at seven different compression forces (ranging from 7 to 45 kN) using NIR transmission spectra of intact tablets and PLS as a regression method. RESULTS: The results show that the developed methods have good trueness, precision and accuracy and are appropriate for direct active content assay in tablets (ranging from 12 to 18 mg/tablet) and also for predicting crushing strength and disintegration time of intact meloxicam tablets. DISCUSSION: The comparative data show that the proposed methods are in good agreement with the reference methods currently used for the characterization of meloxicam tablets (HPLC-UV methods for the assay and European Pharmacopeia methods for determining the crushing strength and disintegration time). CONCLUSION: The results show the possibility to predict both chemical properties (active content) and physical/pharmaceutical properties (crushing strength and disintegration time) directly, without any sample preparation, from the same NIR transmission spectrum of meloxicam tablets.

Mechanochemical Synthesis of New Praziquantel Cocrystals: Solid-State Characterization and Solubility.

New cocrystals of praziquantel with suberic, 3-hydroxybenzoic, benzene-1,2,4,5-tetracarboxylic, trimesic, and 5-hydroxyisophthalic acids were obtained through ball milling experiments. The optimal conditions for the milling process were chosen by changing the solvent volume and the mechanical action time. Supramolecular interactions in the new cocrystals are detailed based on single-crystal X-ray diffraction analysis, confirming the expected formation of hydrogen bonds between the praziquantel carbonyl group and the carboxyl (or hydroxyl) moieties of the coformers. Different structural characterization techniques were performed for all samples, but the praziquantel:suberic acid cocrystal includes a wider range of investigations such as thermal analysis, infrared and X-ray photoelectron spectroscopies, and SEM microscopy. The stability for up to five months was established by keeping it under extreme conditions of temperature and humidity. Solubility studies were carried out for all the new forms disclosed herein and compared with the promising cocrystals previously reported with salicylic, 4-aminosalicylic, vanillic, and oxalic acids. HPLC analyses revealed a higher solubility for most of the new cocrystal forms, as compared to pure praziquantel.

The impact of lutein-loaded poly(lactic-co-glycolic acid) nanoparticles following topical application: An in vitro and in vivo study.

Antioxidant therapies are of interest in the prevention and management of ocular disorders such as cataracts. Although an active area of interest, topical therapy with antioxidants for the treatment of cataracts is complicated by multiple ocular anatomical barriers, product stability, and solubility. Entrapment and delivery of antioxidants with poly(lactic-co-glycolic acid) nanoparticles is a possible solution to these challenges, however, little is known regarding their effects in vitro or in vivo. Our first aim was to investigate the impact of blank and lutein loaded PLGA nanoparticles on viability and development of reactive oxygen species in lens epithelial cells in vitro. Photo-oxidative stress was induced by ultraviolet light exposure with cell viability and reactive oxygen species monitored. Next, an in vivo, selenite model was utilized to induce cataract formation in rodents. Eyes were treated topically with both free lutein and lutein loaded nanoparticles (LNP) at varying concentrations. Eyes were monitored for the development of anterior segment changes and cataract formation. The ability of nanodelivered lutein to reach the anterior segment of the eye was evaluated by liquid chromatography coupled to mass spectrometry of aqueous humor samples and liquid chromatography coupled to tandem mass spectrometry (targeted LC-MS/MS) of lenses. LNP had a minimal impact on the viability of lens epithelial cells during the short exposure timeframe (24 h) and at concentrations < 0.2 µg LNP/µl. A significant reduction in the development of reactive oxygen species was also noted. Animals treated with LNPs at an equivalent lutein concentration of 1,278 µg /mL showed the greatest reduction in cataract scores. Lutein delivery to the anterior segment was confirmed through evaluation of aqueous humor and lens sample evaluation. Topical treatment was not associated with the development of secondary keratitis or anterior uveitis when applied once daily for one week. LNPs may be an effective in the treatment of cataracts.

EC-SERS detection of thiabendazole in apple juice using activated screen-printed electrodes.

Thiabendazole (TBZ), a benzimidazole fungicide used for post-harvest treatment, may be a trace contaminant of food matrices. In this work, we report the first EC-SERS (electrochemical-surface enhanced Raman spectroscopy) detection of TBZ in spiked apple juice using electrochemically (EC) roughened, gold-based screen-printed electrodes (AuSPEs) and portable instrumentation. Polarizing the substrate (-0.8 V vs Ag/AgCl) improves the recorded SERS signal of TBZ, allowing to reach a limit of detection (LOD) in juice of 0.061 ppm with a relatively wide linear range (0.5-10 µM) and good intermediate precision (%RSD < 10). The recovery of TBZ from unprocessed juice was found to be more than 82 %. Furthermore, a proof-of-concept integration of AuSPEs with a miniaturized flow cell for the preconcentration of TBZ and the controlled delivery of sample and reagents has been demonstrated. This approach paves the way for integrated, portable analytical systems applicable for on-site sample collection, processing, and analysis.

Toward microfluidic SERS and EC-SERS applications via tunable gold films over nanospheres.

Many promising applications of surface-enhanced Raman scattering (SERS), such as microfluidic SERS and electrochemical (EC)-SERS, require immersion of plasmonic nanostructured films in aqueous media. Correlational investigations of the optical response and SERS efficiency of solid SERS substrates immersed in water are absent in the literature. This work presents an approach for tuning the efficiency of gold films over nanospheres (AuFoN) as SERS substrates for applications in aqueous environment. AuFoN are fabricated by convective self-assembly of colloidal polystyrene nanospheres of various diameters (300-800 nm), followed by magnetron sputtering of gold films. The optical reflectance of the AuFoN and Finite-Difference Time-Domain simulations in both water and air reveal the dependence of the surface plasmon band on nanospheres' diameter and environment. SERS enhancement of a common Raman reporter on AuFoN immersed in water is analyzed under 785 nm laser excitation, but also using the 633 nm line for the films in air. The provided correlations between the SERS efficiency and optical response in both air and water indicate the best structural parameters for high SERS efficiency and highlight a route for predicting and optimizing the SERS response of AuFoN in water based on the behavior in air, which is more practical. Finally, the AuFoN are successfully tested as electrodes for EC-SERS detection of the thiabendazole pesticide and as SERS substrates integrated in a flow-through microchannel format. The obtained results represent an important step toward the development of microfluidic EC-SERS devices for sensing applications.