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Biomimetic surface coatings based on plant polyphenols and catecholamines have been used broadly in a variety of applications. However, the lack of a rational cost-effective platform for screening these coatings and their properties limits the true potential of these functional materials to be unleashed. Here, we investigated the oxidation behavior and coating formation ability of a library consisting of 45 phenolic compounds and catecholamines. UV-vis spectroscopy demonstrated significant acceleration of oxidation and polymerization under UV irradiation. We discovered that several binary mixtures resulted in non-additive behavior (synergistic or antagonistic effect) yielding much thicker or thinner coatings than individual compounds measured by ellipsometry. To investigate the properties of coatings derived from new combinations, we used a miniaturized high-throughput strategy to screen 2,532 spots coated with single, binary, and ternary combinations of coating precursors in one run. We evaluated the use of machine learning models to learn the relation between the chemical structure of the precursors and the thickness of the nanocoatings. Formation and stability of nanocoatings were investigated in a high-throughput manner via discontinuous dewetting. 30 stable combinations (hits) were used to tune the surface wettability and to form water droplet microarray and spot size gradients of water droplets on the coated surface. No toxicity was observed against eukaryotic HeLa cells and Pseudomonas aeruginosa (strain PA30) bacteria after 24 h incubation at 37 °C. The strategy introduced here for high-throughput screening of nanocoatings derived from combinations of coating precursors enables the discovery of new functional materials for various applications in science and technology in a cost-effective miniaturized manner.

Drug-induced differential gene expression analysis (DGEA) is essential for uncovering the molecular basis of cell phenotypic changes and understanding individual tumor responses to anticancer drugs. Performing high throughput DGEA is challenging due to the high cost and labor-intensive multi-step sample preparation protocols. In particular, performing drug-induced DGEA on cancer cells derived from patient biopsies is even more challenging due to the scarcity of available cells. A novel, miniaturized, nanoliter-scale method for drug-induced DGEA is introduced, enabling high-throughput and parallel analysis of patient-derived cell drug responses, overcoming the limitations and laborious nature of traditional protocols. The method is based on the Droplet Microarray (DMA), a microscope glass slide with hydrophilic spots on a superhydrophobic background, facilitating droplet formation for cell testing. DMA allows microscopy-based phenotypic analysis, cDNA extraction, and DGEA. The procedure includes cell lysis for mRNA isolation and cDNA conversion followed by droplet pooling for qPCR analysis. In this study, the drug-induced DGEA protocol on the DMA platform is demonstrated using patient-derived chronic lymphocytic leukemia (CLL) cells. This methodology is critical for DGEA with limited cell numbers and promise for applications in functional precision oncology. This method enables molecular profiling of patient-derived samples after drug treatment, crucial for understanding individual tumor responses to anticancer drugs., (© 2024 The Author(s). Advanced Healthcare Materials published by Wiley‐VCH GmbH.)

Label-free real-time monitoring of cellular behavior using impedance spectroscopy is important for drug development and toxicological assessments. Parallelization and miniaturization of such experiments are essential for increasing throughput and enabling experiments with low abundant stem or primary cells. Traditional methods are not miniaturized and require large volumes of reagents and number of cells, limiting their suitability for cost effective high-throughput screening of cells of limited availability. Here, the fabrication, optimization, and application of a bioelectrical signaling monitoring system - electrode droplet microarray (eDMA) are demonstrated. The eDMA platform is based on preparation of a hydrophilic-superhydrophobic patterns covering an array of individually addressable microelectrodes, which confines cells to individual microelectrodes, allowing for parallel, real-time, and label-free detection of cellular responses to drug treatments in nanoliter droplets. The real-time monitoring of cytotoxic effect of an anticancer drug is demonstrated over 48 h with real-time calculation of the half-inhibitory concentration (IC50) values through impedance spectroscopy. This demonstrates eDMA's ability to dynamically assess responses to various drugs in parallel at any given time point, which is crucial for functional personalized oncology. Specifically, the platform can be employed for monitoring anticancer drug toxicity using limited patient samples, where the miniaturization provided by eDMA is essential., (© 2024 The Author(s). Advanced Healthcare Materials published by Wiley‐VCH GmbH.)

The development of miniaturized high-throughput in situ screening platforms capable of handling the entire process of drug synthesis to final screening is essential for advancing drug discovery in the future. In this study, an approach based on combinatorial solid-phase synthesis, enabling the efficient synthesis of libraries of proteolysis targeting chimeras (PROTACs) in an array format is presented. This on-chip platform allows direct biological screening without the need for transfer steps.  UV-induced release of target molecules into individual droplets facilitates further on-chip experimentation. Utilizing a mitogen-activated protein kinase kinases (MEK1/2) degrader as a template, a series of 132 novel PROTAC-like molecules is synthesized using solid-phase Ugi reaction. These compounds are further characterized using various methods, including matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) imaging, while consuming only a few milligrams of starting materials in total. Furthermore, the feasibility of culturing cancer cells on the modified spots and quantifying the effect of MEK suppression is demonstrated. The miniaturized synthesis platform lays a foundation for high-throughput in situ biological screening of potent PROTACs for potential anticancer activity and offers the potential for accelerating the drug discovery process by integrating miniaturized synthesis and biological steps on the same array., (© 2024 Wiley‐VCH GmbH.)

Using thiol-ene chemistry, polymerization-induced phase separation, and DLP 3D printing, we present a method for manufacturing reactive macroporous 3D structures. This approach enables the fabrication of structures with tunable physicochemical properties and compressibility. Moreover, it facilitates post-functionalization through thiol-Michael addition reactions, thereby expanding performance and application potential.

In inkjet printing technology, one important factor influencing the printing quality and reliability of printed films is the interaction of the jetted ink with the substrate surface. This short-range interaction determines the wettability and the adhesion of the ink to the solid surface and is hence responsible for the final shape of the deposited ink. Here, we investigate wetting morphologies of inkjet-printed inks on patterned substrates by carefully designed experimental test structures and simulations. The contact angles, the surface properties, and drop shapes, as well as their influence on the device variability, are experimentally and theoretically analyzed. For the simulations, we employ the phase-field method, which is based on the free energy minimization of the two-phase system with the given wetting boundary conditions. Through a systematic investigation of printed drops on patterned substrates consisting of hydrophilic and hydrophobic areas, we report that the printed morphology is related not only to the designed layout and the drop volume but also to the printing strategy and the wettability. Furthermore, we show how one can modify the intrinsic wettability of the patterned substrates to enhance the printing quality and reliability. Based on the present findings, we cast light on the improvement of the fabrication quality of thin film transistors.

The Droplet Microarray (DMA) has emerged as a tool for high-throughput biological and chemical applications by enabling miniaturization and parallelization of experimental processes. Due to its ability to hold hundreds of nanoliter droplets, the DMA enables simple screening and analysis of samples such as cells and biomolecules. However, handling of nanoliter volumes poses a challenge, as manual recovery of nanoliter volumes is not feasible, and traditional laboratory equipment is not suited to work with such low volumes, and small array formats. To tackle this challenge, we developed the Automated Nanoliter Droplet Selection device (ANDeS), a robotic system for automated collection and transfer of nanoliter samples from DMA. ANDeS can automatically collect volumes from 50 to 350 nL from the flat surface of DMA with a movement accuracy of ±30 µm using fused silica capillaries. The system can automatically collect and transfer the droplets from DMA chip into other platforms, such as microtiter plates, conical tubes or another DMA. In addition, to ensure high throughput and multiple droplet collection, the uptake of multiple droplets within a single capillary, separated by air gaps to avoid mixing of the samples within the capillary, was optimized and demonstrated. This study shows the potential of ANDeS in laboratory applications by using it for the collection and transfer of biological samples, contained in nanoliter droplets, for subsequent analysis. The experimental results demonstrate the ability of ANDeS to increase the versatility of the DMA platform by allowing for automated retrieval of nanoliter samples from DMA, which was not possible manually on the level of individual droplets. Therefore, it widens the variety of analytical techniques that can be used for the analysis of content of individual droplets and experiments performed using DMA. Thus, ANDeS opens up opportunities to expand the development of miniaturized assays in such fields as cell screening, omics analysis and combinatorial chemistry., Competing Interests: Declaration of Competing Interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Joaquin E. Urrutia Gomez reports financial support was provided by German Academic Exchange Service. Anna A. Popova reports financial support was provided by Impuls und Vernetzungsfonds der Helmholtz-Gemeinschaft, Heidelberg and Karlsruhe cooperation program. Pavel A. Levkin reports financial support was provided by German Research Foundation. Pavel A. Levkin reports a relationship with Aquarray GmbH that includes: board membership. Anna A. Popova reports a relationship with Aquarray GmbH that includes: board membership., (Copyright © 2023. Published by Elsevier Inc.)

Polymer gel-based pressure sensors offer numerous advantages over traditional sensing technologies, including excellent conformability and integration into wearable devices. However, challenges persist in terms of their performance and manufacturing technology. In this study, a method for fabricating gel pressure sensors using a hydrophobic/hydrophilic patterned surface is introduced. By shaping and fine-tuning the droplets of the polymer gel prepolymerization solution on the patterned surface, remarkable sensitivity improvements compared to unshaped hydrogels have been achieved. This also showcased the potential for tailoring gel pressure sensors to different applications. By optimizing the configuration of the sensor array, an uneven conductive gel array is fabricated, which exhibited a high sensitivity of 0.29 kPa -1 in the pressure range of 0-30 kPa, while maintaining a sensitivity of 0.13 kPa -1 from 30 kPa up to 100 kPa. Furthermore, the feasibility of using these sensors for human motion monitoring is explored and a conductive gel array for 2D force detection is successfully developed. This efficient and scalable fabrication method holds promise for advancing pressure sensor technology and offers exciting prospects for various industries and research fields., (© 2023 The Authors. Small published by Wiley-VCH GmbH.)

The rising costs of pharmaceutical research are currently limiting the productivity of drug discovery and development, but can potentially be diminished via miniaturization of the synthesis and screening of new compounds. As droplet microarrays already present themselves as a versatile tool for highly miniaturized biological screening of various targets, their use for chemical synthesis is still limited. In this study, the influential palladium-catalyzed Suzuki-Miyaura reaction is successfully implemented at the nanoliter scale on droplet microarrays for the synthesis of an 800-compound library of biphenyls. Each reaction is carried out in individual 150 nL droplets. Remarkably, the synthesis of these 800 compounds requires a minimal amount of reagents, totaling 80 µmol, and a solvent volume of 400 µL. Furthermore, the cleavage kinetics and purity of the obtained biphenylic compounds are investigated. Via the solid-phase synthesis approach, the compounds could be purified from excess reactants and catalyst prior to the analysis and a UV-cleavable linker allows for fast and additive-free cleavage of each compound into the individual 100 nL droplet. This novel approach expands the toolbox of the droplet microarray for miniaturized high-throughput chemical synthesis and paves the way for future synthesis and screening of chemical compounds in a single platform., (© 2023 The Authors. Small published by Wiley-VCH GmbH.)

Superabsorbers based on crosslinked sodium polyacrylate polymers cannot be easily recycled, resulting in 2 million tons of superabsorbers being landfilled or burned every year. A fast and efficient strategy to recycle superabsorbers would significantly alleviate environmental pollution and promote a sustainable use of these polymers. Herein, the rapid recycling of crosslinked sodium polyacrylate hydrogels based on their inherent UV degradation is demonstrated without the need for chemicals besides water. A quantitative conversion of crosslinked sodium polyacrylate into soluble sodium polyacrylate is achieved in minutes, almost 200 times faster than a previous approach based on de-esterification. The obtained soluble sodium polyacrylate can be used, for example, as a thickener for aqueous dyes or can be esterified with n -butanol or 2-ethylhexanol to serve as a pressure-sensitive adhesive. The UV photodegradation and esterification of superabsorbers is fast, scalable, safe, and economical and yields polymers with controllable molecular weight in the range of 100-400 kg/mol. It thus offers distinct advantages over the chemical de-crosslinking strategies presented previously.

Label-free fluorescence-based chemosensing has been increasingly brought into focus due to its simplicity and high sensitivity for intracellular monitoring of molecules. Currently used methods, such as conventional indicator displacement assays (IDAs), pose limitations related to dissociation upon dilution, random diffusion of the released indicators, and high sensitivity to interference by agents from the ambient cellular environment ( e.g. , salts, enzymes, and proteins). Herein we report a potentially widely applicable strategy to overcome the limitations of conventional IDAs by employing a macrocyclic cucurbit[7]uril (CB7) host covalently coupled to a nitrobenzoxadiazole (NBD) fluorescent dye (CB7-NBD conjugate). As a proof of concept, we demonstrated that the CB7-NBD unimolecular conjugate responded to various target analytes even in the complex live cell system. Moreover, the sensing system was compatible with fluorescence imaging, fluorescence-assisted cell sorting (FACS), and fluorescence spectrometry with a microplate reader. These experiments demonstrated an application of covalently bound unimolecular CB7-NBD conjugate as a sensor for detecting diverse analytes in the intracellular compartment of live cells., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

Due to the large chemical space, the design of functional and responsive soft materials poses many challenges but also offers a wide range of opportunities in terms of the scope of possible properties. Herein, an experimental workflow for miniaturized combinatorial high-throughput screening of functional hydrogel libraries is reported. The data created from the analysis of the photodegradation process of more than 900 different types of hydrogel pads are used to train a machine learning model for automated decision making. Through iterative model optimization based on Bayesian optimization, a substantial improvement in response properties is achieved and thus expanded the scope of material properties obtainable within the chemical space of hydrogels in the study. It is therefore demonstrated that the potential of combining miniaturized high-throughput experiments with smart optimization algorithms for cost and time efficient optimization of materials properties., (© 2023 The Authors. Small Methods published by Wiley-VCH GmbH.)

To address the challenge of drug resistance and limited treatment options for recurrent gliomas with IDH1 mutations, a highly miniaturized screening of 2208 FDA-approved drugs is conducted using a high-throughput droplet microarray (DMA) platform. Two patient-derived temozolomide-resistant tumorspheres harboring endogenous IDH1 mutations (IDH1 mut ) are utilized. Screening identifies over 20 drugs, including verteporfin (VP), that significantly affected tumorsphere formation and viability. Proteomics analysis reveals that nuclear pore complex may be a potential VP target, suggesting a new mechanism of action independent of its known effects on YAP1. Knockdown experiments exclude YAP1 as a drug target in tumorspheres. Pathway analysis shows that NUP107 is a potential upstream regulator associated with VP response. Analysis of publicly available genomic datasets shows a significant correlation between high NUP107 expression and decreased survival in IDH1 mut astrocytoma, suggesting NUP107 may be a potential biomarker for VP response. This study demonstrates a miniaturized approach for cost-effective drug repurposing using 3D glioma models and identifies nuclear pore complex as a potential target for drug development. The findings provide preclinical evidence to support in vivo and clinical studies of VP and other identified compounds to treat IDH1 mut gliomas, which may ultimately improve clinical outcomes for patients with this challenging disease., (© 2023 The Authors. Advanced Healthcare Materials published by Wiley-VCH GmbH.)

Cancer is a devastating disease and the second leading cause of death worldwide. However, the development of resistance to current therapies is making cancer treatment more difficult. Combining the multi-omics data of individual tumors with information on their in-vitro Drug Sensitivity and Resistance Test (DSRT) can help to determine the appropriate therapy for each patient. Miniaturized high-throughput technologies, such as the droplet microarray, enable personalized oncology. We are developing a platform that incorporates DSRT profiling workflows from minute amounts of cellular material and reagents. Experimental results often rely on image-based readout techniques, where images are often constructed in grid-like structures with heterogeneous image processing targets. However, manual image analysis is time-consuming, not reproducible, and impossible for high-throughput experiments due to the amount of data generated. Therefore, automated image processing solutions are an essential component of a screening platform for personalized oncology. We present our comprehensive concept that considers assisted image annotation, algorithms for image processing of grid-like high-throughput experiments, and enhanced learning processes. In addition, the concept includes the deployment of processing pipelines. Details of the computation and implementation are presented. In particular, we outline solutions for linking automated image processing for personalized oncology with high-performance computing. Finally, we demonstrate the advantages of our proposal, using image data from heterogeneous practical experiments and challenges., (© 2023. The Author(s).)

Limited response rates and frequent relapses during standard of care with hypomethylating agents in myelodysplastic neoplasms (MN) require urgent improvement of this treatment indication. Here, by combining 5-azacytidine (5-AZA) with the pan-lysyl oxidase inhibitor PXS-5505, we demonstrate superior restoration of erythroid differentiation in hematopoietic stem and progenitor cells (HSPCs) of MN patients in 20/31 cases (65%) versus 9/31 cases (29%) treated with 5-AZA alone. This effect requires direct contact of HSPCs with bone marrow stroma components and is dependent on integrin signaling. We further confirm these results in vivo using a bone marrow niche-dependent MN xenograft model in female NSG mice, in which we additionally demonstrate an enforced reduction of dominant clones as well as significant attenuation of disease expansion and normalization of spleen sizes. Overall, these results lay out a strong pre-clinical rationale for efficacy of combination treatment of 5-AZA with PXS-5505 especially for anemic MN., (© 2023. The Author(s).)

In the current drug discovery process, the synthesis of compound libraries is separated from biological screenings both conceptually and technologically. One of the reasons is that parallel on-chip high-throughput purification of synthesized compounds is still a major challenge. Here, on-chip miniaturized high-throughput liquid-liquid extraction in volumes down to 150 nL with efficiency comparable to or better than large-scale extraction utilizing separation funnels is demonstrated. The method is based on automated and programmable merging of arrays of aqueous nanoliter droplets with organic droplets. Multi-step extraction performed simultaneously or with changing conditions as well as handling of femtomoles of compounds are demonstrated. In addition, the extraction efficiency is analyzed with a fast optical readout as well as matrix-assisted laser desorption ionization-mass spectrometry on-chip detection. The new massively parallel and miniaturized purification method adds another important tool to the chemBIOS concept combining chemical combinatorial synthesis with biological screenings on the same miniaturized droplet microarray platform, which will be essential to accelerate drug discovery., (© 2022 The Authors. Small published by Wiley-VCH GmbH.)

Multidrug-resistant (MDR) bacteria is a severe threat to public health. Therefore, it is urgent to establish effective screening systems for identifying novel antibacterial compounds. In this study, a highly miniaturized droplet microarray (DMA) based high-throughput screening system is established to screen over 2000 compounds for their antimicrobial properties against carbapenem-resistant Klebsiella pneumoniae and methicillin resistant Staphylococcus aureus (MRSA). The DMA consists of an array of hydrophilic spots divided by superhydrophobic borders. Due to the differences in the surface wettability between the spots and the borders, arrays of hundreds of nanoliter-sized droplets containing bacteria and different drugs can be generated for screening applications. A simple colorimetric viability readout utilizing a conventional photo scanner is developed for fast single-step detection of the inhibitory effect of the compounds on bacterial growth on the whole array. Six hit compounds, including coumarins and structurally simplified estrogen analogs are identified in the primary screening and validated with minimum inhibition concentration assay for their antibacterial effect. This study demonstrates that the DMA-based high-throughput screening system enables the identification of potential antibiotics from novel synthetic compound libraries, offering opportunities for development of new treatments against multidrug-resistant bacteria., (© 2022 The Authors. Advanced Biology published by Wiley-VCH GmbH.)

Ionically conductive elastomers are necessary for realizing human-machine interfaces, bioelectronic applications, or durable wearable sensors. Current design strategies, however, often suffer from solvent leakage and evaporation, or from poor mechanical properties. Here, we report a strategy to fabricate ionic elastomers (IHPs) demonstrating high conductivity (0.04 S m -1 ), excellent electrochemical stability (>60,000 cycles), ultra-stretchability (up to 1400%), high toughness (7.16 MJ m -3 ), and fast self-healing properties, enabling the restoration of ionic conductivity within seconds, as well as no solvent leakage. The ionic elastomer is composed of in situ formed physically cross-linked poly(2-hydroxyethyl methacrylate) networks and poly(ethylene glycol) (PEG). The long molecular chains of PEG serve as a solvent for dissolving electrolytes, improve its long-term stability, reduce solvent leakage, and ensure the outstanding mechanical properties of the IHP. Surprisingly, the incorporation of ions into PEG simultaneously enhances the strength and toughness of the elastomer. The strengthening and toughening mechanisms were further revealed by molecular simulation. We demonstrate an application of the IHPs as (a) flexible sensors for strain or temperature sensing, (b) skin electrodes for recording electrocardiograms, and (c) a tough and sensing material for pneumatic artificial muscles. The proposed strategy is simple and easily scalable and can further inspire the design of novel ionic elastomers for ionotronics applications.

Human induced pluripotent stem cells (hiPSCs) are crucial for disease modeling, drug discovery, and personalized medicine. Animal-derived materials hinderapplications of hiPSCs in medical fields. Thus, novel and well-defined substrate coatings capable of maintaining hiPSC pluripotency are important for advancing biomedical applications of hiPSCs. Here a miniaturized droplet microarray (DMA) platform to investigate 11 well-defined proteins, their 55 binary and 165 ternary combinations for their ability to maintainpluripotency of hiPSCs when applied as a surface coating, is used. Using this screening approach, ten protein group coatings are identified, which promote significantly higher NANOG expression of hiPSCs in comparison with Matrigel coating. With two of the identified coatings, long-term pluripotency maintenance of hiPSCs and subsequent differentiation into three germ layers are achieved. Compared with conventional high-throughput screening (HTS) in 96-well plates, the DMA platform uses only 83 µL of protein solution (0.83 µg total protein) and only ≈2.8 × 10 5 cells, decreasing the amount of proteins and cells ≈860 and 25-fold, respectively. The identified proteins will be essential for research and applications using hiPSCs, while the DMA platform demonstrates great potential for miniaturized HTS of scarce cells or expensive materials such as recombinant proteins., (© 2022 The Authors. Advanced Healthcare Materials published by Wiley-VCH GmbH.)

Superhydrophobic surfaces with regional functions have widespread applications in biotechnology, diagnostic applications, and micro-chemical synthesis and analysis. However, owing to their chemical inertness, superhydrophobic surfaces with chemical reactivity are difficult to achieve. Superhydrophobic surfaces that can be further modified with varied densities and expanded species of the functional moieties are not readily available. In this study, a single-step approach to achieve a reactive superhydrophobic surface is reported, on which chemical grafting of a library of molecules can be carried out through surface-initiated atom-transfer radical addition or surface-initiated atom-transfer radical polymerization. The excellent spatial and temporal controllability of these chemical processes under visible light enables us to take advantage of programmed liquid-crystal-display (LCD) or Digital Light Processing (DLP) photolithography systems to effortlessly regulate the location, density, and species of the functional molecules on the reactive superhydrophobic surface. The distinctive properties of this surface will provide new insight into intelligent superhydrophobic material development and practical applications, such as aqueous/oil microdroplets array, multi-anti-counterfeiting labels and integrated microfluidic reactors with enzymes for chemical logic learning., (© 2022 Wiley-VCH GmbH.)

Insufficient binding selectivity of chemosensors often renders biorelevant metabolites indistinguishable by the widely used indicator displacement assay. Array-based chemosensing methods are a common workaround but require additional effort for synthesizing a chemosensor library and setting up a sensing array. Moreover, it can be very challenging to tune the inherent binding preference of macrocyclic systems such as cucurbit[ n ]urils (CB n ) by synthetic means. Using a novel cucurbit[7]uril-dye conjugate that undergoes salt-induced adaptation, we now succeeded in distinguishing 14 bioorganic analytes from each other through the facile stepwise addition of salts. The salt-specific concentration-resolved emission provides additional information about the system at a low synthetic effort. We present a data-driven approach to translate the human-visible curve differences into intuitive pairwise difference measures. Ion mobility experiments combined with density functional theory calculations gave further insights into the binding mechanism and uncovered an unprecedented ternary complex geometry for CB7. TThis work introduces the non-selectively binding, salt-adaptive cucurbit[ n ]uril system for sensing applications in biofluids such as urine, saliva, and blood serum.

Desorption/ionization (DI)-mass spectrometric (MS) methods offer considerable advantages of rapidity and low-sample input for the analysis of solid biological matrices such as tissue sections. The concept of desorption electrospray ionization (DESI) offers the possibility to ionize compounds from solid surfaces at atmospheric pressure, without the addition of organic compounds to initiate desorption. However, severe drawbacks from former DESI hardware stability made the development of assays for drug quantification difficult. In the present study, the potential of new prototype source setups (High Performance DESI Sprayer and Heated Transfer Line) for the development of drug quantification assays in tissue sections was evaluated. It was demonstrated that following dedicated optimization, new DESI XS enhancements present promising options regarding targeted quantitative analyses. As a model compound for these developments, ulixertinib, an inhibitor of extracellular signal-regulated kinase (ERK) 1 and 2 was used.

In vitro cell-based experiments are particularly important in fundamental biological research. Microscopy-based readouts to identify cellular changes in response to various stimuli are a popular choice, but gene expression analysis is essential to delineate the underlying molecular dynamics in cells. However, cell-based experiments often suffer from interexperimental variation, especially while using different readout methods. Therefore, establishment of platforms that allow for cell screening, along with parallel investigations of morphological features, as well as gene expression levels, is crucial. The droplet microarray (DMA) platform enables cell screening in hundreds of nanoliter droplets. In this study, a "Cells-to-cDNA on Chip" method is developed enabling on-chip mRNA isolation from live cells and conversion to cDNA in individual droplets of 200 nL. This novel method works efficiently to obtain cDNA from different cell numbers, down to single cell per droplet. This is the first established miniaturized on-chip strategy that enables the entire course of cell screening, phenotypic microscopy-based assessments along with mRNA isolation and its conversion to cDNA for gene expression analysis by real-time PCR on an open DMA platform. The principle demonstrated in this study sets a beginning for myriad of possible applications to obtain detailed information about the molecular dynamics in cultured cells., (© 2022 The Authors. Advanced Healthcare Materials published by Wiley-VCH GmbH.)

The ability of light to remotely control the properties of soft matter materials in a dynamic fashion has fascinated material scientists and photochemists for decades. However, only recently has our ability to map photochemical reactivity in a finely wavelength resolved fashion allowed for different colors of light to independently control the material properties of polymer networks with high precision, driven by monochromatic irradiation enabling orthogonal reaction control. The current concept article highlights the progress in visible light-induced photochemistry and explores how it has enabled the design of polymer networks with dynamically adjustable properties. We will explore current applications ranging from dynamic hydrogel design to the light-driven adaptation of 3D printed structures on the macro- and micro-scale. While the alternation of mechanical properties via remote control is largely reality for soft matter materials, we herein propose the next frontiers for adaptive properties, including remote switching between conductive and non-conductive properties, hydrophobic and hydrophilic surfaces, fluorescent or non-fluorescent, and cell adhesive vs. cell repellent properties., (© 2022 The Authors. Chemistry - A European Journal published by Wiley-VCH GmbH.)

The inverse vulcanization produces high sulfur content polymers from alkenes and elemental sulfur. Control over properties such as the molar mass or the solubility of polymers is not well established, and existing strategies lack predictability or require large variations of the composition. Systematic design principles are sought to allow for a targeted design of materials. Herein, we report on the inverse vulcanization of norbornenylsilanes (NBS), with a different number of hydrolysable groups at the silicon atom. Inverse vulcanization of mixtures of NBS followed by polycondensation yielded soluble high sulfur content copolymers (50 wt % S) with controllable weight average molar mass (M W ), polydispersity (Đ), glass transition temperature (T G ), or zero-shear viscosity (η 0 ). Polycondensation was conducted in the melt with HCl as a catalyst, abolishing the need for a solvent. Purification by precipitation afforded polymers with a greatly reduced amount of low molar mass species., (© 2022 The Authors. Angewandte Chemie International Edition published by Wiley-VCH GmbH.)

Polymer gels, such as hydrogels, have been widely used in biomedical applications, flexible electronics, and soft machines. Polymer network design and its contribution to the performance of gels has been extensively studied. In this study, the critical influence of the solvent nature on the mechanical properties and performance of soft polymer gels is demonstrated. A polymer gel platform based on poly(ethylene glycol) (PEG) as solvent is reported (PEGgel). Compared to the corresponding hydrogel or ethylene glycol gel, the PEGgel with physically cross-linked poly(hydroxyethyl methacrylate-co-acrylic acid) demonstrates high stretchability and toughness, rapid self-healing, and long-term stability. Depending on the molecular weight and fraction of PEG, the tensile strength of the PEGgels varies from 0.22 to 41.3 MPa, fracture strain from 12% to 4336%, modulus from 0.08 to 352 MPa, and toughness from 2.89 to 56.23 MJ m -3 . Finally, rapid self-healing of the PEGgel is demonstrated and a self-healing pneumatic actuator is fabricated by 3D-printing. The enhanced mechanical properties of the PEGgel system may be extended to other polymer networks (both chemically and physically cross-linked). Such a simple 3D-printable, self-healing, and tough soft material holds promise for broad applications in wearable electronics, soft actuators and robotics., (© 2022 The Authors. Advanced Materials published by Wiley-VCH GmbH.)

Simple and rapid imaging and analysis of 2D and 3D cell culture compatible with miniaturized arrays of nanoliter droplets are essential for high-throughput screening and personalized medicine applications. In this study, we have developed a simple one-step, cost-effective and sensitive colorimetric method for the analysis of cell viability in 2D and 3D cell cultures on a nanoliter droplet microarray. The method utilizes a flatbed document scanner that detects a color change in response to cell metabolism in nanoliter droplets with high sensitivity in a single step without the need for expensive specialized equipment. This new nanoliter-based method is faster and more sensitive than equivalent methods using multi-well plate assays. The method detects quantifiable signal from as few as 10 cells and requires only 5 min. This is 2.5 to 10-fold more sensitive and 12 times faster than the same assay in multi-well plates. The method is simple, affordable, fast and sensitive. It can be used for various applications including high-throughput cell-based and biochemical screenings., Competing Interests: Declaration of Competing Interests The authors declare the following potential conflicts of interest with respect to the research, authorship, and/or publication of this article: in addition to being employed by the Karlsruhe Institute of Technology, A.A.P. and P.A.L. are (since March 2018) shareholders of Aquarray GmbH. M.R., D.K., H.C. and T.A. declare that there is no conflict of interest regarding the publication of this article., (Copyright © 2021. Published by Elsevier Inc.)

Hypothesis: Droplet wetting on a solid substrate is affected by the surface heterogeneity. Introducing patterned wettability on the solid substrate is expected to engender anisotropic wetting morphologies, thereby manipulating droplet wetting behaviors. However, when the droplet size is comparable with that of the surface heterogeneity, the wetting morphologies cannot be depicted by the quintessential Cassie's theory but should be possible to be predicted from the perspective of thermodynamics via surface energy minimization., Methods: Here, we investigate the equilibrium droplet shapes on chemically patterned substrates by using an analytical model, phase-field simulations, and experiments. The former two methods are sharp and diffuse interface treatments, respectively, which both are based on minimizing the free energy of the system. The experimental results are obtained by depositing droplets on chemically patterned glass substrates., Findings: Various anisotropic wetting shapes are found from the three methods. Excellent agreement is observed between different methods, showing the possibility to quantify the anisotropic wetting droplet morphologies on patterned substrates by present methods. We also address a series of non-rotationally symmetric droplet shapes, which is the first resport about these special wetting morphologies. Furthermore, we reveal the anisotropic wetting shapes in a quasi-equilibrium evaporation process., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Actuated structures are becoming relevant in medical fields; however, they call for flexible/soft-base materials that comply with biological tissues and can be synthesized in simple fabrication steps. In this work, we extend the palette of techniques to afford soft, actuable spherical structures taking advantage of the biosynthesis process of bacterial cellulose. Bacterial cellulose spheres (BCS) with localized magnetic nanoparticles (NPs) have been biosynthesized using two different one-pot processes: in agitation and on hydrophobic surface-supported static culture, achieving core-shell or hollow spheres, respectively. Magnetic actuability is conferred by superparamagnetic iron oxide NPs (SPIONs), and their location within the structure was finely tuned with high precision. The size, structure, flexibility and magnetic response of the spheres have been characterized. In addition, the versatility of the methodology allows us to produce actuated spherical structures adding other NPs (Au and Pt) in specific locations, creating Janus structures. The combination of Pt NPs and SPIONs provides moving composite structures driven both by a magnetic field and a H 2 O 2 oxidation reaction. Janus Pt/SPIONs increased by five times the directionality and movement of these structures in comparison to the controls.

The rapid development of 3D printing (or additive manufacturing) technologies demands new materials with novel properties and functionalities. Superhydrophobic materials, owing to their ultralow water adhesion, self-cleaning, anti-biofouling, or superoleophilic properties are useful for myriad applications involving liquids. However, the majority of the methods for making superhydrophobic surfaces have been based on surface functionalization and coatings, which are challenging to apply to 3D objects. Additionally, these coatings are vulnerable to abrasion due to low mechanical stability and limited thickness. Here, a new materials concept and methodology for 3D printing of superhydrophobic macroscopic objects with bulk nanostructure and almost unlimited geometrical freedom is presented. The method is based on a specific ink composed of hydrophobic (meth)acrylate monomers and porogen solvents, which undergoes phase separation upon photopolymerization to generate inherently nanoporous and superhydrophobic structures. Using a desktop Digital Light Processing printer, superhydrophobic 3D objects with complex shapes are demonstrated, with ultralow and uniform water adhesion measured with scanning droplet adhesion microscopy. It is shown that the 3D-printed objects, owing to their nanoporous structure throughout the entire volume, preserve their superhydrophobicity upon wear damage. Finally, a superhydrophobic 3D-printed gas-permeable and water-repellent microfluidic device and a hierarchically structured 3D-printed super-oil-absorbent are demonstrated., (© 2021 The Authors. Advanced Materials published by Wiley-VCH GmbH.)

The capacity of human induced pluripotent stem cells (hiPSCs) for indefinite self-renewal warrants their application in disease modeling, drug discovery, toxicity assays and efficacy screening. However, their poor proliferation ability, inability to adhere to surfaces without Matrigel coating and tendency to spontaneously differentiate in vitro hinder the application of hiPSCs in these fields. Here we study the ability to culture hiPSCs inside 200 ​nL droplets on the droplet microarray (DMA) platform. We demonstrate that (1) hiPSCs can attach to the Matrigel (MG)-free surface of DMA and show good viability after 24 h culture; (2) hiPSC do not spontaneously differentiate when cultured on the MG-free surface of DMAs; (3) culturing of hiPSCs in 200 ​nL as compared to 2 ​mL culture leads to higher expression of the Nanog pluripotency marker. Overall, the results demonstrate the possibility to culture undifferentiated hiPSCs in 200 ​nL droplets on DMA, thereby opening the possibility for high-throughput screenings of hiPSCs with various factors without compromising the results through the involvement of animal-derived materials, such as Matrigel., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Both A.A.P. and P.A.L. in addition to being employed by the KIT are also shareholders of Aquarray GmbH. Other co-authors declare no conflict of interests., (© 2020 The Author(s).)

Light-based microfabrication techniques constitute an indispensable approach to fabricate tissue assemblies, benefiting from noncontact spatially and temporarily controlled manipulation of soft matter. Light-triggered degradation of soft materials, such as hydrogels, is important in tissue engineering, bioprinting, and related fields. The photoresponsiveness of hydrogels is generally not intrinsic and requires complex synthetic procedures wherein photoresponsive crosslinking groups are incorporated into the hydrogel. This paper demonstrates a novel biocompatible and inherently photodegradable poly(ethylene glycol) methacrylate (PEGMA)-based gelatin-methacryloyl (GelMA)-containing hydrogel that can be used to culture cells in 3D for at least 14 d. These gels are conveniently and quickly degraded via UV irradiation for 10 min to produce structured hydrogels of various geometries, sizes, and free-standing cell-laden hydrogel particles. These structures can be flexibly produced on demand. In particular, photodegradation can be temporarily delayed from photopolymerization, offering an alternative to hydrogel array production via photopolymerization with a photomask. The paper investigates the influences of hydrogel composition and swelling liquid on both its photodegradability and biocompatibility., (© 2021 The Authors. Advanced Healthcare Materials published by Wiley-VCH GmbH.)

Liquids are traditionally handled and stored in solid vessels. Solid walls are not functional, adaptive, or self-repairing, and are difficult to remove and re-form. Liquid walls can overcome these limitations, but cannot form free-standing 3D walls. Herein, a liquid analogue of a well, termed a "liquid well" is introduced. Water tethered to a surface with hydrophobic-hydrophilic core-shell patterns forms stable liquid walls capable of containing another immiscible fluid, similar to fluid confinement by solid walls. Liquid wells with different liquids, volumes, and shapes are prepared and investigated by confocal and Raman microscopy. The confinement of various low-surface-tension liquids (LSTLs) on surfaces by liquid wells can compete with or be complementary to existing confinement strategies using perfluorinated surfaces, for example, in terms of the shape and height of the confined LSTLs. Liquid wells show unique properties arising from their liquid aggregate state: they are self-healing, dynamic, and functional, that is, not restricted to a passive confining role. Water walls can be easily removed and re-formed, making them interesting as sacrificial templates. This is demonstrated in a process termed water-templated polymerization (WTP). Numerical phase-field model simulations are performed to scrutinize the conditions required for the formation of stable liquid wells., (© 2021 The Authors. Advanced Materials published by Wiley-VCH GmbH.)

Testing the sensitivity of patient-derived tumor cells ex vivo can potentially help determining the appropriate treatment for each patient and spot the development of resistance to a given therapy. The number of cells obtainable from a biopsy is, however, often insufficient for performing ex vivo tests in conventional microtiter plates. Here, we introduce a novel Droplet-Microarray platform based on a hydrophilic-superhydrophobic patterned surface that enables screenings using only 100 cells and 30 picomoles of a drug per individual nanoliter-sized droplet. We demonstrate that the dose-response of as few as 100 primary patient-derived chronic lymphocytic leukemia (CLL) cells to anticancer compounds on the Droplet-Microarray platform resembles the dose-response obtained in 384-well plates requiring 20,000 tumor cells per experiment. The extremely miniaturized Droplet-Microarray platform thus carries great potential for ex vivo drug sensitivity and resistance tests on patient-derived tumor cells and potentially for implementing such tests in medical practice of precision medicine.

Conductive polymers have been intensively investigated as materials for electrodes in flexible electronics due to their favorable biocompatibility and reliable electrochemical stability. Nevertheless, patterning of conductive polymers for the fabrication of devices and in various electronics applications confronts multifarious limitations and challenges. Here, we present a simple but efficient strategy to obtain conductive polymer microelectrodes via utilization of surface-tension-confined liquid patterns. This method shows universality for various oxidizers and conductive polymers, high resolution, stability, and favorable compatibility with different surfaces and materials. The developed method has been demonstrated for creating conductive polymer microelectrodes with a customized reaction process, defined geometry, and flexible substrates. The obtained microelectrodes were assembled into flexible capacitive sensors. Thus, the method realizes a facile approach to conductive polymer microelectrodes for flexible electronics, biomedical applications, human activity monitors, and electronic skin.

The development of novel chemically developed and physically defined surfaces and environments for cell culture and screening is important for various biological applications. The Droplet microarray (DMA) platform based on hydrophilic-superhydrophobic patterning enables high-throughput cellular screening in nanoliter volumes and on various biocompatible surfaces. Here we performed phenotypic and transcriptomic analysis of HeLa-CCL2 cells cultured on DMA, with a goal to analyze cellular response on different surfaces and culture volumes down to 3 nL, compared with conventional cell culture platforms. Our results indicate that cells cultured on four tested substrates: nanostructured nonpolymer, rough and smooth variants of poly(2-hydroxyethyl methacrylate- co -ethylene dimethacrylate) polymer and poly(thioether) dendrimer are compatible with cells grown in Petri dish. Cells cultured on nanostructured nonpolymer coating exhibited the closet transcriptomic resemblance to that of cells grown in Petri dish. Analysis of cells cultured in 100, 9, and 3 nL media droplets on DMA indicated that all but cells grown in 3 nL volumes had unperturbed viability with minimal alterations in the transcriptome compared with 96-well plate. Our findings demonstrate the applicability of DMA for cell-based assays and highlight the possibility of establishing regular cell culture on various biomaterial-coated substrates and in nanoliter volumes, along with routinely used cell culture platforms., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: S.C., V.G., M.B., and J.S. declare no competing financial interests or personal relationships that could have appeared to influence the work reported in this article. A.P. and P.L. in addition to being employed by the KIT are also shareholders of Aquarray GmbH., (© 2021 Published by Elsevier Ltd.)

The ability to fabricate materials with ultrathin architectures enables the breakthrough of low-dimensional structures with high surface area that showcase distinctive properties from their bulk counterparts. They are exploited in a wide range of fields, including energy harvesting, catalysis, and biomedicine. Despite such versatility, the fine tuning of the lateral dimensions and geometry of these structures remains challenging. Prepatterned platforms gain significant attention as enabling technologies to process materials with highly controlled shapes and dimensions. Herein, different nanometer-thick particles of various lateral sizes and geometries (e.g., squares, circles, triangles, hexagons) are processed with high precision and definition, taking advantage of the wettability contrast of oleophilic-oleophobic patterned surfaces. Quasi-2D polymeric microparticles with high shape- and size-fidelity can be retrieved as freestanding objects in a single step. These structures show cell-mediated pliability, and their integration in gravity-enforced human adipose-derived stem cell spheroids leads to an enhanced metabolic activity and a modulated secretion of proangiogenic factors., (© 2021 Wiley-VCH GmbH.)

Biomimetic surface coatings based on plant polyphenols and catecholamines have been used broadly in a variety of applications. However, the lack of a rational cost-effective platform for screening these coatings and their properties limits the true potential of these functional materials to be unleashed. Here, we investigated the oxidation behavior and coating formation ability of a library consisting of 45 phenolic compounds and catecholamines. UV-vis spectroscopy demonstrated significant acceleration of oxidation and polymerization under UV irradiation. We discovered that several binary mixtures resulted in non-additive behavior (synergistic or antagonistic effect) yielding much thicker or thinner coatings than individual compounds measured by ellipsometry. To investigate the properties of coatings derived from new combinations, we used a miniaturized high-throughput strategy to screen 2,532 spots coated with single, binary, and ternary combinations of coating precursors in one run. We evaluated the use of machine learning models to learn the relation between the chemical structure of the precursors and the thickness of the nanocoatings. Formation and stability of nanocoatings were investigated in a high-throughput manner via discontinuous dewetting. 30 stable combinations (hits) were used to tune the surface wettability and to form water droplet microarray and spot size gradients of water droplets on the coated surface. No toxicity was observed against eukaryotic HeLa cells and Pseudomonas aeruginosa (strain PA30) bacteria after 24 h incubation at 37 °C. The strategy introduced here for high-throughput screening of nanocoatings derived from combinations of coating precursors enables the discovery of new functional materials for various applications in science and technology in a cost-effective miniaturized manner., Competing Interests: The authors declare that they have no financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 The Author(s).)

In pharmaceutical research and development, cell-based assays are primarily used with readout that rely on fluorescence-based and other label-dependent techniques for analysis of different cellular processes. Superhydrophobic-hydrophilic droplet microarrays (DMA) and matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) have recently emerged as key technologies for miniaturized high-throughput cell assays and for label-free molecular high-content drug profiling, respectively. Here, nanoliter-scale cell assays are integrated on DMAs with MALDI-MS imaging (MALDI-MSI) approaches to a droplet microarray-mass spectrometry imaging (DMA-MSI) platform. Using A549 lung cancer cells, concentration-response profiling of a pharmaceutical compound, the fatty acid synthase inhibitor GSK2194069, are demonstrated. Direct cell culture on DMAs enables combination of microscopy and high speed, high molecular content analysis using MALDI-MSI. Miniaturization of array spots down to 0.5 mm confining 40 nL droplets allows for MALDI imaging analysis of as few as ten cells per spot. Partial automation ensures a fast sample preparation workflow. Taken together, the integrated DMA-MSI platform that combines MALDI-MSI, as a label-free analytical readout, with the miniaturized droplet microarray platform is a valuable complement to high throughput cell-based assays technologies., (© 2021 The Authors. Advanced Biology published by Wiley-VCH GmbH.)

3D printing offers enormous flexibility in fabrication of polymer objects with complex geometries. However, it is not suitable for fabricating large polymer structures with geometrical features at the sub-micrometer scale. Porous structure at the sub-micrometer scale can render macroscopic objects with unique properties, including similarities with biological interfaces, permeability and extremely large surface area, imperative inter alia for adsorption, separation, sensing or biomedical applications. Here, we introduce a method combining advantages of 3D printing via digital light processing and polymerization-induced phase separation, which enables formation of 3D polymer structures of digitally defined macroscopic geometry with controllable inherent porosity at the sub-micrometer scale. We demonstrate the possibility to create 3D polymer structures of highly complex geometries and spatially controlled pore sizes from 10 nm to 1000 µm. Produced hierarchical polymers combining nanoporosity with micrometer-sized pores demonstrate improved adsorption performance due to better pore accessibility and favored cell adhesion and growth for 3D cell culture due to surface porosity. This method extends the scope of applications of 3D printing to hierarchical inherently porous 3D objects combining structural features ranging from 10 nm up to cm, making them available for a wide variety of applications.

Artificial multicellular systems are gaining importance in the field of tissue engineering and regenerative medicine. Reconstruction of complex tissue architectures in vitro is nevertheless challenging, and methods permitting controllable and high-throughput fabrication of complex multicellular architectures are needed. Here, a facile and high-throughput method is developed based on a tunable droplet-fusion technique, allowing programmed assembly of multiple cell spheroids into complex multicellular architectures. The droplet-fusion technique allows for construction of various multicellular architectures (double-spheroids, multi-spheroids, hetero-spheroids) in a miniaturized high-density array format. As an example of application, the propagation of Wnt signaling is investigated within hetero-spheroids formed from two fused Wnt-releasing and Wnt-reporter cell spheroids. The developed method provides an approach for miniaturized, high-throughput construction of complex 3D multicellular architectures and can be applied for studying various biological processes including cell signaling, cancer invasion, embryogenesis, and neural development., (© 2020 The Authors. Advanced Materials published by Wiley-VCH GmbH.)

Multiplexed detection of viral nucleic acids is important for rapid screening of viral infection. In this study, we present a molybdenum disulfide (MoS 2 ) nanosheet-modified dendrimer droplet microarray (DMA) for rapid and sensitive detection of retroviral nucleic acids of human immunodeficiency virus-1 (HIV-1) and human immunodeficiency virus-2 (HIV-2) simultaneously. The DMA platform was fabricated by omniphobic-omniphilic patterning on a surface-grafted dendrimer substrate. Functionalized MoS 2 nanosheets modified with fluorescent dye-labeled oligomer probes were prepatterned on positively charged amino-modified omniphilic spots to form a fluorescence resonance energy transfer (FRET) sensing microarray. With the formation of separated microdroplets of sample on the hydrophobic-hydrophilic micropattern, prepatterned oligomer probes specifically hybridized with the target HIV genes and detached from the MoS 2 nanosheet surface due to weakening of the adsorption force, leading to fluorescence signal recovery. As a proof of concept, we used this microarray with a small sample size (<150 nL) for simultaneous detection of HIV-1 and HIV-2 nucleic acids with a limit of detection (LOD) of 50 pM. The multiplex detection capability was further demonstrated for simultaneous detection of five viral genes (HIV-1, HIV-2, ORFlab, and N genes of SARS-COV-2 and M gene of Influenza A). This work demonstrated the potential of this novel MoS 2 -DMA FRET sensing platform for high-throughput multiplexed viral nucleic acid screening.

Spatial control of bacteria and biofilms on surfaces is necessary to understand the biofilm formation and the social interactions between bacterial communities, which could provide useful hints to study the biofilm-involved diseases. Here patterned lubricant-infused surfaces (pLIS) are utilized to fabricate connective structures named "bacterial bridges" between bacterial colonies of Pseudomonas aeruginosa by a simple dewetting method. It is demonstrated that the bacteria attached to hydrophilic areas and bacteria precipitated on lubricant infused borders both contribute to the formation of bacterial bridges. The geometry and distribution of bridges can be controlled using predesigned superhydrophobic-hydrophilic patterns. It is demonstrated that bacterial bridges connecting bacteria colonies act as bio-microfluidic channels and can transport liquids, nutrients, and antibacterial substances between neighboring bacteria clusters. Thus, bacterial bridges can be used to study formation, spreading, and development of bacterial colonies, and communication within and between isolated biofilms., (© 2020 The Authors. Small published by Wiley-VCH GmbH.)

Non-viral vectors are a safety tool for gene therapy to deliver therapeutic genes. Among the different non-viral vectors, polyvinylpyrrolidone (PVP), a well-known hydrosoluble, neutral, and non-toxic polymer, satisfies the requirements and becomes a suitable candidate for gene delivery. In this study, we describe the preparation of polyvinylpyrrolidones decorated with pyrrolidine, piperidine, and piperazine groups, and evaluate them in vitro as non-viral gene carriers. The properties of these new systems are compared with those of hyperbranched polyethyleneimine (PEI) used as a positive control. Their ability to complex DNA at different N/P molar ratios, from 1:1 up to 10:1, was studied through agarose gel electrophoresis and dynamic light scattering. The resulting complexes (polyplexes) were characterized and evaluated in vitro with murine fibroblast (Swiss 3T3) as non-viral gene carriers, using luciferase as the reporter gene and a calcein cytocompatibility assay. All the copolymers condensed DNA to a particle average size between 100-400 nm when used at N/P ratios of 4:1 or higher. The copolymers with piperidine groups showed higher transfection efficiency than the pyrrolidine and piperazine modified copolymers, and even higher than the positive control of PEI at N/P ratios of 4:1 or higher. All the synthesized polyplexes from an aminated PVP displayed a general tendency of high cytocompatibility (75-95%) in comparison with the positive control PEI (55%).

Acceleration and unification of drug discovery is important to reduce the effort and cost of new drug development. Diverse chemical and biological conditions, specialized infrastructure and incompatibility between existing analytical methods with high-throughput, nanoliter scale chemistry make the whole drug discovery process lengthy and expensive. Here, we demonstrate a chemBIOS platform combining on-chip chemical synthesis, characterization and biological screening. We developed a dendrimer-based surface patterning that enables the generation of high-density nanodroplet arrays for both organic and aqueous liquids. Each droplet (among > 50,000 droplets per plate) functions as an individual, spatially separated nanovessel, that can be used for solution-based synthesis or analytical assays. An additional indium-tin oxide coating enables ultra-fast on-chip detection down to the attomole per droplet by matrix-assisted laser desorption/ionization mass spectrometry. The excellent optical properties of the chemBIOS platform allow for on-chip characterization and in-situ reaction monitoring in the ultraviolet, visible (on-chip UV-Vis spectroscopy and optical microscopy) and infrared (on-chip IR spectroscopy) regions. The platform is compatible with various cell-biological screenings, which opens new avenues in the fields of high-throughput synthesis and drug discovery.

Sulfur as a side product of natural gas and oil refining is an underused resource. Converting landfilled sulfur waste into materials merges the ecological imperative of resource efficiency with economic considerations. A strategy to convert sulfur into polymeric materials is the inverse vulcanization reaction of sulfur with alkenes. However, the materials formed are of limited applicability, because they need to be cured at high temperatures (>130 °C) for many hours. Herein, we report the reaction of elemental sulfur with styrylethyltrimethoxysilane. Marrying the inverse vulcanization and silane chemistry yielded high sulfur content polysilanes, which could be cured via room temperature polycondensation to obtain coated surfaces, particles, and crosslinked materials. The polycondensation was triggered by hydrolysis of poly(sulfur-r-styrylethyltrimethoxysilane) (poly(S n -r-StyTMS) under mild conditions (HCl, pH 4). For the first time, an inverse vulcanization polymer could be conveniently coated and mildly cured via post-polycondensation. Silica microparticles coated with the high sulfur content polymer could improve their Hg 2+ ion remediation capability., (© 2020 The Authors. Published by Wiley-VCH GmbH.)

Currently, there are no time-saving and cost-effective high-throughput screening methods for the evaluation of bacterial drug-resistance. In this study, a droplet microarray (DMA) system is established as a miniaturized platform for high-throughput screening of antibacterial compounds using the emerging, opportunistic human pathogen Pseudomonas aeruginosa (P. aeruginosa) as a target. Based on the differences in wettability of DMA slides, a rapid method for generating microarrays of nanoliter-sized droplets containing bacteria is developed. The bacterial growth in droplets is evaluated using fluorescence. The new method enables immediate screening with libraries of antibiotics. A novel simple colorimetric readout method compatible with the nanoliter size of the droplets is established. Furthermore, the drug-resistance of P. aeruginosa 49, a multi-resistant strain from an environmental isolate, is investigated. This study demonstrates the potential of the DMA platform for the rapid formation of microarrays of bacteria for high-throughput drug screening., (© 2020 The Authors. Published by Wiley-VCH GmbH.)

Development of non-viral gene vectors which can efficiently and safely transfect plasmid DNA and siRNA into cells is of great importance for gene therapy. Despite lots of efforts spent, it is still imperative to develop suitable gene vectors with better transfection efficiency and low cytotoxicity. To this end, we successfully designed, synthesized and screened a library of 120 polymers (via nucleophilic substitution reaction between dihalides and amines). With cell-based transfection screening assays, 120 polymers were tested to evaluate their transfection efficiency of transporting DNA and siRNA into cells. Our results indicated that hydrophobic modification could greatly enhance cationic polymers' transfection efficiency, and polymers with long linkers usually showed better transfection performance, especially for polymers with the linker of 1, 12-dibromododecane (L3 linker). Besides, polyalkylamines exhibited better transfection efficiency with the polymer particle size around 200 nm and the zeta potential in the range of + 40 mV to +50 mV. Interestingly, polymer particles made from N15HL3 not only exhibited better DNA transfection efficiency in HEK 293T cells but also showed higher siRNA transfection efficiency in U87 Luc-GFP cells together with low cell toxicity than Lipofectamine 2000 (one of commercial transfection reagents). Therefore, it is hoped that our study here not only provides promising gene vector candidates for further evaluation in gene therapy, but also provides valuable insights for better understanding of the relationship between the chemical structures and gene transfection efficiency to rationally design better non-viral gene vectors for gene therapy in the future., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020. Published by Elsevier Ltd.)

Non-covalent chemosensing ensembles of cucurbit[ n ]urils (CB n ) have been widely used in proof-of-concept sensing applications, but they are prone to disintegrate in saline media, e.g. biological fluids. We show here that covalent cucurbit[7]uril-indicator dye conjugates are buffer- (10× PBS buffer) and saline-stable (up to 1.4 M NaCl) and allow for selective sensing of Parkinson's drug amantadine in human urine and saliva, where the analogous non-covalent CB7⊃dye complex is dysfunctional. The in-depth analysis of the covalent host-dye conjugates in the gas-phase, and deionized versus saline aqueous media revealed interesting structural, thermodynamic and kinetic effects that are of general interest for the design of CB n -based supramolecular chemosensors and systems. This work also introduces a novel high-affinity indicator dye for CB7 through which fundamental limitations of indicator displacement assays (IDA) were exposed, namely an impractical slow equilibration time. Unlike non-covalent CB n ⊃dye reporter pairs, the conjugate chemosensors can also operate through a S N 2-type guest-dye exchange mechanism, which shortens assay times and opens new avenues for tailoring analyte-selectivity., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)