Electrostatic adsorption of polyanions onto lipid nanoparticles controls uptake, trafficking, and transfection of RNA and DNA therapies.

Rapid advances in nucleic acid therapies highlight the immense therapeutic potential of genetic therapeutics. Lipid nanoparticles (LNPs) are highly potent nonviral transfection agents that can encapsulate and deliver various nucleic acid therapeutics, including but not limited to messenger RNA (mRNA), silencing RNA (siRNA), and plasmid DNA (pDNA). However, a major challenge of targeted LNP-mediated systemic delivery is the nanoparticles' nonspecific uptake by the liver and the mononuclear phagocytic system, due partly to the adsorption of endogenous serum proteins onto LNP surfaces. Tunable LNP surface chemistries may enable efficacious delivery across a range of organs and cell types. Here, we describe a method to electrostatically adsorb bioactive polyelectrolytes onto LNPs to create layered LNPs (LLNPs). LNP cores varying in nucleic acid cargo and component lipids were stably layered with four biologically relevant polyanions: hyaluronate (HA), poly-L-aspartate (PLD), poly-L-glutamate (PLE), and polyacrylate (PAA). We further investigated the impact of the four surface polyanions on the transfection and uptake of mRNA- and pDNA-loaded LNPs in cell cultures. PLD- and PLE-LLNPs increased mRNA transfection twofold over unlayered LNPs in immune cells. HA-LLNPs increased pDNA transfection rates by more than twofold in epithelial and immune cells. In a healthy C57BL/6 murine model, PLE- and HA-LLNPs increased transfection by 1.8-fold to 2.5-fold over unlayered LNPs in the liver and spleen. These results suggest that LbL assembly is a generalizable, highly tunable platform to modify the targeting specificity, stability, and transfection efficacy of LNPs, as well as incorporate other charged targeting and therapeutic molecules into these systems.

Targeting and monitoring ovarian cancer invasion with an RNAi and peptide delivery system.

RNA interference (RNAi) therapeutics are an emerging class of medicines that selectively target mRNA transcripts to silence protein production and combat disease. Despite the recent progress, a generalizable approach for monitoring the efficacy of RNAi therapeutics without invasive biopsy remains a challenge. Here, we describe the development of a self-reporting, theranostic nanoparticle that delivers siRNA to silence a protein that drives cancer progression while also monitoring the functional activity of its downstream targets. Our therapeutic target is the transcription factor SMARCE1, which was previously identified as a key driver of invasion in early-stage breast cancer. Using a doxycycline-inducible shRNA knockdown in OVCAR8 ovarian cancer cells both in vitro and in vivo, we demonstrate that SMARCE1 is a master regulator of genes encoding proinvasive proteases in a model of human ovarian cancer. We additionally map the peptide cleavage profiles of SMARCE1-regulated proteases so as to design a readout for downstream enzymatic activity. To demonstrate the therapeutic and diagnostic potential of our approach, we engineered self-assembled layer-by-layer nanoparticles that can encapsulate nucleic acid cargo and be decorated with peptide substrates that release a urinary reporter upon exposure to SMARCE1-related proteases. In an orthotopic ovarian cancer xenograft model, theranostic nanoparticles were able to knockdown SMARCE1 which was in turn reported through a reduction in protease-activated urinary reporters. These LBL nanoparticles both silence gene products by delivering siRNA and noninvasively report on downstream target activity by delivering synthetic biomarkers to sites of disease, enabling dose-finding studies as well as longitudinal assessments of efficacy.

Domain-Selective Enzymatic Cross-linking and Etching for Shape-Morphing DNA-Linked Nanoparticle Films.

The highly programmable and responsive molecular recognition properties of DNA provide unparalleled opportunities for fabricating dynamic nanostructures capable of structural transformation in response to various external stimuli. However, they typically operate in tightly controlled environments because certain conditions (ionic strength, pH, temperature, etc.) must be met for DNA duplex formation. In this study, we adopted site-specific enzymatic ligation and DNA-based layer-by-layer thin film fabrication to build shape-morphing DNA-linked nanoparticle films operational in a broad range of environments. The ligated films remained intact in unusual conditions such as pure water and high temperature causing dissociation of DNA duplexes and showed predictable and reversible shape morphing in response to various environmental changes and DNA exchange reactions. Furthermore, domain-selective ligation combined with photoinduced interlayer mixing allowed for the fabrication of unusual edge-sealed double-layered films through midlayer etching, which is difficult to realize by other methods.

Advanced Functionalized Materials Based on Layer-by-Layer Assembled Natural Cellulose Nanofiber for Electrodes: A Review.

The depletion of fossil fuel resources and its impact on the environment provide a compelling motivation for the development of sustainable energy sources to meet the increasing demand for energy. Accordingly, research and development of energy storage devices have emerged as a critical area of focus. The electrode materials are critical in the electrochemical performance of energy storage devices, such as energy storage capacity and cycle life. Cellulose nanofiber (CNF) represents an important substrate with potentials in the applications of green electrode materials due to their environmental sustainability and excellent compatibility. By utilizing the layer-by layer (LbL) process, well-defined nanoscale multilayer structure is prepared on a variety of substrates. In recent years, increasing attention has focused on electrode materials produced from LbL process on CNFs to yield electrodes with exceptional properties, such as high specific surface area, outstanding electrical conductivity, superior electrochemical activity, and exceptional mechanical stability. This review provides a comprehensive overview on the development of functional CNF via the LbL approach as electrode materials.

Intermolecular Interactions, Morphology, and Photovoltaic Patterns in p-i-n Heterojunction Solar Cells With Fluorine-Substituted Organic Photovoltaic Materials.

During the layer-by-layer (LBL) processing of polymer solar cells (PSCs), the swelling and molecule interdiffusion are essential for achieving precise, controllable vertical morphology, and thus efficient PSCs. However, the influencing mechanism of material properties on morphology and correlated device performance has not been paid much attention. Herein, a series of fluorinated/non-fluorinated polymer donors (PBDB-T and PBDB-TF) and non-fullerene acceptors (ITIC, IT-2F, and IT-4F) are employed to investigate the performance of LBL devices. The impacts of fluorine substitution on the repulsion and miscibility between the donor and acceptor, as well as the molecular arrangement of the donor/acceptor and the vertical distribution of the LBL devices are systematically explored by the measurement of donor/acceptor Flory-Huggins interaction parameters, spectroscopic ellipsometry, and neutron reflectivity, respectively. With efficient charge transfer due to the ideal vertical and horizon morphology properties, devices based on PBDB-TF/IT-4F exhibit the highest fill factors (FFs) as well as champion power conversion efficiencies (PCEs). With this guidance, high-performance LBL devices with PCE of 17.2%, 18.5%, and 19.1% are obtained by the fluorinated blend of PBDB-TF/Y6, PBDB-TF/L8-BO, and D18/L8-BO respectively.

PC71BM as Morphology Regulator for Highly Efficient Ternary Organic Solar Cells with Bulk Heterojunction or Layer-by-Layer Configuration.

The ternary strategy is one of the effective methods to regulate the morphology of the active layer in organic solar cells (OSCs). In this work, the ternary OSCs with bulk heterojunction (BHJ) or layer-by-layer (LbL) active layers are prepared by using the polymer donor PM6 and the non-fullerene acceptor L8-BO as the main system and the fullerene acceptor PC71BM as the third component. The power conversion efficiencies (PCEs) of BHJ OSCs and LbL OSCs are increased from 17.10% to 18.02% and from 17.20% to 18.20% by introducing PC71BM into the binary active layer, respectively. The in situ UV-vis absorption spectra indicate that the molecular aggregation and crystallization process can be prolonged by introducing PC71BM into the PM6:L8-BO or PM6/L8-BO active layer. The molecular orientation and molecular crystallinity in the active layer are optimized by introducing the PC71BM into the binary BHJ or LbL active layers, which can be confirmed by the experimental results of grazing incidence wide-angle X-ray scattering. This study demonstrates that the third component PC71BM can be used as a morphology regulator to regulate the morphology of BHJ or LbL active layers, thus effectively improving the performance of BHJ and LbL OSCs.

Reversible Sensing Technologies Using Upcycled TPEE: Crafting pH and Light Responsive Materials towards Sustainable Monitoring.

Multiresponsive materials with reversible and durable characteristics are indispensable because of their promising applications in environmental change detections. To fabricate multiresponsive materials in mass production, however, complex reactions and impractical situations are often involved. Herein, a dual responsive (light and pH) spiropyran-based smart sensor fabricated by a simple layer-by-layer (LbL) assembly process from upcycled thermoplastic polyester elastomer (TPEE) materials derived from recycled polyethylene terephthalate (r-PET) is proposed. Positively charged chitosan solutions and negatively charged merocyanine-COOH (MC-COOH) solutions are employed in the LbL assembly technique, forming the chitosan-spiropyran deposited TPEE (TPEE-CH-SP) film. Upon UV irradiation, the spiropyran-COOH (SP-COOH) molecules on the TPEE-CH-SP film undergo the ring-opening isomerization, along with an apparent color change from colorless to purple, to transform into the MC-COOH molecules. By further exposing the TPEE-CH-MC film to hydrogen chloride (HCl) and nitric acid (HNO3) vapors, the MC-COOH molecules can be transformed into protonated merocyanine-COOH (MCH-COOH) with the simultaneous color change from purple to yellow.

Construction of Hierarchical Films via Layer-by-Layer Assembly of Exfoliated Unilamellar Zeolite Nanosheets.

Zeolites have been widely applied as versatile catalysts, sorbents, and ion exchangers with unique porous structures showing molecular sieving capability. In these years, it is reported that some layered zeolites can be delaminated into molecularly thin 2-dimensional (2D) nanosheets characterized by inherent porous structures and highly exposed active sites. In the present study, two types of zeolite nanosheets with distinct porous structures with MWW topology (denoted mww) and ferrierite-related structure (denoted bifer) are deposited on a substrate through the solution process via electrostatic self-assembly. Alternate deposition of zeolite nanosheets with polycation under optimized conditions allows the layer-by-layer growth of their multilayer films with a stacking distance of 2-3 nm. Furthermore, various hierarchical structures defined at the unit-cell dimensions can be constructed simply by conducting the deposition of mww and bifer nanosheets in a designed sequence. Adsorption of a dye, Rhodamine B, in these films, is examined to show that adsorption is dependent on constituent zeolite nanosheets and their assembled nanostructures. This work has provided fundamental advancements in the fabrication of artificial zeolite-related hierarchical structures, which may be extended to other zeolite nanosheets, broadening their functionalities, applications, and benefits.

Determining Structures of Layer-by-Layer Spin-Coated Zinc Dicarboxylate-Based Metal-Organic Thin Films.

Thin films of crystalline solids with substantial free volume built from organic chromophores and metal secondary building units (SBUs) are promising for engineering new optoelectronic properties through control of interchromophore coupling. Zn-based SBUs are especially relevant in this case because they avoid quenching the chromophore's luminescence. We find that layer-by-layer spin-coating using Zn acetate dihydrate and benzene-1,4-dicarboxylic acid (H2BDC) and biphenyl-4,4'-dicarboxylic acid (H2BPDC) linkers readily produces crystalline thin films. However, analysis of the grazing-incidence wide-angle X-ray scattering (GIWAXS) data reveals the structures of these films vary significantly with the linker, and with the metal-to-linker molar ratio used for fabrication. Under equimolar conditions, H2BPDC creates a type of structure like that proposed for SURMOF-2, whereas H2BDC generates a different metal-hydroxide-organic framework. Large excess of Zn2+ ions causes the growth of layered zinc hydroxides, irrespective of the linker used. Density functional theory (DFT) calculations provide structural models with minimum total energy that are consistent with the experimentally observed diffractograms. In the broader sense, this work illustrates the importance in this field of careful structure determination, e. g., by utilizing GIWAXS and DFT simulations to determine the structure of the obtained crystalline metal-organic thin films, such that properties can be rationally engineered and explained.

pH-response of protein-polysaccharide multilayers adsorbed on a flat gold surface: A surface plasmon resonance study.

Polysaccharide-protein multilayers (PPMLs) consisting of bovine serum albumin (BSA) and chondroitin sulfate (CS) are assembled in acidic solution (pH 4.2) via layer-by-layer deposition method. The formation of PPMLs on gold surface and their responsiveness to pH change from 4.2 to 7 is investigated by Surface Plasmon Resonance Spectroscopy. The buildup of the multilayer at pH 4.2 exhibits non-linear growth while the formation of the first layers is strongly affected by the physicochemical properties of the gold surface. Neutral solution (pH 7) affects the interactions between the biopolymers and results in a partially disassemble (disintegration) of the multilayer film. On one hand, the single pair of layers, BSA-CS and the double pair of layers, (BSA-CS)2, assemblies are stable in neutral pH, a result that will be of interest for biomedical applications. On the other hand, multilayer films consisting of more than four layers that is (BSA-CS)2

Fabrication of Fully Positively Charged Layer-by-Layer Polyelectrolyte Nanofilms with pH-Dependent Swelling Properties.

Nanofilms fabricated by layer-by-layer (LbL) assembly from polyelectrolytes (PEs) are important materials for various applications. However, PE films cannot retain the charges along the polymer chains during fabrication, resulting in a low charge density. In this study, the preparation of LbL nanofilms with preserved positive charges via a controllable and efficient approach was achieved. To fabricate fully positively charged (FPC) LbL nanofilms, a polycation, poly-l-lysine, was partially grafted with azide and alkyne groups. Through copper-catalyzed azide-alkyne cycloaddition and the LbL procedure, nanofilms were fabricated with all of the individual layers covalently bonded, improving the pH stability of the nanofilms. Because the resulting nanofilms had a high charge density with positive charges both inside and on the surface, they showed unique pH-dependent swelling properties and adsorption of negatively charged molecules compared with those of traditional polyelectrolyte LbL nanofilms. This kind of FPC nanofilm has great potential for use in sensors, diagnostics, and filter nanomaterials in the biomedical and environmental fields.

Synthesis, characterization and application of chitosan functionalized and functional graphene oxide membranes for desalination of water by pervaporation.

The use of graphene sheets in water treatment is increasing due to its adsorption capacity, reactivity, catalytic action and surface area. The challenges linked to wastewater treatment are vast due to the constant influx of various pollutants. Can the challenges of water desalination and purification be encountered by graphene-based composites and membranes?.The current work describes the synthesis of graphene oxide (GO) using modified Hummers' method. GO was functionalized with chitosan and used as adsorbents. On the other hand, it was reported that the surface of thin-film-composite (TFC) polyamide membranes was modified in order to desalinate highly saline water using pervaporation. The findings showed that GO synthesized by modified Hummers' method has a greater capacity for the adsorption of sodium ion and have better regeneration performance. Functionalization with chitosan increased adsorption capacity from 680.2 to 740.5 mg/g at the initial concentration of 45,000 mg/l of Na+ ions. On the other hand, modification in membrane comprises the chlorine treatment of surface of polyamide membrane. Layer-by-layer (LbL) deposition of positively charged polyethyleneimine (PEI) and negatively charged graphene oxide (GO) was followed. The PEI/GO LbL membrane's pure water flux was twice as high as compare to the original membrane. The synthesized membrane was tested against the aqueous solutions containing Na2SO4, MgSO4, NaCl and MgCl2 salts for their desalination. At different concentrations, a water flux of 8.9 kg/m2h with a huge salt rejection (>99.9%) was attained for every tested salt. It was observed that CS functionalized GO and GO membrane showed higher adsorption capacity and improved regeneration performance can be measured as an operational and active adsorbent for sea water desalination.

Functionalization and magnetonavigation of T-lymphocytes functionalized via nanocomposite capsules targeting with electromagnetic tweezers.

Modification of T-lymphocytes, which are capable of paracellular transmigration is a promising trend in modern personalized medicine. However, the delivery of required concentrations of functionalized T-cells to the target tissues remains a problem. We describe a novel method to functionalize T-cells with magnetic nanocapsules and target them with electromagnetic tweezers. T-cells were modified with the following magnetic capsules: Parg/DEX (150 nm), BSA/TA (300 nm), and BSA/TA (500 nm). T-cells were magnetonavigated in a phantom blood vessel capillary in cultural medium and in whole blood. The permeability of tumor tissues to captured T-cells was analyzed by magnetic delivery of modified T-cells to spheroids formed from 4T1 breast cancer cells. The dynamics of T-cell motion under a magnetic field gradient in model environments were analyzed by particle image velocimetry. The magnetic properties of the nanocomposite capsules and magnetic T-cells were measured. The obtained results are promising for biomedical applications in cancer immunotherapy.

Near-Infrared Fluorescent Silica Nanoparticles Based on Gold-Silver Alloy Nanoclusters for Clinical Diagnosis.

In clinical diagnosis, fluorescent particles are applied to detect analytes in biofluids, such as blood and saliva. However, current fluorescence detection methods have not been optimized to account for the overlapping autofluorescence peaks of biological substances. Gold and silver nanoclusters are known to the novel fluorescent materials and their emission wavelengths depend on cluster size. In this study, we developed fluorescent silica nanoparticles using gold-silver alloy nanoclusters and chitosan (CS) (NH2-SiO2@Au@CS@AuAg) by the layer-by-layer method. Under UV-light irradiation at 365 nm, the emission wavelength of NH2-SiO2@Au@CS@AuAg reached 750 nm in the near-IR region. Scanning electron microscopy images revealed that the shape of NH2-SiO2@Au@CS@AuAg was uniform and spherical. The fluorescence spectrum of horse blood obtained in the presence of NH2-SiO2@Au@CS@AuAg contained a specific fluorescence peak attributed to NH2-SiO2@Au@CS@AuAg, which was distinguishable from the autofluorescence peaks. These results showed that NH2-SiO2@Au@CS@AuAg has advantageous fluorescence properties for clinical diagnostic applications.

A highly-selective layer-by-layer membrane modified with polyethylenimine and graphene oxide for vanadium redox flow battery.

A selective composite membrane for vanadium redox flow battery (VRFB) was successfully prepared by layer-by-layer (LbL) technique using a perfluorosulfonic sulfonic acid or Nafion 117 (N117). The composite membrane referred as N117-(PEI/GO)n, was obtained by depositing alternating layers of positively charged polyethylenimine (PEI) and negatively charged graphene oxide (GO) as polyelectrolytes. The physicochemical properties and performance of the pristine and composite membranes were investigated. The membrane showed an enhancement in proton conductivity and simultaneously exhibited a notable 90% reduction in vanadium permeability. This, in turn, results in a well-balanced ratio of proton conductivity to vanadium permeability, leading to high selectivity. The highest selectivity of the LbL membranes was found to be 19.2 × 104 S.min/cm3, which is 13 times higher than the N117 membrane (n = 0). This was translated into an improvement in the battery performance, with the n = 1 membrane showing a 4-6% improvement in coulombic efficiency and a 7-15% improvement in voltage efficiency at current densities ranging from 40 to 80 mA/cm2. Furthermore, the membrane displays stable operation over a long-term stability at around 88% at a current density of 40 mA/cm2, making it an attractive option for VRFB applications using the LbL technique. The use of PEI/GO bilayers maintains high proton conductivity and VE of the battery, opening up possibilities for further optimization and improvement of VRFBs.

An innovative N117-(PEI/GO)n layer-by-layer membrane prepared for vanadium redox flow battery improved the balance between the proton conductivity and vanadium permeability, yielding a remarkable selectivity of 19.2 × 10⁴ S.min/cm³.

Layer-by-Layer Immobilization of DNA Aptamers on Ag-Incorporated Co-Succinate Metal-Organic Framework for Hg(II) Detection.

Layer-by-layer (LbL) immobilization of DNA aptamers in the realm of electrochemical detection of heavy metal ions (HMIs) offers an enhancement in specificity, sensitivity, and low detection limits by leveraging the cross-reactivity obtained from multiple interactions between immobilized aptamers and developed material surfaces. In this research, we present a LbL approach for the immobilization of thiol- and amino-modified DNA aptamers on a Ag-incorporated cobalt-succinate metal-organic framework (MOF) (Ag@Co-Succinate) to achieve a cross-reactive effect on the electrochemical behavior of the sensor. The solvothermal method was utilized to synthesize Ag@Co-Succinate, which was also characterized through various techniques to elucidate its structure, morphology, and presence of functional groups, confirming its suitability as a host matrix for immobilizing both aptamers. The Ag@Co-Succinate aptasensor exhibited extraordinary sensitivity and selectivity towards Hg(II) ions in electrochemical detection, attributed to the unique binding properties of the immobilized aptamers. The exceptional limit of detection of 0.3 nM ensures the sensor's suitability for trace-level Hg(II) detection in various environmental and analytical applications. Furthermore, the developed sensor demonstrated outstanding repeatability, highlighting its potential for long-term and reliable monitoring of Hg(II).

Graphene Oxide, Carbon Nanotubes, and Polyelectrolytes-Based Impedanciometric E-Tongue for Estrogen Detection in Complex Matrices.

Currently, it is necessary to maintain the quality of aquifers and water bodies, which means the need for sensors that detect molecules as emerging pollutants (EPs) at low concentrations in aqueous complex solutions. In this work, an electronic tongue (e-tongue) prototype was developed to detect 17ß-estradiol in tap water. To achieve such a prototype, an array of sensors was prepared. Each sensor consists of a solid support with interdigitated electrodes without or with thin films prepared with graphene oxide, nanotubes, and other polyelectrolytes molecules adsorbed on them. To collect data from each sensor, impedance spectroscopy was used to analyze the electrical characteristics of samples of estrogen solutions with different concentrations. To analyze the collected data from the sensors, principal components analysis (PCA) method was used to create a three-dimensional plane using the calculated principal components, namely PC1 and PC2, and the estrogen concentration values. Then, damped least squares (DLS) was used to find the optimal values for the hyperplane calibration, as the sensitivity of this e-tongue was not represented by a straight line but by a surface. For the collected data, from nanotubes and graphene oxide sensors, a calibration curve for concentration given by the 10PC1×0.492-PC2×0.14-14.5 surface was achieved. This e-tongue presented a detection limit of 10-16 M of 17ß-estradiol in tap water.

Oscillatory Order-Disorder Transition during Layer-by-Layer Growth of Indium Selenide.

It is important to understand the polymorph transition and crystal-amorphous phase transition in In2Se3 to tap the potential of this material for resistive memory storage. By monitoring layer-by-layer growth of ß-In2Se3 during molecular beam epitaxy (MBE), we are able to identify a cyclical order-disorder transition characterized by a periodic alternation between a glassy-like metastable subunit cell film consisting of n < 5 sublayers (nth layers = the number of subunit cell layers), and a highly crystalline ß-In2Se3 at n = 5 layers. The glassy phase shows an odd-even alternation between the indium-cluster layer (n = 1, 3) and an In-Se solid solution (n = 2, 4), which suggests the ability of In and Se atoms to diffuse, aggregate, and intermix. These dynamic natures of In and Se atoms contribute to a defect-driven memory resistive behavior in current-voltage sweeps that is different from the ferroelectric switching of α-In2Se3.

Permselective Ionic-Shield for High-Performance Lithium-Sulfur Batteries.

Lithium-sulfur batteries (LiSBs) are promising next-generation batteries because of their low cost and high theoretical energy densities. Despite remarkable advances over the decades, polysulfide (PS) shuttling during battery cycling remains a challenge in the development of commercial LiSBs and is accelerated under practical conditions. Herein, we report a permselective ionic shield between the electrodes that blocks PS shuttles and passes Li ions to high-performance LiSBs. This shield is easily built onto the separator by ionic complexation and intermolecular bonding of functional polymers, thereby improving the battery performance and safety. The LiSB with the developed shield delivers a remarkable discharge capacity of 917 mAh g-1 after 1000 cycles at 2 C. In addition, the behavior of LiSBs under practical conditions that can realize a high energy density is investigated to achieve the optimal balance in this system. This study provides new insights into the imminent development of separators for practical LiSBs.

Layer-by-Layer-Processed All-Polymer Solar Cells with Enhanced Performance Enabled by Regulating the Microstructure of Upper Layer.

The layer-by-layer (LBL) fabrication method allows for controlled microstructure morphology and vertical component distribution, and also offers a reproducible and efficient technique for fabricating large-scale organic solar cells (OSCs). In this study, the polymers D18 and PYIT-OD are employed to fabricate all-polymer solar cells (all-PSCs) using the LBL method. Morphological studies reveal that the use of additives optimizes the microstructure of the active layer, enhancing the cells' crystallinity and charge transport capability. The optimized device with 2% CN additive significantly reduces bimolecular recombination and trap-assisted recombination. All-PSCs fabricated by the LBL method based on D18/PYIT-OD deliver a power conversion efficiency (PCE) of 15.07%. Our study demonstrates the great potential of additive engineering via the LBL fabrication method in regulating the microstructure of active layers, suppressing charge recombination, and enhancing the photovoltaic performance of devices.