Stimuli-responsive Nanostructures Research Articles

Genetically Fused Resilin-like Polypeptide-Coiled Coil Bundlemer Conjugates Exhibit Tunable Multistimuli-Responsiveness and Undergo Nanofibrillar Assembly.

Peptide-based materials are diverse candidates for self-assembly into modularly designed and stimuli-responsive nanostructures with precisely tunable compositions. Here, we genetically fused computationally designed coiled coil-forming peptides to the N- and C-termini of compositionally distinct multistimuli-responsive resilin-like polypeptides (RLPs) of various lengths. The successful expression of these hybrid polypeptides in bacterial hosts was confirmed through techniques such as gel electrophoresis, mass spectrometry, and amino acid analysis. Circular dichroism spectroscopy and ultraviolet-visible turbidimetry demonstrated that despite the fusion of disparate structural and responsive units, the coiled coils remained stable in the hybrid polypeptides, and the sequence-encoded differences in thermoresponsive phase separation of the RLPs were preserved. Cryogenic transmission electron microscopy and coarse-grained modeling showed that after thermal annealing in solution, the hybrid polypeptides adopted a closed loop conformation and assembled into nanofibrils capable of further hierarchically organizing into cluster structures and ribbon-like structures mediated by the self-association tendency of the RLPs.

Scintillating and magnetic stimuli-responsive nanostructures as adjuvants in cancer therapy

We present an overview of recent advances in stimuli-responsive nanostructures for medicine and drug delivery aiming at improving the efficacy and minimizing side effects of cancer therapies. We focus on two complementary system: Self-Lighted Photodynamic Therapy (SLPDT), and magnetic systems for hyperthermia and for controlled drug release. SPLDT, also known as X-ray triggered PDT, bases on hybrid nanostructures such as functionalized SiC/SiOx nanowires and CeF3/ZnO nanostructures. We emphasize the importance of discriminating the various Reactive Oxygen Species (ROS) produced, i.e., singlet oxygen, peroxide, superoxide, and hydroxyl radicals, which have different effects on cells and different therapeutic efficacy. Singlet oxygen is quantified by an ad hoc NIR fluorimeter. SLPDT can be usefully complemented by smart systems responsive to magnetic fields. As examples, we discuss the use of superparamagnetic iron oxide nanoparticles for magnetic hyperthermia and magnetic nanocapsules for triggered drug release. Finally, we highlight promising directions for future developments.

PH-Responsive and Reversible A-Motif-Based DNA Hydrogel: Synthesis and Biosensing Application.

Functional DNA hydrogels with various motifs and functional groups require perfect sequence design to avoid cross-bonding interference with themselves or other structural sequences. This work reports an A-motif functional DNA hydrogel that does not require any sequence design. A-motif DNA is a noncanonical parallel DNA duplex structure containing homopolymeric deoxyadenosines (poly-dA) strands that undergo conformation changes from single strands at neutral pH to a parallel duplex DNA helix at acidic pH. Despite this and other advantages over other DNA motifs like no cross-bonding interference with other structural sequences, the A-motif has not been explored much. We successfully synthesized a DNA hydrogel by using an A-motif as a reversible handle to polymerize a DNA three-way junction. The A-motif hydrogel was initially characterized by electrophoretic mobility shift assay, and dynamic light scattering, which showed the formation of higher-order structures. Further, we used imaging techniques like atomic force microscopy and scanning electron microscope to validating its hydrogel like highly branched morphology. pH-induced conformation transformation from monomers to gel is quick and reversible, and was analysed for multiple acid-base cycles. The sol-to-gel transitions and gelation properties were further examined in rheological studies. The use of the A-motif hydrogel in the visual detection of pathogenic target nucleic acid sequence was demonstrated for the first time in a capillary assay. Moreover, pH-induced hydrogel formation was observed in situ as a layer over the mammalian cells. The proposed A-motif DNA scaffold has enormous potential in designing stimuli-responsive nanostructures that can be used for many biological applications.

Smart Nanostructured Materials for SARS-CoV-2 and Variants Prevention, Biosensing and Vaccination.

The coronavirus disease 2019 (COVID-19) pandemic, caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has raised great concerns about human health globally. At the current stage, prevention and vaccination are still the most efficient ways to slow down the pandemic and to treat SARS-CoV-2 in various aspects. In this review, we summarize current progress and research activities in developing smart nanostructured materials for COVID-19 prevention, sensing, and vaccination. A few established concepts to prevent the spreading of SARS-CoV-2 and the variants of concerns (VOCs) are firstly reviewed, which emphasizes the importance of smart nanostructures in cutting the virus spreading chains. In the second part, we focus our discussion on the development of stimuli-responsive nanostructures for high-performance biosensing and detection of SARS-CoV-2 and VOCs. The use of nanostructures in developing effective and reliable vaccines for SARS-CoV-2 and VOCs will be introduced in the following section. In the conclusion, we summarize the current research focus on smart nanostructured materials for SARS-CoV-2 treatment. Some existing challenges are also provided, which need continuous efforts in creating smart nanostructured materials for coronavirus biosensing, treatment, and vaccination.

Photo-controlled reversible assemblies from rod-coil molecules with azobenzene group

Rod-coil amphiphilic molecules composed of hydrophilic polyethylene glycol chains and an aromatic domain based on biphenyl and azobenzene groups self-assembled into two-dimensional columnar and lamellar nanostructures in the solid state. In the aqueous solution, these molecules produced wormlike micelles, nanoribbons, and large sheets, depending on their chain length. Interestingly, the quenched fluorescence emission of these molecules was observed in methylene chloride; nevertheless, the intensity of fluorescence emission in aqueous solution significantly increased relative to that observed in methylene chloride because the molecular self-assembly strongly suppressed the energy transfer in aqueous solution. Notably, UV/vis illumination resulted in a trans/cis isomerism of the azobenzene unit of these rod-coil molecules. However, these molecules did not respond to light in the liquid crystal phase because the transformation temperature of the liquid crystal phase was higher than the trans/cis isomer critical temperature. In aqueous solution, the trans/cis isomerism of the azobenzene unit induced the conversion of corresponding nanostructures into dispersed wormlike micelles, wormlike micelles and small fragments, and sheets. These results revealed that the trans/cis isomerism of azobenzene could be used as an external stimulus for amphiphilic molecular self-assembling systems to build various stimuli-responsive nanostructures.

Oppositely Charged, Stimuli-Responsive Anisotropic Nanoparticles for Colloidal Self-Assembly.

Anisotropic nanoparticles (ANPs) composed of distinct compartments are of interest as advanced materials because they offer unique physicochemical properties controlled by polymer composition, distribution of functional groups, and stimuli responsiveness of each compartment. Furthermore, colloidal self-assembly of ANPs via noncovalent interactions between compartments can create superstructures with additional functionality. In this study, ANPs with two compartments composed of oppositely charged and thermally responsive ternary copolymers were prepared using electrohydrodynamic cojetting. One compartment was composed of poly( N-isopropylacrylamide- co-stearyl acrylate- co-allylamine), which is positively charged in aqueous solution at pH 7, and the other compartment was composed of poly( N-isopropylacrylamide- co-stearyl acrylate- co-acrylic acid), which is negatively charged. The ANPs were stabilized in aqueous solution by physical cross-linking because of hydrophobic interactions between the 18-carbon alkyl chains of their stearyl acrylate moieties and self-assembled into supracolloidal nanostructures via electrostatic interactions. Colloidal self-assembly and thermal responsiveness were controlled by compartment charge density and solution ionic strength. The supracolloidal nanostructures exhibited both the intrinsic temperature-responsive properties of the ANPs and collective properties from self-assembly. These multifunctional, stimuli-responsive nanostructures will be useful in a variety of applications, including switchable displays, drug delivery carriers, and ion-sensitive gels.

Enzyme-Responsive Polymer Nanoparticles via Ring-Opening Metathesis Polymerization-Induced Self-Assembly.

Open-to-air aqueous-phase ring-opening metathesis polymerization-induced self-assembly (ROMPISA) is reported for forming well-defined peptide polymer nanoparticles at room temperature and with high solids concentrations (10 w/w%). For these materials, ROMPISA is shown to provide control over molecular weight with high conversion while open-to-air. Moreover, these peptide polymer nanoparticles can spontaneously rearrange into larger aggregate scaffolds in the presence of the proteolytic enzyme, thermolysin. This work demonstrates the robust nature of ROMPISA, highlighted here for the preparation of stimuli-responsive nanostructures in one pot, in air.

Light-Responsive Nanoparticles for Highly Efficient Cytoplasmic Delivery of Anticancer Agents.

Stimuli-responsive nanostructures have shown great promise for intracellular delivery of anticancer compounds. A critical challenge remains in the exploration of stimuli-responsive nanoparticles for fast cytoplasmic delivery. Herein, near-infrared (NIR) light-responsive nanoparticles were rationally designed to generate highly efficient cytoplasmic delivery of anticancer agents for synergistic thermo-chemotherapy. The drug-loaded polymeric nanoparticles of selenium-inserted copolymer (I/D-Se-NPs) were rapidly dissociated in several minutes through reactive oxygen species (ROS)-mediated selenium oxidation upon NIR light exposure, and this irreversible dissociation of I/D-Se-NPs upon such a short irradiation promoted continuous drug release. Moreover, I/D-Se-NPs facilitated cytoplasmic drug translocation through ROS-triggered lysosomal disruption and thus resulted in highly preferable distribution to the nucleus even in 5 min postirradiation, which was further integrated with light-triggered hyperthermia for achieving synergistic tumor ablation without tumor regrowth.

Single-wavelength-controlled in situ dynamic super-resolution fluorescence imaging for block copolymer nanostructures via blue-light-switchable FRAP.

Photoswitchable fluorophores are promising in single-molecule optical devices and super-resolution fluorescence imaging, especially in single-molecule photo-activated localization microscopy (PALM) or stochastic optical reconstruction microscopy (STORM). However, the scarcity of current photoswitchable fluorophores stimulates researchers to develop complicated optical systems and processing software, in accordance with the limited photoswitchable fluorescent proteins and organic fluorophores. Previous efforts to develop synthetic photoswitchable fluorophores have exhibited their promising potential in super-resolution fluorescence imaging. Here, we have designed and synthesized a fluorescence molecular switch with reversible green emission, a napthalimide-hexaarylbiimidazole conjugate (NI-N-HABI), which exhibits strong fluorescence in the emissive state, with fast thermal fading of the photochromism and spontaneous fluorescence recovery after photobleaching (FRAP) induced by blue-light. The photoswitchable fluorophore enables the red-edge wavelength of the optical response to red-shift from the initial near-UV region at less than 400 nm, to 500 nm. The relatively fast fading speed of NI-N-HABI and its sensitivity to longer blue-light irradiation (400-500 nm) have allowed simplification of the optical microscopic system from a two-wavelength laser source to a single-wavelength laser. We applied NI-N-HABI in single-wavelength-controlled in situ dynamic super-resolution fluorescence imaging for the self-assembly and solvent annealing of amphiphilic block polymers, with 50 nm of optical resolution. Single-wavelength-controlled dynamic super-resolution fluorescence imaging facilitates nanoscale optical visualization for the dynamic physical and chemical fluctuation processes of stimuli-responsive nanostructures.

Thermoreversible Morphology and Conductivity of a Conjugated Polymer Network Embedded in Block Copolymer Self-Assemblies.

Self-assembly of block copolymers provides numerous opportunities to create functional materials, utilizing self-assembled microdomains with a variety of morphology and periodic architectures as templates for functional nanofillers. Here new progress is reported toward the fabrication of thermally responsive and electrically conductive polymeric self-assemblies made from a water-soluble poly(thiophene) derivative with short poly(ethylene oxide) side chains and Pluronic L62 block copolymer solution in water. The structural and electrical properties of conjugated polymer-embedded self-assembled architectures are investigated by combining small-angle neutron and X-ray scattering, coarse-grained molecular dynamics simulations, and impedance spectroscopy. The L62 solution template organizes the conjugated polymers by stably incorporating them into the hydrophilic domains thus inhibiting aggregation. The changing morphology of L62 during the micellar-to-lamellar phase transition defines the embedded conjugated polymer network. As a result, the conductivity is strongly coupled to the structural change of the templating L62 phase and exhibits thermally reversible behavior with no signs of quenching of the conductivity at high temperature. This study shows promise for enabling more flexibility in processing and utilizing water-soluble conjugated polymers in aqueous solutions for self-assembly based fabrication of stimuli-responsive nanostructures and sensory materials.

Noncovalent Modulation of the Inverse Temperature Transition and Self-Assembly of Elastin-b-Collagen-like Peptide Bioconjugates.

Stimuli-responsive nanostructures produced with peptide domains from the extracellular matrix offer great opportunities for imaging and drug delivery. Although the individual utility of elastin-like (poly)peptides and collagen-like peptides in such applications has been demonstrated, the synergistic advantages of combining these motifs in short peptide conjugates have surprisingly not been reported. Here, we introduce the conjugation of a thermoresponsive elastin-like peptide (ELP) with a triple-helix-forming collagen-like peptide (CLP) to yield ELP-CLP conjugates that show a remarkable reduction in the inverse transition temperature of the ELP domain upon formation of the CLP triple helix. The lower transition temperature of the conjugate enables the facile formation of well-defined vesicles at physiological temperature and the unexpected resolubilization of the vesicles at elevated temperatures upon unfolding of the CLP domain. Given the demonstrated ability of CLPs to modify collagens, our results not only provide a simple and versatile avenue for controlling the inverse transition behavior of ELPs, but also suggest future opportunities for these thermoresponsive nanostructures in biologically relevant environments.

Synthesis of poly(2-hydroxyethyl methacrylate)-grafted poly(aminoamide) dendrimers as polymeric nanostructures

Fourth-generation poly(aminoamide) (PAMAM) dendrimer was synthesized via a divergent method with iterative sequence Michael addition or alkylation and amidation reactions with ethylenediamine as primary core and methyl acrylate. Then, its peripheral primary amine groups were conjugated with S-(thiobenzoyl)thioglycolic acid as reversible addition-fragmentation chain transfer (RAFT) agent and poly(2-hydroxyethyl methacrylate) (P(HEMA)) was grafted onto surface via RAFT polymerization. Fourier transform infrared spectroscopy (FT-IR), proton nuclear magnetic resonance (1H NMR), and dynamic light scattering (DLS) were used to approve the successful synthesis of different nanostructures. Then, fourth-generation, RAFT-conjugated fourth-generation, and P(HEMA)-grafted fourth-generation PAMAM dendrimers were examined as stimuli-responsive nanostructures. Finally, in vitro cellular cytotoxicity was applied to evaluate the biocompatibility of synthesized dendrimers and investigate the cytotoxic effect of grafted polymer using HeLa cells. As a results, fourth-generation and RAFT-conjugated fourth-generation samples showed no stimuli-responsive behavior while P(HEMA)-grafted fourth-generation PAMAM dendrimer had a LCST of 26 °C. Also, fourth-generation and RAFT-conjugated fourth-generation samples were characterized as toxic compounds while grafting P(HEMA) decreased cytotoxicity significantly.

Surface segregation of polypeptide-based block copolymer micelles: An approach to engineer nanostructured and stimuli responsive surfaces

We describe the surface segregation of polypeptide-based block copolymer micelles to produce stimuli-responsive nanostructures at the polymer blend/air interface. Such structures were obtained by simultaneous surface migration and self assembly at the surface of diblock copolymer/homopolymer blends. We employed blends composed of homopolymer (PS) and an amphiphilic block copolymer polystyrene-b-poly(l-glutamic acid) (PS-b-PGA). The surface was functionalized based on the preferential segregation to the polymer blend/air interface of the hydrophilic PGA block of the diblock copolymer upon annealing to water vapor. The surface migration of the diblock copolymer to the interface was demonstrated both by XPS and contact angle measurements. As a consequence, the PGA interfacial attraction leads to a large surface excess on diblock copolymer which in turn, through macrophase and microphase separation, produced separated domains at the surface with regions composed either of homo or block copolymer. Herein we demonstrate that the use of asymmetric diblock copolymers with a higher content in PS lead to spherical micellar assemblies randomly distributed at the surface. As observed by AFM imaging the blend composition, i.e. the amount of block copolymer within the blend influences the density of micelles at the surface. Finally, when exposed to water, the pH affects the surface morphology. The PGA segments are collapsed at low pH values and extended at pH values above 4.8, thus inducing variations on the topography of the films at the nanometer scale.

Design of a Thermocontrollable Protein Complex

It is widely recognized that stimuli-responsive nanostructures play a promising role in nanodevices for medical treatments and experimental tools. We have designed and constructed a basic structure which controls the distance between two termini domains through temperature reversibility. Our structure, shaped like a bouquet, is composed of two proteins, alpha-helix and elastin-like protein (ELP). Alpha-helices align and bundle the ELP while ELP twists and forms a fiber-like structure at warm temperatures. This ELP conformational change alters the distance between the structure termini at the site opposite the alpha-helix. We connected enhanced yellow fluorescent protein (EYFP) and enhanced cyan fluorescent protein (ECFP) at the structure's two termini to evaluate the distance using fluorescence resonance energy transfer (FRET) efficiency. These proteins spontaneously formed a complex which decreased the distance between the two fluorescent proteins located at its termini, at physiologically relevant temperatures. This change was repeated with complete reversibility (n = 5).