Hydrogen-bonded cytosine-endowed supramolecular polymeric nanogels: Highly efficient cancer cell targeting and enhanced therapeutic efficacy.

We demonstrate that cytosine moieties within physically cross-linked supramolecular polymers not only manipulate drug delivery and release, but also confer specific targeting of cancer cells to effectively enhance the safety and efficacy of chemotherapy-and thus hold significant potential as a new perspective for development of drug delivery systems. Herein, we successfully developed physically cross-linked supramolecular polymers (PECH-PEG-Cy) comprised of hydrogen-bonding cytosine pendant groups, hydrophilic poly(ethylene glycol) side chains, and a hydrophobic poly(epichlorohydrin) main chain. The polymers spontaneously self-assemble into a reversibly hydrogen-bonded network structure induced by cytosine and directly form spherical nanogels in aqueous solution. Nanogels with a high hydrogen-bond network density (i.e., a higher content of cytosine moieties) exhibit outstanding long-term structural stability in cell culture substrates containing serum, whereas nanogels with a relatively low hydrogen-bond network density cannot preserve their structural integrity. The nanogels also exhibit numerous unique physicochemical characteristics in aqueous solution, such as a desirable spherical size, high biocompatibility with normal and cancer cells, excellent drug encapsulation capacity, and controlled pH-responsive drug release properties. More importantly, in vitro experiments conclusively indicate the drug-loaded PECH-PEG-Cy nanogels can selectively induce cancer cell-specific apoptosis and cell death via cytosine receptor-mediated endocytosis, without significantly harming normal cells. In contrast, control drug-loaded PECH-PEG nanogels, which lack cytosine moieties in their structure, can only induce cell death in cancer cells through non-specific pathways, which significantly inhibits the induction of apoptosis. This work clearly demonstrates that the cytosine moieties in PECH-PEG-Cy nanogels confer selective affinity for the surface of cancer cells, which enhances their targeted cellular uptake, cytotoxicity, and subsequent induction of programmed cell death in cancer cells.

Spontaneous Self-Assembly of Single-Chain Amphiphilic Polymeric Nanoparticles in Water.

Single-chain polymeric nanoparticles (SCPNs) have great potential as functional nanocarriers for drug delivery and bioimaging, but synthetic challenges in terms of final yield and purification procedures limit their use. A new concept to modify and improve the synthetic procedures used to generate water-soluble SCPNs through amphiphilic interactions has been successfully exploited. We developed a new ultrahigh molecular weight amphiphilic polymer containing a hydrophobic poly(epichlorohydrin) backbone and hydrophilic poly(ethylene glycol) side chains. The polymer spontaneously self-assembles into SCPNs in aqueous solution and does not require subsequent purification. The resulting SCPNs possess a number of distinct physical properties, including a uniform hydrodynamic nanoparticle diameter of 10-15 nm, extremely low viscosity and a desirable spherical-like morphology. Concentration-dependent studies demonstrated that stable SCPNs were formed at high concentrations up to 10 mg/mL in aqueous solution, with no significant increase in solution viscosity. Importantly, the SCPNs exhibited high structural stability in media containing serum or phosphate-buffered saline and showed almost no change in hydrodynamic diameter. The combination of these characteristics within a water-soluble SCPN is highly desirable and could potentially be applied in a wide range of biomedical fields. Thus, these findings provide a path towards a new, innovative route for the development of water-soluble SCPNs.

Self-Assembled Supramolecular Micelles with pH-Responsive Properties for More Effective Cancer Chemotherapy.

pH-Responsive hydrogen-bonded supramolecular micelles, composed of a water-soluble poly(ethylene glycol) polymer with two terminal sextuple hydrogen bonding groups, can spontaneously organize in aqueous media to give well-defined, uniformly sized spherical micelles. The supramolecular micelles exhibit a number of unique physical characteristics, such as interesting amphiphilic behavior, desirable micellar size and nanospherical morphology, excellent biocompatibility, tailorable drug-loading capacities, and high structural stability in media containing serum or red blood cells. In addition, the drug release kinetics of drug-loaded micelles can be easily manipulated to achieve the desired release profile by regulating the environmental pH, thus these micelles are highly attractive candidates as an intelligent drug carrier system for cancer therapy. Cytotoxicity assays showed that the drug-loaded micelles induced pH-dependent intracellular drug release and exerted strong antiproliferative and cytotoxic activities toward cancer cells. Importantly, cellular uptake and flow cytometric analyses confirmed that a mildly acidic intracellular environment significantly increased cellular internalization of the drug-loaded micelles and subsequent drug release in the cytoplasm and nucleus of cancer cells, resulting in more effective induction of apoptotic cell death. Thus, this system may provide an efficient route toward achieving the fundamental properties and practical realization of pH-sensitive drug-delivery systems for chemotherapy.

Dual stimuli-responsive supramolecular boron nitride with tunable physical properties for controlled drug delivery.

The new concept of modifying and tailoring the properties of existing two-dimensional (2D) nanomaterials by invoking the assembly of supramolecular networks upon association with a adenine-functionalized macromer (A-PPG) has significant potential to facilitate the development of highly water-dispersible few-layered 2D nanosheets. In this study, we propose that water-soluble A-PPG directly self-assembles into a long-period stacking-ordered lamellar structure over the surface of hexagonal boron nitride (BN) in aqueous solution, due to the efficient non-covalent interactions between A-PPG and BN nanosheets. The layer number of BN nanosheets can be easily tuned by altering the mass ratio of the A-PPG and BN blend, and the resulting exfoliated nanosheets also exhibit excellent temperature/pH-responsive behavior, biocompatibility and extremely high drug-loading capacity (up to 36.2%), features that are highly desirable yet exceedingly rare in traditional 2D nanomaterials. Importantly, in vitro drug release studies showed the drug-loaded nanosheets function as a stable nanocarrier with excellent stability and drug entrapment under normal physiological conditions. Increasing the environmental temperature to 40 °C or decreasing the pH to 5.5 triggered rapid release of the encapsulated drug from the drug-loaded nanosheets, suggesting this newly developed material has potential as a novel multi-responsive 2D nanocarrier to safely deliver drugs and effectively facilitate controlled drug release under specific microenvironmental conditions. This study provides new insight towards the promising application of this system in controlled release drug delivery systems.

Entrapment of an adenine derivative by a photo-irradiated uracil-functionalized micelle confers controlled self-assembly behavior.

HYPOTHESIS: Invoking cooperative assembly of the uracil-functionalized supramolecular polymer BU-PPG [uracil end-capped poly(propylene glycol)] upon association with the nucleobase adenine derivative A-MA [methyl 3-(6-amino-9H-purin-9-yl)propanoate] as a model drug provides a new concept to control and tune the properties of supramolecular complexes and holds significant potential for the development of safer, more effective drug delivery systems. EXPERIMENTS: BU-PPG and A-MA were successfully developed and exhibited specific recognition and high affinity, which enabled reversible complementary adenine-uracil (A-U) hydrogen bonding-induced formation of spherical micelles in aqueous solution. The self-assembly and controllable A-MA release behavior of BU-PPG/A-MA micelles were studied using morphological analysis and optical and light scattering techniques to investigate the effect of photoirradiation and temperature on the complementary hydrogen bond interactions between BU-PPG and A-MA. FINDINGS: The resulting micelles possess unusual physical properties, including controlled photoreactivity kinetics, controllable self-assembled morphology and low cytotoxicity in vitro, as well as reversible temperature-responsive behavior. Importantly, irradiated micelles exhibited excellent long-term structural stability under normal physiological conditions and serum disturbance. Increasing the temperature triggered rapid release of A-MA by disrupting A-U complexes. These findings represent an entirely new, promising strategy for the development of multi-controlled release drug delivery nanocarriers based on complementary hydrogen bonding interactions.

Highly Effective Photocontrollable Drug Delivery Systems Based on Ultrasensitive Light-Responsive Self-Assembled Polymeric Micelles: An in Vitro Therapeutic Evaluation.

An ultrasensitive light-responsive block copolymer, a combination of a multiarmed poly(ethylene glycol)-b-poly(caprolactone) polymer as a water-soluble element and maleimide-anthracene linkers as a photosensitive group, was successfully synthesized and rapidly self-assembled to form spherical micellar nanoparticles in aqueous media and phosphate-buffered saline. Their unique characteristics, such as extremely low critical micelle concentration, desirable micellar stability, well-controlled light-responsiveness, tailorable drug-loading content, and ultrasensitive light-induced drug release, make these micelles potential candidates for development of a more effective, safer drug delivery platform for cancer treatment. In vitro studies revealed that the drug-loaded micelles exhibited high structural stability in serum-containing media and very low toxicity toward normal and cancer cells under physiological conditions. Irradiation of cancer cells incubated with the drug-loaded micelles with ultraviolet light at 254 nm for only 10 s triggered rapid and complete release of the drug in the intracellular environment and induced strong antiproliferative/cytotoxic activity. Importantly, real-time cytotoxic assays and fluorescence imaging analysis further demonstrated that the drug-loaded micelles were rapidly taken up into the cytosol or nuclei of the cells, and subsequent ultraviolet exposure induced drug release and apoptotic cell death. Given their simplicity of design, high reliability, and performance, this new light-sensitive micelle may provide a promising route for developing a multifunctional therapeutic nanocarrier system.

Dynamic tungsten diselenide nanomaterials: supramolecular assembly-induced structural transition over exfoliated two-dimensional nanosheets.

A simple and effective method for direct exfoliation of tungsten diselenide (WSe2) into few-layered nanosheets has been successfully developed by employing a low molecular weight adenine-functionalized supramolecular polymer (A-PPG). In this study, we discover A-PPG can self-assemble into a long-range, ordered lamellar microstructure on the surface of WSe2 due to the efficient non-covalent interactions between A-PPG and WSe2. Morphological and light scattering studies confirmed the dynamic self-assembly behavior of A-PPG has the capacity to efficiently manipulate the transition between contractile and extended lamellar microstructures on the surface of metallic 1T-phase and semiconducting 2H-phase WSe2 nanosheets, respectively. The extent of WSe2 exfoliation can be easily controlled by systematically adjusting the amount of A-PPG in the composites, to obtain nanocomposites with the desired functional characteristics. In addition, the resulting composites possess unique liquid-solid phase transition behavior and excellent thermoreversible properties, revealing the self-assembled lamellar structure of A-PPG functions as a critical factor to manipulate and tailor the physical properties of exfoliated WSe2. This newly developed method of producing exfoliated WSe2 provides a useful conceptual and potential framework for developing WSe2-based multifunctional nanocomposites to extend their application in solution-processed semiconductor devices.

Self-Assembled pH-Responsive Polymeric Micelles for Highly Efficient, Noncytotoxic Delivery of Doxorubicin Chemotherapy To Inhibit Macrophage Activation: In Vitro Investigation.

Self-assembled pH-responsive polymeric micelles, a combination of hydrophilic poly(ethylene glycol) segments and hydrogen bonding interactions within a biocompatible polyurethane substrate, can spontaneously self-assemble into highly controlled, nanosized micelles in aqueous solution. These newly developed micelles exhibit excellent pH-responsive behavior and biocompatibility, highly controlled drug (doxorubicin; DOX) release behavior, and high drug encapsulation stability in different aqueous environments, making the micelles highly attractive potential candidates for safer, more effective drug delivery in applications such as cancer chemotherapy. In addition, in vitro cell studies revealed the drug-loaded micelles possessed excellent drug entrapment stability and low cytotoxicity toward macrophages under normal physiological conditions (pH 7.4, 37 °C). When the pH of the culture media was reduced to 6.0 to mimic the acidic tumor microenvironment, the drug-loaded micelles triggered rapid release of DOX within the cells, which induced potent antiproliferative and cytotoxic effects in vitro. Importantly, fluorescent imaging and flow cytometric analyses confirmed the DOX-loaded micelles were efficiently delivered into the cytoplasm of the cells via endocytosis and then subsequently gradually translocated into the nucleus. Therefore, these multifunctional micelles could serve as delivery vehicles for precise, effective, controlled drug release to prevent accumulation and activation of tumor-promoting tumor-associated macrophages in cancer tissues. Thus, this unique system may offer a potential route toward the practical realization of next-generation pH-responsive therapeutic delivery systems.