Gradient Redox-Responsive and Two-Stage Rocket-Mimetic Drug Delivery System for Improved Tumor Accumulation and Safe Chemotherapy.

Recent drug delivery nanosystems for cancer treatment still suffer from the poor tumor accumulation and low therapeutic efficacy due to the complex in vivo biological barriers. To resolve these problems, in this work, a novel gradient redox-responsive and two-stage rocket-mimetic drug nanocarrier is designed and constructed for improved tumor accumulation and safe chemotherapy. The nanocarrier is constructed on the basis of the disulfide-doped organosilica-micellar hybrid nanoparticles and the following dual-functional modification with disulfide-bonded polyethylene glycol (PEG) and amido-bonded polyethylenimine (PEI). First, prolonged circulation duration in the bloodstream is guaranteed due to the shielding of the outer PEG chains. Once the nanocarrier accumulates at the tumoral extracellular microenvironment with low glutathione (GSH) concentrations, the first-stage redox-responsive behavior with the separation of PEG and the exposure of PEI is triggered, leading to the improved tumor accumulation and cellular internalization. Furthermore, with their endocytosis by tumor cells, a high concentration of GSH induces the second-stage redox-responsiveness with the degradation of silsesquioxane framework and the release of the encapsulated drugs. As a result, the rocket-mimetic drug carrier displays longer circulation duration in the bloodstream, higher tumor accumulation capability, and improved antitumor efficacy (which is 2.5 times higher than that with inseparable PEG). It is envisioned that the rocket-mimetic strategy can provide new solutions for improving tumor accumulation and safety of nanocarriers in further cancer chemotherapy.

Aqueous Synthesis of Multi-Carbon Dot Cross-Linked Polyethyleneimine Particles with Enhanced Photoluminescent Properties.

Heavy-metal-free fluorescent nanoparticles with high photostability and low toxicity are highly desirable as imaging probes for biological applications. Here, a novel synthetic strategy to prepare ultrabright multi-carbon dot cross-linked PEI particles, namely CDs@PEI, through self-assembly of hydrophobically modified PEI and in situ generations of carbon dots within residual monomer-swollen micelles is reported. The resulting particles consist of numerous carbon dots, which are individually and homogeneously embedded within the PEI network, and have an average hydrodynamic diameter of approximately 100 nm with ζ-potential above +35 mV. The CDs@PEI particles possess the synergistic effect of photoluminescent carbon dot and crosslink-enhanced emission of PEI, giving the particles superior optical properties such as high fluorescence quantum yield (up to 66%) in the aqueous system, excitation-dependent emission phenomenon, stable fluorescence in a wide pH range, and resistance to photobleaching.

Synthesis of a Pillar[5]arene-Based Polyrotaxane for Enhancing the Drug Loading Capacity of PCL-Based Supramolecular Amphiphile as an Excellent Drug Delivery Platform.

A pillar[5]arene-based nonionic polyrotaxane (PR) with star-poly(ε-caprolactone) ( S-PCL) as the axle, pillar[5]arene (DEP5) as the wheel and adamantane as the end-capped group is designed and synthesized. The resulting PR is subsequently assembled with ß-cyclodextrin end-capped pH-stimulated poly(acrylic acid) (CD-PAA) via a host-guest interaction to form the supramolecular pseudoblock polymer PR-PAA. This supramolecular pseudoblock polymer could self-assemble in aqueous solution to produce PR-PAA-based supramolecular vesicular nanoparticles (PR-SVNPs), which present significantly enhanced drug loading capacity (DLC, 45.6%) of DOX, much higher than those of superamphiphiles (PCL-PAA, 17.1%). Such a high DLC of PR-SVNPs can be most probably attributed to the greatly decreased crystallinity of PCL in PR. Moreover, the loaded drugs could be selectively released in an acidic microenvironment-responsive manner. Compared to free DOX, the DOX-loaded PR-SVNPs (DOX@PR-SVNPs) shows much enhanced cellular uptake and cytotoxicity against the SMMC-7721. More importantly, thanks to the enhanced permeability and retention (EPR) effect, DOX@PR-SVNPs exhibits appealing features such as extremely low toxicity, highly efficient intratumoral accumulation and substantial antitumor efficacy in vivo.

A confined crosslinking strategy towards an intelligent organosilica-micellar hybrid drug delivery system.

An ideal drug delivery system must have a high level of stability to ensure effective circulation and passive aggregation, good retention performance, and dynamic delivery and treatment monitoring. Thus, the development of a smart drug delivery carrier with both precise drug release and real-time detection remains a challenge. Herein, we propose a confined crosslink protocol to prepare an intelligent hybrid delivery system for auto-fluorescent monitoring, protonation-induced retention and precise drug release. The construction of this system involves the hydrolysis and condensation of (3-aminopropyl)triethoxysilane (APTES) silanes inside the Pluronic polymer micelles and thereafter a confined Schiff base crosslinking between glutaraldehyde (GA) and residual silane amino groups. The size of the intelligent docetaxel (DTX)-loaded nanosystem changes from ∼25 nm in blood circulation or normal tissues (pH ∼ 7.4) to ∼250 nm in slightly acidic environments (pH ∼ 6.5-7.0) owing to intra-molecular hydrogen bond-induced aggregation and imine cleavage-induced disintegration in the endosome (pH ∼ 5.0-6.2) along with auto-fluorescent monitoring contributing to the high-efficiency chemotherapy. This work provides a new method to construct smart, acid-responsive and fluorescent-guided drug-delivery carrier systems for efficient and safe tumor chemotherapy.

Peroxisome inspired hybrid enzyme nanogels for chemodynamic and photodynamic therapy.

Peroxisome, a special cytoplasmic organelle, possesses one or more kinds of oxidases for hydrogen peroxide (H2O2) production and catalase for H2O2 degradation, which serves as an intracellular H2O2 regulator to degrade toxic peroxides to water. Inspired by this biochemical pathway, we demonstrate the reactive oxygen species (ROS) induced tumor therapy by integrating lactate oxidase (LOx) and catalase (CAT) into Fe3O4 nanoparticle/indocyanine green (ICG) co-loaded hybrid nanogels (designated as FIGs-LC). Based on the O2 redistribution and H2O2 activation by cascading LOx and CAT catalytic metabolic regulation, hydroxyl radical (·OH) and singlet oxygen (1O2) production can be modulated for glutathione (GSH)-activated chemodynamic therapy (CDT) and NIR-triggered photodynamic therapy (PDT), by manipulating the ratio of LOx and CAT to catalyze endogenous lactate to produce H2O2 and further cascade decomposing H2O2 into O2. The regulation reactions of FIGs-LC significantly elevate the intracellular ROS level and cause fatal damage to cancer cells inducing the effective inhibition of tumor growth. Such enzyme complex loaded hybrid nanogel present potential for biomedical ROS regulation, especially for the tumors with different redox state, size, and subcutaneous depth.

Synthesis of core-shell structured dual-mesoporous silica spheres with tunable pore size and controllable shell thickness.

Core-shell structured dual-mesoporous silica spheres (DMSS) that possess smaller pores (2.0 nm) in the shell and larger tunable pores (12.8-18.5 nm) in the core have been successfully synthesized by utilizing an amphiphilic block copolymer (polystyrene-b-poly (acrylic acid), PS-b-PAA) and cetyl trimethyl ammonium bromide (CTAB) as cotemplates. The thickness of the shells and the larger pore size in the core could be easily tuned by changing the amounts of TEOS and the hydrophobic block (PS) length during synthesis, respectively. By encapsulating hydrophobic magnetite nanoparticles into the cores, superparamagnetic dual-mesoporous silica spheres were obtained. Drug storage and release testing results showed that the diffusing rate of the stored drug could be efficiently controlled by changing the shell thickness of DMSS.

Biodegradable organosilica magnetic micelles for magnetically targeted MRI and GSH-triggered tumor chemotherapy.

Recently, block copolymer micelles have attracted widespread attention due to their controlled biodegradability and excellent loading capability. Unfortunately, the poor in vivo stability and low delivery efficiency of drug-loaded micelles greatly hampered their biomedical applications. Herein, we develop a new kind of biodegradable magnetite/doxorubicin (Fe3O4/DOX) co-loaded PEGylated organosilica micelles (designated as FDPOMs) with both high circulating stability and smart GSH-triggered biodegradability for magnetically targeted magnetic resonance imaging (MRI) and tumor chemotherapy. The FDPOMs are prepared by the self-assembly of biodegradable polycaprolactone-block-poly(glutamic acid) (PCL-b-PGA), a chemotherapeutic DOX drug and Fe3O4 nanoparticles in an oil/water system, subsequent organosilica cross-linking with 3-mercaptopropyltrimethoxysilane (MPTMS) molecules and surface PEGylation. The resultant FDPOMs exhibit excellent dispersity and stability in biological media, remarkable T2-weighted MR imaging capability, unique GSH-responsive release behavior and selective toxicity to tumor cells. The in vivo experiments show that the FDPOMs not only have improved MR tumor imaging capability, but also exhibit high anti-tumor efficacy due to the strong magnetic targeting ability under an external magnetic field. Consequently, the FDPOMs are promising candidates for magnetically targeted MR imaging and imaging-guided tumor chemotherapy.

Effective gene delivery of shBMP-9 using polyethyleneimine-based core-shell nanoparticles in an animal model of insulin resistance.

Bone morphogenetic protein (BMP)-9 has been associated with insulin resistance and type 2 diabetes mellitus. However, methods for delivering exogenous BMP-9 genes in vivo are lacking. In this study, we developed a gene delivery system using polyethyleneimine (PEI)-based core-shell nanoparticles (PCNs) as gene delivery carriers, and investigated the effectiveness and safety for delivery of the shBMP-9 gene. PCNs possessed a well-defined core-shell nanostructure with hydrophobic polymer cores and dense PEI shells of uniform particle size and highly positively charged surfaces. In vitro evaluation suggested that PCNs had high loading capacity for exogenous genes and low cytotoxicity toward hepatocytes. The transfection efficiency of PCNs/pENTR-shBMP9 complexes was higher than that of commercial lipofectamine 2000/shBMP9. In vivo studies showed that PCNs/pENTR-shBMP9 transfection led to a significant decrease in hepatic BMP9 expression compared with pENTR-shBMP9 transfection. Under high fat diet (HFD) feeding, PCNs/pENTR-shBMP9 mice exhibited aggravated glucose and insulin tolerance. At a molecular level, PCNs/pENTR-shBMP9 mice displayed elevated PEPCK protein levels and lower levels of InsR and Akt phosphorylation than pENTR-shBMP9 mice. These results suggest that the biological effects of PCNs/pENTR-shBMP9 in vivo are much more effective than those of pENTR-shBMP9. Therefore, the polyethyleneimine (PEI)-based core-shell nanoparticle can be applied as promising nanocarriers for effective and safe gene delivery.

A facile approach to fabricate functionalized superparamagnetic copolymer-silica nanocomposite spheres.

Novel amino- or thiol-functionalized superparamagnetic copolymer-silica nanospheres (NH2-SMCSNs/SH-SMCSNs), which consist of a magnetic core and a silica cross-linked block copolymer shell, have been fabricated.

Amphiphilic core-shell nanoparticles containing dense polyethyleneimine shells for efficient delivery of microRNA to Kupffer cells.

Efficient and targeted delivery approach to transfer exogenous genes into macrophages is still a great challenge. Current gene delivery methods often result in low cellular uptake efficiency in vivo in some types of cells, especially for the Kupffer cells (KCs). In this article, we demonstrate that amphiphilic core-shell nanoparticles (NPs) consisting of well-defined hydrophobic poly(methyl methacrylate) (PMMA) cores and branched polyethyleneimine (PEI) shells (denoted as PEI@PMMA NPs) are efficient nanocarriers to deliver microRNA (miRNA)-loaded plasmid to the KCs. Average hydrodynamic diameter of PEI@ PMMA NPs was 279 nm with a narrow size distribution. The NPs also possessed positive surface charges up to +30 mV in water, thus enabling effective condensation of negatively charged plasmid DNA. Gel electrophoresis assay showed that the resultant PEI@PMMA NPs were able to completely condense miRNA plasmid at a weight ratio of 25:1 (N/P ratio equal to 45:1). The Cell Counting Kit-8 assay and flow cytometry results showed that the PEI@PMMA/miRNA NPs displayed low cytotoxicity and cell apoptosis activity against the KCs. The maximum cell transfection efficiency reached 34.7% after 48 hours, which is much higher than that obtained by using the commercial Lipofectamine™ 2000 (1.7%). Bio-transmission electron microscope observation revealed that the PEI@PMMA NPs were mainly distributed in the cytoplasm of the KCs. Furthermore, when compared to the control groups, the protein expression of target nuclear factor κB P65 was considerably inhibited (P<0.05) both in vitro and in vivo. These results demonstrate that the PEI@PMMA NPs with a unique amphiphilic core-shell nanostructure are promising nanocarriers for delivering miRNA plasmid to KCs.

Polyethyleneimine-Based Nanocarriers for Gene Delivery.

There is a tremendous progress in the design and synthesis of nano-scaled, non-viral carriers in the previous two decades. The nanodelivery systems can significantly improve biopharmaceutical features, pharmacokinetic properties and therapeutic efficacy of entrapped drugs. Branched polyethylenimine (PEI) is a cationic polymer that contains primary, secondary and tertiary amino groups. Such type of water-soluble polymer having high density of amines is one of the most promising cationic vectors for gene delivery. Hence, constructing nanocarriers that contain PEI have attracted much research effort in gene therapy because of the synergy effects of PEI molecules for their efficient transfection and the multi-functionality of nanoparticles in delivery. In this review, we focus on the recent development on the design and synthesis of four types of PEI-based nanocarriers: 1) PEI-based polymeric micelles systems; 2) PEI-based polymeric nanoparticle system; 3) PEI/silica nanoparticle systems; and 4) PEI/metal nanoparticle systems. Their in vitro gene transfaction and in vivo gene therapy will be also discussed. Results from these studies have demonstrated that PEI-based nanocarriers are promising delivery systems because of their efficient gene transfection, negligible toxicity, capability to co-deliver nucleic acids and chemotherapy drugs, ease of modification with the targeting molecules, and good responsiveness to external stimulus.

Multifunctional gold nanostar-based nanocomposite: Synthesis and application for noninvasive MR-SERS imaging-guided photothermal ablation.

We report here the design and facile synthesis of multifunctional gold nanostars based nanocomposites (MGSNs) through direct organosilica coating onto anisotropic gold nanostars followed by the conjugation of Gd chelates. The as-synthesized MGSNs possess strong NIR absorbance, SERS signal and enhanced T1-MR imaging capability with excellent dispersivity and uniform size, as well as great photothermal stability and Raman stability under photothermal conditions. Importantly, MGSNs present excellent performance in vivo after their intravenous injection for both MR and SERS imaging and the high efficiency for killing tumor cells through photothermal ablation with NIR irradiation. A combination of the high spatial resolution of MR and the exciting sub-cell-level sensitivity and resolution of SERS can provide comprehensive information about the tumor to achieve the optimized therapeutic outcome. Therefore, MGSNs are of great potential as a multifunctional nanoplatform for MR-SERS bimodal imaging-guided, focused photothermal tumor therapy.

Dual-Enzyme-Loaded Multifunctional Hybrid Nanogel System for Pathological Responsive Ultrasound Imaging and T2-Weighted Magnetic Resonance Imaging.

A dual-enzyme-loaded multifunctional hybrid nanogel probe (SPIO@GCS/acryl/biotin-CAT/SOD-gel, or SGC) has been developed for dual-modality pathological responsive ultrasound (US) imaging and enhanced T2-weighted magnetic resonance (MR) imaging. This probe is composed of functionalized superparamagnetic iron oxide particles, a dual enzyme species (catalase and superoxide dismutase), and a polysaccharide cationic polymer glycol chitosan gel. The dual-modality US/MR imaging capabilities of the hybrid nanogel for responsive US imaging and enhanced T2-weighted MR imaging have been evaluated both in vitro and in vivo. These results show that the hybrid nanogel SGC can exhibit efficient dual-enzyme biocatalysis with pathological species for responsive US imaging. SGC also demonstrates increased accumulation in acidic environments for enhanced T2-weighted MR imaging. Further research on these nanogel systems may lead to the development of more efficient US/MR contrast agents.

Monodispersed and ordered large-pore mesoporous silica nanospheres with tunable pore structure for magnetic functionalization and gene delivery.

It can be larger: A facile self-assembly/solvothermal approach to synthesize monodispersed, large-pore (>12 nm) silica nanospheres (LPSNs) with ordered, accessible, and interconnected pore channels has been successfully developed by utilizing an amphiphilic block copolymer (polystyrene-b-poly (acrylic acid), PS-b-PAA) as pore template and cetyltrimethylammonium bromide (CTAB) as structure-stabilizing agent.

Manganese-loaded dual-mesoporous silica spheres for efficient T1- and T2-weighted dual mode magnetic resonance imaging.

A novel class of manganese-based dual-mode contrast agents (DMCAs) based on the core-shell structured manganese-loaded dual-mesoporous silica spheres (Mn-DMSSs) for simultaneous T1- and T2-weighted magnetic resonance imaging (MRI) has been successfully reported. The in vitro MR tests demonstrate that the Mn-based DMCAs display an excellent simultaneous T1-weighted and T2-weighted MR imaging effect with a noticeably high T1 relaxivity (r1) of 10.1 mM(-1) s(-1) and a moderately high T2 relaxivity (r2) of 169.7 mM(-1) s(-1). The Mn-based DMCAs exhibit negligible cytotoxicity with >80% cell viability at a concentration of up to 200 µg/mL in human liver carcinoma (HepG2) and mouse macrophage (RAW264.7) cells after 24 h. Confocal laser scanning microscopy (CLSM) results show that the Mn-DMSSs were internalized via endocytosis and located in the cytoplasm but not in the nucleus. The in vivo experiment shows that the signals of rat liver increased by 29% under T1-weighted imaging mode and decreased by 28% under T2-weighted imaging mode in 5 min postinjection of Mn-DMSSs, which reveal that the novel Mn-loaded DMSSs can be used as both positive (T1-weighted) and negative (T2-weighted) MR contrast agents in further biomedical applications.

A pH-responsive hybrid fluorescent nanoprober for real time cell labeling and endocytosis tracking.

Hydrophilic, fluorescent hybrid nanoprobes (NDI@HNPs) encapsulated with the hydrophobic pH-responsive fluorophore (N,N'-di-n-dodecyl-2,6-di(4-methyl-piperazin-1-yl)naphthalene-1,4,5,8-tetracarboxylic acid diimide, NDI) for recognizing and mapping the route of cell phagocytosis have been fabricated based on the self-assembly of amphiphilic diblock copolymer PS-b-PAA and the subsequent shell cross-linking with 3-mercaptopropyltrimethoxy silane (MPTMS). The as-synthesized NDI@HNPs has a typical spherical morphology of 46 nm in diameter with excellent monodispersity in aqueous solution. The NDI@HNPs probe exhibits extremely low cytotoxicity, fast real time pH response and enhanced fluorescence intensity under acidic environment with respect to the corresponding free dye in highly polar aqueous system because of the encapsulation of NDI molecules inside nanoparticle cores with weak polarity environment. The fluorescence intensity of NDI@HNPs is enhanced by 55-fold upon changing from neutral (pH = 7.4) or basic (pH = 8.4) to acid (pH = 3.4) in aqueous system, in contrast to the serious fluorescence quenching of free NDI in the same medium, which can exactly meet the physiological pH range in cells. The favorably long emission wavelength is beneficial to the low scattering and minimal interfering requirements to fluorescent bioimaging. Moreover, functionalization with rapid cell-penetrating peptides (HIV-1 TAT) allows them to overcome the physiological and biological barriers during the phagocytosis process. Its characteristic fluorescent response to pH benefits the intracellular labeling and organelle targeting, realizing the real time tracking of the probe entry into cancer cells, the accumulation into the endolysosome and the further escape.