pH-responsive Vesicles Research Articles

Self-Assembled Recombinant Elastin and Globular Protein Vesicles with Tunable Properties for Diverse Applications.

Vesicles are self-assembled structures comprised of a membrane-like exterior surrounding a hollow lumen with applications in drug delivery, artificial cells, and micro-bioreactors. Lipid or polymer vesicles are the most common and are made of lipids or polymers, respectively. They are highly useful structures for many applications but it can be challenging to decorate them with proteins or encapsulate proteins in them, owing to the use of organic solvent in their formation and the large size of proteins relative to lipid or polymer molecules. By utilization of recombinant fusion proteins to make vesicles, specific protein domains can be directly incorporated while also imparting tunability and stability. Protein vesicle assembly relies on the design and use of self-assembling amphiphilic proteins. A specific protein vesicle platform made in purely aqueous conditions of a globular, functional protein fused to a glutamate-rich leucine zipper (ZE) and a thermoresponsive elastin-like polypeptide (ELP) fused to an arginine-rich leucine zipper (ZR) is discussed here. The hydrophobic conformational change of the ELP above its transition temperature drives assembly, and strong ZE/ZR binding enables incorporation of the desired functional protein. Mixing the soluble proteins on ice induces zipper binding, and then warming above the ELP transition temperature (Tt) triggers the transition to and growth of protein-rich coacervates and, finally, reorganization of proteins into vesicles. Vesicle size is tunable based on salt concentration, rate of heating, protein concentration, size of the globular protein, molar ratio of the proteins, and the ELP sequence. Increasing the salt concentration decreases vesicle size by decreasing the Tt, resulting in a shorter coacervation transition stage. Likewise, directly changing the heating rate also changes this time and increasing protein concentration increases coalescence. Increasing globular protein size decreases the size of the vesicle due to steric hindrance. By changing the ELP sequence, which consists of (VPGXG)n, through the guest residue (X) or number of repeats (n), Tt is changed, affecting size. Additionally, the chemical nature of X variation has endowed vesicles with stimuli responsiveness and stability at physiological conditions.Protein vesicles have been used for biocatalysis, biomacromolecular drug delivery, and vaccine applications. Photo-cross-linkable vesicles were used to deliver small molecule cargo to cancer cells in vitro and antigen to immune cells in vivo. pH-responsive vesicles effectively delivered functional protein cargo, including cytochrome C, to the cytosol of cancer cells in vitro, using hydrophobic ion pairing to improve cargo distribution in the vesicles and release. The globular protein used to make the vesicles can be varied to achieve different functions. For example, enzyme vesicles exhibit biocatalysis, and antigen vesicles induce antibody and cellular immune responses after vaccination in mice. Collectively, the development and engineering of the protein vesicle platform has employed amphiphilic self-assembly strategies and rational protein engineering to control physical, chemical, and biological properties for biotechnology and nanomedicine applications.

Light and pH responsive catanionic vesicles based on a chalcone/flavylium photoswitch for smart drug delivery: From molecular design to the controlled release of doxorubicin

Spatially and temporally localized delivery is a promising strategy to circumvent adverse effects of traditional drug therapy such as drug toxicity and prolonged treatments. Stimuli-responsive colloidal nanocarriers can be crucial to attain such goals. Here, we develop a delivery system based on dual light and pH responsive vesicles having a cationic bis-quat gemini surfactant, 12–2-12, and a negatively charged amphiphilic chalcone, C4SCh. The premise is to exploit the chalcone/flavylium interconversion to elicit a morphological change of the vesicles leading to the controlled release of an encapsulated drug. First, the phase behavior of the catanionic system is studied and the desirable composition yielding stable unilamellar vesicles identified and selected for further studies. The solutions containing vesicles (Dh ≈ 200 nm, ζ-potential ≈ 80 mV) are in-depth characterized by light microscopy, cryo-transmission electron microscopy (cryo-TEM), dynamic light scattering (DLS) and surface tension measurements. Upon subjecting the vesicles to UV irradiation (λ = 365 nm) at near neutral pH (≈ 6.0), no morphological effects are observed, yet when irradiation is coupled with pH = 3.0, the majority of the vesicles are disrupted into bilayer fragments. The anticancer drug doxorubicin (DOX) is successfully entrapped in the non-irradiated vesicles, yielding an encapsulation efficiency of ≈ 25% and a loading capacity of ≈ 3%. The release profile of the drug-loaded vesicles is then studied in vitro in four conditions: i) no stimuli (pH = 6.0); ii) irradiation, pH = 6.0; iii) no irradiation and adjusted pH = 3.0; iv) irradiation and adjusted pH = 3.0 Crucially, irradiation at pH = 3.0 leads to a sustained release of DOX to ca. 80% (within 4 h), whereas cases i) and ii) lead to only ≈ 25 % release and case iii) to 50% release but precipitation of the vesicles. Thus, our initial hypothesis is confirmed: we present a proof of concept delivery system where light and pH act as inputs of an AND logic gate mechanism for the controlled release of a relevant biomedical drug (output). This may prove useful if the irradiated nanocarriers meet acidified physiological environments such as tumors sites, endosomes or lysosomes.

PH-Responsive Self-Assembled Compartments as Tuneable Model Protocellular Membrane Systems.

Prebiotically plausible single-chain amphiphiles are enticing as model protocellular compartments to study the emergence of cellular life, owing to their self-assembling properties. Here, we investigated the self-assembly behaviour of mono-N-dodecyl phosphate (DDP) and mixed systems of DDP with 1-dodecanol (DDOH) at varying pH conditions. Membranes composed of DDP showed pH-responsive vesicle formation in a wide range of pH with a low critical bilayer concentration (CBC). Further, the addition of DDOH to DDP membrane system enhanced vesicle formation and stability in alkaline pH regimes. We also compared the high-temperature behaviour of DDP and DDP:DDOH membranes with conventional fatty acid membranes. Both, DDP and DDP:DDOH mixed membranes possess packing that is similar to decanoic acid membrane. However, the micropolarity of these systems is similar to phospholipid membranes. Finally, the pH-dependent modulation of different phospholipid membranes doped with DDP was also demonstrated to engineer tuneable membranes with potential translational implications.

Amino acid induced self-assembled vesicles of choline oleate: pH responsive nano-carriers for targeted and localized delivery of doxorubicin for breast cancer

• Choline Oleate ionic liquids was synthesized and characterized. • Glutamic Acid induced micellar transition was studied. • DLS, SANS and FRET were used to measure the size of the aggregates. • Stability of the vesicles were studied over dilution, pH and temperature. • The vesicles were studied for their encapsulation and release study for the anticancer drug. Traditionally administered chemotherapeutic drugs via oral or parental routes for the treatment of breast cancer often results in serious systemic side effects. To ameliorate this, efforts have been made for the advanced drug carrier system that precisely releases the drug at the targeted site and increases the therapeutic efficiency with reduced side effects. The advanced nano-carrier system with high permeability can penetrate or absorbed through the mammary fat layers of the breast and deliver the drug judiciously. In our efforts towards addressing the proof of concept of drug delivery for the treatment of breast cancer, herein we had designed the pH-responsive vesicles with high permeability within the mammary fat layers. The vesicles were designed as a result of synergistic interaction between the surface active ionic liquid (SAIL), choline oleate ([Ch][Ol]) and glutamic acid (GA). [Ch][Ol] have the ability to permeate through the mammary fat layers whereas GA is a known α-amino acid and chemotherapeutic agent that can modulate the cancer cell development, proliferation and metastatic through regulating the cell signalling pathways. As such designed vesicles show good biocompatibility and pH-responsive behaviour and are formed through structurally transitioned from the micelles. Various state of the art analytical techniques were used to characterized the formation of the vesicles. These vesicles were explored for the encapsulation as well as sustained and targeted release of anticancer drug, doxorubicin. The proposed study would pave the way for the preparation of biocompatible and stimuli responsive drug carrier for targeted and localized delivery of anticancer drug to treat the breast cancer.

Antibacterial and Biofilm-Eradicating Activities of pH-Responsive Vesicles against Pseudomonas aeruginosa.

The formation of biofilms by a microcolony of bacteria is a significant burden on the healthcare industry due to difficulty eradicating it. In this study, pH-responsive vesicles capable of releasing apramycin (APR), a model aminoglycoside antibiotic, in response to the low pH typical of establishedPseudomonas aeruginosa biofilms resulted in improved eradication of existing biofilms in comparison to the free drug. The amphiphilic polymeric vesicle (PV) comprised of block polymer poly (ethylene glycol)-block-poly 2-(dimethylamino) ethyl methacrylate (mPEG-b-pDEAEMA) averaged 128 nm. The drug encapsulation content of APR in PV/APR was confirmed to be 28.2%, and the drug encapsulation efficiency was confirmed to be 51.2%. At pH 5.5, PV/APR released >90% APR after 24 h compared to <20% at pH 7.4. At pH 5.5, protonation of the pDEAEMA block results in a zeta potential of +23 mV compared to a neutral zeta potential of +2.2 mV at pH 7.4. Confocal microscopy, flow cytometry, and scanning electron microscopy reveal that the positively charged vesicles can compromise the integrity of the planktonic bacterial membrane in a pH-dependent manner. In addition, PV/APR is able to diffuse into mature biofilms to release APR in the acidic milieu of biofilm bacteria, and PV/APR was more efficient at eliminating preexisting biofilms compared to free APR at 128 and 256 μg/mL. This study reveals that dynamic charge density in response to pH can lead to differential levels of interactions with the biofilm and bacterial membrane. This effectively results in enhanced antibacterial and antibiofilm properties against both planktonic and difficult-to-treat biofilm bacteria at concentrations significantly lower than those of the free drug. Overall, this pH-responsive vesicle could be especially promising for treating biofilm-associated infectious diseases.

PH-Responsive Vesicle Formation by PEGylated Cholesterol Derivatives: Physicochemical Characterization, Stability, Encapsulation, and Release Study.

PEGylated vesicles are known to serve as blood-persistent drug-delivery systems (DDSs) with potential applications in intravenous drug administration. pH-responsive PEGylated vesicles are also among the most promising stimuli-responsive carriers for drug delivery and controlled release for cancer chemotherapy. Herein, we report design and synthesis of two novel pH-responsive amphiphiles by coupling a cholesterol (Chol) and poly(ethylene glycol) chain with l-cysteine amino acid through hydrolysable linkages. The objective of this work is to physicochemically characterize the nanoaggregates of the amphiphiles under different experimental conditions. We have demonstrated spontaneous vesicle formation by the amphiphiles in water using various spectroscopic, calorimetric, and microscopic techniques. The size of vesicles was observed to increase on reduction of solution pH and increase in amphiphile concentration. The vesicles were found to be sufficiently stable under physiological conditions and were shown to be able to encapsulate not only hydrophilic dyes in their aqueous core but also hydrophobic guest molecules in the bilayer membrane constituted by the Chol units. These nanosized vesicles exhibit pH-triggered release of encapsulated dye molecules in acidic pH. Thus, these spontaneously formed stable vesicles might hold potential as biocompatible DDSs in cancer chemotherapy.

Janus macromolecular brushes for synergistic cascade-amplified photodynamic therapy and enhanced chemotherapy

The aggregation-caused quenching (ACQ) effect of photosensitizers and multidrug resistance are the major obstacles in photodynamic therapy (PDT) and chemotherapy, respectively. Synergistic photo-chemotherapy is a promising cancer treatment to overcome the short boards of each single therapy. However, the fabrication of nanocarriers acting as both photosensitizers in PDT and the vehicle of drug release is a key challenge. Herein, we constructed a well-defined porphyrin-containing Janus macromolecular brush and used it as both a photosensitizer and a pH-responsive vehicle for DOX release. The Janus macromolecular brush with pH-responsive side chains and porphyrin units linked covalently in each repeat unit was synthesized by the combination of reversible addition-fragmentation chain transfer (RAFT) polymerization and click chemistry. The high grafting content of porphyrin units in the macromolecular brush improved the DOX loading capability by π–π stacking and therefore reduced the total treatment dose of DOX-loaded macromolecular brush nanoparticles (NPs). The pH-responsive side chains played triple roles in synergistic cascade-amplified PDT and enhanced chemotherapy including an executor of controlled drug release, a ligand with a mitochondria-targeting feature, and a barrier to reduce the ACQ effect of porphyrin units. In vitro and in vivo studies confirmed that the DOX-loaded macromolecular brush NPs exhibited high phototoxicity and significant tumor inhibition efficacy. Statement of SignificanceSynergistic photodynamic therapy (PDT) and chemotherapy has emerged as a promising cancer treatment to overcome the challenges of a single modality. Herein, we constructed new pH-responsive vesicles using porphyrin-containing Janus macromolecular brushes as theranostic nanocarriers to encapsulate high-loading doxorubicin (DOX) for synergistic cascade-amplified PDT and enhanced chemotherapy. The high grafting content of porphyrin units in Janus macromolecular brushes improved DOX loading capability by π–π stacking for enhanced chemotherapy. Moreover, pH-responsive side chains subsequently enhanced the suppression of the aggregation-caused quenching (ACQ) effect of porphyrins for cascade-amplified PDT. In vitro and in vivo studies confirmed that DOX-loaded macromolecular brush nanoparticles exhibited high phototoxicity and significant tumor inhibition efficacy.

Catalyst-Free One-Step Preparation of Self-Crosslinked pH-Responsive Vesicles.

The fabrication of block copolymer (BCP) vesicles with controlled membrane permeability and promising stability remains a considerable challenge. Herein, a new type of pH-responsive and self-crosslinked vesicle based on a hydrolytically hindered urea bond is reported. This kind of vesicle is formed by the self-assembly of a pH-responsive and hydrolytically self-crosslinkable copolymer poly(ethylene glycol)-block-poly[2-(3-(tert-butyl)-3-ethylureido)ethyl methacrylate-co-2-(diethylamino)ethyl methacrylate] (PEG-b-P(TBEU-co-DEA)). The BCP can be easily synthesized by reversible addition-fragmentation chain transfer (RAFT) polymerization of 2-(3-(tert-butyl)-3-ethylureido)ethyl methacrylate (TBEU) and 2-(diethylamino)ethyl methacrylate (DEA) using PEG-based macro-chain transfer agent. The copolymer could self-assemble into stable vesicles by the hydrophobic interaction and in situ cross-linking between amines and isocyanates after the hydrolysis of the hindered urea bonds without any catalyst. Dynamic light scattering (DLS) studies show that the vesicles exhibit enhanced stability against the dilution of organic solvent, and the size can be adjusted through the change of pH values. Moreover, the alkaline phosphatase-loaded vesicles can act as nano-reactor and enable free diffusion of small molecules into the vesicles, followed by the significantly improved fluorescence intensity of phosphate-caged fluorescein. This self-crosslinking and pH-sensitive vesicles may serve as a smart platform in controlled drug delivery and molecular reactor.

PH responsive vesicles with tunable size formed by single-tailed surfactants with a dendritic headgroup

Vesicles of variable sizes are obtained by changing pH with single-tailed dendritic surfactants.

PH-Responsive Vesicles with Tunable Membrane Permeability and Hydrodynamic Diameters from a Cross-Linkable Amphiphilic Block Copolymer

A series of cross-linked, pH responsive vesicles with tunable diameters were prepared by self-assembly of poly (ethyleneoxide)-b-poly [2-(diethylamino) ethyl methacrylate-co-glycidyl methacrylate, ...

PH-responsive vesicles from supra-amphiphiles based on dynamic imine bond

Abstract pH-responsive vesicles have been prepared from the mixture of 1-(10-(4-formyl-phenoxy) decyl)-pyridinium (FDP) or 1-methyl-3-(10-(4-formyl-phenoxy) decyl) imidazolium (FDI) with 1-octyl amine at a molar ratio of 1:1 in alkaline aqueous solutions. Their structures and morphologies were characterized by transmission electron microscopy with sample preparations by negative staining or freeze-fracture. Basically, the spherical unilamellar vesicles were formed with the mean diameters respectively of about 71 and 107 nm for two supra-amphiphile systems, which were based on the formation of dynamic imine bonds between amine and FDP (i-FDP) or FDI (i-FDI). The loading and release studies of a fluorophore, Nile red (NR), demonstrated the capabilities of these vesicles to encapsulate NR and a rapid release at low pH environment. The vesicles from i-FDP behaved relatively more sensitive to acidic environment than those from i-FDI. The dynamic transitions of supra-amphiphiles were characterized by nuclear magnetic resonance or fluorescence emission spectroscopy measurements. A lower critical micelle concentration and a higher aldehyde conversion efficiency of i-FDI than those of i-FDP were observed. The characteristics of facile formation, fast dissociation, and switchable morphologies of studied systems should be referable for designing novel smart supra-amphiphile systems with potential applications as controlled drug delivery systems.

PH-Responsive Vesicles with Tunable MembranePermeabilityand Hydrodynamic Diameters from Self-Catalyzed Cross-Linkable Amphiphilic BlockCopolymer

A series of cross-linked pH responsive vesicles with tunable diameters were prepared by self-assembly of poly (ethyleneoxide)-b-poly [2-(diethylamino) ethyl methacrylate-co-glycidyl methacrylate] [PEO-b-P(DEA-co-GMA)] inTHF/water mixture. The copolymers were synthesized by random copolymerization of DEA and GMA initiated by PEO-Br.With epoxy group and tertiary amine in its structure, self-catalyzed cross-linking was triggered easily by mild heating. The vesicles showed pH responsiveness as characterized by DLS and tunable permeability can be understood accordingly. As the pH of the solvent changed from 9.8 to 7.4, the diameters of the vesicles increased. It was observed that the diameters at pH = 6.0 were comparable to that at pH = 7.4. This alteration may have happened because of the protonation of PDEA segment.This particular property has great potential and applications since most drug loading and controlled release are conducted within this pH range.

Multifunctional Homopolymer Vesicles for Facile Immobilization of Gold Nanoparticles and Effective Water Remediation

Homopolymers have been considered as a nonideal building block for creating well-defined nanostructures due to their fuzzy boundary between hydrophobic and hydrophilic moieties. However, this unique fuzzy boundary may provide some opportunities for fabricating functional nanomaterials. Presented in this paper is a pH-responsive multifunctional homopolymer vesicle based on poly[2-hydroxy-3-(naphthalen-1-ylamino)propyl methacrylate] (PHNA). This vesicle is confirmed to be an excellent supporter for gold nanoparticles (AuNPs) to facilitate the reduction reaction of 4-nitrophenol (4-NP). The pH-responsive vesicle membrane favors the effective embedding and full immobilization of AuNPs because it is kinetically frozen under neutral and basic environments, preventing AuNPs from aggregation. Meanwhile, there is a synergistic effect between the AuNPs and the supporter (PHNA vesicle). Due to the π-π interaction between the naphthalene pendants in every repeat unit of PHNA and the extra aromatic compounds, a substrate-rich (high concentration of 4-NP) microenvironment can be created around AuNPs, which can dramatically accelerate the AuNPs-catalyzed reactions. In addition, we proposed a method for more accurately determining the membrane thickness of rigid polymer vesicles from TEM images based on "stack-up" vesicles, which may overturn the measuring method commonly used by far. Moreover, proof-of-concept studies showed that those homopolymer vesicles may be used as a powerful adsorbent for effective water remediation to remove trace carcinogenic organic pollutants such as polycyclic aromatic hydrocarbons to below parts per billion (ppb) level at a very fast rate based on the π-π interaction between the naphthalene pendants in PHNA vesicle and polycyclic aromatic hydrocarbons. Overall, this multifunctional homopolymer vesicle provides an alternative insight on preparing effective recyclable AuNPs-decorated nanoreactor and powerful water remediation adsorbent.

Peptide-based lipid mimetics with tunable core properties via thiol–alkyne chemistry

Thiol–alkyne chemistry is utilized to produce peptide-based A2B star polymers. This topography resembles a phospholipid where the polypeptide B block represents a functional, polar head group and the A blocks represent lipophilic units. We utilize a convergent, modular approach to produce lipid mimetics through conjugation of poly(L-glutamic acid) with three different lipophilic moieties (octadecane, cholesterol and polyhedral oligomeric silsesquioxane). All samples are shown to self-assemble in aqueous solution into pH-responsive vesicles. The self-assembly and pH-responsiveness were characterized using circular dichroism spectroscopy and light scattering. Detailed light scattering experiments determined that the aggregation number of the vesicles remains nearly constant as a function of pH, suggesting that the pH-responsiveness is a result of both the helix–coil transition as well as a change in chain packing at the vesicle interface.

PH-Responsive Vesicles Based on a Hydrolytically Self-Cross-Linkable Copolymer

A new type of shape-persistent, pH-responsive vesicle was prepared by the self-assembly of a novel poly(ethylene oxide)-block-poly[2-(diethylamino)ethyl methacrylate-stat-3-(trimethoxysilyl)propyl methacrylate], PEO-b-P(DEA-stat-TMSPMA), copolymer. Vesicles were formed spontaneously in aqueous THF solution, with the hydrophilic PEO chains forming the corona and the pH-sensitive P(DEA-stat-TMSPMA) blocks being located in the membrane walls. Hydrolytic cross-linking within the hydrophobic membrane walls fixed the vesicle morphology. The resulting colloidally stable vesicles were characterized by 1H NMR, transmission electron microscopy (TEM), dynamic laser light scattering (DLS), and stopped-flow fluorescence experiments. The latter technique indicated that the permeability of the vesicle walls was sensitive to the pH of the aqueous solution, as expected. Gold-decorated vesicles were obtained by in situ reduction of AuCl4- anions to produce gold nanoparticles within the vesicle walls. (Yellow, hydrophilic PEO; green, pH-responsive DEA residues; blue, hydrolytically self-cross-linkable TMSPMA residues.)

On the Characterization of pH-sensitive Liposome/Polymer Complexes

A randomly alkylated copolymer of N-isopropylacrylamide, methacrylic acid and N-vinyl-2-pyrrolidone was characterized with regard to its pH- and temperature-triggered conformational change. It was then complexed to liposomes to produce pH-responsive vesicles. Light scattering and differential scanning calorimetry experiments performed at neutral pH revealed that the polymer underwent coil-to-globule phase transition over a wide range of temperatures. At 37 degrees C and pH 7.4, although the polymer was water-soluble, Fourier transform infrared spectroscopy analysis showed that it was partly dehydrated. At acidic pH, the decrease in the lower critical solution temperature was accompanied by an increase in cooperativity degree of the phase transition. Complexation of copolymer to liposomes did not substantially influence its phase transition. The liposome/copolymer complexes were stable at neutral pH but rapidly released their contents under acidic conditions. The copolymer slightly increased liposome circulation time following intravenous administration to rats. The addition of poly(ethylene glycol) to the formulation had a detrimental effect on pH-sensitivity but enhanced substantially the circulation time.