Polyelectrolyte Complex Micelles Research Articles

Enzyme-Triggered Polyelectrolyte Complex for Responsive Delivery of α-Helical Polypeptides to Optimize Antibacterial Therapy.

Responsive nanomaterials hold significant promise in the treatment of bacterial infections by recognizing internal or external stimuli to achieve stimuli-responsive behavior. In this study, we present an enzyme-responsive polyelectrolyte complex micelles (PTPMN) with α-helical cationic polypeptide as a coacervate-core for the treatment of Escherichia coli (E. coli) infection. The complex was constructed through electrostatic interaction between cationic poly(glutamic acid) derivatives and phosphorylation-modified poly(ethylene glycol)-b-poly(tyrosine) (PEG-b-PPTyr) by directly dissolving them in aqueous solution. The cationic polypeptide adopted α-helical structure and demonstrated excellent broad-spectrum antibacterial activity against both Gram-negative and Gram-positive bacteria, with a minimum inhibitory concentration (MIC) as low as 12.5 μg mL-1 against E. coli. By complexing with anionic PEG-b-PPTyr, the obtained complex formed β-sheet structures and exhibited good biocompatibility and low hemolysis. When incubated in a bacterial environment, the complex cleaved its phosphate groups triggered by phosphatases secreted by bacteria, exposing the highly α-helical conformation and restoring its effective bactericidal ability. In vivo experiments confirmed accelerated healing in E. coli-infected wounds.

Preparation and stability of pegylated poly(S-alkyl-L-homocysteine) coacervate core micelles in aqueous media.

We report development and preparation of synthetic polypeptide based, coacervate core polyelectrolyte complex micelles, PCMs, in aqueous media, which were characterized and evaluated for the encapsulation and in vitro release of a model single-stranded RNA, polyadenylic acid, poly(A). Cationic, α-helical polypeptides pegylated at their N-termini, PEG113-b-5bn and PEG113-b-5cn, were designed to form coacervate core PCMs upon mixing with multivalent anions in aqueous media. Sodium tripolyphosphate (TPP) and poly(A) were used as model multivalent anions that allowed optimization of polypeptide composition and chain length for formation of stable, nanoscale PCMs. PEG113-b-5c27 was selected for preparation of PCMs that were characterized under different environmental conditions using dynamic light scattering, atomic force microscopy and cryoelectron microscopy. The PCMs were found to efficiently encapsulate poly(A), were stable at physiologically relevant pH and solution ionic strength, and were able to release poly(A) in the presence of excess polyvalent anions. These PCMs were found to be a promising model system for further development of polypeptide based therapeutic delivery vehicles.

Effect of Charged Block Length Mismatch on Double Diblock Polyelectrolyte Complex Micelle Cores.

Polyelectrolyte complex micelles are hydrophilic nanoparticles that self-assemble in aqueous environments due to associative microphase separation between oppositely charged blocky polyelectrolytes. In this work, we employ a suite of physical characterization tools to examine the effect of charged block length mismatch on the equilibrium structure of double diblock polyelectrolyte complex micelles (D-PCMs) by mixing a diverse library of peptide and synthetic charged-neutral block polyelectrolytes with a wide range of charged block lengths (25-200 units) and chemistries. Early work on D-PCMs suggested that this class of micelles can only be formed from blocky polyelectrolytes with identical charged block lengths, a phenomenon referred to as chain length recognition. Here, we use salt annealing to create PCMs at equilibrium, which shows that chain length recognition, a longstanding hurdle to repeatable self-assembly from mismatched polyelectrolytes, can be overcome. Interestingly, D-PCM structure-property relationships display a range of values that vary systematically with the charged block lengths and chemical identity of constituent polyelectrolyte pairings and cannot be described by generalizable scaling laws. We discuss the interdependent growth behavior of the radius, ionic pair aggregation number, and density in the micelle core for three chemically distinct diblock pairings and suggest a potential physical mechanism that leads to this unique behavior. By comparing the results of these D-PCMs to the scaling laws recently developed for single diblock polyelectrolyte complex micelles (S-PCMs: diblock + homopolymer), we observe that D-PCM design schemes reduce the size and aggregation number and restrict their growth to a function of charged block length relative to S-PCMs. Understanding these favorable attributes enables more predictive use of a wider array of charged molecular building blocks to anticipate and control macroscopic properties of micelles spanning countless storage and delivery applications.

Abstract 139: Precision Nanomedicine Targeting Novel Endothelial Mechano-sensing Mechanisms To Treat Vascular Diseases

Endothelial mechano-transduction mechanisms are instrumental to vascular health and disease but targeting disease-causing mechano-sensing pathways remains extremely challenging. Atherosclerosis preferentially develops at arterial curvatures and bifurcations where disturbed blood flow activates endothelium, leading to peripheral artery disease, carotid artery disease and ischemic stroke. Nevertheless; current atherosclerosis therapies mainly target systematic risk factors but not the vasculature per se . This underscores the significance and unique opportunity to identify and target novel mechanosensitive mechanisms in activated endothelium subjected to disturbed flow. Our objectives are to (1) delineate novel endothelial mechano-sensing mechanisms and (2) devise innovative precision nanomedicine approaches targeting these disease-causing mechano-sensitive pathways. Using multiple new transgenic mouse lines, our studies demonstrated that disturbed flow (DF) significantly increases the endothelial expression of TXNDC5 (thioredoxin domain containing 5) and microRNA-92a (miR-92a), two atherogenic molecules driving atherogenesis. Mechanistically, DF-induced TXNDC5 increases proteasome-mediated degradation of heat shock factor 1 and consequently promotes eNOS/NOS3 protein degradation. Meanwhile, DF induces endothelial miR-92a to suppress key anti-inflammatory molecules such as KLF2, KLF4, and PLPP3. We successfully formulated nanoparticles to deliver TXNDC5-targeting CRISPR/Cas9 plasmids driven by an endothelium-specific promoter (CDH5), which effectively deleted endothelial TXNDC5 expression and reduced atherosclerosis in Apoe -/- mice. Moreover, VCAM1-tageting polyelectrolyte complex micelles were engineered to selectively deliver miR-92a inhibitors to inflamed endothelium, which markedly decreased arterial stenosis and atherosclerosis in Apoe -/- mice. These results elucidate novel endothelial mechano-sensing mechanisms and provide a proof of concept of innovative targeted nanomedicine approaches, addressing an unmet medical need in vascular therapies.

Targeted polyelectrolyte complex micelles treat vascular complications in vivo

Vascular disease is a leading cause of morbidity and mortality in the United States and globally. Pathological vascular remodeling, such as atherosclerosis and stenosis, largely develop at arterial sites of curvature, branching, and bifurcation, where disturbed blood flow activates vascular endothelium. Current pharmacological treatments of vascular complications principally target systemic risk factors. Improvements are needed. We previously devised a targeted polyelectrolyte complex micelle to deliver therapeutic nucleotides to inflamed endothelium invitro by displaying the peptide VHPKQHR targeting vascular cell adhesion molecule 1 (VCAM-1) on the periphery of the micelle. This paper explores whether this targeted nanomedicine strategy effectively treats vascular complications invivo. Disturbed flow-induced microRNA-92a (miR-92a) has been linked to endothelial dysfunction. We have engineered a transgenic line (miR-92aEC-TG /Apoe-/- ) establishing that selective miR-92a overexpression in adult vascular endothelium causally promotes atherosclerosis in Apoe-/- mice. We tested the therapeutic effectiveness of the VCAM-1-targeting polyelectrolyte complex micelles to deliver miR-92a inhibitors and treat pathological vascular remodeling invivo. VCAM-1-targeting micelles preferentially delivered miRNA inhibitors to inflamed endothelial cells invitro and invivo. The therapeutic effectiveness of anti-miR-92a therapy in treating atherosclerosis and stenosis in Apoe-/- mice is markedly enhanced by the VCAM-1-targeting polyelectrolyte complex micelles. These results demonstrate a proof of concept to devise polyelectrolyte complex micelle-based targeted nanomedicine approaches treating vascular complications invivo.

Advances in the Structural Design of Polyelectrolyte Complex Micelles.

Polyelectrolyte complex micelles (PCMs) are a unique class of self-assembled nanoparticles that form with a core of associated polycations and polyanions, microphase-separated from neutral, hydrophilic coronas in aqueous solution. The hydrated nature and structural and chemical versatility make PCMs an attractive system for delivery and for fundamental polymer physics research. By leveraging block copolymer design with controlled self-assembly, fundamental structure–property relationships can be established to tune the size, morphology, and stability of PCMs precisely in pursuit of tailored nanocarriers, ultimately offering storage, protection, transport, and delivery of active ingredients. This perspective highlights recent advances in predictive PCM design, focusing on (i) structure–property relationships to target specific nanoscale dimensions and shapes and (ii) characterization of PCM dynamics primarily using time-resolved scattering techniques. We present several vignettes from these two emerging areas of PCM research and discuss key opportunities for PCM design to advance precision medicine.

Micellization through complexation of oppositely charged diblock copolymers: Effects of composition, polymer architecture, salt of different valency, and thermoresponsive block

AbstractThe structural and electrical characteristics of polyelectrolyte complex micelles (PCMs) formed by mixing of oppositely charged double hydrophilic copolymers are studied by means of molecular dynamics simulations. In mixtures of linear diblock copolymers we found that the preferential aggregation number Np of PCMs is a universal function of the ratio γ± of the total positive to total negative charges of the mixture. The addition of divalent salts ions induces a secondary micellization. In mixtures of copolymers bearing a common neutral thermoresponsive block, micelles with contracted corona consisting of thermoresponsive blocks and complex polyelectrolyte core are formed at low salt concentration and temperature far away the biphasic regime. At high salt concentration and temperature in the biphasic regime, reversed micelles are obtained. In equimolar mixtures of linear copolymers with miktoarm stars we found that Np of PCMs decreases as the number of charged branches of miktoarm copolymer increases. The shape of micelles progressively changes from spherical to worm‐like with the increase of number of branches of miktoarm copolymers. Our findings are in full agreement with existing experimental and theoretical predictions and provides new and additional insights.

Spatiotemporal Formation and Growth Kinetics of Polyelectrolyte Complex Micelles with Millisecond Resolution.

We have directly observed the in situ self-assembly kinetics of polyelectrolyte complex (PEC) micelles by synchrotron time-resolved small-angle X-ray scattering, equipped with a stopped-flow device that provides millisecond temporal resolution. A synthesized neutral-charged diblock polycation and homopolyanion that we have previously investigated as a model charge-matched, core-shell micelle system were selected for this work. The initial micellization of the oppositely charged polyelectrolytes was completed within the dead time of mixing of 100 ms, followed by micelle growth and equilibration up to several seconds. By combining the structural evolution of the radius of gyration (Rg) with complementary molecular dynamics simulations, we show how the self-assemblies evolve incrementally in size over time through a two-step kinetic process: first, oppositely charged polyelectrolyte chains pair to form nascent aggregates that immediately assemble into spherical micelles, and second, these PEC micelles grow into larger micellar entities. This work has determined one possible kinetic pathway for the initial formation of PEC micelles, which provides useful physical insights for increasing fundamental understanding self-assembly dynamics, driven by polyelectrolyte complexation that occurs on ultrafast time scales.

Comparing Zwitterionic and PEG Exteriors of Polyelectrolyte Complex Micelles.

A series of model polyelectrolyte complex micelles (PCMs) was prepared to investigate the consequences of neutral and zwitterionic chemistries and distinct charged cores on the size and stability of nanocarriers. Using aqueous reversible addition-fragmentation chain transfer (RAFT) polymerization, we synthesized a well-defined diblock polyelectrolyte system, poly(2-methacryloyloxyethyl phosphorylcholine methacrylate)-block-poly((vinylbenzyl) trimethylammonium) (PMPC-PVBTMA), at various neutral and charged block lengths to compare directly against PCM structure–property relationships centered on poly(ethylene glycol)-block-poly((vinylbenzyl) trimethylammonium) (PEG-PVBTMA) and poly(ethylene glycol)-block-poly(l-lysine) (PEG-PLK). After complexation with a common polyanion, poly(sodium acrylate), the resulting PCMs were characterized by dynamic light scattering (DLS) and small angle X-ray scattering (SAXS). We observed uniform assemblies of spherical micelles with a diameter ~1.5–2× larger when PMPC-PVBTMA was used compared to PEG-PLK and PEG-PVBTMA via SAXS and DLS. In addition, PEG-PLK PCMs proved most resistant to dissolution by both monovalent and divalent salt, followed by PEG-PVBTMA then PMPC-PVBTMA. All micelle systems were serum stable in 100% fetal bovine serum over the course of 8 h by time-resolved DLS, demonstrating minimal interactions with serum proteins and potential as in vivo drug delivery vehicles. This thorough study of the synthesis, assembly, and characterization of zwitterionic polymers in PCMs advances the design space for charge-driven micelle assemblies.

Assembly and Characterization of Polyelectrolyte Complex Micelles.

Polyelectrolyte complex micelles (PCMs), core-shell nanoparticles formed by self-assembly of charged polymers in aqueous solution, provide a powerful platform for exploring the physics of polyelectrolyte interactions and also offer a promising solution to the pressing problem of delivering therapeutic oligonucleotides in vivo. Developing predictive structure-property relationships for PCMs has proven difficult, in part due to the presence of strong kinetic traps during nanoparticle self-assembly. This article discusses criteria for choosing polymers for PCM construction and provides protocols based on salt annealing that enable assembly of repeatable, low-polydispersity nanoparticles. We also discuss PCM characterization using light scattering, small-angle X-ray scattering, and electron microscopy.

Assembly and Characterization of Polyelectrolyte Complex Micelles

Assembly and Characterization of Polyelectrolyte Complex Micelles

Mechanism of Dissociation Kinetics in Polyelectrolyte Complex Micelles

Polyelectrolyte-based nanoscale self-assemblies, such as micelles, possess diverse desirable attributes such as the capability for sequestering and protecting biomacromolecules against inhospitable environments, responsiveness to external stimuli, and tunability of physical behavior. However, little is known on the mechanisms of dissociation when micelles encounter and respond to environmental changes. Using salt-jump, time-dependent light scattering, the pathway of dissociation is observed in polyelectrolyte complex micelles that have complex cores and neutral coronas. The micelle dissociation kinetics appear to be a three-stage process, in good agreement with the scattering data. Using kinetic models of amphiphilic block copolymer micelles in polyelectrolyte complexation-driven micelles, we derive an analytical expression for dissociation relaxation rates as a function of solvent temperature, salt concentration, and the length of the charged polymer blocks. The theoretical predictions are compatible with the experimental data from light scattering experiments. This study experimentally demonstrates the relaxation kinetics of polyelectrolyte complex micelle dissociation and illustrates the underlying mechanism governing the dissociation kinetics. It is anticipated that these findings can be generalized to other electrostatic interaction-driven self-assemblies to better understand the relationship among the kinetics of dissociation, constituent polymer properties, and environmental parameters.

Response of metal-coordination-based polyelectrolyte complex micelles to added ligands and metals.

Polyelectrolyte complex based micelles have attracted significant attention due to their potential regarding bio-applications. Although the morphology and functions have been studied extensively, dynamic properties, particularly component exchange with other surrounding molecules, have remained elusive to date. Here, we show how micelles based on metal-ligand coordination complex coacervate-core micelles (M-C3Ms) respond to addition of extra ligand and metal ions. The micelles are prepared from a polycationic-neutral diblock copolymer and an anionic coordination polyelectrolyte, which is obtained by coordination between metal ions (lanthanides Ln3+ and Zn2+) and a bis-ligand (LEO) containing two dipicolinic acid (DPA) groups connected by a tetra-ethylene oxide spacer (4EO). Our findings show that the bis-ligand LEO is essential for the growth of coordination polymers and consequently the formation of micelles, leading to equilibrium structures with the same micellar composition and structure independent of the order of mixing. In other words, adding single DPA has no effect on the formed M-C3Ms. As for metal exchange, we find that added Zn2+ can replace some of the Ln3+ from Ln-C3Ms, leading to a hybrid coordination structure with both Ln3+ and Zn2+. We find that component exchange occurs in these coordination polyelectrolyte micelles, but it is more favorable in the direction of replacing the weak binding components with strong ones. Hence, the designed M-C3Ms based on the strong binding components, such as Ln-C3Ms, shall be relatively stable in biological surroundings, paving the way for the application of such particles as bio-imaging probes.

Protein-Polyelectrolyte Complexes and Micellar Assemblies.

In this review, we highlight the recent progress in our understanding of the structure, properties and applications of protein–polyelectrolyte complexes in both bulk and micellar assemblies. Protein–polyelectrolyte complexes form the basis of the genetic code, enable facile protein purification, and have emerged as enterprising candidates for simulating protocellular environments and as efficient enzymatic bioreactors. Such complexes undergo self-assembly in bulk due to a combined influence of electrostatic interactions and entropy gains from counterion release. Diversifying the self-assembly by incorporation of block polyelectrolytes has further enabled fabrication of protein–polyelectrolyte complex micelles that are multifunctional carriers for therapeutic targeted delivery of proteins such as enzymes and antibodies. We discuss research efforts focused on the structure, properties and applications of protein–polyelectrolyte complexes in both bulk and micellar assemblies, along with the influences of amphoteric nature of proteins accompanying patchy distribution of charges leading to unique phenomena including multiple complexation windows and complexation on the wrong side of the isoelectric point.

Interparticle Interactions in Dilute Solutions of Polyelectrolyte Complex Micelles.

The application of dilute solutions of polyelectrolyte complex (PEC) micelles for delivering therapeutic nucleic acids into disease sites has gained momentum. This Letter reports a detailed characterization of PEC micelles in dilute solutions including their internal structures and the determination of the interparticle interactions. The polymer concentration ranges from 0.1 to 0.5 wt %, a regime where micelle-micelle interactions are infrequent. We employ synchrotron small-angle X-ray scattering (SAXS) to simultaneously probe the morphology, internal structure, and radius of gyration (Rg) of the self-assemblies formed by charged diblock polyelectrolytes and homopolyelectrolytes. The emerging appearance of the structure factor in SAXS profiles with the increasing polymer concentration demonstrates the presence of the repulsive intermicellar correlations, which is further confirmed by the differences between the "reciprocal Rg" estimated by Guinier approximation and the "real space Rg" determined by pair distribution functions. We find that the soft corona chains tethered on the surface of phase-separated complex domains are compressed when micelles come closeto thepointwhere a hard-sphere interaction takes over. These findings contribute to the fundamental understanding of the structure and space-filling constraints in the complexation-driven self-assemblies and advance the rational design of cationic polymer-based nonviral gene delivery vectors.

Polyelectrolyte Complexation of Oligonucleotides by Charged Hydrophobic-Neutral Hydrophilic Block Copolymers.

Polyelectrolyte complex micelles (PCMs, core-shell nanoparticles formed by complexation of a polyelectrolyte with a polyelectrolyte-hydrophilic neutral block copolymer) offer a solution to the critical problem of delivering therapeutic nucleic acids, Despite this, few systematic studies have been conducted on how parameters such as polycation charge density, hydrophobicity, and choice of charged group influence PCM properties, despite evidence that these strongly influence the complexation behavior of polyelectrolyte homopolymers. In this article, we report a comparison of oligonucleotide PCMs and polyelectrolyte complexes formed by poly(lysine) and poly((vinylbenzyl) trimethylammonium) (PVBTMA), a styrenic polycation with comparatively higher charge density, increased hydrophobicity, and a permanent positive charge. All of these differences have been individually suggested to provide increased complex stability, but we find that PVBTMA in fact complexes oligonucleotides more weakly than does poly(lysine), as measured by stability versus added salt. Using small angle X-ray scattering and electron microscopy, we find that PCMs formed from both cationic blocks exhibit very similar structure-property relationships, with PCM radius determined by the cationic block size and shape controlled by the hybridization state of the oligonucleotides. These observations narrow the design space for optimizing therapeutic PCMs and provide new insights into the rich polymer physics of polyelectrolyte self-assembly.

Structural transitions and encapsulation selectivity of thermoresponsive polyelectrolyte complex micelles.

Polyelectrolyte complex (PEC) micelles are formed by mixing a block copolymer composed of a neutral block and a charged block, with an oppositely charged polymer. The micelles formed have a PEC core, capable of encapsulating charged molecules like nucleic acids or proteins, while the neutral block(s) forms the corona that offers protection that can prevent cargo from being degraded under physiological conditions. This work explores using a thermosensitive polymer, poly(N-isopropyl acrylamide) (pNIPAM), as the primary corona-forming block and how that can be leveraged in the context of drug delivery. pNIPAM has a lower critical solubility temperature (LCST), above which a hydrophilic to hydrophobic transition occurs. We are characterizing micelles formed using a diblock copolymer of pNIPAM-b-poly(acrylic acid) with (1) poly(lysine) and (2) poly(ethylene glycol)-b-poly(lysine) using dynamic light scattering, small angle X-ray scattering, absorbance and fluorescence spectroscopy, and transmission electron microscopy. Our results indicate that mixing pNIPAM-b-poly(acrylic acid) and poly(lysine) creates worm-like micelles, while mixing the same diblock with poly(ethylene glycol)-b-poly(lysine) forms spherical micelles. At temperatures above 35 °C, the transition temperature of pNIPAM-b-poly(acrylic acid), the worm-like micelles lose their structure, while the spherical micelles retain their structure, but both aggregate into larger assemblies. Lastly, we evaluate the ability of these micelles to encapsulate and release model charged therapeutics, using a cationic monoion, methylene blue and a cationic polyion, polylysine conjugated rhodamine. We find that methylene blue is not encapsulated by the micelle, and instead exhibits counterion-like behaviour upon polyelectrolyte complex formation. Conversely, fluorescence measurements of rhodamine-labelled polylysine show fluorescence quenching indicating that the polyion is encapsulated. By mixing quenched and unlabelled micelles, we show that both systems are immune to molecular exchange with their environment and instead the cargo remains trapped in the quenched micellar cores. Measurement of the micelle fluorescence above the LCST, decreases indicating no substantial release for either system. However, the increase in micelle quenching above the LCST and its persistence after cooling may offer an additional protective environment for cargo that can be triggered by temperature.

Structure-Property Relationships of Oligonucleotide Polyelectrolyte Complex Micelles.

Polyelectrolyte complex micelles (PCMs), nanoparticles formed by electrostatic self-assembly of charged polymers with charged-neutral hydrophilic block copolymers, offer a potential solution to the challenging problem of delivering therapeutic nucleic acids into cells and organisms. Promising results have been reported in vitro and in animal models but basic structure-property relationships are largely lacking, and some reports have suggested that double-stranded nucleic acids cannot form PCMs due to their high bending rigidity. This letter reports a study of PCMs formed by DNA oligonucleotides of varied length and hybridization state and poly(l)lysine-poly(ethylene glycol) block copolymers with varying block lengths. We employ a multimodal characterization strategy combining small-angle X-ray scattering (SAXS), multiangle light scattering (MALS), and cryo-electron microscopy (cryo-TEM) to simultaneously probe the morphology and internal structure of the micelles. Over a wide range of parameters, we find that nanoparticle shape is controlled primarily by the hybridization state of the oligonucleotides with single-stranded oligonucleotides forming spheroidal micelles and double-stranded oligonucleotides forming wormlike micelles. The length of the charged block controls the radius of the nanoparticle, while oligonucleotide length appears to have little impact on either size or shape. At smaller length scales, we observe parallel packing of DNA helices inside the double-stranded nanoparticles, consistent with results from condensed genomic DNA. We also describe salt- and thermal-annealing protocols for preparing PCMs with high repeatability and low polydispersity. Together, these results provide a capability to rationally design PCMs with desired sizes and shapes that should greatly assist development of this promising delivery technology.

Synthesis and Self-Assembly of the pH-Responsive Anionic Copolymers for Enhanced Doxorubicin-Loading Capacity.

Polyelectrolyte complex micelles self-assembled from an ionic polymer and oppositely charged small molecules are a promising drug delivery system. In this study, the anionic block copolymers composed of poly(ethylene glycol), poly(ε-caprolactone), and carboxyl modified poly(ε-caprolactone), COOH-PCEC, were designed to encapsulate doxorubicin (DOX) via electrostatic and hydrophobic interactions to form spherical micelles with a particle size of 90-140 nm. The higher payload capacity of these micelles than noncharged micelles of PCL-poly(ethylene glycol)-PCL (PCEC) was achieved, and it was strongly dependent on the composition of the micelles. In vitro drug release studies showed that the release of DOX from the micelles was faster at pH 5.5 than at pH 7.4, which was mainly due to the protonation of carboxyl groups and the solubility of DOX. Studies of intracellular uptake demonstrated that the DOX-loaded micelles could be internalized effectively by HeLa cells. In vitro cytotoxicity revealed that the blank COOH-PCEC micelles had a low cytotoxicity against both L929 and HeLa cells. However, the DOX-loaded micelles inhibited the growth of HeLa cells remarkably, demonstrating their potential for use as an efficient carrier for the delivery of DOX.

Kinetics of Morphological Transition between Cylindrical and Spherical Micelles in a Mixture of Anionic–Neutral and Cationic–Neutral Block Copolymers Studied by Time-Resolved SAXS and USAXS

This study is concerned with the morphological transition kinetics of polyelectrolyte complex micelles formed from an anionic–neutral block copolymer and a cationic–neutral block copolymer in aqueous NaCl solution. The transformation was induced by changing the mixing ratio of the anionic and cationic monomer units in the copolymers. The kinetics of the morphological transition was directly tracked by time-resolved (ultra) small-angle X-ray scattering coupled with rapid mixing of the block copolymer components by a stopped flow apparatus. The transformation from cylindrical to spherical micelles upon changing the mixing ratio of the copolymers was reversible, and the process occurred via the random scission of the cylindrical micelles along their contours. The reverse transition from spherical to cylindrical micelles was found to be a slow process with high activation energy.