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Due to the escalating threat of the pathogens' capability of quick adaptation to antibiotics, finding new alternatives is crucial. Although antimicrobial peptides (AMPs) are highly potent and effective, their therapeutic use is limited' as they are prone to enzymatic degradation, are cytotoxic and have low retention. To overcome these challenges, we investigate the complexation of the cationic AMP colistin with diblock copolymers poly(ethylene oxide)- b -poly(methacrylic acid) (PEO- b -PMAA) forming colistin-complex coacervate core micelles (colistin-C3Ms). We present long-term stable kinetically controlled colistin-C3Ms that can be prepared from several block lengths of PEO- b -PMAA polymers, where the polymerisation degree governs the overall micellar size. To achieve precise control over size and polydispersity, which are crucial for drug delivery applications, we investigate the hybridisation of PEO- b -PMAA polymers with varying chain lengths or PMAA homopolymers in ternary complex coacervation systems with colistin. This results in size-tunable colistin-C3Ms, ranging, depending on the mixing ratios, from micellar sizes of 26 nm to 100 nm. With size tunability at rather narrow size distributions and high stability, ternary colistin-C3Ms offer potential advancements in C3M drug delivery, paving the way for more effective and targeted treatments for bacterial infections in precision medicine.

Colistin methanesulfonate (CMS) is the less-toxic prodrug of highly nephrotoxic colistin. To develop and understand highly necessary new antibiotic formulations, the hydrolysis of CMS to colistin must be better understood. Herein, with the addition of poly(ethylene oxide)-b-poly(methacrylic acid) (PEO-b-PMAA) to CMS, we show that we can follow the hydrolysis kinetics, employing small-angle X-ray scattering (SAXS) through complex coacervation. During this hydrolysis, hydroxy methanesulfonate (HMS) groups from CMS are cleaved, while the newly formed cationic amino groups complex with the anionic charge from the PMAA block. As the hydrolysis of HMS groups is slow, we can follow the complex coacervation process by the gradual formation of complex micelles containing activated antibiotics. Combining mass spectrometry (MS) with SAXS, we quantify the hydrolysis as a function of pH. Upon modeling the kinetic pathways, we found that complexation only happens after complete hydrolysis into colistin and that the process is accelerated under acidic conditions. At pH = 5.0, effective charge switching was identified as the slowest step in the CMS conversion, constituting the rate-limiting step in colistin formation.

Understanding the pathways of solubilization of lipid membranes is of high importance for their use in biotechnology and industrial applications. Although lipid vesicle solubilization by classical detergents has been widely investigated, there are few systematic structural and kinetic studies where different detergents are compared under varying conditions. This study used small-angle X-ray scattering to determine the structures of lipid/detergent aggregates at different ratios and temperatures and studied the solubilization in time using the stopped-flow technique. Membranes composed of either of two zwitterionic lipids, DMPC or DPPC, and their interactions with three different detergents, sodium dodecyl sulfate (SDS), n -dodecyl-beta-maltoside (DDM), and Triton X-100 (TX-100), were tested. The detergent TX-100 can cause the formation of collapsed vesicles with a rippled bilayer structure that is highly resistant to TX-100 insertion at low temperatures, while at higher temperatures, it partitions and leads to the restructuring of vesicles. DDM also causes this restructuring into multilamellar structures at subsolubilizing concentrations. In contrast, partitioning of SDS does not alter the vesicle structure below the saturation limit. Solubilization is more efficient in the gel phase for TX-100 but only if the cohesive energy of the bilayer does not prevent sufficient partitioning of the detergent. DDM and SDS show less temperature dependence compared to TX-100. Kinetic measurements reveal that solubilization of DPPC largely occurs through a slow extraction of lipids, whereas DMPC solubilization is dominated by fast and burst-like solubilization of the vesicles. The final structures obtained seem to preferentially be discoidal micelles where the detergent can distribute in excess along the rim of the disc, although we do observe the formation of worm- and rodlike micelles in the case of solubilization of DDM. Our results are in line with the suggested theory that bilayer rigidity is the main factor influencing which aggregate is formed.

3-Helix micelles have demonstrated excellent in vitro and in vivo stability. Previous studies showed that the unique design of the peptide-polymer conjugate based on protein tertiary structure as the headgroup is the main design factor to achieve high kinetic stability. In this contribution, using amphiphiles with different alkyl tails, namely, C16 and C18, we quantified the effect of alkyl length on the stability of 3-helix micelles to delineate the contribution of the micellar core and shell on the micelle stability. Both amphiphiles form well-defined micelles, <20 nm in size, and show good stability, which can be attributed to the headgroup design. C18-micelles exhibit slightly higher kinetic stability in the presence of serum proteins at 37 °C, where the rate constant of subunit exchange is 0.20 h(-1) for C18-micelles vs 0.22 h(-1) for C16-micelles. The diffusion constant for drug release from C18-micelles is approximately half of that for C16-micelles. The differences between the two micelles are significantly more pronounced in terms of in vivo stability and extent of tumor accumulation. C18-micelles exhibit significantly longer blood circulation time of 29.5 h, whereas C16-micelles have a circulation time of 16.1 h. The extent of tumor accumulation at 48 h after injection is ∼43% higher for C18-micelles. The present studies underscore the importance of core composition on the biological behavior of 3-helix micelles. The quantification of the effect of this key design parameter on the stability of 3-helix micelles provides important guidelines for carrier selection and use in complex environment.

Here we present an extensive small-angle neutron scattering (SANS) structural characterization of micelles formed by poly(ethylene oxide)-mono-n-alkyl ethers (Cn-PEOx) in dilute aqueous solution. Chemically, Cn-PEOx can be considered as a hybrid between a low-molecular weight surfactant and an amphiphilic block copolymer. The present system, prepared through anionic polymerization techniques, is better defined than other commercially available polymers and allows a very precise and systematic testing of the theoretical predictions from thermodynamical models. The equilibrium micellar properties were elaborated by systematically varying the n-alkyl chain length (n) at constant PEO molecular weight or increasing the soluble block size (x), respectively. The structure was reminiscent of typical spherical star-like micelles i.e. a constant core density profile, ∼r(0), and a diffuse corona density profile, ∼r(-4/3). Through a careful quantitative analysis of the scattering data, it is found that the aggregation number, Nagg initially rapidly decreases with increasing PEO length until it becomes independent at higher PEO molecular weight as expected for star-like micelles. On the other hand, the dependency on the n-alkyl length is significantly stronger than that expected from the theories for star-like block copolymer micelles, Nagg ∼ n(2) similar to what is expected for surfactant micelles. Hence the observed aggregation behavior suggests that the Cn-PEOx micelles exhibit a behavior that can be considered as a hybrid between low-molecular weight surfactant micelles and diblock copolymer micelles.

We present a study of equilibrium chain-exchange kinetics of a well-defined model system for starlike polymeric micelles. The results show that the kinetics follows a logarithmic time dependence. The same feature has been confirmed for two other micellar systems. This is in sharp contrast to theoretical predictions and hints towards strongly coupled chain dynamics within the micellar cores induced by geometrical constraints.

Antimicrobial peptides (AMPs) are promising pharmaceutical candidates for the prevention and treatment of infections caused by multidrug-resistant ESKAPE pathogens, which are responsible for the majority of hospital-acquired infections. Clinical translation of AMPs has been limited, in part by apparent toxicity on systemic dosing and by instability arising from susceptibility to proteolysis. Peptoids (sequence-specific oligo-N-substituted glycines) resist proteolytic digestion and thus are of value as AMP mimics. Only a few natural AMPs such as LL-37 and polymyxin self-assemble in solution; whether antimicrobial peptoids mimic these properties has been unknown. Here, we examine the antibacterial efficacy and dynamic self-assembly in aqueous media of eight peptoid mimics of cationic AMPs designed to self-assemble and two nonassembling controls. These amphipathic peptoids self-assembled in different ways, as determined by small-angle X-ray scattering; some adopt helical bundles, while others form core-shell ellipsoidal or worm-like micelles. Interestingly, many of these peptoid assemblies show promising antibacterial, antibiofilm activity in vitro in media, under host-mimicking conditions and antiabscess activity in vivo. While self-assembly correlated overall with antibacterial efficacy, this correlation was imperfect. Certain self-assembled morphologies seem better-suited for antibacterial activity. In particular, a peptoid exhibiting a high fraction of long, worm-like micelles showed reduced antibacterial, antibiofilm, and antiabscess activity against ESKAPE pathogens compared with peptoids that form ellipsoidal or bundled assemblies. This is the first report of self-assembling peptoid antibacterials with activity against in vivo biofilm-like infections relevant to clinical medicine.

The shape and size of self‐assembled structures upon local organization of their molecular building blocks are hard to predict in the presence of long‐range interactions. Combining small‐angle X‐ray/neutron scattering data, theoretical modelling, and computer simulations, sodium dodecyl sulfate (SDS), over a broad range of concentrations and ionic strengths, was investigated. Computer simulations indicate that micellar shape changes are associated with different binding of the counterions. By employing a toy model based on point charges on a surface, and comparing it to experiments and simulations, it is demonstrated that the observed morphological changes are caused by symmetry breaking of the irreducible building blocks, with the formation of transient surfactant dimers mediated by the counterions that promote the stabilization of cylindrical instead of spherical micelles. The present model is of general applicability and can be extended to all systems controlled by the presence of mobile charges. [ABSTRACT FROM AUTHOR]

The kinetics of micelles involving photosensitive surfactants is still not well understood. In this work, we unravel the mechanistic pathways involved in the micelle formation and dissolution of photocontrollable micelles. We focus on the fast self-assembly processes of photosensitive cationic azobenzene-containing surfactants (AzoTMA) that display a change in hydrophobicity induced by a reversible cis-trans conformational transition upon exposure to light. By combining both in situ time-resolved small-angle X-ray scattering (SAXS) and light scattering, we characterized the detailed structure and phase behavior of AzoTMA in mixtures of water and dimethylformamide (DMF). Time-resolved synchrotron SAXS with monochromatic light as a trigger enabled us to observe the nonequilibrium formation and dissolution process of micelles (demicellization) directly on the nanoscale with a time resolution starting from milliseconds. The structural results show that in pure water UV-light illumination leads to a 12% reduction of the aggregation number of the micelles and more than a 50% increase in the critical micelle concentration (CMC). Close to the CMC, adjusted by the addition of DMF, UV light illumination leads to a complete dissolution of the micelles, while shining blue light reverses the process and leads to the reformation of micelles. The UV-triggered dissolution follows a two-step mechanism; the first and rapid (second time scale) release of unimers is followed by a slower decomposition of the micelles (over tens of seconds) as a result of an increase in temperature due to optical absorption. Similarly, the reverse process, i.e., micelle formation, occurs rapidly upon photoconversion to trans conformers under blue light, and micelles are disrupted at long exposure time due to the optical absorption and corresponding increase in temperature. Interestingly, the coexistence of unimers with regular micelles is found at all times, and no other transient assemblies could be detected by SAXS. [ABSTRACT FROM AUTHOR]

The micellization behavior of a temperature and pH responsive pentablock terpolymer, poly(2-( N , N -diethylamino)ethylmethacrylate)- block -poly( N -isopropylacrylamide)- block -poly(ethylene glycol)- block -poly( N -isopropylacrylamide)- block -poly(2-( N , N -diethylamino)ethylmethacrylate, abbreviated as PDEAEMA x - b -PNIPAAM y - b -PEG z - b -PNIPAAM y - b -PDEAEMA x , was studied. Two variants of the polymer, with difference in block lengths, were examined using light scattering and small angle neutron scattering (SANS) as the main tools, with the aim of following nanostructural changes in the system with varying temperature and pH. With a sufficiently long PDEAEMA block, micelles of overall sizes around 50 nm and with a PDEAEMA-rich corona are formed at room temperature and at low pH. A change to a system with a PDEAEMA-rich core can be obtained via a jump in pH from 3 to 9. An increase in temperature will, at low pH, lead to formation of micelles also for the polymer with a short PDEAEMA block, with a transition around 42 °C. If the temperature increase is instead performed at pH 9, the transition to compact PDEAEMA-core micelles occurs at a slightly lower temperature (39 °C). This difference is attributed to the increased hydrophobic character for a system with deprotonated PDEAEMA-blocks. [ABSTRACT FROM AUTHOR]

Nanoparticles, 10-30 nm in size, have shown great prospects as nanocarriers for drug delivery. We designed amphiphiles based on 3-helix peptide-PEG conjugate forming 15 nm micelles (defined as "3-helix micelles") with good in vivo stability. Here, we investigated the effect of the site of PEG conjugation on the kinetic stability and showed that the conjugation site affects the PEG chain conformation and the overall molecular architecture of the subunit. Compared to the original design where the PEG chain is located in the middle of the 3-helix bundle, micelle kinetic stability was reduced when the PEG chain was attached near the N-terminus (t1/2 = 35 h) but was enhanced when the PEG chain was attached near the C-terminus (t 1/2 = 80 h). Quantitative structural and kinetic analysis suggest that the kinetic stability was largely dictated by the combined effects of entropic repulsion associated with PEG chain conformation and the geometric packing of the trimeric subunits. The modular design approach coupled with a variety of well-defined protein stucture and functional polymers will significantly expand the utility of these materials as nanocarriers to meet current demands in nanomedine. [ABSTRACT FROM AUTHOR]

Despite over a century of modern surfactant science, the kinetic pathways of morphological transitions in micellar systems are still not well understood. This is mainly as a result of the lack of sufficiently fast methods that can capture the structural changes of such transitions. Herein, a simple surfactant system consisting of sodium dodecyl sulfate (SDS) in aqueous NaCl solutions is investigated. Combining synchrotron radiation small-angle X-ray scattering (SAXS) with fast stopped-flow mixing schemes allows monitoring the process where polymer-like micelles are formed from globular micelles when the salt concentration is suddenly increased. The results show that "worm-like" micelles are formed by fusion of globular micelles and short cylinders in a fashion that bears similarities to a step-like polymerization process. [ABSTRACT FROM AUTHOR]

Self-assembly of amphiphilic molecules into micelles occurs on very short times scales of typically some milliseconds, and the structural evolution is therefore very challenging to observe experimentally. While rate constants of surfactant micelle kinetics have been accessed by spectroscopic techniques for decades, so far no experiments providing detailed information on the structural evolution of surfactant micelles during their formation process have been reported. In this work we show that by applying synchrotron small-angle X-ray scattering (SAXS) in combination with the stopped-flow mixing technique, the entire micelle formation process from single surfactants to equilibrium micelles can be followed in situ. Using a sugar-based surfactant system of dodecyl maltoside (DDM) in dimethylformamide (DMF), micelle formation can be induced simply by adding water, and this can be followed in situ by SAXS. Mixing of water and DMF is an exothermic process where the micelle formation process occurs under nonisothermal conditions with a temperature gradient relaxing from about 40 to 20 °C. A kinetic nucleation and growth mechanism model describing micelle formation by insertion/expulsion of single molecules under nonisothermal conditions was developed and shown to describe the data very well. [ABSTRACT FROM AUTHOR]

Hypothesis: A key question in the kinetics of surfactant self-assembly is whether exchange of unimers or fusion/fission of entire micelles is the dominant pathway. In this study, an isomerizable surfactant is used to explore fundamental out-of-equilibrium kinetics and mechanisms for growth and dissolution of micelles. Experiments: The kinetics of cationic surfactant 4-butyl-4'-(3-trimethylammoniumpropoxy)-phenylazobenzene was studied using molecular dynamics simulations. The fusion and exchange processes were investigated using umbrella sampling. Equilibrium states were validated by comparison with small-angle X-ray scattering data. The photo-isomerization event was simulated by modifying the torsion potential of the photo-responsive group to emulate the trans -to- cis transition. Findings: Micelle growth is dominated by unimer exchange processes, whereas, depending on the conditions, dissolution can occur both through fission and unimer expulsion. Fusion barriers increase steeply with the aggregation number making this an unlikely pathway to equilibrium for micelles of sizes that fit with the experimental data. The barriers for unimer expulsion remain constant and are much lower for unimer insertion, making exchange more likely at high aggregation. When simulating photo-conversion events, both fission and a large degree of unimer expulsion can occur depending on the extent of the out-of-equilibrium stress that is put on the system. [ABSTRACT FROM AUTHOR]

In this paper, we present a study of micellar structures formed by poly(styrene)-poly(butadiene) (PS10-PB10; the numbers indicate the molecular weight in kg mol−1) diblock copolymers and PB10-PS20-PB10 triblock copolymers in different n-alkane solvents. Particular emphasis is placed on the dynamic properties of these micelles under equilibrium which are studied using a novel time-resolved small-angle neutron scattering technique. The results show that the structures of the micelles are very similar for both the diblock and triblock copolymers, which allows a direct comparison of the dynamic properties. A novel logarithmic relaxation is found for both the triblock and the diblock micelles which is not consistent with theoretical expectations. However, for the diblock micelles, the relaxation kinetics seem to approach the rate and the single exponential decay predicted by Halperin & Alexander [ Macromolecules, (1989), 22, 2403–2412] when the micellar cores are strongly swollen with solvent. For the triblock micelles a logarithmic relaxation is found for all cases as an effect of additional topological knots present even in highly swollen micellar cores. This behavior is assigned to an increased coupling of chain motion within the dense confined core – an effect which seems to vanish in diblock micelles when the core is sufficiently swollen. [ABSTRACT FROM AUTHOR]

3-Helix micelles (3HM) formed by self-assembly of peptide-polymer conjugate amphiphiles have shown promise as a nanocarrier platform due to their long-circulation, deep tumor penetration, selective accumulation in tumor, and ability to cross the blood-brain barrier (BBB) for glioblastoma therapy. There is a need to understand the structural contribution to the high in vivo stability and performance of 3HM. Using selective deuteration, the contrast variation technique in small-angle neutron scattering, and coarse-grained molecular dynamics simulation, we determined the spatial distribution of each component within 3HM. Our results show a slightly deformed polyethylene glycol (PEG) conformation within the micelle that is radially offset from its conjugation site toward the exterior of the micelle and a highly solvated shell. Surprisingly, ∼85 v/v % of 3HM is water, unusually higher than any micellar nanocarrier based on our knowledge. The result will provide important structural insights for future studies to uncover the molecular origin of 3HM's in vivo performance, and development of the nanocarriers. [ABSTRACT FROM AUTHOR]

Telechelic polymers contain two chain ends that are able to promote self-assembly into "flowerlike" or interconnected micellar structures. Here, we investigate the molecular exchange kinetics of such micelles using time-resolved small-angle neutron scattering. We show that the activation energies of monofunctional and telechelic chain exchange are identical. This demonstrates that the two chain ends are not simultaneously released in a single event. Instead, the results show that, contrary to regular micelles, the kinetics occurs in a multistep process involving a collision-induced single-molecule exchange mechanism where the exchange rate is directly proportional to the polymer concentration. We show that this novel mechanism can be quantitatively explained by a simple kinetic model. [ABSTRACT FROM AUTHOR]

Molecular exchange processes are important equilibration and transport mechanisms in both synthetic and biological self-assembled systems such as micelles, vesicles, and membranes. Still, these processes are not entirely understood, in particular the effect of crystallinity and the interplay between cooperative melting processes and chain exchange. Here we focus on a set of simple polymer micelles formed by binary mixtures of poly(ethylene oxide)-mono-n-alkyl-ethers (Cn-PEO5) which allows the melting point to be tuned over a wide range. We show that the melting transition is cooperative in the confined 4-5 nm micellar core, whereas the exchange process is widely decoupled and unimeric in nature. As confirmed by differential scanning calorimetry, the total activation energy for ejecting a molecule out of the micellar core below the melting point is the sum of the enthalpy of fusion and the corresponding activation energy in the melt state. This suggests that a "local, single-chain melting process" preludes the molecular diffusion out of the micelle during chain exchange. [ABSTRACT FROM AUTHOR]