Understanding associative polymer self-assembly with shrinking gate fluorescence correlation spectroscopy.

The self-assembly of polymers is integral to their role in liquid formulations. In this study, we combine a dye whose lifetime is sensitive to the nanoviscosity of its local environment with shrinking gate fluorescence correlation spectroscopy (sgFCS) to study the self-assembly of a model telechelic polymer, hydrophobically modified ethoxylated urethane (HEUR). Fluorescence lifetime measurements show a monotonic increase in average lifetime with increasing HEUR concentration driven by a small fraction of dye (<1%) with long lifetimes strongly bound to HEUR. Despite this small fraction, sgFCS isolates the diffusional dynamics of the bound fraction with no a priori assumptions as to the distribution of lifetimes. Sensitivity is greatly enhanced compared to standard FCS, revealing micellar aggregates forming between 0.2 and 1 wt% followed by formation of a percolated network. This sgFCS approach, which we apply for the first time to polymers in this work, is readily extendable to any dye that changes lifetime on binding.

One Step Closer to Coatings Applications Utilizing Self-Stratification: Effect of Rheology Modifiers.

Self-stratification of model blends of colloidal spheres has recently been demonstrated as a method to form multifunctional coatings in a single pass. However, practical coating formulations are complex fluids with upward of 15 components. Here, we investigate the influence of three different rheology modifiers (RMs) on the stratification of a 10 wt % 7:3 w:w blend of 270 and 96 nm anionic latex particles that do not stratify without RM. However, addition of a high molar mass polysaccharide thickener, xanthan gum, raises the viscosity and corresponding Péclet number enough to achieve small-on-top stratification as demonstrated by atomic force microscopy (AFM) measurements. Importantly, this was possible due to minimal particle-rheology modifier interactions, as demonstrated by the bulk rheology. In contrast, Carbopol 940, a microgel-based RM, was unable to achieve small-on-top stratification despite a comparable increase in viscosity. Instead, pH-dependent interactions with latex particles lead to either laterally segregated structures at pH 3 or a surface enrichment of large particles at pH 8. Strong RM-particle interactions are also observed when the triblock associative RM HEUR10kC12 is used. Here, small-on-top, large-enhanced, and randomly mixed structures were observed at respectively 0.01, 0.1, and 1 wt % HEUR10kC12. Combining rheology, dynamic light scattering, and AFM results allows the mechanisms behind the nonmonotonic stratification in the presence of associative RMs to be elucidated. Our results highlight that stratification can be predicted and controlled for RMs with weak particle interactions, while a strong RM-particle interaction may afford a wider range of stratified structures. This takes a step toward successfully harnessing stratification in coatings formulations.

Investigating the Sequence Specific Adsorption Behavior of Polypeptides at the Solid/Liquid Interface.

Polymer adsorption at the solid/liquid interface depends not only on the chemical composition of the polymer but also on the specific placement of the monomers along the polymer sequence. However, challenges in designing polymers with well-controlled sequences have limited explorations into the role of polymer sequence on adsorption behavior to molecular simulations. Here, we demonstrate how the sequence control offered by polypeptide synthesis can be utilized to study the effects small changes in polymer sequence have on polymer adsorption behavior at the solid/liquid interface. Through a combination of quartz crystal microbalance with dissipation monitoring and total internal reflection ellipsometry, we study the adsorption behavior of three polypeptides, consisting of 90% lysine and 10% cysteine, onto a gold surface. We find different mechanisms are responsible for the adsorption of polypeptides and the resulting conformation on the surface. The initial adsorption of the polypeptides is driven by electrostatic interactions between the polylysine and the gold surface. Once adsorbed, the cysteine undergoes a thiol-Au reaction with the surface, altering the conformation of the polymer layer. Our findings suggest the conformation of the polypeptide layer is dependent on the placement of the cysteines within the sequence; polypeptide chains with evenly spaced cysteine groups adopt a more tightly bound "train" conformation as compared to polypeptides with closely grouped cysteine groups. We envision that the methodologies presented here to study sequence specific adsorption behaviors using polypeptides could be a valuable tool to complement molecular simulations studies.

Helper lipid structure influences protein adsorption and delivery of lipid nanoparticles to spleen and liver.

Nucleic acids, such as messenger RNAs, antisense oligonucleotides, and short interfering RNAs, hold great promise for treating previously 'undruggable' diseases. However, there are numerous biological barriers that hinder nucleic acid delivery to target cells and tissues. While lipid nanoparticles (LNPs) have been developed to protect nucleic acids from degradation and mediate their intracellular delivery, it is challenging to predict how alterations in LNP formulation parameters influence delivery to different organs. In this study, we utilized high-throughput in vivo screening to probe for structure-function relationships of intravenously administered LNPs along with quartz crystal microbalance with dissipation monitoring (QCM-D) to measure the binding affinity of LNPs to apolipoprotein E (ApoE), a protein implicated in the clearance and uptake of lipoproteins by the liver. High-throughput in vivo screening of a library consisting of 96 LNPs identified several formulations containing the helper lipid 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) that preferentially accumulated in the liver, while identical LNPs that substituted DOPE with the helper lipid 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) preferentially accumulated in the spleen. Using QCM-D, it was found that one DOPE-containing LNP formulation (LNP 42) had stronger interactions with ApoE than an identical LNP formulation that substituted DOPE with DSPC (LNP 90). In order to further validate our findings, we formulated LNP 42 and LNP 90 to encapsulate Cy3-siRNA or mRNA encoding for firefly luciferase. The DSPC-containing LNP (LNP 90) was found to increase delivery to the spleen for both siRNA (two-fold) and mRNA (five-fold). In terms of liver delivery, the DOPE-containing LNP (LNP 42) enhanced mRNA delivery to the liver by two-fold and improved liver transfection by three-fold. Understanding the role of the helper lipid in LNP biodistribution and ApoE adsorption may aid in the future design of LNPs for nucleic acid therapeutics.

Enrichment of Charged Monomers Explains Non-monotonic Polymer Volume Fraction Profiles of Multi-stimulus Responsive Copolymer Brushes.

Multi-stimulus responsive poly(2-(2-methoxyethoxy)ethyl methacrylate-co-2-(diethylamino)ethyl methacrylate) [P(MEO2MA-co-DEA)] 80:20 mol % copolymer brushes were synthesized on planar silica substrates via surface-initiated activators continuously regenerated via electron transfer atom transfer radical polymerization. Brush thickness was sensitive to changes in pH and temperature as monitored with ellipsometry. At low pH, the brush is charged and swollen, while at high pH, the brush is uncharged and more collapsed. Clear thermoresponsive behavior is also observed with the brush more swollen at low temperatures compared to high temperatures at both high and low pH. Neutron reflectometry was used to determine the polymer volume fraction profiles (VFPs) at various pH values and temperatures. A region of lower polymer content, or a depletion region, near the substrate is present in all of the experimental polymer VFPs, and it is more pronounced at low pH (high charge) and less so at high pH (low charge). Polymer VFPs calculated through numerical self-consistent field theory suggest that enrichment of DEA monomers near the substrate results in the experimentally observed non-monotonic VFPs. Adsorption of DEA monomers to the substrate prior to initiation of polymerization could give rise to DEA segment-enriched region proximal to the substrate.

Temperature dependent specific ion effects in mixed salt environments on a thermoresponsive poly(oligoethylene glycol methacrylate) brush.

The temperature induced swelling/collapse transition of poly(oligoethylene glycol methacrylate) (POEGMA) brushes has been investigated in electrolyte solutions comprised of multiple anions. The behaviour of a POEGMA brush in mixed salt environments of potassium acetate (KCH3COO, causes collapse) and thiocyanate (KSCN, causes swelling), two ions at opposite ends of the Hofmeister series, has been monitored with neutron reflectometry (NR) and quartz crystal microbalance with dissipation (QCM-D). These techniques revealed that the balance of the swelling/collapse influence of the two ions on the structure of the brush is temperature dependent. At low temperatures in mixed salt environments, the influence of the acetate and thiocyanate ions appears additive, antagonistic and approximately equal in magnitude, with brush thickness and dissipation similar to the brush in the absence of electrolyte. At higher temperatures, the influence of the acetate ion diminishes, resulting in an increase in the relative influence of the thiocyanate ion on the brush conformation. These temperature dependent specific ion effects are attributed to increased steric crowding in the brush, along with an increased affinity of the thiocyanate ion for the polymer at higher temperatures.

Influence of molecular weight on PNIPAM brush modified colloidal silica particles.

The effect of molecular weight and temperature on the phase transition and internal structure of poly(N-isopropylacrylamide) brush modified colloidal silica particles was investigated using dynamic light scattering (DLS) and small angle neutron scattering (SANS) between 15 and 45 °C. Dry particle analysis utilising transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR) and thermogravimetric analysis (TGA) all confirmed the thickness of the polymer brush shell increased as a function of polymerisation time. Hydrodynamic diameter and electrophoretic mobility results revealed that the brush modified particles transitioned from swollen shells to a collapsed conformation between 15 and 35 °C. The dispersions were electrosterically stabilised over the entire temperature range investigated, with minimal thermal hysteresis recorded. Modelling of the hydrodynamic diameter enabled the calculation of a lower critical solution temperature (LCST) which increased as a function of brush thickness. The internal structure determined via SANS showed a swollen brush at low temperatures (18 and 25 °C) which decayed radially away from the substrate, while a collapsed block-like conformation with 60% polymer volume fraction was present at 40 °C. Radial phase separation was evident at intermediate temperatures (30 and 32.5 °C) with the lower molecular weight sample having a greater volume fraction of polymer in the dense inner region at these temperatures.

Specific ion effects on thermoresponsive polymer brushes: Comparison to other architectures.

Thermoresponsive polymers have received significant research attention as smart materials with particular interest in biomedical applications. The composition and architecture are known to strongly influence the thermoresponsive properties of the materials. For example, the strong overlap of end-grafted polymer chains in polymer brushes leads to a broader collapse transition relative to linear ungrafted chains as well as temperature dependent adhesion. The temperature response of free polymer has been widely reported to depend on the concentration and identity of ions in solution and is further modified by the composition of the solvent and presence of cosolutes. However, the influence of polymer architecture on these specific ion effects is relatively unknown. Herein, we compare the current understanding of specific ion effects on free polymer chains and gels with recent studies of polymer brushes. Further studies on mixed salt systems are found to be the next step to predicting the behaviour of these materials in biological systems.

Behavior of Weak Polyelectrolyte Brushes in Mixed Salt Solutions.

Hydrophilic and hydrophobic weak polybasic brushes immersed in aqueous solutions of mixed salt counterions are considered using a mean-field numerical self-consistent field approach. On top of the solvent quality of the polymer, the counterion-solvent interactions are accounted for by implementing Flory-Huggins interaction parameters. We show that ion specificity within the brush can bring about large changes in conformation. It is found that the collapse transition of hydrophobic, weak polyelectrolyte brushes features an intermediate two-phase state wherein a subset of chains are collapsed in a dense layer at the substrate, while the remainder of chains are well-solvated and strongly stretched away from the it. Besides pH and ionic strength, solvent quality of counterions and the composition of ions in the solvent are important control parameters for the behavior of polyelectrolyte brushes. Increasingly hydrophobic counterions penetrate deeper within the brush and stabilize the collapsed region, while hydrophilic counterions do the opposite.

Enhanced specific ion effects in ethylene glycol-based thermoresponsive polymer brushes.

The thermoresponse of poly(di(ethyleneglycol) methyl ether methacrylate) (PMEO2MA) brushes has been investigated in the presence of monovalent anions at either end of the Hofmeister series using ellipsometry, neutron reflectometry (NR) and colloid probe atomic force microscopy (AFM). NR measurements in deuterium oxide showed no evidence of vertical phase separation perpendicular to the grafting substrate with a gradual transition between a block-like, dense structure at 45°C and an extended, dilute conformation at lower temperatures. All three techniques revealed a shift to a more collapsed state for a given temperature in kosmotropic potassium acetate solutions, while more swollen structures were observed in chaotropic potassium thiocyanate solutions. No difference was observed between 250mM and 500mM thiocyanate for a 540Å brush studied by ellipsometry, while the lower molecular weight ∼200Å brushes used for NR and AFM measurements continued to respond with increasing salt concentration. The effect of thiocyanate on the temperature response was greatly enhanced relative to PNIPAM with the shift in temperature response at 250mM being five times greater than a PNIPAM brush of similar thickness and grafting density.

Specific ion modulated thermoresponse of poly(N-isopropylacrylamide) brushes.

The influence of specific anions on the equilibrium thermoresponse of poly(N-isopropylacrylamide) (pNIPAM) brushes has been studied using in situ ellipsometry, quartz crystal microbalance with dissipation (QCM-D) and static contact angle measurements between 20 and 45 °C in the presence of up to 250 mM acetate and thiocyanate anions in water. The thickness and changes in dissipation exhibited a broad swelling transition spanning approximately 15 °C from collapsed (high temperatures) to swollen conformation (low temperatures) while the brush surface wettability changed over approximately 2 °C. In the presence of the kosmotropic acetate anions, the measured lower critical solution temperature (LCST) by the three techniques was very similar and decreased linearly as a function of ionic strength. Conversely, increasing the concentration of the chaotropic thiocyanate anions raised the LCST of the pNIPAM brushes with variation in the measured LCST between the three techniques increasing with ionic strength. The thickness of the pNIPAM brush was seen to progressively increase with increasing thiocyanate concentration at all temperatures. It is proposed that specific ion binding of the chaotropic thiocyanate anion with pNIPAM amide moieties increases the electrostatic intra- and intermolecular repulsion within and between pNIPAM chains. This allows the brush to begin to swell at higher temperatures and to an overall greater extent.

Anion-specific effects on the behavior of pH-sensitive polybasic brushes.

The anion-specific solvation and conformational behavior of weakly basic poly(2-dimethylamino)ethyl methacrylate (poly(DMA)), poly(2-diethylamino)ethyl methacrylate (poly(DEA)), and poly(2-diisopropylamino)ethyl methacrylate (poly(DPA)) brushes, with correspondingly increasing inherent hydrophobicity, have been investigated using in situ ellipsometric and quartz crystal microbalance with dissipation (QCM-D) measurements. In the osmotic brush regime, as the initial low concentration of salt is increased, the brushes osmotically swell by the uptake of solvent as they become charged and the attractive hydrophobic inter- and intrachain interactions are overcome. With increased ionic strength, the brushes move into the salted brush regime where they desolvate and collapse as their electrostatic charge is screened. Here, as the brushes collapse, they transition to more uniform and rigid conformations, which dissipate less energy, than similarly solvated brushes at lower ionic strength. Significantly, in these distinct regimes brush behavior is not only ionic strength dependent but is also influenced by the nature of the added salt based on its position in the well-known Hofmeister or lyotropic series, with potassium acetate, nitrate, and thiocyanate investigated. The strongly kosmotropic acetate anions display low affinity for the hydrophobic polymers, and largely unscreened electrosteric repulsions allow the brushes to remain highly solvated at higher acetate concentrations. The mildly chaotropic nitrate and strongly chaotropic thiocyanate anions exhibit a polymer hydrophobicity-dependent affinity for the brushes. Increasing thiocyanate concentration causes the brushes to collapse at lower ionic strength than for the other two anions. This study of weak polybasic brushes demonstrates the importance of all ion, solvent, and polymer interactions.

Hydrophobic effects within the dynamic pH-response of polybasic tertiary amine methacrylate brushes.

The solvation and swelling behaviour of three dialkylaminoethyl methacrylate polymer brushes, of varying hydrophobicity, have been investigated using a combination of in situ ellipsometry and a quartz crystal microbalance with dissipation (QCM-D). At low pH the tertiary amine groups of the three polymers are protonated and all three brushes are significantly solvated and swell by adopting an extended conformation. As the pH is increased the weak polybasic brushes become increasingly deprotonated and collapse via solvent expulsion. By employing high temporal resolution measurements we have found that monomer hydrophobicity has a direct influence on the dynamics of this pH-response. The most hydrophobic poly(2-diisopropylamino)ethyl methacrylate (poly(DPA)) brush exhibits the fastest maximum swelling rate. This maximum swelling rate is reduced with decreasing monomer hydrophobicity for the 2-diethylamino, and even further for the 2-dimethylamino analogues. For all three brushes, the corresponding collapse transition is slower and compounded by an induction time that decreases with monomer hydrophobicity. Here also, the maximum collapse rate is greatest for the most hydrophobic polymer. This domination of the pH-response kinetics by monomer hydrophobicity is attributed to attractive hydrophobic forces between polymer segments overcoming the repulsive electrostatic forces between the tertiary amine residues.

Critical salt effects in the swelling behavior of a weak polybasic brush.

The swelling behavior of poly(2-(diethylamino)ethyl methacrylate) (PDEA) brushes in response to changes in solution pH and ionic strength has been investigated. The brushes were synthesized by ARGET ATRP methodology at the silica-aqueous solution interface via two different surface-bound initiator approaches: electrostatically adsorbed cationic macroinitiator and covalently anchored silane-based ATRP initiator moieties. The pH-response of these brushes is studied as a function of the solvated brush thickness in a constant flow regime that elucidates the intrinsic behavior of polymer brushes. In situ ellipsometry equilibrium measurements show the pH-induced brush swelling and collapse transitions are hysteretic in nature. Furthermore, high temporal resolution kinetic studies demonstrate that protonation and solvent ingress during swelling occur much faster than the brush charge neutralization and solvent expulsion during collapse. This hysteresis is attributed to the formation of a dense outer region or skin during collapse that retards solvent egress. Moreover, at a constant pH below its pKa, the PDEA brush exhibited a critical conformational change in the range 0.5-1 mM electrolyte, a range much narrower than predicted by the theory of the osmotic brush regime. This behavior is attributed to the hydrophobicity of the collapsed brush. The swelling and collapse kinetics for this salt-induced transition are nearly identical. This is in contrast to the asymmetry in the rate of the pH-induced response, suggesting an alternative mechanism for the two processes dependent on the nature of the environmental trigger.

Polyelectrolyte brush pH-response at the silica-aqueous solution interface: a kinetic and equilibrium investigation.

Polymer brushes are commonly used to modify the properties of solid surfaces. Here a family of polybasic poly(2-(diethylamino)ethyl methacrylate) brushes have been grown using ARGET ATRP from a cationic macroinitiator adsorbed on two types of silica surfaces: QCM crystals and oxidised silicon wafers. The pH-response of these brushes is investigated as a function of brush thickness in a constant flow environment in order to focus on the intrinsic dynamics of the polymer brushes. Independent QCM-D and in situ ellipsometry kinetic studies demonstrate the swelling process of protonation and solvent uptake is typically eight times faster than the corresponding neutralisation and solvent expulsion from the collapsing brush. However, the maximum rate of these processes is independent of brush thickness. The initial pH response of the brushes is hysteretic due to brush entanglement, which once overcome allows highly reversible pH-induced conformational changes. Multiple pH cycles demonstrate that the viscoelastic nature of the swollen state relative to the collapsed brush is independent of brush thickness.