Flower Micelles Research Articles

Poly(L-lysine)-block-poly(ethylene glycol)-block-poly(L-lysine) triblock copolymers for the preparation of flower micelles and their irreversible hydrogel formation

ABSTRACT Poly(L-lysine)-block-poly(ethylene glycol)-block-poly(L-lysine) (PLys-block-PEG-block-PLys) triblock copolymers formed polyion complex (PIC) with poly(acrylic acid) (PAAc) or sodium poly(styrenesulfonate) (PSS), leading to the formation of flower micelle-type nanoparticles (NanoLys/PAAc or NanoLys/PSS) with tens of nanometers size in water at a polymer concentration of 10 mg/mL. The flower micelles exhibited irreversible temperature-driven sol-gel transitions at physiological ionic strength, even at low polymer concentrations such as 40 mg/mL, making them promising candidates for injectable hydrogel applications. Rheological studies showed that the chain length of PLys segments and the choice of polyanions significantly impacted irreversible hydrogel formation, with PSS being superior to PAAc for the formation. The incorporation of silica gel nanoparticles into the PIC flower micelles also resulted in irreversible gelation phenomena. The highest storage modulus exceeded 10 kPa after gelation, which is sufficient for practical applications. This study demonstrates the potential of these PIC-based hydrogels as biomaterials with tunable properties for biomedical applications.

Micellar structure of decenyl succinic anhydride modified pullulan with degree of substitution dependence in aqueous solution

A broad distribution pullulan (PUL) sample was hydrophobically modified by a relative longer decenyl succinic anhydride (DCSA) side chains with seven different degree of substitution (DS) to obtain the PUL-DCSA. The micellar structure of PUL-DCSA with DS dependence was investigated by pyrene-probe fluorescence and small-angle X-ray scattering (SAXS) in 0.05 M aqueous NaCl. While PUL-DCSA with DS > 0.26, both the SAXS and fluorescence results can be explained by the flower necklace model, when DS < 0.26, PUL-DCSA forms the loose flower necklace model. Moreover, the micellar structure of PUL-DCSA characterized by SAXS and fluorescence was different from that of the PUL-bearing shorter alkyl (octenyl or nonenyl) chains. The smaller chain stiffness qFN and larger number of monomer (glucose residues) per unit flower micelle N0u of PUL-DCSA, mainly due to the difference in the different hydrophobic alkyl side chains. This research will provide a theoretical guidance for designing biomedical materials with appropriate scale and characteristic functions in drug delivery system.

Pegylated surfactants based on fatty acids: 12-hydroxystearic acid versus stearic acid

We describe the aqueous behavior of short Polyethylene glycol (PEG) chains of Mw 4 kDa that are either mono-functionalized or di-functionalized by 12-hydroxy stearic acid (HSA), and stearic acid (SA), respectively. The end capping of the chains by the fatty acids is achieved by Steglich esterification, after a step of protection of the 12-OH functional group in the case of HSA. Small Angle Neutron Scattering (SANS) shows that mono-functionalized chains with both SA or HSA sticky ends self-assemble in star micelles in aqueous solution, and interact through repulsive steric interactions. The di-functionalized telechelic chains self-assemble in flower micelles that interact through interactions that are attractive on average due to the formation of bridges by chains that share their both ends in two different micelles. Solutions of telechelic chains undergo a phase separation between a dilute solution of micelles and a dense hexagonal crystalline network of flower micelles above a given polymer concentration threshold. This threshold is lower for SA-PEG-SA than for HSA-PEG-HSA. We postulate that this difference of behavior arises from the formation of hydrogen bonds between the OH groups at the C12 of the alkyl chain in case of HSA that increase the stability of the hydrophobic core of the micelles.

Orientational Dynamics of Magnetic Iron Oxide Nanoparticles in a Hydrogel: Observation by Magnetic Linear Dichroism under Oscillating Field.

For the success of biomedical applications of magnetic iron oxide nanoparticles (MION), such as magnetic hyperthermia and magnetic particle imaging, it is essential to understand the orientational dynamics of MION in a complex fluid under an alternating field. Here, using the magnetic linear dichroism (MLD) measurement, we directly observed the orientational behavior of MION in a hydrogel under a damped oscillating magnetic field (DOMF) of 33 kHz in frequency. Hydrophobically modified ethoxylated urethane (HEUR) is examined as the network polymer because the mesh size of the network is controllable with its concentration. We used two MIONs: a bare MION (MION1) and a MION coated with an amphiphilic polymer (MION2). Where the mesh size of the gel network is larger than the particle's hydrodynamic diameter, MION1 in the hydrogel rotates in the same manner in a simple solution, although the macroscopic rheological property of the medium is quite different. Meanwhile, the orientational behavior of MION2 is dramatically changed by the addition of HEUR molecules even below the minimum gelation concentration, indicating that MION2 is associated with the flower micelles of HEUR. By analyzing the MLD waveform, the orientational behavior of MION1 in the HEUR gel under a DOMF can be explained with single-mode relaxation, whereas that of MION2 is complicated; a rapid partial rotation near the particle and a whole slow rotation of the particle-flower micelle associate are superimposed. It is hard to distinguish this difference in orientational behaviors from the dynamic magnetization curve because the dominant magnetization reversal process is Néel rotation, the rotation of the magnetic moment in the particle. The MLD measurement is a potential tool for optimizing biomedical techniques utilizing MIONs and for nanorheology or colloid science in a complex matrix such as a hydrogel or cytoplasmic matrix.

Association Behavior of Amphiphilic ABA Triblock Copolymer Composed of Poly(2-methoxyethyl acrylate) (A) and Poly(ethylene oxide) (B) in Aqueous Solution.

Poly(2-methoxyethyl acrylate) (PMEA) and poly(ethylene oxide) (PEO) have protein-antifouling properties and blood compatibility. ABA triblock copolymers (PMEAl-PEO11340-PMEAm (MEOMn; n is average value of l and m)) were prepared using single-electron transfer-living radical polymerization (SET-LRP) using a bifunctional PEO macroinitiator. Two types of MEOMn composed of PMEA blocks with degrees of polymerization (DP = n) of 85 and 777 were prepared using the same PEO macroinitiator. MEOMn formed flower micelles with a hydrophobic PMEA (A) core and hydrophilic PEO (B) loop shells in diluted water with a similar appearance to petals. The hydrodynamic radii of MEOM85 and MEOM777 were 151 and 108 nm, respectively. The PMEA block with a large DP formed a tightly packed core. The aggregation number (Nagg) of the PMEA block in a single flower micelle for MEOM85 and MEOM777 was 156 and 164, respectively, which were estimated using a light scattering technique. The critical micelle concentrations (CMCs) for MEOM85 and MEOM777 were 0.01 and 0.002 g/L, respectively, as determined by the light scattering intensity and fluorescence probe techniques. The size, Nagg, and CMC for MEOM85 and MEOM777 were almost the same independent of hydrophobic DP of the PMEA block.

Thermoresponsive Self-Assembly of Twofold Fluorescently Labeled Block Copolymers in Aqueous Solution and Microemulsions.

A nonionic double hydrophilic block copolymer with a long permanently hydrophilic and a small thermoresponsive block is synthesized by reversible addition-fragmentation chain-transfer polymerization (RAFT). By employing a specifically designed chain-transfer agent, the polymer is functionalized with complementary end groups which are suited for Förster resonance energy transfer (FRET). The end group attached to the permanently hydrophilic block of poly(N,N-dimethylacrylamide) pDMAm is designed as a permanently hydrophobic segment ("sticker") comprising a long alkyl chain and the 4-aminonaphthalimide fluorophore. The other end attached to the thermoresponsive block of poly(N-isopropylacrylamide) pNiPAm incorporates a coumarin fluorophore. The temperature-dependent self-assembly of the twofold fluorescently labeled copolymer is studied in pure aqueous solution as well as in an o/w microemulsion by several techniques including turbidimetry, dynamic light scattering (DLS), and fluorescence spectroscopy. It is compared to the behaviors of the analogous twofold-labeled pDMAm and pNiPAm homopolymer references. The findings indicate that the block copolymer behaves as a polymeric surfactant at low temperatures, with one relatively small hydrophobic end block and an extended hydrophilic chain forming "hairy micelles". At elevated temperatures above the LCST phase transition of the pNiPAm block, however, the copolymer behaves as an associative telechelic polymer with two nonsymmetrical hydrophobic end blocks, which do not mix. Thus, instead of a network of bridged "flower micelles", large dynamic aggregates are formed. These are connected alternatingly by the original micellar cores as well as by clusters of the collapsed pNiPAm blocks. This type of structure is even more favored in the o/w microemulsion than in pure aqueous solution, as the microemulsion droplets constitute an attractive anchoring point for the hydrophobic dodecyl sticker but not for the collapsed pNiPAm chains.

The effect of topology of PEG chain on the stability of micelles in brine and serum

Polymeric micelles are promising vehicles for drug delivery these years. The stability of micelles in blood system is very important for the encapsulated therapeutic agents reaching the targeting area. However, generally, micelles are not stable in the presence of proteins and ions after being injected intravenously. The topology of polymer chain on the surface of micelles plays a key role on the stability of micelles. Here we have prepared flower micelles and the traditional micelles via self-assembly of triblock and diblock copolymer of polyethylene glycol (PEG) and poly(ε-caprolactone) (PCL) in water. The special PEG shell of flower micelles prevents micelles from ions and proteins, which endows the micelles with the ability to resist aggregation. The experiment results show that flower micelles are much more stable in the presence of serum and salt than traditional micelles, which indicates these micelles would be promising in drug delivery in the future.

Transition from the random coil to the flower necklace of a hydrophobically modified pullulan in aqueous solution by changing the degree of substitution

The degree of substitution (DS) dependence of the micellar structure of a hydrophobically modified polysaccharide, octenyl succinic anhydride modified pullulan (PUL-OSA), in aqueous solution was investigated by small angle X-ray scattering and pyrene-probe fluorescence. While PUL-OSA with DS > 0.9 forms the flower necklace in aqueous solution as demonstrated in the previous study, some of unit flower micelles composing the flower necklace are unfolded to the random coil, and the transition from the flower necklace to the random coil occurs via the loose flower necklace, when DS is decreased below 0.9. The fraction of the unit flower micelle in the loose flower necklace decreases gradually with decreasing DS from 0.9 to 0.

Synthesis and self-assembly of amphiphilic comb-copolymers possessing polyethyleneimine and its derivatives: Site-selective formation of loop-cluster covered vesicles and flower micelles

Amphiphilic block copolymers have attracted a considerable amount of attention in the past two decades because of their ability to self-organize into various complex nanostructures. Most of these studies concentrated on linear amphiphilic block copolymers. In this work, we focused our attention on two type unique comb-like copolymers which grafting amphiphilic diblocked side chains onto main chain of polystyrenic backbone. One is Type-I in which a block of hydrophilic polymers is located inside near to the backbone while other block of hydrophobic poly (phenyl-2-oxazoline) [PPOZ] is in outside far away the backbone. The other one is Type-O in which the hydrophobic block of PPOZ is grafted (inside) onto the main chain backbone then the hydrophilic block is joined (outside) on the PPOZ. A series of five kinds of Type-I and five kinds of Type-O comb-like block copolymers with different components of the amphiphiles were prepared. We performed self-assembly of them in a DMF/water or methanol/water (v/v, 1/9) media. All of Type-O comb-like block copolymers formed micelle. In contrast, in the case of Type-I, interestingly, the comb-like block copolymers possessing hydrophilic blocks with crystalline feature formed loop-cluster covered vesicles, while the others having non-crystalline hydrophilic blocks formed flower-like micelle. We proposed the self-organizing mechanism of these two types comb-like block copolymers and provides new molecular design strategy for the site-selective fabrication of vesicle/micelle possessing special surface structures.

Flower Necklaces of Controllable Length Formed From N-(2-Hydroxypropyl) Methacrylamide-Based Amphiphilic Statistical Copolymers.

N-(2-Hydroxypropyl)methacrylamide (HPMA)-based statistical copolymers bearing anticancer drugs have attracted attention for their efficacy in cancer treatments. However, controlling the size and morphology of aggregates of this type of polymer has been challenging and is far from being understood. In this study, small-angle X-ray scattering and asymmetric-flow field-flow fractionation with multiangle light scattering were used to investigate the structure of aggregates formed in aqueous solutions of HPMA-based statistical copolymers of different molecular weights with the model drug pyrene borne in different amounts. The analysis revealed that spherical objects (flower micelles) were formed by the assembly of pyrene moieties in low-molecular-weight copolymers, and the flower micelles connected linearly to form string-of-pearls assemblies (flower necklaces) in high-molecular-weight copolymers. The number of pyrene moieties per polymer chain likely dominates the size and morphology of the copolymer micelles. This study shows how to alter the aggregate structure by changing the molecular weight and composition of copolymers.

The Two Phase Transitions of Hydrophobically End-Capped Poly(N-isopropylacrylamide)s in Water.

High-sensitivity differential scanning calorimetry (HS-DSC) thermograms of aqueous poly(N-isopropylacrylamide) (PNIPAM) solutions present a sharp unimodal endotherm that signals the heat-induced dehydration/collapse of the PNIPAM chain. Similarly, α,ω-di-n-octadecyl-PNIPAM (C18-PN-C18) aqueous solutions exhibit a unimodal endotherm. In contrast, aqueous solutions of α,ω-hydrophobically modified PNIPAMs with polycyclic terminal groups, such as pyrenylbutyl (Py-PN-Py), adamantylethyl (Ad-PN-Ad), and azopyridine- (C12-PN-AzPy) moieties, exhibit bimodal thermograms. The origin of the two transitions was probed using microcalorimetry measurements, turbidity tests, variable temperature 1H NMR (VT-NMR) spectroscopy, and 2-dimensional NOESY experiments with solutions of polymers of molar mass (Mn) from 5 to 20 kDa and polymer concentrations of 0.1 to 3.0 mg/mL. The analysis outcome led us to conclude that the difference of the thermograms reflects the distinct self-assembly structures of the polymers. C18-PN-C18 assembles in water in the form of flower micelles held together by a core of tightly packed n-C18 chains. In contrast, polymers end-tagged with azopyridine, pyrenylbutyl, or adamantylethyl form a loose core that allows chain ends to escape from the micelles, to reinsert in them, or to dangle in surrounding water. The predominant low temperature (T1) endotherm, which is insensitive to polymer concentration, corresponds to the dehydration/collapse of PNIPAM chains within the micelles, while the higher temperature (T2) endotherm is attributed to the dehydration of dangling chains and intermicellar bridges. This study of the two phase transitions of telechelic PNIPAM homopolymer highlights the rich variety of morphologies attainable via responsive hydrophobically modified aqueous polymers and may open the way to a variety of practical applications.

Relationships between Diffusion and Viscoelasticity of Associative Polymer Networks

Hydrophobically ethoxylated modified urethane (HEUR) form the flower micelles and transient network structures through the aggregation of end groups depending on the polymer concentration. The HEUR networks show the viscoelastic relaxation which is described by the Maxwell model. Although there have been many attempts to understand and predict the molecular mechanism, the full understanding remains incomplete. To understand the relaxation mechanism of associative polymers, we measured the linear viscoelasticity and the diffusibility using the fluorescence recovery after photobleaching method. With increasing the polymer concentration, the self-diffusion coefficient decreased, while the relaxation time increased, suggesting that the viscoelastic relaxation is correlated with the diffusion of the polymers. The calculated diffusion distance of the HEUR chains within the viscoelastic relaxation time is 100 times larger than the size of a HEUR chain. This significant deviation suggests that some of the HEUR chains diffuse quickly through the unimer and flower micelles at low concentrations and through the recombination process of the network strands at high concentrations.

Redox-active injectable gel using polyion complex to achieve sustained release of exenatide and enhance therapeutic efficacy for the treatment of type 2 diabetes.

To provide sustained release of exenatide and enhance therapeutic efficacy for the treatment of type 2 diabetes compared to the existing products for exenatide, we developed an exenatide-loaded, redox-active, injectable gel (Exe@RIG). This injectable gel is formed by a polyion complex (PIC) comprising three components, (1) cationic polyamine-poly(ethylene glycol)-polyamine triblock copolymer possessing reactive oxygen species (ROS)-scavenging moieties as side chains, (2) anionic poly(acrylic acid), and (3) exenatide. The mixture formed exenatide-loaded PIC flower micelles at room temperature, which immediately converted to a gel under physiological conditions. Owing to electrostatic interactions between exenatide and the PIC gel network, RIG was able to provide sustained release of exenatide without a significant initial burst. Subcutaneous injection of Exe@RIG once a week prevented the increase in glucose concentration significantly in db/db mice compared to those in control groups. In addition, Exe@RIG suppressed the degeneration of pancreatic islets, which is reported to be caused by increased ROS. Our result indicates that Exe@RIG has the potential to provide a long acting exenatide as well as enhanced efficacy in the treatment of type 2 diabetes compared to the existing products. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1107-1113, 2019.

Self-assembly of amphiphilic block pendant polymers as microphase separation materials and folded flower micelles

Herein, we created amphiphilic polymers bearing hydrophilic/hydrophobic block pendants as a new class of self-assembled materials for microphase separation in the solid state and folded flower micelles in water.

Theory of the Flower Micelle Formation of Amphiphilic Random and Periodic Copolymers in Solution.

The mixing Gibbs energy Δgm for the flower-micelle phase of amphiphilic random and periodic (including alternating) copolymers was formulated on the basis of the lattice model. The formulated Δgm predicts (1) the inverse proportionality of the aggregation number to the degree of polymerization of the copolymer, (2) the increase of the critical micelle concentration with decreasing the hydrophobe content, and (3) the crossover from the micellization to the liquid–liquid phase separation as the hydrophobe content increases. The transition from the uni-core flower micelle to the multi-core flower necklace as the degree of polymerization increases was also implicitly indicated by the theory. These theoretical results were compared with experimental results for amphiphilic random and alternating copolymers reported so far.

Development of a local anesthetic lidocaine-loaded redox-active injectable gel for postoperative pain management.

1. We have been working on nanomaterials, which effectively eliminate ROS, avoiding dysfunction of mitochondria in healthy cells. 2. We designed redox injectable gel using polyion complexed flower type micelle, which can eliminates ROS locally. 3. We could prepare local anesthesia-loaded redox injectable gel (lido@RIG). 4. Drug release could be extended by local administration of lido@RIG. 5. Deprotonation of lidocaine improved anesthetic effect because ROS were eliminated locally by RIG. 6. Local inflammation could be also suppressed by lido@RIG.

1,2-Dithiolane-Derived Dynamic, Covalent Materials: Cooperative Self-Assembly and Reversible Cross-Linking.

The use of dithiolane-containing polymers to construct responsive and dynamic networks is an attractive strategy in material design. Here, we provide a detailed mechanistic study on the self-assembly and gelation behavior of a class of ABA triblock copolymers containing a central poly(ethylene oxide) block and terminal polycarbonate blocks with pendant 1,2-dithiolane functionalities. In aqueous solution, these amphiphilic block copolymers self-assemble into bridged flower micelles at high concentrations. The addition of a thiol initiates the reversible ring-opening polymerizations of dithiolanes in the micellar cores to induce the cross-linking and gelation of the micellar network. The properties of the resulting hydrogels depend sensitively on the structures of 1,2-dithiolanes. While the methyl asparagusic acid-derived hydrogels are highly dynamic, adaptable, and self-healing, those derived from lipoic acid are rigid, resilient, and brittle. The thermodynamics and kinetics of ring-opening polymerization of the two dithiolanes were investigated to provide important insights on the dramatically different properties of the hydrogels derived from the two different dithiolanes. The incorporation of both dithiolane monomers into the block copolymers provides a facile way to tailor the properties of these hydrogels.

Structural Analysis of Hydrophobe-Uptake Micelle of an Amphiphilic Alternating Copolymer in Aqueous Solution.

We investigated the structure of the hydrophobe-uptake micelle of an alternating amphiphilic copolymer in aqueous solutions, by combining light scattering and small-angle X-ray scattering (SAXS). When the copolymer micelle includes the hydrophobe (1-dodecanol), the unicore flower micelle transforms into the multicore flower necklace, and the flower necklace is slightly stiffer than the hydrophobe-free flower necklace of the same copolymer. Moreover, the hydrophobe is included not in the hydrophobic core region but in the intermingled region of the hydrophobic group and the loop chain of the unit flower micelle. Therefore, the structure of the hydrophobe-uptake micelle of the amphiphilic alternating copolymer is quite different from that of hydrophobe-uptake spherical micelles of low molar mass surfactants and of amphiphilic block copolymers, where the hydrophobe is included in the hydrophobic region of the micelles.

Local and global conformations of flower micelles and flower necklaces formed by an amphiphilic alternating copolymer in aqueous solution

The local and global conformations of flower micelles and flower necklaces formed by the amphiphilic alternating copolymer of sodium maleate and dodecyl vinyl ether P(MAL/C12) in an aqueous solution were investigated using small-angle X-ray scattering and light scattering. Both local and global conformations for micelles of P(MAL/C12) with N01 (the degree of polymerization) 300 have been explained consistently by the flower micelle and flower necklace models with minimum-sized loops, respectively, as previously proposed.

Development of a long-acting, protein-loaded, redox-active, injectable gel formed by a polyion complex for local protein therapeutics

Although cancer immunotherapies are attracting much attention, it is difficult to develop bioactive proteins owing to the severe systemic toxicity. To overcome the issue, we designed new local protein delivery system by using a protein-loaded, redox-active, injectable gel (RIG), which is formed by a polyion complex (PIC) comprising three components, viz., cationic polyamine-poly(ethylene glycol)-polyamine triblock copolymer possessing ROS-scavenging moieties as side chains; anionic poly(acrylic acid); and a protein. The mixture formed the protein-loaded PIC flower micelles at room temperature, which immediately converted to a gel with high mechanical strength upon exposure to physiological conditions. Because the protein electrostatically interacts with the PIC gel network, RIG provided a sustained release of the protein without a significant initial burst, regardless of the types of proteins in vitro, and much longer retention of the protein at the local injection site in mice than that of the naked protein. Subcutaneous injections of IL-12@RIG in the vicinity of tumor tissue showed remarkable tumor growth inhibition in tumor-bearing mice, compared to that observed with injection of IL-12 alone, suppressing adverse events caused by IL-12–induced ROS. Our results indicate that RIG has potential as a platform technology for an injectable sustained-release carrier for proteins.