Formation of Unimer Nanoparticles by Controlling the Self-Association of Hydrophobically Modified Poly(amino acid)s

Amphiphilic block or graft copolymers have been demonstrated to form a variety of self-assembled nano/microstructures in selective solvents. In this study, the self-association behavior of biodegradable graft copolymers composed of poly­(γ-glutamic acid) (γ-PGA) as the hydrophilic segment and l-phenylalanine (Phe) as the hydrophobic segment in aqueous solution was investigated. The association behavior and unimer nanoparticle formation of these γ-PGA-<i>graft</i>-Phe (γ-PGA-Phe) copolymers in aqueous solution were characterized with a focus on the effect of the Phe grafting degree on the intra- and interpolymer association of γ-PGA-Phe. The particle size and number of polymer aggregates (<i>N</i>_agg) in one particle of the γ-PGA-Phe depended on the Phe grafting degree. The size of γ-PGA-Phe with 12, 27, 35, or 42% Phe grafting (γ-PGA-Phe-12, -27, -35, or -42) was about 8–14 nm and the <i>N</i>_agg was about 1, supporting the presence of a unimolecular graft copolymer in PBS. The pyrene fluorescence data indicated that γ-PGA-Phe-35 and -42 have hydrophobic domains formed by the intrapolymer association of Phe attached to γ-PGA. These results suggest that the Phe grafting degree is critical to the association behavior of γ-PGA-Phe and that γ-PGA-Phe-35 and -42 could form unimer nanoparticles. Moreover, when γ-PGA-Phe-42 dissolved in DMSO was added to various concentrations of NaCl solution, the particle size and <i>N</i>_agg could be easily controlled by changing the NaCl concentration during the formation of the particles. These results suggest that biodegradable γ-PGA-Phe is useful for the fabrication of very small nanoparticles. It is expected that γ-PGA-Phe nanoparticles, including unimer particles, will have great potential as multifunctional carriers for pharmaceutical and biomedical applications, such as drug and vaccine delivery systems

Inkjet Printing of Layer-by-Layer Assembled Poly(lactide) Stereocomplex with Encapsulated Proteins

Inkjet printing, a technique that precisely deposits liquid droplets in picoliter-volume ranges on a substrate, has received increased attention for its novelty and ability to produce functional materials. This technology is considered one of the most promising methods for the controlled deposition of different polymers. In our previous study, a poly­(lactide) (PLA) stereocomplex was fabricated using inkjet printing on a substrate. The stereocomplex was formed by the layer-by-layer (LbL) stepwise deposition of poly­(l-lactide) (PLLA) and poly­(d-lactide) (PDLA). Multiple inkjet passes could conclusively improve the PLAs crystal structure with solvent evaporation (solidification) and dissolution of PLA. We suggested that this technique may also be applicable for fabricating polymer composites with drugs, such as peptides, proteins, and nanoparticles, which is incompatible with the PLA. Here, we report the utilization of this technique to create a PLA stereocomplex with drugs as a drug carrier/reservoir. The three components of PLLA, PDLA, and model drugs (an 8-mer peptide, ovalbumin, and protein-encapsulating nanoparticles) were alternately overprinted onto the substrate without an intermediate rinsing step. Inkjet printing was used successfully to form PLA stereocomplex composites with drugs by the LbL deposition of polymers and functioned as drug carriers/reservoirs. The sustained release of the drugs was observed from the PLLA/PDLA/drug composites. By varying the crystalline structure of PLAs-drug composites, the release kinetics of drugs could be altered and controlled efficiently. Moreover, a high drug loading content (wt %) of PLA stereocomplex composites was achieved up to 100 wt % loading, and the composites with 50 wt % of drug loading content were available for sustained-release formulation. This fabrication technique would provide a platform for creating protein/vaccine/gene delivery carriers with the desired release profiles by controlling the microphase-separated structure and drug distribution within the composites

Structural Analysis of Unimer Nanoparticles Composed of Hydrophobized Poly(amino acid)s

Amphiphilic random copolymers of poly­(γ-glutamic acid)-<i>graft</i>-l-phenylalanine (γ-PGA-Phe), with various lengths of γ-PGA main chains (molecular weight of 70, 140, and 220 kDa) plus hydrophobic moieties of Phe groups (grafting degree 12–60%), self-assembled in aqueous media to form nanoparticles (NPs). The aggregation number (<i>N</i>_agg) could be adjusted according to their molecular structures as well as the preparative methods/conditions. The γ-PGA-Phe NPs were further characterized by means of dynamic and static light scattering, small-angle neutron scattering, as well as steady-state fluorescence measurements/quenching techniques. The single chain state had hydrophobic domains, and formed spherical structures, herein called unimer NPs, and were obtained using molecular weights of γ-PGA higher than 140 kDa conjugated with Phe at 27–42% due to the balance of hydrophobicity/hydrophilicity along the single polymer chain. The number of hydrophobic domains in one NP (<i>N</i>_domain), estimated by means of fluorescence quenching techniques and the rigidity of the inner particles detected by dipyrene fluorescence demonstrated that the <i>N</i>_domain and the rigidity were affected by the particle size and preparative methods. In addition, the effect of pH on the stability of the unimer NPs indicated a reduction of the <i>N</i>_domain upon the pH, supporting a loose packing due to hydrophobic association under alkaline conditions