Fluorinated Peptide Hydrogels Result in Longer In Vivo Residence Time after Subcutaneous Administration.

Peptide-based hydrogels are of interest to biomedical applications. Herein, we have explored the introduction of fluorinated amino acids in hydrogelator H-FQFQFK-NH2 (P1) to design a series of fluorinated peptide hydrogels and evaluate the in vitro and in vivo properties of the most promising analogues. The impact of fluorinated groups on peptide gelation, secondary structure, and self-assembly processes was assessed. We show that fluorine can significantly improve hydrogel stiffness, compared to the nonfluorinated reference P1. For P15 (H-FQFQF(o-CF3)K-NH2), P18 (H-FQFQF(F5)K-NH2), and P19 (H-FQFQM(CF3)K-NH2), microscopy studies scrutinized fiber morphologies and alignment in the network. In vitro release studies of hydrogels loaded with an opioid cargo suggested improved hydrogel stability for P15 and P18. This improved stability was further validated in vivo, notably for P15, giving the most significant increased gel residence time, with more than 20% of hydrogel still present 9 days post-injection, as monitored by nuclear SPECT-CT imaging.

Impact of doubling peptide length on in vivo hydrogel stability and sustained drug release.

Peptide-based hydrogels represent promising systems for the sustained release of different types of drugs, ranging from small molecules to biologicals. Aiming at subcutaneous injection, which is a desirable parenteral administration route, especially for biologicals, we herein focus on physically crosslinked systems possessing thixotropic behaviour. The purpose of this study was to evaluate the in vitro and in vivo properties of hydrogels based on the amphipathic hexapeptide H-FQFQFK-NH2, which served as the lead sequence. Upon doubling the length of this peptide, the dodecapeptide H-FQFQFKFQFQFK-NH2 gave a significant improvement in terms of in vivo stability of the hydrogel post-injection, as monitored by nuclear SPECT/CT imaging. This increased hydrogel stability also led to a more prolonged in vivo release of encapsulated peptide cargoes. Even though no direct link with the mechanical properties of the hydrogels before injection could be made, an important effect of the subcutaneous medium was noticed on the rheological properties of the hydrogels in post in vivo injection measurements. The results were validated in vivo for a therapeutically relevant analgesic peptide using the hot-plate test as an acute pain model. It was confirmed that elongation of the hydrogelator sequence induced more extended antinociceptive effects. Altogether, this simple structural modification of the hydrogelating peptide could provide a basis for reaching longer durations of action upon use of these soft biomaterials.

Hydrogen-Bond-Assisted Diels-Alder Kinetics or Self-Healing in Reversible Polymer Networks? A Combined Experimental and Theoretical Study.

Diels-Alder (DA) cycloadditions in reversible polymer networks are important for designing sustainable materials with self-healing properties. In this study, the DA kinetics of hydroxyl-substituted bis- and tetrafunctional furans with bis- and tris-functional maleimides, both containing ether-functionalized spacers, is investigated by modelling two equilibria representing the endo and exo cycloadduct formation. Concretely, the potential catalysis of the DA reaction through hydrogen bonding between hydroxyl of the furans and carbonyl of the maleimides or ether of the spacers is experimentally and theoretically scrutinized. Initial reaction rates and forward DA rate constants are determined by microcalorimetry at 20 °C for a model series of reversible networks, extended with (i) a hydroxyl-free network and hydroxyl-free linear or branched systems, and (ii) polypropylene glycol additives, increasing the hydroxyl concentration. A computational density-functional theory study is carried out on the endo and exo cycloadditions of furan and maleimide derivatives, representative for the experimental ones, in the absence and presence of ethylene glycol as additive. Additionally, an ester-substituted furan was investigated as a hydroxyl-free system for comparison. Experiment and theory indicate that the catalytic effect of H-bonding is absent or very limited. While increased concentration of H-bonding could in theory catalyze the DA reaction, the experimental results rule out this supposition.

Self-Healing in Mobility-Restricted Conditions Maintaining Mechanical Robustness: Furan-Maleimide Diels-Alder Cycloadditions in Polymer Networks for Ambient Applications.

Two reversible polymer networks, based on Diels-Alder cycloadditions, are selected to discuss the opportunities of mobility-controlled self-healing in ambient conditions for which information is lacking in literature. The main methods for this study are (modulated temperature) differential scanning calorimetry, microcalorimetry, dynamic rheometry, dynamic mechanical analysis, and kinetic simulations. The reversible network 3M-3F630 is chosen to study the conceptual aspects of diffusion-controlled Diels-Alder reactions from 20 to 65 °C. Network formation by gelation is proven and above 30 °C gelled glasses are formed, while cure below 30 °C gives ungelled glasses. The slow progress of Diels-Alder reactions in mobility-restricted conditions is proven by the further increase of the system's glass transition temperature by 24 °C beyond the cure temperature of 20 °C. These findings are employed in the reversible network 3M-F375PMA, which is UV-polymerized, starting from a Diels-Alder methacrylate pre-polymer. Self-healing of microcracks in diffusion-controlled conditions is demonstrated at 20 °C. De-gelation measurements show the structural integrity of both networks up to at least 150 °C. Moreover, mechanical robustness in 3M-F375PMA is maintained by the poly(methacrylate) chains to at least 120 °C. The self-healing capacity is simulated in an ambient temperature window between -40 and 85 °C, supporting its applicability as self-healing encapsulant in photovoltaics.