Enzymatically Cross-Linked Hydrogel Beads Based on a Novel Poly(aspartamide) Derivative

In recent years, hydrogel beads and in situ hydrogels have gained wide attention in various fields such as biomedicine. In this study, 3-(4-hydroxyphenyl) propionic acid (HP) was introduced into the side chain of poly(α,β-[N-(2-hydroxyethyl)-D,L-aspartamide]) (PHEA) to synthesize phenolic hydroxyl-functionalized poly(aspartamide) derivative PHEA-HP with enzyme-catalyzed cross-linking potential. First, the chemical structure of PHEA-HP was characterized by FT-IR, UV and ¹H NMR, and the results of in vitro cytotoxicity against L929 cell line and hemolysis experiment showed that PHEA-HP did not have toxicity to cells (viability > 90%) and had good blood compatibility. Then, rheological measurement confirmed the formation of PHEA-HP-based in situ hydrogel with a high storage modulus (G') around 10⁴ Pa, and the vial-tilting method revealed that the gelation time of PHEA-HP aqueous solution could be tuned in the wide range of 5-260 s by varying the concentrations of hydrogen peroxide (H₂O₂) and horseradish peroxidase (HRP). Finally, hydrogel beads of different diameters containing methylene blue (for easy observation) were prepared using a coaxial needle and syringe pumps, and the effect of the flow rate of the outer phase on the diameters of the hydrogel beads was also investigated. Therefore, PHEA-HP may be a promising and safe poly(aspartamide) derivative that can be used to prepare in situ hydrogels and hydrogel beads for applications closely related to the human body.