Honey-mimetic Antibacterial ROS in situ forming Hydrogel Wound Dressing

Due to increasing antibiotic resistance, alternative antimicrobial strategies are under exploration, such as the use of plant based products like honey. This study aims to mimic the ability of honey to produce antibacterial Reactive Oxygen Species (ROS) in the form of hydrogen peroxide (H2O2) within the hydrogel. This was achieved using two components found in honey: the glucose oxidase enzyme and glucose, which were incorporated into the quick forming hydrogel. For this study Hyperbranched Polyethylene Glycol Diacrylate (HB PEGDA) was synthesized using Reversible Addition Chain Transfer Fragmentation (RAFT) polymerization. With the mechanism of thiol-ene click chemistry, HB PEGDA is able to form a hydrogel with thiolated Hyaluronic Acid (HA-SH). For a hydrogel with a composition of 5 w/w% HBPEGDA and 1 w/w% HA-SH, an average gelation time of 585 seconds was obtained. For the 10 w/w% HBPEGDA and 1 w/w% HA-SH gelation time was 62 seconds.The study characterized the chemical characteristics of the polymer and the hydrogels' rheological properties, swelling and degradation properties, and enzyme encapsulation properties. The ROS produced with varying glucose and enzyme concentrations at different time points were quantified. These same enzyme and glucose concentrations were then tested for their biocompatibility in the 3T3 mouse fibroblast cell line using an Alamar Blue Assay, and were found to have positive viability. Given its quick forming property, biocompatibility, and ability to continuously produce antibacterial ROS, this type of hydrogel system makes a promising material for wound dressing applications and should be further characterized and studied. This work has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie (Grant Agreement No. 643050)