Heparin-incorporated whey protein isolate-derived hydrogels with an intended dual function as snakebite wound dressings and drug delivery systems inhibit spitting cobra venom-induced cytotoxicity

Grey, Matthew and Baines, Daniel and De Castro, Gabrielle and Cobb, Thomas and Tyrrell, Layla and Hyam, Sam and Smith, Alan and Casewell, Nicholas R. and Allinson, Sarah and Douglas, Timothy and Hall, Steve (2026) Heparin-incorporated whey protein isolate-derived hydrogels with an intended dual function as snakebite wound dressings and drug delivery systems inhibit spitting cobra venom-induced cytotoxicity. Toxicon, 275: 109061. ISSN 0041-0101

Text (2026-03-09 - Full revised hydrogel submission - TOXCON-D-25-01026_R1)
2026-03-09_-_Full_revised_hydrogel_submission_-_TOXCON-D-25-01026_R1.pdf - Accepted Version
Download (10MB)

Abstract

Snakebite envenoming affects millions of people annually, with current treatments limited to animal-derived antivenoms. Repurposed drug-inhibitors of toxin families offer an exploitable avenue to improve snakebite treatment, including heparins which can inhibit cytotoxic three-finger toxins. However, to be effective therapies in-the-field such treatments must be engineered into drug delivery devices capable of rapidly administering drug(s) to the envenomation site. Herein we introduce the concept of integrating heparins, specifically unfractionated heparin (H) and its low molecular weight heparinoid variant, tinzaparin (T), into hydrogels composed of whey protein isolate (WPI), an inexpensive byproduct of the dairy industry that is cytocompatible, stiff, sterilizable by autoclaving, and that has the dual function of being able to locally deliver drugs and act as wound dressings. The aims of this research were to investigate whether heparin-containing WPI hydrogels displayed physical characteristics suitable for wound dressings and could effectively release drug in sufficient quantities to inhibit the cytotoxic activity of spitting cobra venom. To do so, five hydrogel prototypes were produced: 40% WPI no drug control, and 5% H, 10% H, 5% T, and 10% T all within 40% WPI hydrogels. It was determined that heparins successfully integrated into WPI hydrogels and heparin-containing WPI hydrogels exhibited improved swelling versus the 40% WPI no drug control, suggesting improved absorption of wound exudate; however, this heparin integration also increased the hydrogel degradation rate in simulated wound environments. Most importantly, sufficiently high concentrations of T were released into phosphate buffered saline from the 10% T hydrogels to neutralise Naja nigricollis venom cytotoxicity in a HaCaT cell cytotoxicity model. Together, our results suggest that the integration and diffusion of venom-inhibiting drugs in WPI hydrogels is possible and that the development of such drug-integrated hydrogels into snakebite wound dressings warrants further research.