In situ forming and self-crosslinkable protein hydrogels for localized cancer therapy and topical wound healing

Proteins, inherently biocompatible and biodegradable, face a challenge in forming stable hydrogels without external chemical crosslinkers. Here, we construct a ring-shaped trimeric SpyTag-fused Proliferating Cell Nuclear Antigen Protein (ST-PCNA) as a core protein building block, and a dumbbell-shaped tandem dimeric SpyCatcher (SC-SC) as a bridging component. Self-crosslinked PCNA/SC-SC Protein (2SP) hydrogels are successfully formed by simply mixing the solutions of ST-PCNA and SC-SC, without chemical crosslinkers. During their formation by mixing, various cargo molecules, including anti-cancer drugs, photosensitizers, and functional proteins, are efficiently incorporated, producing cargo@2SP hydrogels. Most of the entrapped cargo molecules gradually release as the hydrogels erode. Anti-cancer drug- or photosensitizer-incorporated 2SP hydrogels are successfully formed through localized injection beneath the 4 T1 tumor in mice. The localized gradual release of drugs in physiological microenvironment substantially suppresses tumor growth, and highly localized photosensitizers retained in the 2SP hydrogels raises the local temperature above 45 °C upon laser irradiation, resulting in a significant suppression of tumor growth. Additionally, the topical administration of growth factor proteins-incorporated 2SP hydrogels to the incision wound area effectively regenerates the skin, with rapid reconstruction of extracellular matrix. The injectable and self-crosslinkable 2SP hydrogels developed here offer promise as novel biocompatible scaffolds for local therapy.