Construction Of A Novel Peptide Hrh-Based Nanomaterial And Its Inhibition Of Corneal Neovascularization

This study aims to explore the modification of the HRH peptide sequence to enhance its bioavailability and provide new methods and strategies for the clinical treatment of corneal neovascularization.

Prospective animal study.

NapFFHRH NFs were formed by conjugating HRH with NapFF molecules and self-assembled into nanofibers. Their structure was analyzed by material characterization. In vitro anti-angiogenic activity was assessed via cytotoxicity, proliferation, migration, and tube formation assays, with Western blot detecting p-PI3K and p-AKT in the PI3K-AKT pathway. In vivo, a rat alkali-burn model was used to evaluate anti-neovascularization via neovascularization area, immunofluorescence, and VEGF/α-SMA detection. Biocompatibility was tested by acute ocular reactions, fundus photography, ERG, and hematoxylin staining.

Material characterization confirmed NapFFHRH NFs preparation. Cytotoxicity tests showed no significant toxicity at 400 μM. After 72 hours, cell proliferation decreased to ~60%. Migration was delayed, and tube formation was significantly inhibited. Western blot revealed downregulated p-PI3K and p-AKT in HUVECs. In a rat corneal alkali-burn model, neovascularization length and area were significantly reduced in the NapFFHRH NFs group vs. HRH and NS groups after 14 days. VEGFA and α-SMA expression were also decreased. Biocompatibility tests showed no acute or chronic ocular damage in vitro or in vivo.

The peptide nanofibers NapFFHRH NFs enhanced the stability and surface multivalency of the HRH peptide, effectively improving its bioactivity and therapeutic efficacy for corneal neovascularization. The nanofibers exhibited minimal toxicity and provided a more effective strategy and safer administration method for enhancing the clinical efficacy of various peptide-based drugs.