Design and <i>In Vitro</i> Application of Bioactive Cell-Penetrating Peptides as Effective Therapeutic Agents for Corneal Wound Healing

The management of corneal injuries remains a formidable global health challenge, yet current therapeutic strategies remain supportive rather than actively regenerative, primarily because current clinical standards fail to address the delicate biological tension between rapid re-epithelialization and the risk of permanent fibrotic scarring. To address this unmet need, we report the design, applications, and mechanisms of novel collagen-derived cell-penetrating peptides that function as dual-action bioactive agents. In human corneal epithelial cells, these peptides demonstrated efficient cell-penetrating capabilities and macromolecular cargo delivery. All-atom molecular dynamics simulations revealed that these peptides interact with membrane interfaces through distinct energetic profiles, facilitating the rapid translocation. Beyond their utility as delivery vectors, the peptides exhibited intrinsic, dose-dependent wound healing activity in vitro. Mechanistic investigations revealed that this capacity is driven by targeted transcriptional modulation. Both peptides suppressed fibrotic and matrix-degradation genes while simultaneously upregulating pathways governing migration, epithelial identity, and proliferation. These results suggest that rationally engineered peptides can navigate complex microenvironments to actively enforce tissue integrity. Furthermore, they offer a promising peptide-based strategy that promotes corneal epithelial repair through an intrinsic, nonfibrotic mechanism while offering a potential platform for intracellular cargo delivery.