Matrix-tuned hyaluronic nanofibrils promoting regenerative ocular surfacing with minimal scarring

Abstract Corneal epithelial damage remains a considerable clinical challenge, often leading to delayed wound healing and impaired vision. Hyaluronic acid (HA)-based nanofibers face fabrication and stability challenges, reducing their translational potential for corneal regeneration. The fabrication and characterization of aminolyzed polycaprolactone (PCL) nanofibrils (aNFs) with manipulated HA conjugation was investigated to enhance human corneal epithelial cell (hCEC) cell adhesion and proliferation. HA was chemically immobilized on aNF via aza -Michael addition using varying FeCl ₃ concentrations over 24 h. Quantitative, turbidimetric, and colorimetric analyses confirmed HA conjugation to aNFs after 24 h of reaction. Additionally, HA-conjugated NFs exhibited increased swelling and attenuated protein adsorption compared to aNFs. HA-conjugated nanofibrils (HA@NF) exhibited no detectable cytotoxicity toward hCECs. hCECs adhesion gradually increased proportionally with the degree of HA conjugation on HA@NFs. HA@NF24 induced the highest gene expression of hCEC-specific and proliferation markers, suggesting improved identity maintenance and hCEC proliferation in 3D cell sheet cultivation. Furthermore, in vivo corneal wound healing studies using a rabbit model demonstrated that topically applied HA@NFs not only accelerated epithelial defect closure but also ensured high-quality, scarless regeneration by effectively suppressing inflammation and fibrotic remodeling. Collectively, these results demonstrate that HA-optimized PCL nanofibrils are promising biofunctional scaffolds for superior corneal regeneration and ocular tissue repair.