3D-printed artificial cornea featuring aligned fibrous structure and enhanced mechanical strength

The global shortage of donor eye bank tissue has significantly impeded advancements in biomaterial for corneal implantation. To address this issue, we have developed a three-dimensional (3D)-printed artificial cornea using a composite scaffolds of sodium alginate (SA) and cellulose nanofibers (CNF), crosslinked with poly-L-lysine-co-L-glutamic acid (PLL₈₀GA₂₀, PG) and calcium chloride (CaCl₂, CC). The 2 wt% SA/CNF composite scaffolds offer several advantages, including low toxicity, cost-effectiveness, excellent printability, and high mechanical strength, even with low crosslinker concentrations. The PG was synthesized via ring-opening polymerization of L-glutamate N-carboxyl anhydride (BGNCA) and L-lysine N-carboxyl anhydride (CBZNCA). The purity of the monomers was verified through differential scanning calorimetry analysis, revealing a melting point of 97°C. The molecular weight of the synthesized PG was determined to be 47 kDa. A dual crosslinking strategy was employed, starting with electrostatic crosslinking, followed by ionic crosslinking using varying concentrations of PG and CC at different effective charge concentrations of 6.25 12.25, and 25 mM.The hydrogel and 3D-printed cornea were comprehensively evaluated for chemical structure, surface functional groups, water content, mechanical strength, orientation, cytotoxicity, biocompatibility, and transparency. Notably, the inclusion of PG significantly enhanced the mechanical properties of the 3D-printed cornea, with the hydrogel achieving a storage modulus of 2,360 kPa at 6.25 mM of PG/CC, while maintaining over 95% water content. The artificial cornea demonstrated 86% transparency, and the cell viability showed 96% viable on day 7 with degradation rate of 35.9% in 28 days. The superior hydrophilicity, transparency, and mechanical strength of the printed scaffolds highlights their potential for the development of full-thickness corneal structures, making them a promising candidate for future corneal implants.