This study aimed to investigate the effectiveness of two-photon induced collagen cross-linking (CXL) using femtosecond lasers in human corneal stroma. An 800-nm femtosecond laser optical path for CXL was established. Corneal samples that received two-photon induced CXL and ultraviolet-A (UVA) CXL underwent uniaxial stretching experiments, proteolytic resistance assays and observation of collagen fiber structure changes. Two-photon induced CXL can achieve corneal stiffening effects comparable to UVA CXL and showed better advantages at low strains. The cornea after two-photon induced CXL exhibited high enzymatic resistance and tight collagen fiber arrangement. Two-photon induced CXL promises to be a new option for keratoconus.
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