Comparative Study of Physicochemical Properties on Corneal Stromal Lenticules Following Four Decellularization Methods.

Abstract

This study compares the physicochemical properties of corneal stromal lenticules following decellularization via four methods. Human corneal stromal lenticules, derived from small incision lenticule extraction surgery, underwent decellularization with sodium dodecyl sulfate (SDS), Triton X-100 (Tx) combined with SDS, trypsin-ethylenediaminetetraacetic acid (TE), or NaCl with deoxyribonuclease (DNase), respectively. Lenticule DNA and glycosaminoglycan (GAG) contents, immunofluorescence staining intensity of cell nuclei and collagen, transparency, biomechanics, histological structure, and immunogenicity were examined in each group and compared with fresh lenticules. All decellularized groups exhibited effective cell removal, with no significant decrease in GAG contents (all P > 0.05). DNA contents decreased in all decellularization groups (all P < 0.01), most notably in the SDS and Tx+SDS groups. Additionally, collagen I and IV fluorescence intensity was reduced in the TE group only (P < 0.0001). Histological staining revealed close similarity in collagen arrangement between the Tx+SDS group and fresh lenticules. Collagen fiber density increased while spacing and diameter decreased in all decellularized groups (all P < 0.05), with partial collagen degradation in the TE group. Light transmittance remained above 60% in the visible light spectrum in all groups. The Young's modulus or elastic modulus did not decrease significantly among decellularized lenticules (all P > 0.05). Human leukocyte antigen (HLA)-DR, HLA-ABC, and CD45 expression decreased in the Tx+SDS and NaCl+DNase groups (all P < 0.001). Although all four decellularization methods showed varying decellularization efficacy, Tx+SDS effectively removed cells without damaging corneal morphology, extracellular matrix, or biomechanics, indicating its potential for lenticule storage, transplantation, and bio-scaffold fabrication.