Characterization of the lamellar rearrangement induced by cross-linking treatment in keratoconic corneal samples imaged by SHG microscopy

Year: 2017

Authors: Mercatelli R., Ratto F., Tatini F., Rossi F., Menabuoni L., Nicoletti R., Pini R., Pavone F.S., Cicchi R.

Autors Affiliation: Consiglio Nazionale delle Ricerche, Istituto Nazionale di Ottica (INO-CNR) Largo E Fermi 6, I-50125 Florence, Italy; Consiglio Nazionale delle Ricerche, Istituto di Fisica Applicata N. Carrara, Via Madonna del Piano 10, I-50019 Sesto Fiorentino, FI, Italy; Publ Hosp Prato, Dept Ophthalmol, Piazza Osped 5, I-59100 Prato, PO, Italy; CSO Srl, Via Stagnacci 12-E, I-50018 Scandicci, FI, Italy; Univ Florence, European Lab Nonlinear Spect LENS, Via Nello Carrara 1, I-50019 Sesto Fiorentino, Italy

Abstract: Keratoconus is an eye disorder that features a reduced stiffness of the cornea and its consequent pathological deformation. Cross-Linking (CXL) treatment has proven useful in hindering the progression of keratoconus, offering a minimally-invasive alternative to corneal surgical transplantation. In this study, the biomechanical characteristics of a human keratoconic cornea were clinically examined in vivo soon before keratoplasty, and the morphological alterations of the collagen scaffold in the same cornea were examined ex vivo by means of Second-Harmonic Generation (SHG) microscopy. A healthy cornea and a CXL-treated keratoconus were compared. In particular, the lamellar organization in the three corneal samples was characterized in different stromal layers by detecting both forward- and backwardscattered SHG signal and then considering the forward/backward (F/B) ratio as parameter. The F/B ratio was used to characterize the morphological organization of collagen lamellae within different stromal layers, finding an increased disorder at the level of Bowman

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More Information: The research leading to these results has received funding from Tuscany Region and EU FP7 BiophotonicsPlus projects \”LITE\” (Laser Imaging of The Eye) and \”LighTPatcH\” (Led Technology in Photo Haemostasis), from the Italian Ministry for Education, University and Research in the framework of the Flagship Project NANOMAX, from the Italian Ministry of Health (GR-2011-02349626), from the European Union\’s Horizon 2020 research and innovation programme under grant agreement No 654148 Laserlab-Europe and No. 720270 HBP SGA1, from Fondazione Pisa and from Ente Cassa di Risparmio di Firenze.
KeyWords: Collagen; Nonlinear optics; Transplantation (surgical), Biomechanical characteristics; Collagen scaffolds; Fibrillar collagens; Keratoconus; Lamellar organizations; Minimally invasive; Morphological alteration; Second harmonic generation microscopies (SHG), Harmonic generation

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