June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Cross-linking Wound Healing In Animal Models: A Complementary Study Using Two-photon Microscopy And Histological Analysis
Author Affiliations & Notes
  • Carmen Martinez-Garcia
    Cell Biology, University of Valladolid, Valladolid, Spain
  • Francisco J. Avila
    Laboratorio de Optica, Universidad de Murcia, Murcia, Spain
  • Lucía Ibares-Frías
    Oftalmologia, Hospital Clinico Universitario, Valladolid, Spain
  • Raquel Palacios
    Laboratorio de Optica, Universidad de Murcia, Murcia, Spain
  • Patricia Gallego-Muñoz
    Cell Biology, University of Valladolid, Valladolid, Spain
  • Roberto Cantalapiedra
    Cell Biology, University of Valladolid, Valladolid, Spain
  • Juan M Bueno
    Laboratorio de Optica, Universidad de Murcia, Murcia, Spain
  • Footnotes
    Commercial Relationships Carmen Martinez-Garcia, None; Francisco Avila, None; Lucía Ibares-Frías, None; Raquel Palacios, None; Patricia Gallego-Muñoz, None; Roberto Cantalapiedra, None; Juan Bueno, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 1629. doi:
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      Carmen Martinez-Garcia, Francisco J. Avila, Lucía Ibares-Frías, Raquel Palacios, Patricia Gallego-Muñoz, Roberto Cantalapiedra, Juan M Bueno, ; Cross-linking Wound Healing In Animal Models: A Complementary Study Using Two-photon Microscopy And Histological Analysis. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):1629.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: <br /> To analyze and compare the wound healing after cross-linking (CXL) treatment in two animal models (hens and rabbits) using second harmonic generation (SHG) microscopy imaging and histological analysis.

Methods: Monolateral CXL treatment was performed on 10 rabbits and 10 hens. The central 7-mm of the corneal epithelium was removed and 0.125% riboflavin+20% dextran solution was instilled during 30 minutes, then every 5 min during of UVA radiation. The eye was illuminated with UV-light (370 nm, 3 w/cm2) during 30 minutes. Animals were euthanized after 30 days of CXL treatment, the eyes enucleated and the corneas excised. The unstained ex-vivo corneas were imaged using a custom two-photon microscope (Bueno et al., 2010), which allows optical sectioning. Later, a histological analysis was done under bright-field microscopy after hematoxylin-eosin staining. Treated and contralateral corneas were compared using both techniques

Results: SHG images showed sets of collagen bundles running parallel to each other with some orthogonal interweaving. One month after the CXL treatment, central and posterior stromal organization seemed not to be affected by the treatment. However, for the anterior stroma the effects of CXL differed between hens and rabbits. In the former, the collagen distribution recovered its usual arrangement and did not differ from control corneas. In rabbits, this part of the stroma presented changes that remain one month after CXL treatment. Collagen bundles lost their regular orientation and appeared less delineated and less interwoven after CXL compared to the control samples. Histology from hens showed corneas depleted of cells in the anterior stroma and light hipercellularity in the medium stroma. In rabbits, treated corneas were similar to control.

Conclusions: The wound healing after CXL was different in both animal models. SHG imaging and histological analyses provided complementary information on corneal structures: collagen organization and cell distribution respectively. The former revealed that the regular stromal arrangement was recovered faster in hens than in rabbits. The later showed that the cellular repopulation was slower in chickens than in rabbits.

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