June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Wound healing in rabbit corneas after flapless refractive lenticule extraction with a novel 345nm ultraviolet femtosecond laser
Author Affiliations & Notes
  • Christian Hammer
    Department of Ophthalmology, University of Erlangen-Nuernberg, Erlangen, Germany
  • Johannes Menzel-Severing
    Department of Ophthalmology, University of Erlangen-Nuernberg, Erlangen, Germany
  • Corinna Petsch
    Department of Ophthalmology, University of Erlangen-Nuernberg, Erlangen, Germany
  • Naresh Polisetti
    Department of Ophthalmology, University of Erlangen-Nuernberg, Erlangen, Germany
  • Bjoern Bachmann
    Department of Ophthalmology, University of Erlangen-Nuernberg, Erlangen, Germany
  • Jörg Klenke
    WaveLight GmbH (Alcon), Erlangen, Germany
  • Katrin Skerl
    WaveLight GmbH (Alcon), Erlangen, Germany
  • Christian Wüllner
    WaveLight GmbH (Alcon), Erlangen, Germany
  • Christof Donitzky
    WaveLight GmbH (Alcon), Erlangen, Germany
  • Friedrich Kruse
    Department of Ophthalmology, University of Erlangen-Nuernberg, Erlangen, Germany
  • Footnotes
    Commercial Relationships Christian Hammer, None; Johannes Menzel-Severing, None; Corinna Petsch, RESORBA Wundversorgung GmbH &Co.KG, Nuremberg, Germany (F); Naresh Polisetti, None; Bjoern Bachmann, None; Jörg Klenke, WaveLight GmbH (E); Katrin Skerl, WaveLight GmbH (E); Christian Wüllner, WaveLight GmbH (E); Christof Donitzky, WaveLight GmbH (E), WaveLIght GmbH (P); Friedrich Kruse, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 3889. doi:
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      Christian Hammer, Johannes Menzel-Severing, Corinna Petsch, Naresh Polisetti, Bjoern Bachmann, Jörg Klenke, Katrin Skerl, Christian Wüllner, Christof Donitzky, Friedrich Kruse; Wound healing in rabbit corneas after flapless refractive lenticule extraction with a novel 345nm ultraviolet femtosecond laser. Invest. Ophthalmol. Vis. Sci. 2013;54(15):3889.

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

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Abstract

Purpose: To preclinically characterize corneal wound healing in New Zealand White (NZW) rabbits after flapless refractive lenticule extraction (FLE) with a novel 345nm ultraviolet femtosecond laser (UV-FSL) developed by Alcon-WaveLight.

Methods: A total of 20 NZW rabbits was included in this study. One eye received FLE while the contralateral eye served as untreated control. The lenticules (-5dpt) had a diameter of 5.5mm and were cut with 80nJ and a spot separation of 4µmx4µm and were removed via two extraction canals. Four animals were sacrificed after 48 hours, 1 week, 2 weeks, 4 weeks and 3 months, respectively. Lenticular beds and extraction canals were prepared for standard histology and fluorescence microscopy. To investigate cell death rate, proliferation, myofibroblastic transdifferentiation of keratocytes and inflammation, TUNEL assay (Roche) as well as immunostaining for Ki67, αSMA and CD11b was performed, respectively, on sagittal sections of kryoembedded specimens and subjected to qualitative and quantitative analysis.

Results: Standard histological analysis revealed a zone of keratocyte depletion of approximately 50µm thickness at the extraction site. Epithelial and endothelial cells appeared unharmed. At the sites of epithelial incision, a callus of epithelial cells formed. At 48h, TUNEL-assay analysis showed pronounced staining of keratocytes near the extraction site (159.9±18.4cells/mm), which steadily decreased to 74.9±19.8cells/mm at 1 week and 5.7±4.8 cells/mm at 2 weeks. At 4 weeks, no TUNEL-positive keratocytes were detected anymore. Marked Ki67 staining of keratocytes was evident at 48h (10.0±3.8 cells/mm), which had decreased conspicuously at one week (5.2±1.7 cells/mm) and 2 weeks (0.4±0.5cells/mm). At 4 weeks, no Ki67 staining was found anymore. The corneal stroma was free of αSMA- and CD11b-positive cells at all times. Only in close proximity to the epithelial callus, some isolated keratocytes were αSMA-positive and very few CD11b-positive cells were observed.

Conclusions: Preclinical in-vivo assessment of corneal wound healing after application of the novel 345nm UV-FSL showed promising results after FLE in NZW rabbits. Together with the benign development of cell death rate and keratocyte proliferation, the absence of inflammation and transdifferentiated keratocytes advocates an initiation of the clinical phase.

Keywords: 479 cornea: clinical science • 578 laser • 765 wound healing  
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