May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Epithelial and keratocyte cell kinetics in human corneas in vitro with corneal refractive surgery (PRK, LASEK& LASIK) as the wound healing model
Author Affiliations & Notes
  • M. Soundararajan
    Dept of Academic Ophthalmology, Rayne Institute –St Thomas Hospital, London, United Kingdom
  • W. Watters
    Dept of Academic Ophthalmology, Rayne Institute –St Thomas Hospital, London, United Kingdom
  • J. Marshall
    Dept of Academic Ophthalmology, Rayne Institute –St Thomas Hospital, London, United Kingdom
  • Footnotes
    Commercial Relationships  M. Soundararajan, None; W. Watters, None; J. Marshall, None.
  • Footnotes
    Support  IRIS Fund, London, U.K.
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 4880. doi:
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      M. Soundararajan, W. Watters, J. Marshall; Epithelial and keratocyte cell kinetics in human corneas in vitro with corneal refractive surgery (PRK, LASEK& LASIK) as the wound healing model . Invest. Ophthalmol. Vis. Sci. 2004;45(13):4880.

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

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Abstract

Abstract: : Purpose: To develop an in vitro human corneal model to investigate differential wound healing responses following PRK, LASEK and LASIK and to study the effect of modulating factors on corneal wound healing. Methods: Human donor corneas were mounted on specialized acrylic corneal holders and were placed in an air–interface organ culture. The epithelial, keratocyte and endothelial cell viability were assessed in vitro in corneas that acted as controls. Wound healing responses in the model following a – 6 D myopic PRK, LASEK and LASIK were compared. Epithelial regeneration and keratocyte kinetics were studied using serial digital imaging and sequential imaging by confocal microscopy respectively for up to 4 weeks. The effect of Mitomycin C (0.01%, 0.02%, 0.04%, 0.06%, 0.08%, 0.1%) on stromal–epithelial interactions following PRK was further analysed in this study Results: Epithelial regeneration was completed by four days following PRK as compared to six days in LASEK. Epithelial latency was significantly increased in LASEK, which was further delayed by the use of 20% alcohol. Thereafter the epithelial flap got fully replaced by regenerating epithelium from the periphery. In LASEK, keratocyte repopulation of the anterior stroma was significantly delayed (18 – 20 days) when compared to PRK (12–14 days) that correlated with delayed re–epithelialisation. The keratocyte density of the ablated stroma was significantly elevated following PRK and LASEK (37065 +/– 3989 cells/mm3) in comparison to LASIK (17,872 +/– 4512 cells/mm3). Mitomycin C had a dose dependant effect on epithelial latency, epithelial differentiation and keratocyte repopulation at 0.01 and 0.02%. MMC Concentrations greater than 0.02% led to significantly delayed re–epithelialisation and reduced keratocyte density. Conclusion: Keratocyte activation following excimer laser injury demonstrates a temporal and spatial relationship with the regenerating epithelium that could potentially explain the differential wound healing responses following PRK, LASEK and LASIK. The in vitro human corneal model is effective in simulating in vivo wound healing responses following refractive surgery. It avoids unnecessary animal experimentation and inter–species bias while testing newer strategies and wound healing modulators to improve predictability and safety of refractive surgical procedures.

Keywords: cornea: stroma and keratocytes • refractive surgery 
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