May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Nitric oxide correlates with the quality of human corneas cultured for keratoplasty
Author Affiliations & Notes
  • M.H. Groppe
    Experimental Ophthalmology,
    Eye Bank,
    University Eye Hospital Muenster, Muenster, Germany
  • R. Naskar
    Experimental Ophthalmology,
    University Eye Hospital Muenster, Muenster, Germany
  • C. Harmsen
    Eye Bank,
    University Eye Hospital Muenster, Muenster, Germany
  • P. Vogt
    Eye Bank,
    University Eye Hospital Muenster, Muenster, Germany
  • S. Thanos
    Experimental Ophthalmology,
    Eye Bank,
    University Eye Hospital Muenster, Muenster, Germany
  • Footnotes
    Commercial Relationships  M.H. Groppe, None; R. Naskar, None; C. Harmsen, None; P. Vogt, None; S. Thanos, None.
  • Footnotes
    Support  none
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 4864. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      M.H. Groppe, R. Naskar, C. Harmsen, P. Vogt, S. Thanos; Nitric oxide correlates with the quality of human corneas cultured for keratoplasty . Invest. Ophthalmol. Vis. Sci. 2004;45(13):4864.

      Download citation file:


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

      ×
  • Supplements
Abstract

Abstract: : Purpose: The purpose of this study was to quantify nitric oxide release by corneas under organ culture conditions and to investigate the correlation between nitric oxide release and age, gender, cause of death of the donor and the outcome of organ culture after three days. In addition the expression of nitric oxide synthase (NOS) in corneas was investigated. Methods: Samples of standard corneal culture media were taken at one–day intervals from ten chambers containing a corneal button with an attached scleral rim (14.5 mm) and from 83 chambers at the time of media change after 3, 14 and 24 days of culture. The total amount of nitrite and nitrate in each sample was determined by a spectophotometric method based on the Griess reaction. On day four and the last day of culture the endothelial cell density was documented. Corneas were stained with antibodies against the three isoforms of NOS after 4 and 25 days in culture. Results: The average nitrite and nitrate production of the ten corneas measured daily followed an s–shaped curve with an initial high production until day 4–6, followed by decreased production until day 12–16 and a final marked increase. A mean nitrite and nitrate production of 47.6 µM ± 15.1 µM after 3 days (n=10), 156.5 µM ± 40.6 µM after 13.5 days (n=10) and 309.9 µM ± 35.0 µM after 23 days (n=6) was measured. The average nitrite and nitrate production of 28 corneas, which lacked sufficient endothelial cells for transplantation after three days in culture, was 20.2 µM/day ± 8.5 µM/day compared to 15.8 µM/day ± 5.6 µM/day of 55 corneas suitable for transplantation (p<0.01). Donor's age, gender and cause of death had no significant influence on nitrite and nitrate production of corneas. There was no detectable change in NOS–expression as determined by immunohistochemistry at different time points of culture. Conclusions: The increase in nitrite and nitrate in corneal culture media over time suggests that nitric oxide is released continuously by corneas during cultivation at 36°C. The production rate of nitric oxide seems to influence the quality of the endothelial cell layer. Given the potentialtoxic, free–radical properties of nitric oxide, corneas may benefit from lowering the nitric oxide concentration by introducing a NOS inhibitor into the culture medium.

Keywords: cornea: storage • nitric oxide • cornea: endothelium 
×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×