Journal Mobile Options
Table of Contents
Vol. 35, No. 6, 2003
Issue release date: November–December 2003
Ophthalmic Res 2003;35:324–328

Corneal Endothelial Cytotoxicity of Riboflavin/UVA Treatment in vitro

Wollensak G. · Spörl E. · Reber F. · Pillunat L. · Funk R.
Departments of aOphthalmology and bAnatomy, Technical University of Dresden, Dresden, Germany

Individual Users: Register with Karger Login Information

Please create your User ID & Password

Contact Information

I have read the Karger Terms and Conditions and agree.

To view the fulltext, please log in

To view the pdf, please log in


Recently, we have developed collagen crosslinking induced by combined riboflavin/UVA treatment, thus increasing the biomechanical rigidity of the cornea to treat progressive keratoconus. The present safety study was performed to evaluate possible cytotoxic effects of combined riboflavin/UVA treatment on the corneal endothelium in vitro. Endothelial cell cultures from porcine corneas were treated with 500 µM riboflavin solution, exposed to various endothelial UVA irradiances (370 nm) ranging from 0.1 to 1.6 mW/cm2 for 30 min and evaluated 24 h later using trypan blue staining and Yopro fluorescence staining. The effect of either treatment alone (UVA irradiation ranging from 0.2 to 6 mW/cm2) was also tested. An abrupt cytotoxic threshold irradiance level was found at 0.35 mW/cm2 after combined treatment with riboflavin plus UVA irradiation and at 4 mW/cm2 with UVA irradiation alone. Riboflavin alone was not toxic. A cytotoxic effect of the combined riboflavin/UVA treatment on corneal endothelial cells is to be expected with a corneal thickness of less than 400 µm. Therefore, pachymetry should be routinely performed before riboflavin/UVA treatment to exclude patients at risk.

Copyright / Drug Dosage

Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.


  1. Spoerl E, Huhle M, Seiler T: Induction of cross-links in corneal tissue. Exp Eye Res 1998;66:97–103.
  2. Spoerl E, Schreiber J, Hellmund K, Seiler T, Knuschke P: Untersuchungen zur Verfestigung der Hornhaut am Kaninchen. Ophthalmologe 2000;97:203–206.
  3. Wollensak G, Spoerl E, Seiler T: Stress-strain measurements of human and porcine cornea after riboflavin/UVA-induced crosslinking. J Cataract Refract Surg, accepted.
  4. Spoerl E, Wollensak G, Seiler T: Increased resistance of crosslinked cornea against enzymatic digestion. Curr Eye Res, submitted.
  5. Wollensak G, Spoerl E, Seiler T: Treatment of keratoconus using riboflavin/UVA-induced crosslinking of the cornea. Am J Ophthalmol 2003;135:620–627.
  6. Schnitzler E, Spoerl E, Seiler T: Bestrahlung der Hornhaut mit UV-Licht und Riboflavingabe als neuer Behandlungsversuch bei einschmelzenden Hornhautprozessen, erste Ergebnisse bei vier Patienten. Klin Monatsbl Augenheilkd 2000;217:190–193.
  7. Seiler T: Iatrogenic keratectasia: Academic anxiety or serious risk (editorial)? J Cataract Refract Surg 1999;25:1307–1308.
  8. Rieck PW, Cholidis S, Hartmann C: Intracellular signaling pathway of FGF-2-modulated corneal endothelial cell migration during wound healing in vitro. Exp Eye Res 2001;73:639–650.
  9. Idziorek T, Estaquier J, De Bels F, Ameisen J-C: YOPRO-1 permits cytofluorometric analysis of programmed cell death (apoptosis) without interfering with cell viability. J Immunol Methods 1995;185:249–258.
  10. Crawford KM, Ernst SA, Meyer RF, MacCallum DK: NaK-ATPase pump sites in cultured bovine corneal endothelium of varying cell density at confluence. Invest Ophthalmol Vis Sci 1995;36:1317–1326.
  11. MacCallum DK, Lillie JH, Scaletta LJ, Occhino JC, Frederick WG, Ledbetter SR: Bovine corneal endothelium in vitro. Elaboration and organization of a basement membrane. Exp Cell Res 1982;139:1–13.
  12. Araie M, Maurice D: The rate of diffusion of fluorophores through the corneal epithelium and stroma. Exp Eye Res 1987;44:73–87.
  13. Cho K-S, Lee EH, Choi J-S, Joo C-K: Reactive oxygen species-induced apoptosis and necrosis on bovine corneal endothelial cells. Invest Ophthalmol Vis Sci 1999;40:911–919.
  14. Olsen EG, Ringvold A: Human cornea endothelium and ultraviolet radiation. Acta Ophthalmol 1982;60:54–56.
  15. Ringvold A, Davanger M, Olsen EG: Changes of the cornea endothelium after ultraviolet radiation. Acta Ophthalmol 1982;60:41–53.
  16. Pitts DG, Cullen AP, Hacker PD: Ocular effects of ultraviolet radiation from 295–365 nm. Invest Ophthalmol Vis Sci 1977;16:932–939.
  17. Podskochy A, Gan L, Fagerholm P: Apoptosis in UV-exposed rabbit corneas. Cornea 2000;19:99–103.
  18. Dollery C.: Therapeutic Drugs. Edinburgh, Churchill Livingstone, 1991, vol 2, pp 24–25.
  19. Kolozsvári L, Nógrádi A, Hopp B, Bor Z: UV absorbance of the human cornea in the 240- to 400-nm range. Invest Ophthalmol Vis Sci 2002;43:2165–2168.
  20. Watters G, Owens H: Evaluation of mild, moderate, and advanced keratoconus using ultrasound pachymetry and the EyeSys videokeratoscope. Optom Vis Sci 1998;75:640–646.

Pay-per-View Options
Direct payment This item at the regular price: USD 38.00
Payment from account With a Karger Pay-per-View account (down payment USD 150) you profit from a special rate for this and other single items.
This item at the discounted price: USD 26.50