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Effects of daunorubicin, mitomycin C, azathioprine and cyclosporin A on human retinal pigmented epithelial, corneal endothelial and conjunctival cell lines

  • Laboratory Investigation
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Abstract

Background

We wished to investigate the toxicity of four immunosuppressant and antimetabolic drugs, which are known to influence postoperative wound healing, on three different human ocular cell lines.

Methods

Acute toxicity to cyclosporin A, azathioprine, mitomicyn C and daunorubicin was assessed in Chang cells by monitoring their uptake of propidium iodide during a 3-h period. Chronic toxicity was assessed by monitoring the proliferation and viability of subconfluent cultures of Chang cells, human corneal endothelial cells (HCECs) and retinal pigmented epithelial (RPE) cells after continuous exposure to the drugs for 7 days.

Results

Acute toxicity testing revealed no obvious effects. However, the chronic toxicity tests disclosed a narrow concentration range over which cell proliferation decreased dramatically but calcein metabolism was sustained. Although the three lines reacted similarly to each agent, HCECs were the most vulnerable to daunorubicin and mitomycin. At a daunorubicin concentration of 0.05 μg/ml, a 75% decrease in calcein metabolism (P<0.001) and a ≥95% cell loss (P<0.001) were observed. At a mitomycin concentration of 0.01 μg/ml, cell density decreased by 61% (P<0.001) without a change in calcein metabolism, but at 0.1 μg/ml, the latter parameter decreased to 12% (P=0.00014). At this concentration the proliferation of Chang and RPE cells decreased by more than 50%, whilst calcein metabolism was largely sustained. Cyclosporin inhibited cell proliferation moderately at lower concentrations (<5 μg/ml; P=0.05) and substantially at higher ones, with a corresponding decline in calcein metabolism. Azathioprine induced a profound decrease in both parameters at concentrations above 5 μg/ml.

Conclusion

Daunorubicin, cyclosporin and azathioprine could be used to inhibit excessive intraocular scarring after glaucoma and vitreoretinal surgery without overly reducing cell viability. The attributes of immunosuppressants lie in their combined antiproliferative and immunomodulatory effects.

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References

  1. Ben-Simon GJ, Glovinsky Y (2003) Needle revision of failed filtering blebs augmented with subconjunctival injection of mitomycin C. Ophthalmic Surg Lasers 34:94–99

    Google Scholar 

  2. Berger AS, Cheng CK, Pearson A, Ashton P, Crooks PA, Cynkowski T, Cynkowska G, Jaffe GJ (1996) Intravitreal sustained release corticosteroid–5-fluorouracil conjugate in the treatment of experimental proliferative vitreoretinopathy. Investig Ophthalmol Vis Sci 37:2318–2325

    CAS  Google Scholar 

  3. Bindlish R, Condon GP, Schlosser JD, D'Antonio J, Lauer KB, Lehrer R (2002) Efficacy and safety of mitomycin-C in primary trabeculectomy. Ophthalmology 109:1336–1341

    Article  PubMed  Google Scholar 

  4. Blumenkranz M, Hernandez E, Ophir A, Norton EWD (1984) 5-Fluorouracil: new applications in complicated retinal detachment for an established antimetabolite. Ophthalmology 91:122–129

    PubMed  CAS  Google Scholar 

  5. Daniels JT, Occleston NL, Crowston JG, Khaw PT (1999) Effects of antimetabolite induced cellular growth arrest on fibroblast–fibroblast interactions. Exp Eye Res 69:117–127

    Article  PubMed  CAS  Google Scholar 

  6. Hui YN, Liang HC, Cai AS, Kirchhof B, Heidemann K (1993) Corticosteroids and daunomycin in the prevention of experimental proliferative vitreoretinopathy induced by macrophages. Graefe Arch Clin Exp Ophthalmol 231:109–114

    Article  CAS  Google Scholar 

  7. Khaw PT, Sherwood MB, MacKay SL, Rossi MJ, Schultz G (1992) Five-minute treatments with fluorouracil, floxuridine, and mitomycin have long-term effects on human Tenon's capsule fibroblasts. Arch Ophthalmol 110:1150–1154

    PubMed  CAS  Google Scholar 

  8. Khaw PT, Doyle JW, Sherwood MB, Grierson I, Schultz G, McGorray S (1993) Prolonged localized tissue effects from 5-minute exposures to fluorouracil and mitomycin C. Arch Ophthalmol 111:263–267

    PubMed  CAS  Google Scholar 

  9. Khawly JA, Saloupis P, Hatchell DL, Machemer R (1991) Daunorubicin treatment in a refined experimental model of proliferative vitreoretinopathy. Graefe Arch Clin Exp Ophthalmol 229:464–467

    Article  CAS  Google Scholar 

  10. Leonardi A, DeFranchis G, Fregona IA, Violato D, Plebani M, Secchi AG (2001) Effects of cyclosporin A on human conjunctival fibroblasts. Arch Ophthalmol 10:1512–1517

    Google Scholar 

  11. Machemer R, Sugita G, Tano Y (1979) Treatment of intraocular proliferations with intravitreal steroids. Trans Am Ophthalmol Soc 77:171–180

    PubMed  CAS  Google Scholar 

  12. Mandal AK, Bagga H, Nutheti R, Gothwal VK, Nanda AK (2003) Trabeculectomy with or without mitomycin-C for paediatric glaucoma in aphakia and pseudophakia following congenital cataract surgery. Eye 17:53–62

    Article  PubMed  CAS  Google Scholar 

  13. Mietz H, Jacobi PC, Krieglstein GK (2002) Intraoperative episcleral versus postoperative topical application of mitomycin C for trabeculectomies. Ophthalmology 109:1343–1349

    Article  PubMed  Google Scholar 

  14. Neri S, Mariani E, Meneghetti A, Cattini L, Facchini A (2001) Calcein-acetyoxymethyl cytotoxicity assay: standardization of a method allowing additional analyses on recovered effector cells and supernatants. Clin Diagn Lab Immunol 8:1131–1135

    Article  PubMed  CAS  Google Scholar 

  15. Nuzzi R, Cerruti A, Finazzo C (1998) Cyclosporine C: a study of wound-healing modulation after trabeculectomy in rabbit. Acta Ophthalmol Scand Suppl 227:48–49

    PubMed  Google Scholar 

  16. Occlston NL, Daniels JT, Tarnuzzer RW, Sethi KK, Alexander RA, Bhattacharya SS, Schultz GS, Khaw PT (1997) Single exposures to antiproliferatives. Investig Ophthalmol Vis Sci 38:1998–2007

    Google Scholar 

  17. Rubinfeld RS, Pfister RR, Stein RM (1992) Serious complications of topical mitomycin-C after pterygium surgery. Ophthalmology 99:1647–1654

    PubMed  CAS  Google Scholar 

  18. Salah-Eldin M, Peyman GA, El-Aswad M, Bandok G, Bahgat MM, Niesman MR (1994) Evaluation of toxicity and efficacy of antineoplastic agents in the prevention of PVR. Int Ophthalmol 18:53–60

    Article  PubMed  CAS  Google Scholar 

  19. Schipper I, Suppelt C, Gebbers JO (1997) Mitomycin C reduces scar formation after excimer laser (193 nm) photorefractive keratectomy in rabbits. Eye 11:649–655

    PubMed  Google Scholar 

  20. Shin DH, Kim YY, Ginda SY, Kim PH, Eliassi-Rad B, Khatana AK, Keole NS (2001) Risk factors for failure of 5-fluorouracil needling revision for failed conjunctival filtration blebs. Am J Ophthalmol 132:875–880

    Article  PubMed  CAS  Google Scholar 

  21. Shin DH, Iskander NG, Ahee JA, Singal IP, Kim C, Hughes GA, Eliassi-Rad B, Kim YY (2002) Long-term filtration and visual field outcomes after primary glaucoma triple procedure with and without mitomycin-C. Ophthalmology 109:1607–1611

    Article  PubMed  Google Scholar 

  22. Suñer IJ, Greenfield DS, Miller MP, Nicolela MT, Palmberg PF (1997) Hypotony maculopathy after filtering surgery with mitomycin C. Ophthalmology 104:207–214

    PubMed  Google Scholar 

  23. Talamo JH, Gollamundi S, Green WR (1991) Modulation of corneal wound healing after excimer laser keratomileusis using topical mitomycin C and steroids. Arch Ophthalmol 109:1141–1146

    PubMed  CAS  Google Scholar 

  24. Tano Y, Sugita G, Abrams G, Machemer R (1989) Inhibition of intraocular proliferation with intravitreal corticosteroids. Am J Ophthalmol 89:131–136

    Google Scholar 

  25. The Fluorouracil Filtering Surgery Study Group (1989) Fluorouracil filtering surgery study one-year follow-up. Am J Ophthalmol 108:625–635

    Google Scholar 

  26. Tung IC, Wu WC, Kao YH, Chang YC, Chung CH, Hu DN (2001) The effect of combined 5-fluorouracil and dexamethasone on cultured human retinal pigment epithelial cells. Kaohsiung J Med Sci 17:524–529

    PubMed  CAS  Google Scholar 

  27. Turaçli E, Gündüz K, Aktan G, Tamer C (1996) A comparative clinical trial of mitomycin C and cyclosporin A in trabeculectomy. Eur J Ophthalmol 6:398–401

    PubMed  Google Scholar 

  28. Ventura AC, Böhnke M (1999) Toxicity of pentoxifylline on monolayers of highly proliferative cells of epithelial origin. J Ocul Pharmacol Ther 15:525–535

    Article  PubMed  CAS  Google Scholar 

  29. Wiedemann P, Lemmen K, Schmiedl R, Heimann K (1987) Intraocular daunorubicin for the treatment and prophylaxis of traumatic proliferative vitreoretinopathy. Am J Ophthalmol 104:10–14

    PubMed  CAS  Google Scholar 

  30. Wiedemann P, Hilgers RD, Bauer P, Heimann K (1998) Adjunctive daunorubicin in the treatment of proliferative vitreoretinopathy: results of a multicenter clinical trial. Am J Ophthalmol 126:550–559

    Article  PubMed  CAS  Google Scholar 

  31. Wu WC, Kao YH, Hu DN (2002) A comparative study of effects of antiproliferative drugs on human retinal pigment epithelial cells in vitro. J Ocul Pharmacol Ther 18:251–264

    Article  PubMed  CAS  Google Scholar 

  32. Yang A, Cardona DL, Barile FA (2002) In vitro cytotoxicity testing with fluorescence-based assays in cultured human lung and dermal cells. Cell Biol Toxicol 18:97–108

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Ophthalmology, Inselspital, University of Bern, 3010, Berne, Switzerland

    Justus G. Garweg, Maria Wegmann-Burns & David Goldblum

  2. Service d'ophtalmologie, Hôpital Cantonal de Fribourg, Fribourg, Switzerland

    Maria Wegmann-Burns

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  1. Justus G. Garweg
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Correspondence to Justus G. Garweg.

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The authors have no financial interest in this study and received no funding.

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Garweg, J.G., Wegmann-Burns, M. & Goldblum, D. Effects of daunorubicin, mitomycin C, azathioprine and cyclosporin A on human retinal pigmented epithelial, corneal endothelial and conjunctival cell lines. Graefe's Arch Clin Exp Ophthalmo 244, 382–389 (2006). https://doi.org/10.1007/s00417-005-0017-4

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