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Synergistic effects of combined cytotoxic and apoptosis-inducing drugs on Tenon’s capsule fibroblasts in vitro and in vivo

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Abstract

Background

Highly toxic antimetabolites have gained access to routine clinical use to modulate and reduce the amount of postoperative scarring following glaucomatous filtering procedures. It could be speculated that by combining two different antiproliferative substances with different mechanisms of action total amounts of the substances could be decreased and side effects reduced.

Methods

Twenty-two substances were tested that had antiproliferative effects by acting cytotoxically, inhibiting growth factors, or inducing apoptosis. With combinations of each two substances, cell culture experiments using 3T3 and human Tenon’s capsule fibroblasts were performed evaluating cell toxicity, proliferation and migration, the extent of free radicals, and the amount of apoptosis (TUNEL, electron microscopy). The five most potent combinations were used in an animal experiment with rabbits performing filtering procedures. The extent of episcleral scarring was evaluated by histopathology.

Results

The results of the various assays revealed consistently strong effects in 5 of the 462 combinations. Of these five combinations, two were highly effective in the rabbit model. Substances with strong effects when applied in combination included staurosporine, mitomycin, and CD95L.

Conclusions

We found synergistic effects in assays that evaluated different aspects of cell function. The amount of scarring in an animal experiment was inhibited to a level comparable with a high single dose of mitomycin. Combination therapy of two antiproliferative acting substances may be a promising concept.

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References

  1. Addicks EM, Quigley HA, Green WR, Robin AL (1983) Histologic characteristics of filtering blebs in glaucomatous eyes. Arch Ophthalmol 101:795–798

    PubMed  CAS  Google Scholar 

  2. Border WA, Noble NA (1994) Transforming growth factor β in tissue fibrosis. N Engl J Med 331:1286–1292

    Article  PubMed  CAS  Google Scholar 

  3. Buskirk van EM (1982) Cysts of Tenon’s capsule following filtration surgery. Am J Ophthalmol 94:522–527

    Google Scholar 

  4. Cairns JE (1968) Trabeculectomy: preliminary report of a new method. Am J Ophthalmol 66:673–679

    PubMed  CAS  Google Scholar 

  5. Cordeiro MF, Bhattacharya SS, Schultz GS, Khaw PT (2000) TGF-β1, -β2, -β3 in vitro: biphasic effects on Tenon’s fibroblast contraction, proliferation, and migration. Invest Ophthalmol Vis Sci 41:756–763

    PubMed  CAS  Google Scholar 

  6. Cordeiro MF, Reichel MB, Gay JA, D’Esposita F, Alexander RA, Khaw PT (1999) Transforming growth factor-beta1, -beta2, and -beta3 in vivo: effects on normal and mitomycin C-modulated conjunctival scarring. Invest Ophthalmol Vis Sci 40:1975–1982

    PubMed  CAS  Google Scholar 

  7. Costa VP, Spaeth GL, Eiferman RA, Orengo-Nania S (1993) Wound healing modulation in glaucoma filtration surgery. Ophthalmic Surg 24:152–170

    PubMed  CAS  Google Scholar 

  8. Costa VP, Wilson RP, Moster MR, Schmidt CM, Gandham S (1993) Hypotony maculopathy following the use of topical mitomycin C in glaucoma filtration surgery. Ophthalmic Surg 24:389–394

    PubMed  CAS  Google Scholar 

  9. Cunliffe IA, Richardson PS, Rees RC, Rennie IG (1995) Effect of TNF, IL-1, and IL-6 on the proliferation of human Tenon’s capsule fibroblasts in tissue culture. Br J Ophthalmol 79:590–595

    PubMed  CAS  Google Scholar 

  10. DeBry PW, Perkins TW, Heatley G, Kaufman P, Brumback LC (2002) Incidence of late-onset bleb-related complications following trabeculectomy with mitomycin. Arch Ophthalmol 120:297–300

    PubMed  CAS  Google Scholar 

  11. Doxey DL, Ng MC, Dill RE, Iacopino AM (1995) Platelet-derived growth factor levels in wounds of diabetic rats. Life Sci 57:1111–1123

    Article  PubMed  CAS  Google Scholar 

  12. Drance SM, Vargas E (1973) Trabeculectomy and thermosclerostomy: a comparison of two procedures. Can J Ophthalmol 8:413–415

    PubMed  CAS  Google Scholar 

  13. Fankhauser F (1992) Wound healing in glaucoma filtering surgery. Kugler, Amsterdam

    Google Scholar 

  14. Grisanti S, Gralla A, Maurer P, Diestelhorst M, Krieglstein GK, Heimann K (2000) Cellular photoablation to control postoperative fibrosis in filtration surgery: in vitro studies. Exp Eye Res 70:145–152

    Article  PubMed  CAS  Google Scholar 

  15. Grisanti S, Szurman P, Warga M, Kaczmarek R, Ziemssen F, Tatar O, Bartz-Schmidt KU (2005) Decorin modulates wound healing in experimental filtration surgery: a pilot study. Invest Ophthalmol Vis Sci 46:191–196

    Article  PubMed  Google Scholar 

  16. Hitchings RA, Grierson I (1983) Clinicopathological correlation in eyes with failed fistulizing surgery. Trans Ophthalmol Soc U K 103:84–88

    PubMed  Google Scholar 

  17. Hueber A, Esser P, Heimann K, Kociok N, Winter S, Weller M (1998) The topoisomerase I inhibitors, camptothecin and beta-lapachone, induce apoptosis of human retinal pigment epithelial cells. Exp Eye Res 67:525–530

    Article  PubMed  CAS  Google Scholar 

  18. Jampel HD, McGuigan LJB, Dunkelberger GR, L’Herault NL, Quigley HA (1988) Cellular proliferation after experimental glaucoma filtration surgery. Arch Ophthalmol 106:89–94

    PubMed  CAS  Google Scholar 

  19. Kano Y, Suzuki K, Akutsu M, Suda K, Inoue Y, Yoshida M, Sakamoto S, Miura Y (1992) Effects of CPT-11 in combination with other anti-cancer agents in culture. Int J Cancer 20:604–610

    Article  Google Scholar 

  20. Khaw PT, Occleston NL, Schultz G, Grierson I, Sherwood MB, Larkin G (1994) Activation and suppression of fibroblast function. Eye 8:188–195

    PubMed  Google Scholar 

  21. Khaw PT, Sherwood MB, MacKay LD, 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 

  22. Khaw PT, Ward S, Grierson I, Rice NSC (1991) Effect of β radiation on proliferating human Tenon’s capsule fibroblasts. Br J Ophthalmol 75:580–583

    PubMed  CAS  Google Scholar 

  23. Krott R, Lebek J, Grisanti S, Esser P, Heimann K (2000) Antiproliferative effect of genistein on pig retinal pigment epithelial cells in culture. Ophthalmologica 214:296–300

    Article  PubMed  CAS  Google Scholar 

  24. Lee DA, Lee TC, Corres AE, Kitada S (1990) Effects of mithramycin, mitomycin, daunorubicin, and bleomycin on human subconjunctival fibroblast attachment and proliferation. Invest Ophthalmol Vis Sci 31:2136–2144

    PubMed  CAS  Google Scholar 

  25. Lin RY, Sullivan KM, Argenta PA et al (1995) Exogenous transforming growth factor-beta amplifies its own expression and induces scar formation in a model of human fetal skin repair. Ann Surg 222:146–154

    Article  PubMed  CAS  Google Scholar 

  26. Maumenee AE (1960) External filtering operations for glaucoma: the mechanism of function and failure. Trans Am Ophthalmol Soc 58:319–328

    PubMed  CAS  Google Scholar 

  27. Mermoud A, Salmon JF, Murray AND (1993) Trabeculectomy with mitomycin C for refractory glaucoma in blacks. Am J Ophthalmol 116:72–78

    PubMed  CAS  Google Scholar 

  28. Mietz H, Chévas-Barrios P, Feldman RM, Lieberman MW (1998) Suramin inhibits wound healing following filtering procedures for glaucoma. Br J Ophthalmol 82:816–820

    PubMed  CAS  Google Scholar 

  29. Mietz H, Chévaz-Barrios P, Lieberman MW, Wendt M, Gross R, Basinger SF (1997) Decorin and suramin inhibit ocular fibroblast collagen production. Graefes Arch Clin Exp Ophthalmol 235:399–403

    Article  PubMed  CAS  Google Scholar 

  30. Mietz H, Krieglstein GK (1994) Mitomycin C for trabeculectomy in complicated glaucoma: preliminary results after 6 months. Ger J Ophthalmol 3:164–167

    PubMed  CAS  Google Scholar 

  31. Nouri-Mahdavi K, Brigatti L, Weitzman M, Caprioli J (1995) Outcomes of trabeculectomy for primary open-angle glaucoma. Ophthalmology 102:1760–1769

    PubMed  CAS  Google Scholar 

  32. Palmer SS (1991) Mitomycin as adjunct chemotherapy with trabeculectomy. Ophthalmology 98:317–321

    PubMed  CAS  Google Scholar 

  33. Pejnovic N, Lilic D, Zunic G et al (1995) Aberrant levels of cytokines within the healing wound after burn injury. Arch Surg 130:999–1006

    PubMed  CAS  Google Scholar 

  34. Pierce GF, Tarpley JE, Tseng J et al (1995) Detection of platelet-derived growth factor (PDGF)-AA in actively healing human wounds treated with recombinant PDGF-BB and absence of PDGF in chronic nonhealing wounds. J Clin Invest 96:1336–1350

    PubMed  CAS  Google Scholar 

  35. Reber U, Wullner U, Trepel M, Baumgart J, Seyfried J, Klockgether T, Dichgans J, Weller M (1998) Potentiation of treosulfan toxicity by the glutathione-depleting agent buthionine sulfoximine in human malignant glioma cells: the role of bcl-2. Biochem Pharmacol 55:349–359

    Article  PubMed  CAS  Google Scholar 

  36. Rensing-Ehl A, Frei K, Flury R et al (1995) Local Fas/APO-1 (CD95) ligand-mediated cell killing in vivo. Eur J Immunol 25:2253–2258

    Article  PubMed  CAS  Google Scholar 

  37. Schulz JB, Weller M, Klockgether T (1996) Potassium deprivation-induced apoptosis of cerebellar granule neurons: a sequential requirement for new mRNA and protein synthesis, ICE-like protease activity, and reactive oxygen species. J Neurosci 16:4696–4706

    PubMed  CAS  Google Scholar 

  38. Shields MB (1980) Trabeculectomy vs. full-thickness filtering operation for control of glaucoma. Ophthalmic Surg 11:498–505

    PubMed  CAS  Google Scholar 

  39. Siriwardena D, Khaw PT, King AJ, Donaldson ML, Overton BM, Migdal C, Cordeiro MF (2002) Human antitransforming growth factor beta(2) monoclonal antibody—a new modulator of wound healing in trabeculectomy: a randomized placebo controlled clinical study. Ophthalmology 109:427–431

    Article  PubMed  Google Scholar 

  40. Spaeth GL, Joseph NH, Fernandez E (1975) Trabeculectomy: a re-evaluation after three years and a comparison with Scheie’s procedure. Trans Am Acad Ophthalmol Otolaryngol 79:349–361

    Google Scholar 

  41. Sullivan KM, Lorenz HP, Meuli M, Lin RY, Adzick NS (1995) A model of scarless human fetal wound repair is deficient in transforming growth factor beta. J Pediatr Surg 30:198–203

    Article  PubMed  CAS  Google Scholar 

  42. Tahery MM, Lee DA (1989) Pharmacologic control of wound healing in glaucoma filtration surgery. J Ocul Pharmacol 5:155–179

    Article  PubMed  CAS  Google Scholar 

  43. Teng CC, Chi HH, Katzin HM (1959) Histology and mechanism of filtering operations. Am J Ophthalmol 47:16–34

    PubMed  CAS  Google Scholar 

  44. The fluorouracil filtering study group (1996) Five-year follow-up of the Fluorouracil Filtering Surgery Study. The Fluorouracil Filtering Surgery Study Group. Am J Ophthalmol 121:349–366

    Google Scholar 

  45. Thomas DW, O’Neill ID, Harding KG, Shepherd JP (1995) Cutaneous wound healing: a current perspective. J Oral Maxillofac Surg 53:442–447

    Article  PubMed  CAS  Google Scholar 

  46. Watson P (1970) Trabeculectomy. A modified ab externo technique. Ann Ophthalmol 2:199–205

    Google Scholar 

  47. Webb JL (1963) Effect of more than one inhibitor. In: Webb JL (ed) Enzyme and metabolic inhibitors, Vol. 1. Academic, New York, pp 66–79, 487–512

    Google Scholar 

  48. Weller M, Frei K, Groscurth P, Krammer PH, Yonekawa Y, Fontana A (1994) J Clin Invest 94:954–964

    Article  PubMed  CAS  Google Scholar 

  49. Yamamoto T, Varani J, Soong HK, Lichter PR (1990) Effects of 5-fluorouracil and mitomycin C on cultured rabbit subconjunctival fibroblasts. Ophthalmology 97:1204–1210

    PubMed  CAS  Google Scholar 

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Acknowledgement

This study was supported in part by DFG, Bonn, Germany Mi 347/5-1, and Koeln fortune program.

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Authors and Affiliations

  1. Department of Ophthalmology, University of Cologne, Cologne, Germany

    Holger Mietz, Gerhard Welsandt, Arno Hueber, Christina Esser & Günter K. Krieglstein

  2. Eye Clinic Aschaffenburg, Elisenstrasse 32, 63739, Aschaffenburg, Germany

    Holger Mietz

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  1. Holger Mietz
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  2. Gerhard Welsandt
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Correspondence to Holger Mietz.

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Mietz, H., Welsandt, G., Hueber, A. et al. Synergistic effects of combined cytotoxic and apoptosis-inducing drugs on Tenon’s capsule fibroblasts in vitro and in vivo. Graefes Arch Clin Exp Ophthalmol 245, 1367–1375 (2007). https://doi.org/10.1007/s00417-007-0547-z

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  • DOI: https://doi.org/10.1007/s00417-007-0547-z

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