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Molecular Medicine

, Volume 17, Issue 5–6, pp 557–567 | Cite as

Validation of the Glaucoma Filtration Surgical Mouse Model for Antifibrotic Drug Evaluation

  • Li-Fong Seet
  • Wing Sum Lee
  • Roseline Su
  • Sharon N Finger
  • Jonathan G Crowston
  • Tina T Wong
Research Article

Abstract

Glaucoma Is a progressive optic neuropathy, which, If left untreated, leads to blindness. The most common and most modifiable risk factor in glaucoma is elevated intraocular pressure (IOP), which can be managed surgically by filtration surgery. The postoperative subconjunctival scarring response, however, remains the major obstacle to achieving long-term surgical success. Antiproliferatives such as mitomycin C are commonly used to prevent postoperative scarring. Efficacy of these agents has been tested extensively on monkey and rabbit models of glaucoma filtration surgery. As these models have inherent limitations, we have developed a model of glaucoma filtration surgery in the mouse. We show, for the first time, that the mouse model typically scarred within 14 d, but when augmented with mitomycin C, more animals maintained lower intraocular pressures for a longer period of time concomitant with prolonged bleb survival to beyond 28 d. The morphology of the blebs following mitomycin C treatment also resembled well-documented clinical observations, thus confirming the validity and clinical relevance of this model. We demonstrate that the antiscarring response to mitomycin C is likely to be due to its effects on conjunctival fibroblast proliferation, apoptosis and collagen deposition and the suppression of inflammation. Indeed, we verified some of these properties on mouse conjunctival fibroblasts cultured in vitro. These data support the suitability of this mouse model for studying the wound healing response in glaucoma filtration surgery, and as a potentially useful tool for the in vivo evaluation of antifibrotic therapeutics in the eye.

Notes

Acknowledgments

We thank Hla Myint Htoon (Singapore Eye Research Institute) for help with the statistical analysis and the Department of Pathology, Yong Loo Lin School of Medicine, National University of Singapore, for help with the polarizing microscopy. This work was supported by the National Research Foundation Council Translational and Clinical Research (TCR) Programme Grant (NMRC/TCR/002-SERI/2008) and a research grant from the National Medical Research Council (NMRC/EDG/0019/2008) to TT Wong.

References

  1. 1.
    Hitchings RA, Grierson I. (1983) Clinico-pathological correlation in eyes with failed fistulizing surgery. Trans. Ophthalmol. Soc. U. K. 103:84–8.PubMedGoogle Scholar
  2. 2.
    Starita RJ, Fellman RL, Spaeth GL. (1985) Shortand long-term effects of postoperative corticosteroids on trabeculectomy. Ophthalmol. 92:938–46.CrossRefGoogle Scholar
  3. 3.
    Roth SM, Spaeth GL, Starita RJ, Birbillis EM, Steinmann WC. (1991) Effects of postoperative corticosteroids on trabeculectomy and the clinical course of glaucoma. The five-year follow up study. Ophthalmic Surg. 22:724–9.PubMedGoogle Scholar
  4. 4.
    The Fluorouracil Filtering Surgery Study Group. (1989) Fluorouracil filtering surgery study one-year follow-up. Am. J. Ophthalmol. 108:625–35.CrossRefGoogle Scholar
  5. 5.
    Bergstrom TJ, Wilkinson WS, Skuta GL, Watnick RL, Elner VM. (1991) The effects of subconjuctival mitomycin C on glaucoma filtration surgery in rabbits. Arch. Ophthalmol. 109:1725–30.CrossRefGoogle Scholar
  6. 6.
    Chabner BA, et al. (2006) Antineoplastic Agents. In: Goodman and Gilman’s The Pharmacological Basis of Therapeutics. 11th ed. Brunton LL (ed.) McGraw-Hill, New York, pp. 1315–403.Google Scholar
  7. 7.
    Jampel HD, Pasquale LR, DiBernardo C. (1992) Hypotony maculopathy following trabeculectomy with mitomycin C. Arch. Ophthalmol. 110:1049–50.CrossRefGoogle Scholar
  8. 8.
    Greenfield DS, et al. (1996) Endophthalmitis after filtering surgery with mitomycin. Arch. Ophthalmol. 114:943–9.CrossRefGoogle Scholar
  9. 9.
    Higginbotham EJ, et al. (1996) Bleb-related endophthalmitis after trabeculectomy with mitomycin C. Ophthalmol. 103:650–6.CrossRefGoogle Scholar
  10. 10.
    Sheridan CM, et al. (1996) Macrophages during fibrosis following scleral fistulising surgery in a rat model. Curr. Eye Res. 15:559–68.CrossRefGoogle Scholar
  11. 11.
    Sherwood MB, Esson DW, Neelakantan A, Samuelson DA. (2004) A new model of glaucoma filtering surgery in the rat. J. Glaucoma. 13:407–12.CrossRefGoogle Scholar
  12. 12.
    Miller MH, Grierson I, Unger WI, Hitchings RA. (1989) Wound healing in an animal model of glaucoma fistulizing surgery in the rabbit. Ophthalmic Surg. 20:350–7.PubMedGoogle Scholar
  13. 13.
    Doyle JW, Sherwood MB, Khaw PT, McGrory S, Smith MF. (1993) Intraoperative 5-fluorouracil for filtration surgery in the rabbit. Invest. Ophthalmol. Vis. Sci. 34:3313–9.PubMedGoogle Scholar
  14. 14.
    Esson DW, et al. (2004) Expression of connective tissue growth factor after glaucoma filtration surgery in a rabbit model. Invest. Ophthalmol. Vis. Sci. 45:485–91.CrossRefGoogle Scholar
  15. 15.
    Wong TT, Mead AL, Khaw PT. (2005) Prolonged antiscarring effects of ilomastat and MMC after experimental glaucoma filtration surgery. Invest. Ophthalmol. Vis. Sci. 46:2018–22.CrossRefGoogle Scholar
  16. 16.
    Hasty B, Heuer DK, Minckler DS. (1990) Primate trabeculectomies with 5-fluorouracil collagen implants. Am. J. Ophthalmol. 109:721–5.CrossRefGoogle Scholar
  17. 17.
    Bair JS, Chen CW. (1997) Trabeculectomy with multiple applications of mitomycin-C in monkeys with experimental glaucoma. J. Ocul. Pharmacol. Ther. 13:115–28.CrossRefGoogle Scholar
  18. 18.
    Mietz H, Chevez-Barrios P, Lieberman MW. (1998) A mouse model to study the wound healing response following filtration surgery. Graefes Arch. Clin. Exp. Ophthalmol. 236:467–75.CrossRefGoogle Scholar
  19. 19.
    Reichel MB, et al. (1998) New model of conjunctival scarring in the mouse eye. Br. J. Ophthalmol. 82:1072–7.CrossRefGoogle Scholar
  20. 20.
    Haddadin RI, et al. (2009) SPARC-null mice exhibit lower intraocular pressures. Invest. Ophthalmol. Vis. Sci. 50:3771–7.CrossRefGoogle Scholar
  21. 21.
    Seet LF, et al. (2010) SPARC deficiency results in improved surgical survival in a novel mouse model of glaucoma filtration surgery. PLoS One. 5:e9415.CrossRefGoogle Scholar
  22. 22.
    Junqueira LCU, Bignolas G, Brentani RR. (1979) Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections. Histochem. J. 11:447–55.CrossRefGoogle Scholar
  23. 23.
    Goldberg I. (2003) Relationship between intraocular pressure and preservation of visual field in glaucoma. Surv. Ophthalmol. 48(Suppl 1):S3–7.CrossRefGoogle Scholar
  24. 24.
    Labbé A, Dupas B, Hamard P, Baudouin C. (2005) In vivo confocal microscopy study of blebs after filtering surgery. Ophthalmol. 112:1979–86.CrossRefGoogle Scholar
  25. 25.
    Gerdes J, et al. (1984) Cell cycle analysis of a cell proliferation-associated human nuclear antigen defined by the monoclonal antibody Ki-67. J. Immunol. 133:1710–5.PubMedGoogle Scholar
  26. 26.
    Hermiston ML, Xu Z, Weiss A. (2003) CD45: a critical regulator of signaling thresholds in immune cells. Annu. Rev. Immunol. 21:107–37.CrossRefGoogle Scholar
  27. 27.
    Ross GD, Vetvicka V. (1993) CR3 (CD11b, CD18): a phagocyte and NK cell membrane receptor with multiple ligand specificities and functions. Clin. Exp. Immunol. 92:181–4.CrossRefGoogle Scholar
  28. 28.
    Darby I, Skalli O, Gabbiani G. (1990) Alpha-smooth muscle actin is transiently expressed by myofibroblasts during experimental wound healing. Lab. Invest. 63:21–9.PubMedGoogle Scholar
  29. 29.
    Hsu WC, Spilker MH, Yannas IV, Rubin PAD. (2000) Inhibition of conjunctival scarring and contraction by a porous collagen-glycosamino-glycan implant. Invest. Ophthalmol. Vis. Sci. 41:2404–11.PubMedGoogle Scholar
  30. 30.
    Reber F, Gersch U, Funk RW. (2003) Blockers of carbonic anhydrase can cause increase of retinal capillary diameter, decrease of extracellular and increase of intracellular pH in rat retinal organ culture. Graefe’s Arch. Clin. Exp. Ophthalmol. 241:140–8.CrossRefGoogle Scholar
  31. 31.
    Scheuermann T, Sun Y, Sagner S, Rueger B. (2009) Gene expression analysis of paclitaxel-treated HT29 cells using the xCELLigence™ System and RealTime ready™ Panels. Biotechniques. 46:557–8.CrossRefGoogle Scholar
  32. 32.
    Kim TH, et al. (2008) Co-treatment of suberoy-lanilide hydroxamic acid and mitomycin-C induces the apoptosis of rabbit tenon’s capsule fibroblast and improves the outcome of glaucoma filtration surgery. Curr. Eye Res. 33:237–45.CrossRefGoogle Scholar
  33. 33.
    Okada K, et al. (2009) Effects of mitomycin C on the expression of chymase and mast cells in the conjunctival scar of a monkey trabeculectomy model. Mol. Vis. 15:2029–36.PubMedPubMedCentralGoogle Scholar
  34. 34.
    Mietz H, Arnold G, Kirchhof B, Diestelhorst M, Krieglstein GK. (1996) Histopathology of episcleral fibrosis after trabeculectomy with and without mitomycin C. Graefes Arch. Clin. Exp. Ophthalmol. 234:364–8.CrossRefGoogle Scholar
  35. 35.
    Jampel HD. (1992) Effect of brief exposure to mitomycin C on viability and proliferation of cultured human Tenon’s capsule fibroblasts. Ophthalmol. 99:1471–6.CrossRefGoogle Scholar
  36. 36.
    Khaw PT, Sherwood MB, Mackay SLD, Rossi MJ, Schultz G. (1992) Five minute treatments with fluorouracil, floxuidine, mitomycin have long-term effects on human Tenon’s capsule fibroblasts. Arch. Ophthalmol. 110:1150–4.CrossRefGoogle Scholar
  37. 37.
    Smith S, D’Amore PA, Dreyer EB. (1994) Comparative toxicity of mitomycin C and 5-fluo-rouracil in vitro. Am. J. Ophthalmol. 118:332–7.CrossRefGoogle Scholar
  38. 38.
    Crowston JG, et al. (1998) Antimetabolite-induced apoptosis in Tenon’s capsule fibroblasts. Invest. Ophthalmol. Vis. Sci. 39:449–54.PubMedGoogle Scholar
  39. 39.
    Khaw PT, et al. (1993) Prolonged localized tissue effects from 5-minute exposure to fluorouracil and mitomycin C. Arch. Ophthalmol. 111:263–7.CrossRefGoogle Scholar
  40. 40.
    Francis BA, et al. (2005) Histopathologic features of conjunctival filtering blebs. Arch. Ophthalmol. 123:166–70.CrossRefGoogle Scholar
  41. 41.
    Anand N, Arora S, Clowes M. (2006) Mitomycin C augmented glaucoma surgery: evolution of filtering bleb avascularity, transconjunctival oozing, and leaks. Br. J. Ophthalmol. 92:175–80.CrossRefGoogle Scholar

Copyright information

© The Feinstein Institute for Medical Research 2011

Authors and Affiliations

  • Li-Fong Seet
    • 1
  • Wing Sum Lee
    • 1
  • Roseline Su
    • 1
  • Sharon N Finger
    • 1
  • Jonathan G Crowston
    • 2
  • Tina T Wong
    • 1
    • 3
    • 4
    • 5
  1. 1.Ocular Wound Healing and TherapeuticsSingapore Eye Research InstituteSingaporeSingapore
  2. 2.Centre for Eye Research AustraliaRoyal Victorian Eye and Ear HospitalMelbourneAustralia
  3. 3.Glaucoma ServiceSingapore National Eye CenterSingaporeSingapore
  4. 4.Department of Ophthalmology, Yong Loo Lin School of MedicineNational University of SingaporeSingaporeSingapore
  5. 5.School of Materials Science and EngineeringNanyang Technological UniversityNanyangSingapore

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