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Effect of Bevacizumab on Human Tenon's Fibroblasts Cultured from Primary and Recurrent Pterygium

  • Park, Young Min (Department of Pharmacology, Pusan National University College of Medicine, and MRC for Ischemic Tissue Regeneration) ;
  • Kim, Chi Dae (Department of Pharmacology, Pusan National University College of Medicine, and MRC for Ischemic Tissue Regeneration) ;
  • Lee, Jong Soo (Department of Ophthalmology, School of Medicine, Pusan National University & Medical Research Institute, Pusan National University Hospital)
  • Received : 2015.03.24
  • Accepted : 2015.06.02
  • Published : 2015.07.01

Abstract

The purpose of this study was to compare the inhibitory effect of bevacizumab on human Tenon's fibroblasts (HTFs) cultured from primary and recurrent pterygium. Cultured HTFs were exposed to 2.0, 5.0, 7.5, and 15.0 mg/mL concentration of bevacizumab for 24 hours. The 3-[4,5-dimethylthiazol- 2-yl]-2,5-diphenyl tetrazolium bromide and lactate dehydrogenase leakage assays were then performed to assess fibroblast metabolism and viability. The matrix metalloproteinase (MMP), procollagen type I C terminal propeptide (PIP), and laminin immunoassays were performed to examine extracellular matrix production. Changes in cellular morphology were examined by phase-contrast and transmission electron microscopy. Both metabolic activity and viability of primary and recurrent pterygium HTFs were inhibited by bevacizumab in a dose-dependent manner, especially at concentrations greater than 7.5 mg/mL. Both types of HTFs had significant decreases in MMP-1, PIP, and laminin levels. Distinctly, the inhibitory effect of bevacizumab on MMP-1 level related with collagenase in primary pterygium HTFs was significantly higher than that of recurrent pterygium. Significant changes in cellular density and morphology both occurred at bevacizumab concentrations greater than 7.5 mg/mL. Only primary pterygium HTFs had a reduction in cellular density at a bevacizumab concentration of 5.0 mg/mL. Bevacizumab inhibits primary and recurrent pterygium HTFs in a dose-dependent manner, especially at concentrations greater than 7.5 mg/mL. As the primary HTFs produces larger amounts of MMP-1 compared to recurrent HTFs, significant reduction in MMP-1 level in primary pterygium HTFs after exposure to bevacizumab is likely to be related to the faster cellular density changes in primary pterygium HTFs.

Acknowledgement

Supported by : Pusan National University Hospital

References

  1. Cameron ME. Histology of pterygium: an electron microscopic study. Br J Ophthalmol. 1983;67:604-608. https://doi.org/10.1136/bjo.67.9.604
  2. Hill JC, Maske R. Pathogenesis of pterygium. Eye (Lond). 1989;3:218-226. https://doi.org/10.1038/eye.1989.31
  3. Turan-Vural E, Torun-Acar B, Kivanc SA, Acar S. The effect of topical 0.05% cyclosporine on recurrence following pterygium surgery. Clin Ophthalmol. 2011;5:881-885.
  4. Yalcin Tok O, Burcu Nurozler A, Ergun G, Akbas Kocaoglu F, Duman S. Topical cyclosporine A in the prevention of pterygium recurrence. Ophthalmologica. 2008;222:391-396. https://doi.org/10.1159/000151740
  5. Ibanez M, Eugarrios MF, Calderon DI. Topical cyclosporin A and mitomycin C injection as adjunctive therapy for prevention of primary pterygium recurrence. Ophthalmic Surg Lasers Imaging. 2009;40:239-244. https://doi.org/10.3928/15428877-20090430-03
  6. Prabhasawat P, Tesavibul N, Leelapatranura K, Phonjan T. Efficacy of subconjunctival 5-fluorouracil and triamcinolone injection in impending recurrent pterygium. Ophthalmology. 2006;113:1102-1109. https://doi.org/10.1016/j.ophtha.2006.02.026
  7. Hu Q, Qiao Y, Nie X, Cheng X, Ma Y. Bevacizumab in the treatment of pterygium: a meta-analysis. Cornea. 2014;33:154-160. https://doi.org/10.1097/ICO.0000000000000037
  8. Ang LP, Chua JL, Tan DT. Current concepts and techniques in pterygium treatment. Curr Opin Ophthalmol. 2007;18:308-313. https://doi.org/10.1097/ICU.0b013e3281a7ecbb
  9. Paris Fdos S, de Farias CC, Melo GB, Dos Santos MS, Batista JL, Gomes JA. Postoperative subconjunctival corticosteroid injection to prevent pterygium recurrence. Cornea. 2008;27:406-410. https://doi.org/10.1097/ICO.0b013e318162af90
  10. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983;65:55-63. https://doi.org/10.1016/0022-1759(83)90303-4
  11. Jin J, Guan M, Sima J, Gao G, Zhang M, Liu Z, Fant J, Ma JX. Decreased pigment epithelium-derived factor and increased vascular endothelial growth factor levels in pterygia. Cornea. 2003;22:473-477. https://doi.org/10.1097/00003226-200307000-00015
  12. Aspiotis M, Tsanou E, Gorezis S, Ioachim E, Skyrlas A, Stefaniotou M, Malamou-Mitsi V. Angiogenesis in pterygium: study of microvessel density, vascular endothelial growth factor, and thrombospondin-1. Eye (Lond). 2007;21:1095-1101. https://doi.org/10.1038/sj.eye.6702495
  13. Di Girolamo N, Coroneo MT, Wakefield D. Active matrilysin (MMP-7) in human pterygia: potential role in angiogenesis. Invest Ophthalmol Vis Sci. 2001;42:1963-1968.
  14. Lekhanont K, Patarakittam T, Thongphiew P, Suwan-apichon O, Hanutsaha P. Randomized controlled trial of subconjunctival bevacizumab injection in impending recurrent pterygium: a pilot study. Cornea. 2012;31:155-161. https://doi.org/10.1097/ICO.0b013e3182151e0e
  15. Ozgurhan EB, Agca A, Kara N, Yuksel K, Demircan A, Demirok A. Topical application of bevacizumab as an adjunct to recurrent pterygium surgery. Cornea. 2013;32:835-838. https://doi.org/10.1097/ICO.0b013e3182772d4e
  16. van Wijngaarden P, Coster DJ, Williams KA. Inhibitors of ocular neovascularization: promises and potential problems. JAMA. 2005;293:1509-1513. https://doi.org/10.1001/jama.293.12.1509
  17. Chalam KV, Agarwal S, Brar VS, Murthy RK, Sharma RK. Evaluation of cytotoxic effects of bevacizumab on human corneal cells. Cornea. 2009;28:328-333. https://doi.org/10.1097/ICO.0b013e31818b8be0
  18. Yoeruek E, Spitzer MS, Tatar O, Aisenbrey S, Bartz-Schmidt KU, Szurman P. Safety profile of bevacizumab on cultured human corneal cells. Cornea. 2007;26:977-982. https://doi.org/10.1097/ICO.0b013e3180de1d0a
  19. Miguel NC, Matsuda M, Portes AL, Allodi S, Mendez-Otero R, Puntar T, Sholl-Franco A, Krempel PG, Monteiro ML. In vitro effects of bevacizumab treatment on newborn rat retinal cell proliferation, death, and differentiation. Invest Ophthalmol Vis Sci. 2012;53:7904-7911. https://doi.org/10.1167/iovs.12-10283
  20. O'Neill EC, Qin Q, Van Bergen NJ, Connell PP, Vasudevan S, Coote MA, Trounce IA, Wong TT, Crowston JG. Antifibrotic activity of bevacizumab on human Tenon's fibroblasts in vitro. Invest Ophthalmol Vis Sci. 2010;51:6524-6532. https://doi.org/10.1167/iovs.10-5669
  21. Yong VW, Krekoski CA, Forsyth PA, Bell R, Edwards DR. Matrix metalloproteinases and diseases of the CNS. Trends Neurosci. 1998;21:75-80. https://doi.org/10.1016/S0166-2236(97)01169-7
  22. Kahari VM, Saarialho-Kere U. Matrix metalloproteinases and their inhibitors in tumour growth and invasion. Ann Med. 1999;1:34-45.
  23. Dushku N, John MK, Schultz GS, Reid TW. Pterygia pathogenesis: corneal invasion by matrix metalloproteinase expressing altered limbal epithelial basal cells. Arch Ophthalmol. 2001;119:695-706. https://doi.org/10.1001/archopht.119.5.695
  24. Li DQ, Lee SB, Gunja-Smith Z, Liu Y, Solomon A, Meller D, Tseng SC. Overexpression of collagenase (MMP-1) and stromelysin (MMP-3) by pterygium head fibroblasts. Arch Ophthalmol. 2001;119:71-80.
  25. Zeng J, Jiang D, Liu X, Tang L. Expression of matrix metalloproteinase in human pterygia. Yan Ke Xue Bao. 2004;20:242-245.
  26. Di Girolamo N, Wakefield D, Coroneo MT. Differential expression of matrix metalloproteinases and their tissue inhibitors at the advancing pterygium head. Invest Ophthalmol Vis Sci. 2000;41:4142-4149.
  27. Lee JS, Oum BS, Lee SH. Mitomycin c influence on inhibition of cellular proliferation and subsequent synthesis of type I collagen and laminin in primary and recurrent pterygia. Ophthalmic Res. 2001;33:140-146. https://doi.org/10.1159/000055660
  28. Nimmi ME. Fibrillar collagens: their biosynthesis, molecular structure, and mode of assembly. In: Zern MA, Reid LM, editors. Extracellular matrix. New York, NY: Marcel Decker; 1993. p.121-148.
  29. Risteli L, Risteli J. Noninvasive methods for detection of organ fibrosis. In: Rojkind M, editor. Focus on connective tissue in health and disease. Boca Raton, FL: CRC Press; 1990. p.61-68.
  30. Timpl R, Rohde H, Robey PG, Rennard SI, Foidart JM, Martin GR. Laminin--a glycoprotein from basement membranes. J Biol Chem. 1979;254:9933-9937.
  31. Parsian H, Rahimipour A, Nouri M, Somi MH, Qujeq D, Fard MK, Agcheli K. Serum hyaluronic acid and laminin as biomarkers in liver fibrosis. J Gastrointestin Liver Dis. 2010;19:169-174.