Bevacizumab accelerates corneal wound healing by inhibiting TGF-βexpression in alkali-burned mouse cornea

  • Published : 2009.12.31


This study investigated the effect of subconjunctival injections of bevacizumab, an anti-VEGF antibody, on processes involved in corneal wound healing after alkali burn injury. Mice were divided into three groups: Group 1 was the saline-treated control, group 2 received subconjunctival injection of bevacizumab 1hr after injury and group 3 received bevacizumab 1 hr and 4 days after injury. Cornea neovascularization and opacity were observed using a slit lamp microscope. Corneal repair was assessed through histological analysis and immunostaining for CD31, $\alpha$-SMA, collagen I, and TGF-$\beta$2 7 days post-injury. In group 3, injection of bevacizumab significantly lowered neovascularization and improved corneal transparency. Immunostaining analysis demonstrated a reduction in CD31, $\alpha$-SMA and TGF-$\beta$2 levels in stroma compared to group 1. These results indicate that bevacizumab may be useful in reducing neovascularization and improving corneal transparency following corneal alkali burn injury by accelerating regeneration of the basement membrane.


  1. Stoltz, R. A., Conners, M. S., Gerritsen, M. E., Abraham, N. G. and Laniado-Schwartzman, M. (1996) Direct stimulation of limbal microvessel endothelial cell proliferation and capillary formation in vitro by a corneal-derived eicosanoid. Am. J. Pathol. 148, 129-139
  2. Cursiefen, C., Cao, J., Chen, L., Liu, Y., Maruyama, K. Jackson, D., Kruse, F. E., Wiegand, S. J., Dana, M. R. and Streilein, J. W. (2004) Inhibition of hemangiogenesis and lymphangiogenesis after normal-risk corneal ransplantation by neutralizing VEGF promotes graft survival. Invest. Ophthalmol. Vis. Sci. 45, 2666-2673
  3. Casey, R. and Li, W. W. (1997) Factors controlling ocular angiogenesis. Am. J. Ophthalmol. 124, 521-529
  4. Chang, J. H., Gabison, E. E., Kato, T. and Azar, D. T. (2001) Corneal neovascularization. Curr. Opin. Ophthalmol. 12, 242-249
  5. Dawson, D. W., Volpert, O. V., Gillis, P., Crawford, S. E., Xu, H., Benedict, W. and Bouck, N. P. (1999) Pigment epithelium-derived factor: a potent inhibitor of angiogenesis. Science 285, 245-248
  6. Zhang, P., Wu, D., Ge, J., Zhu, Z., Feng, G., Yue, T., Lin, J. and Zheng, H. (2003) Experimental inhibition of corneal neovascularization by endostatin gene transfection in vivo. Chin. Med. J. (Engl.) 116, 1869-1874
  7. Ormerod, L. D., Abelson, M. B. and Kenyon, K. R. (1989) Standard models of corneal injury using alkali-immersed filter discs. Invest. Ophthalmol. Vis. Sci. 30, 2148-2153
  8. Ormerod, L. D., Garsd, A., Reddy, C. V., Gomes, S. A., Abelson, M. B. and Kenyon, K. R. (1989) Dynamics of corneal epithelial healing after an alkali burn. A statistical analysis. Invest. Ophthalmol. Vis. Sci. 30, 1784-1793
  9. Chung, J. H. and Fagerholm, P. (1989) Corneal alkali wound healing in the monkey. Acta. Ophthalmol. (Copenh.) 67, 685-693
  10. Ferrara, N., Hillan, K. J. and Novotny, W. (2005) Bevacizumab (Avastin), a humanized anti-VEGF monoclonal antibody for cancer therapy. Biochem. Biophys. Res. Commun. 333, 328-335
  11. Rosenfeld, P. J., Fung, A. E. and Puliafito, C. A. (2005) Optical coherence tomography findings after an intravitreal injection of bevacizumab (avastin) for macular edema from central retinal vein occlusion. Ophthalmic. Surg. Lasers Imaging 36, 336-339
  12. Kao, W. W., Ebert, J., Kao, C. W., Covington, H. and Cintron, C. (1986) Development of monoclonal antibodies recognizing collagenase from rabbit PMN; the presence of this enzyme in ulcerating corneas. Curr. Eye Res. 5, 801-815
  13. Burns, F. R., Gray, R. D. and Paterson, C. A. (1990) Inhibition of alkali-induced corneal ulceration and perforation by a thiol peptide. Invest. Ophthalmol. Vis. Sci. 31,107-114
  14. Manzano, R. P., Peyman, G. A, Khan, P., Carvounis, P. E., Kivilcim, M., Ren, M., Lake, J. C. and Chevez-Barrios, P. (2007) Inhibition of experimental corneal neovascularisation by bevacizumab (Avastin). Br. J. Ophthalmol. 91, 804-807
  15. Moromizato, Y., Stechschulte, S., Miyamoto, K., Murata, T., Tsujikawa, A., Joussen, A. M. and Adamis, A. P. (2000) CD18 and ICAM-1-dependent corneal neovascularization and inflammation after limbal injury. Am. J. Pathol. 157, 1277-1281
  16. Hosseini, H. and Nejabat, M. (2007) A potential therapeutic strategy for inhibition of corneal neovascularization with new anti-VEGF agents. Med. Hypotheses 68, 799-801
  17. Lee, J. Y. and Lee, K. J. (2008) Histological changes on the wound healing process of alkali burned mouse cornea. J. Korean Oph. Opt. Soc. 13, 161-169
  18. Desmouliere, A., Darby, I. A. and Gabbiani, G. (2003) Normal and pathologic soft tissue remodeling: role of the myofibroblast, with special emphasis on liver and kidney fibrosis. Lab. Invest. 83, 1689-1707
  19. Jester, J. V., Huang, J., Petroll, W. M. and Cavanagh, H. D. (2002) TGFbeta induced myofibroblast differentiation of rabbit keratocytes requires synergistic TGFbeta, PDGF and integrin signaling. Exp. Eye Res. 75, 645-657
  20. Tomasek, J. J., Gabbiani, G., Hinz, B., Chaponnier, C. and Brown, R. A. (2002) Myofibroblasts and mechano-regulation of connective tissue remodelling. Nat. Rev. Mol. Cell Biol. 3, 349-363
  21. Piek, E., Ju, W. J., Heyer, J., Escalante-Alcalde, D., Stewart, C. L., Weinstein, M., Deng, C., Kucherlapati, R., Bottinger, E. P. and Roberts, A. B. (2001) Functional characterization of transforming growth factor beta signaling in Smad2- and Smad3-deficient fibroblasts. J. Biol. Chem. 276, 19945-19953
  22. Evans, R. A., Tian, Y. C., Steadman, R. and Phillips, A. O. (2003) TGF-beta1-mediated fibroblast-myofibroblast terminal differentiation-the role of Smad proteins. Exp. Cell Res. 282, 90-100
  23. Wilson, S. E., Mohan, R. R., Ambrosio, R., Hong, J. and Lee, J. (2001) The corneal wound healing response: cytokine-mediated interaction of the epithelium, stroma, and inflammatory cells. Prog. Retin. Eye Res. 20, 625-637
  24. Stramer, B. M., Zieske, J. D., Jung, J. C., Austin, J. S. and Fini, M. E. (2003) Molecular mechanisms controlling the fibrotic repair phenotype in cornea: implications for surgical outcomes. Invest. Ophthalmol. Vis. Sci. 44, 4237-4246
  25. Bascom, C. C., Wolfshohl, J. R., Coffey, R. J., Madisen, L., Webb, N. R., Purchio, A. R., Derynck, R. and Moses, H. L. (1989) Complex regulation of transforming growth factor beta 1, beta 2, and beta 3 mRNA expression in mouse fibroblasts and keratinocytes by transforming growth factors beta 1 and beta 2. Mol. Cell Biol. 9, 5508-5515
  26. Dwivedi, D. J., Pontoriero, G. F., Ashery-Padan, R., Sullivan, S., Williams, T. and West-Mays, J. A. (2005) Targeted deletion of AP-2alpha leads to disruption in corneal epithelial cell integrity and defects in the corneal stroma. Invest. Ophthalmol. Vis. Sci. 46, 3623-3630
  27. Yoeruek, E., Ziemssen, F., Henke-Fahle, S., Tatar, O., Tura, A., Grisanti, S., Bartz-Schmidt, K. U. and Szurman, P. (2008) Safety, penetration and efficacy of topically applied bevacizumab: evaluation of eyedrops in corneal eovascularization after chemical burn. Acta. Ophthalmol. 86, 322-328

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