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Connective tissue growth factor in tear film of the horse: detection, identification and origin

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Abstract

Background

Healing of corneal ulcers in horses is often associated with profound corneal stromal fibrosis and scar formation resulting in visual impairment. Connective tissue growth factor (CTGF) is a fibrogenic cytokine involved in wound healing and scarring. The purpose of this study was to determine whether CTGF was present in the tear fluid of normal horse eyes and the eyes of horses with corneal ulcers in order to evaluate the role of CTGF in corneal wound healing and corneal scar formation.

Methods

Tear fluid samples were collected from 65 eyes of 44 horses; 32 samples from normal eyes, 21 samples from eyes with corneal ulceration, and 12 samples from the unaffected contralateral eyes of horses with ulcers. CTGF levels in the tears were determined by enzyme immunoassay using goat IgG against human CTGF. Antigenetic similarity of human and horse CTGF was established in a bio-equivalence assay. The identity of horse CTGF was confirmed by western blot. Lacrimal and nictitating membrane glands were investigated by immunohistochemistry in the attempt to clarify the origin of tear fluid CTGF.

Results

CTGF was detected in tear film of 23 normal unaffected eyes (72%) and 8 normal contralateral eyes (67%), with the mean CTGF levels (± SEM) being 51.5±19.2 and 13.4±3.9 ng/ml respectively. CTGF was found in 8 eyes with corneal ulcers (38%) with the mean CTGF concentration of 26.3±14.8 ng/ml. Western blot identified the protein detected as CTGF. The identification of CTGF in lacrimal glands suggests a major role of these glands in the presence of CTGF in tears.

Conclusions

CTGF is present in horse tear fluid and derives, at least partly, from the lacrimal gland. Equine CTGF has strong antigenic similarity with human CTGF. Corneal disease leads to a decrease of CTGF concentrations in tears. The possible role of CTGF in the healing process of ocular surface requires further investigation.

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References

  1. Berman MB, Kerza-Kwiatecki AP, Davison PF (1973) Characterization of human corneal collagenase. Exp Eye Res 15:367–373

    Article  CAS  PubMed  Google Scholar 

  2. Berman MB, Kenyon K, Hayashi K, L’Hernault N (1988) The pathogenesis of epithelial defects and stromal ulceration. In: Cavanagh HD (ed) The cornea: transactions of the World Congress on the Cornea III. Raven, New York, pp 35–43

  3. Blalock TD, Duncan MR, Varela JC, Goldstein MH, Tuli SS, Grotendorst GR, Schultz GS (2003) Connective tissue growth factor expression and action in human corneal fibroblast cultures and rat corneas after photorefractive keratectomy. Invest Ophthalmol Vis Sci 44:1879–1887

    Article  PubMed  Google Scholar 

  4. Brooks D (1999) Equine ophthalmology. In: Gelatt KN (ed) Veterinary ophthalmology, 3rd edn. Lippincott Williams & Wilkins, Philadelphia, pp 31–150

  5. Chen MM, Lam A, Abraham JA, Schreiner GF, Joly AH (2000) CTGF expression is induced by TGF-beta in cardiac fibroblasts and cardiac myocytes: a potential role in heart fibrosis. J Mol Cell Cardiol 32:1805–1819

    Article  CAS  PubMed  Google Scholar 

  6. Frazier K, Williams S, Kothapalli D, Klapper H, Grotendorst GR (1996) Stimulation of fibroblast cell growth, matrix production, and granulation tissue formation by connective tissue growth factor. J Invest Dermatol 107:404–411

    Article  CAS  PubMed  Google Scholar 

  7. Gupta A, Monroy D, Ji Z, Yoshino K, Huang A, Pflugfelder SC (1996) Transforming growth factor beta-1 and beta-2 in human tear fluid. Curr Eye Res 15:605–614

    Article  CAS  PubMed  Google Scholar 

  8. Gupta S, Clarkson MR, Duggan J, Brady HR (2000) Connective tissue growth factor: potential role in glomerulosclerosis and tubulointerstitial fibrosis. Kidney Int 58:1389–1399

    Article  CAS  PubMed  Google Scholar 

  9. Igarashi A, Nashiro K, Kikuchi K, Sato S, Ihn H, Fujimoto M (1996) Connective tissue growth factor gene expression in tissue sections from localized scleroderma, keloid, and other fibrotic skin disorders. J Invest Dermatol 106:729–733

    Article  CAS  PubMed  Google Scholar 

  10. Kueppers F (1971) Proteinase inhibitor in human tears. Biochim Biophys Acta (Amst) 229:845–849

    Article  Google Scholar 

  11. Matrisian LM (1990) Metalloproteinases and their inhibitors in matrix remodeling. Trends Genet 6:121–125

    Article  CAS  PubMed  Google Scholar 

  12. Matsubara M, Zieske JD, Fini ME (1991) Mechanism of basement membrane dissolution preceding corneal ulceration. Invest Ophthalmol Vis Sci 32:3221–3237

    CAS  PubMed  Google Scholar 

  13. Sato S, Nagaoka T, Hasegawa M, Tamatani T, Nakanishi T, Takigawa M (2000) Serum levels of connective tissue growth factor are elevated in patients with systemic sclerosis: association with extent of skin sclerosis and severity of pulmonary fibrosis. J Rheumatol 27:149–154

    CAS  PubMed  Google Scholar 

  14. Schultz G, Nasser C, Grant M, Khaw P, Mackay S (1992) Effects of growth factors on corneal wound healing. Acta Ophthalmol 70:60–66

    Article  Google Scholar 

  15. Schultz G, Rotatori DS, Clark W (1991) EGF and TGF-alpha in wound healing and repair. J Cell Biochem 45:346–352

    Article  CAS  PubMed  Google Scholar 

  16. Shi-wen X, Pennington D, Holmes A, Leask A, Bradham D, Beauchamp JR (2000) Autocrine overexpression of CTGF maintains fibrosis: RDA analysis of fibrosis genes in systemic sclerosis. Exp Cell Res 259:213–224

    Article  CAS  PubMed  Google Scholar 

  17. Takehara K (2001) Growth regulation of skin fibroblasts. J Dermatol Sci 24:S70–S77

    Article  Google Scholar 

  18. Tamatani T, Kobayashi H, Tezuka K, Sakamoto S, Suzuki K, Nakanishi T (1998) Establishment of the enzyme-linked immunosorbent assay for connective tissue growth factor (CTGF) and its detection in the sera of biliary atresia. Biochem Biophys Res Commun 251:748–752

    Article  CAS  PubMed  Google Scholar 

  19. Tervo T, Salonen E-M, Vaheri A, Immonen I, van Setten G-B, Himberg JJ, Tarkkanen A (1988) Elevation of tear fluid plasmin in corneal disease. Acta Ophthalmol (Copenh) 66:393–399

    Google Scholar 

  20. Thörig L, Wijngaards G, van Haeringen NJ (1983) Immunological characterization and possible origin of plasminogen activator in human tear fluid. Ophthalmic Res 15:268–276

    Article  PubMed  Google Scholar 

  21. Thörig L, van Haeringen NJ, Wijngaards G (1984) Comparison of enzymes of tears, lacrimal gland fluid and lacrimal gland tissue in rat. Exp Eye Res 38:605–609

    Article  PubMed  Google Scholar 

  22. Tsung PK, Holly FJ (1981) Protease activities in human tears. Curr Eye Res 1:351–355

    Article  CAS  PubMed  Google Scholar 

  23. Twining SS (1994) Regulation of proteolytic activity in tissues. Crit Rev Biochem Mol Biol 29:315–383

    Article  CAS  PubMed  Google Scholar 

  24. Twining SS, Fukuchi T, Yue BY, Wilson PM, Zhou X, Loushin G (1994) Alpha 2-macroglobulin is present in and synthesized by the cornea. Invest Ophthalmol Vis Sci 35:3226–3233

    CAS  PubMed  Google Scholar 

  25. Twining SS, Fukuchi T, Yue BY, Wilson PM, Zhou X (1994) Alpha 1-antichymotrypsin is present in and synthesized by the cornea. Curr Eye Res 13:433–439

    Article  CAS  PubMed  Google Scholar 

  26. Twining SS, Fukuchi T, Yue BY, Wilson PM, Boskovic G (1994) Corneal synthesis of alpha 1-proteinase inhibitor (alpha 1-antitrypsin). Invest Ophthalmol Vis Sci 35:458–462

    CAS  PubMed  Google Scholar 

  27. van Haeringen NJ, Thörig L (1986) Enzymatic composition of tears. In: Holly FJ (ed) The preocular tear film. Dry Eye Institute Inc., Lubbock, TX, pp 522–527

  28. van Setten GB (1990) Epidermal growth factor in human tear fluid: increased release but decreased concentrations during reflex tearing. Curr Eye Res 9:79–83

    Article  Google Scholar 

  29. van Setten GB (1998) Tear film deficiency in dry eye disease: altered presence of growth factors to the ocular surface. In: Ojo seco—dry eye (ed. Juan Murube del Castillo) in: Mesa Redonda: 73 Congress of the Spanish Society of Ophthalmology, Granada, pp105–109

  30. van Setten GB, Tervo T, Tervo K, Tarkkanen (1992) Epidermal growth factor (EGF) in ocular fluids:presence, origin and therapeutical considerations. Acta Ophthalmol 202:54–9

    Google Scholar 

  31. van Setten GB, Macauley S, Humphreys-Beher M, Chegini N, Schultz G (1996) Detection of transforming growth factor-alpha mRNA and protein in rat lacrimal glands and characterization of transforming growth factor-alpha in human tears. Invest Ophthalmol Vis Sci 37:166–73

    Google Scholar 

  32. van Setten GB, Blalock TD, Grotendorst G, Schultz GS (2002) Detection of connective tissue growth factor (CTGF) in human aqueous humor. Ophthalmic Res 34:306–8

    Article  Google Scholar 

  33. van Setten GB, Blalock TD, Grotendorst G, Schultz GS (2003) Detection of connective tissue growth factor (CTGF) in human tear fluid. Acta Ophthalmologica (Scand) 81:51–53

    Google Scholar 

  34. Vesaluoma M, Teppo AM, Gronhagen-Riska C, Tervo T (1997) Release of TGF-beta 1 and VEGF in tears following photorefractive keratectomy. Curr Eye Res 16:19–25

    Article  CAS  PubMed  Google Scholar 

  35. Wang S, Denichilo M, Brubaker C, Hirschberg R (2001) Connective Tissue growth factor in tubulointerstitial injury of diabetic nephropathy.. Kidney Int 60:96–105

    Article  CAS  PubMed  Google Scholar 

  36. Wunderlich K, Senn BC, Reiser P, Pech M, Flammer J, Meyer P (2000) Connective tissue growth factor in retrocorneal membranes and corneal scars. Ophthalmologica 214:341–346

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The generous financial support of the Karolinska Institutes Forskningsfonder, Stockholm, Sweden and of the Synfrämjandets Forskningsfond, Sweden is most gratefully acknowledged. This work was also supported by NIH grant EY05587.

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Authors and Affiliations

  1. Departments of Large and Small Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32601-0126, USA

    F. J. Ollivier, D. E. Brooks, S. E. Andrew, A. M. Komaromy, T. J. Cutler, M. E. Lassaline & M. E. Kallberg

  2. Department of Obstetrics/Gynecology, Institute of Wound Healing, University of Florida, Gainesville, Florida, USA

    G. S. Schultz & T. D. Blalock

  3. St. Eriks Eyes Clinic, Karolinska Institute, Stockholm, Sweden

    G. B. Van Setten

Authors
  1. F. J. Ollivier
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  2. D. E. Brooks
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  3. G. S. Schultz
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  4. T. D. Blalock
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  5. S. E. Andrew
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  7. T. J. Cutler
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  8. M. E. Lassaline
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  9. M. E. Kallberg
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  10. G. B. Van Setten
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Correspondence to D. E. Brooks.

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Ollivier, F.J., Brooks, D.E., Schultz, G.S. et al. Connective tissue growth factor in tear film of the horse: detection, identification and origin. Graefe's Arch Clin Exp Ophthalmol 242, 165–171 (2004). https://doi.org/10.1007/s00417-003-0802-x

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  • DOI: https://doi.org/10.1007/s00417-003-0802-x

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