Journal of Molecular Histology

, Volume 42, Issue 1, pp 47–58

Keratinocyte growth factor 1, fibroblast growth factor 2 and 10 in the healing tympanic membrane following perforation in rats

  • Peter L. Santa Maria
  • Sharon L. Redmond
  • Marcus D. Atlas
  • Reza Ghassemifar
Original Paper

Abstract

The aim of this study was to provide a transcriptome profile of Keratinocyte Growth Factor (KGF)-1, Fibroblast Growth Factor (FGF) 2 and FGF10 (KGF2) in the healing rat tympanic membrane (TM) over 7 days and an immunohistochemical account over 14 days following perforation. KGF1, FGF2, and FGF10 play important roles in TM wound healing. The tympanic membranes of rats were perforated and sacrificed at time points over a 14-day period following perforation. The normalized signal intensities and immunohistochemical protein expression patterns at each time point for KGF1, FGF2, and FGF10 are presented. The primary role of both KGF1 and FGF2 appeared to be in the proliferation and migration of keratinocytes. Whereas the role of KGF1 appeared to be exclusively concerned with increased proliferation and migration at the perforation site, the continued expression of FGF2, beyond perforation closure, suggested it has an additional role to play. FGF10 (KGF2), whilst possessing the highest sequence homologous to KGF1, has a different role in TM wound healing. The effect of FGF10 on keratinocytes in wound healing appeared to emanate from the connective tissue layer.

Keywords

KGF FGF Keratinocyte Growth Factor Fibroblast Growth Factor Tympanic Membrane Rat Wound healing Histology Microarray 

References

  1. Braun S, auf dem Keller U, Steiling H, Werner S (2004) Fibroblast growth factors in epithelial repair and cytoprotection. Philos Trans R Soc Lond B Biol Sci 359(1445):753–757PubMedCrossRefGoogle Scholar
  2. Chauvin K, Bratton C, Parkins C (1999) Healing large tympanic membrane perforations using hyaluronic acid, basic fibroblast growth factor, and epidermal growth factor. Otolaryngol Head Neck Surg 121(1):43–47PubMedCrossRefGoogle Scholar
  3. Chole RA, Kodama K (1989) Comparative histology of the tympanic membrane and its relationship to cholesteatoma. Ann Otol Rhinol Laryngol 98(10):761–766PubMedGoogle Scholar
  4. Clymer MA, Schwaber MK, Davidson JM (1996) The effects of keratinocyte growth factor on healing of tympanic membrane perforations. Laryngoscope 106(3 Pt 1):280–285PubMedCrossRefGoogle Scholar
  5. Fina M, Bresnick S, Baird A, Ryan A (1991) Improved healing of tympanic membrane perforations with basic fibroblast growth factor. Growth Factors 5(4):265–272PubMedCrossRefGoogle Scholar
  6. Fina M, Baird A, Ryan A (1993) Direct application of basic fibroblast growth factor improves tympanic membrane perforation healing. Laryngoscope 103(7):804–809PubMedCrossRefGoogle Scholar
  7. Finch PW, Rubin JS, Miki T, Ron D, Aaronson SA (1989) Human KGF is FGF-related with properties of a paracrine effector of epithelial cell growth. Science 245(4919):752–755PubMedCrossRefGoogle Scholar
  8. Friedman NR, Wright CG, Pawlowski KS, Meyerhoff WL (1997) Effect of basic fibroblast growth factor on perforated chinchilla tympanic membranes. Ear Nose Throat J 76(8):559–564PubMedGoogle Scholar
  9. Hom DB (1995) Growth factors in wound healing. Otolaryngol Clin North Am 28(5):933–953PubMedGoogle Scholar
  10. Ishibashi T, Shinogami M, Ishimoto SI, Yoshida K, Kaga K (1998) Induction of KGF, basic FGF, and TGFalpha mRNA expression during healing of experimental TM perforations. Acta Otolaryngol 118(5):701–704PubMedCrossRefGoogle Scholar
  11. Ishimoto S, Ishibashi T (2002) Induction of growth factor expression is reduced during healing of tympanic membrane perforations in glucocorticoid-treated rats. Ann Otol Rhinol Laryngol 111(10):947–953PubMedGoogle Scholar
  12. Ishimoto S, Ishibashi T, Bottaro DP, Kaga K (2002) Direct application of keratinocyte growth factor, basic fibroblast growth factor and transforming growth factor-alpha during healing of tympanic membrane perforation in glucocorticoid-treated rats. Acta Otolaryngol 122(5):468–473PubMedCrossRefGoogle Scholar
  13. Jimenez PA, Rampy MA (1999) Keratinocyte growth factor-2 accelerates wound healing in incisional wounds. J Surg Res 81(2):238–242PubMedCrossRefGoogle Scholar
  14. Kato M, Jackler RK (1996) Repair of chronic tympanic membrane perforations with fibroblast growth factor. Otolaryngol Head Neck Surg 115(6):538–547PubMedCrossRefGoogle Scholar
  15. Lim DJ (1968a) Tympanic membrane. Electron microscopic observation. I: pars tensa. Acta Otolaryngol 66(3):181–198PubMedCrossRefGoogle Scholar
  16. Lim DJ (1968b) Tympanic membrane. II. Pars flaccida. Acta Otolaryngol 66(6):515–532PubMedCrossRefGoogle Scholar
  17. Lim DJ (1995) Structure and function of the tympanic membrane: a review. Acta Otorhinolaryngol Belg 49(2):101–115PubMedGoogle Scholar
  18. Lynch SE, Nixon JC, Colvin RB, Antoniades HN (1987) Role of platelet-derived growth factor in wound healing: synergistic effects with other growth factors. Proc Natl Acad Sci U S A 84(21):7696–7700PubMedCrossRefGoogle Scholar
  19. Ma Y, Zhao H, Zhou X (2002) Topical treatment with growth factors for tympanic membrane perforations: progress towards clinical application. Acta Otolaryngol 122(6):586–599PubMedCrossRefGoogle Scholar
  20. Marchese C, Felici A, Visco V, Lucania G, Igarashi M, Picardo M, Frati L, Torrisi MR (2001) Fibroblast growth factor 10 induces proliferation and differentiation of human primary cultured keratinocytes. J Invest Dermatol 116(4):623–628PubMedCrossRefGoogle Scholar
  21. Mondain M, Ryan A (1993) Histological study of the healing of traumatic tympanic membrane perforation after basic fibroblast growth factor application. Laryngoscope 103(3):312–318PubMedCrossRefGoogle Scholar
  22. Mondain M, Ryan A (1994) Effect of basic fibroblast growth factor on normal tympanic membrane. Am J Otolaryngol 15(5):344–350PubMedCrossRefGoogle Scholar
  23. Mondain M, Ryan A (1995) Epidermal growth factor and basic fibroblast growth factor are induced in guinea-pig tympanic membrane following traumatic perforation. Acta Otolaryngol 115(1):50–54PubMedCrossRefGoogle Scholar
  24. Mondain M, Saffiedine S, Uziel A (1991) Fibroblast growth factor improves the healing of experimental tympanic membrane perforations. Acta Otolaryngol 111(2):337–341PubMedCrossRefGoogle Scholar
  25. Mondain M, Renard N, Ryan A (1995) Neutralizing antibodies against basic fibroblast growth factor influence the healing of traumatic tympanic membrane perforations. ORL J Otorhinolaryngol Relat Spec 57(1):28–32PubMedGoogle Scholar
  26. Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29(9):e45PubMedCrossRefGoogle Scholar
  27. Rasband WS (1997–2004) ImageJ software, National Institutes of Health, Bethesda, MD, USA. http://rsb.info.nih.gov/ij/
  28. Redmond SL, Levin B, Heel KA, Atlas MD, Marano RJ (2010) Phenotypic and genotypic profile of human tympanic membrane derived cultured cells. J Mol Histol. doi:10.1007/s10735-010-9303-5
  29. Santa Maria PL, Redmond SL, Atlas MD, Ghassemifar R (2010a) Histology of the healing tympanic membrane following perforation in rats. Laryngoscope 120(10):2061–2070PubMedCrossRefGoogle Scholar
  30. Santa Maria PL, Redmond SL, Atlas MD, Ghassemifar R (2010b) The role of epidermal growth factor in the healing tympanic membrane following perforation in rats. J Mol Histol 41 (6):309–314PubMedCrossRefGoogle Scholar
  31. Schmidt SH, Hellstrom S (1991) Tympanic-membrane structure–new views. A comparative study. ORL J Otorhinolaryngol Relat Spec 53(1):32–36PubMedGoogle Scholar
  32. Schroeder A, Mueller O, Stocker S, Salowsky R, Leiber M, Gassmann M, Lightfoot S, Menzel W, Granzow M, Ragg T (2006) The RIN: an RNA integrity number for assigning integrity values to RNA measurements. BMC Mol Biol 7(3):3PubMedCrossRefGoogle Scholar
  33. Somers T, Goovaerts G, Schelfhout L, Peeters S, Govaerts PJ, Offeciers E (1998) Growth factors in tympanic membrane perforations. Am J Otol 19(4):428–434PubMedGoogle Scholar
  34. Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 3(7):RESEARCH0034PubMedCrossRefGoogle Scholar
  35. Vrabec JT, Schwaber MK, Davidson JM, Clymer MA (1994) Evaluation of basic fibroblast growth factor in tympanic membrane repair. Laryngoscope 104(9):1059–1064PubMedCrossRefGoogle Scholar
  36. Werner S, Grose R (2003) Regulation of wound healing by growth factors and cytokines. Physiol Rev 83(3):835–870PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Peter L. Santa Maria
    • 1
    • 2
    • 3
  • Sharon L. Redmond
    • 1
    • 2
  • Marcus D. Atlas
    • 1
    • 2
  • Reza Ghassemifar
    • 4
    • 5
  1. 1.Ear Sciences Centre, School of SurgeryThe University of Western AustraliaPerthAustralia
  2. 2.Ear Science Institute AustraliaPerthAustralia
  3. 3.Department of ENT, E Block OutpatientsSir Charles Gairdner HospitalNedlandsAustralia
  4. 4.Department of Haematology, Experimental Haematology Research UnitPathWest Laboratory MedicineNedlandsAustralia
  5. 5.The School of Pathology and Laboratory MedicineThe University of Western AustraliaPerthAustralia

Personalised recommendations