Abstract
Hypertrophic scarring (HSc) which frequently develops in patients following severe thermal injury is characterized by accumulation of extracellular matrix (ECM) proteins including type I and type III collagen. In this study, we examined the presence and quantity of IGF-1 mRNA transcripts in post-burn HSc. The results of dot blot experiments showed a 77.5% (100±8.15 vs 177.5±19, p<0.01) increase in expression of IGF-1 IIIRNA in HSc tissue relative to normal dermis obtained from the same patients. A Northern blot analysis confirmed the specificity of the IGF-1 cDNA. This cDNA visualized four different transcripts with apparent sizes of 7.0, 3.9, 1.8 and 1.0 kb, similar to those previously reported. The possible fibrogenic role of IGF-1 was examined by analyzing the effect of this growth factor on the expression of mRNA for the pro α1(I) chain of type I procollagen and the pro α1(III) chain of type III procollagen in dermal fibroblasts. IGF-1 increased the expression of these transcripts as early as 6 h and the effect was maximal at 24 h. Quantitative analysis by densitometry showed a 149 and 166% increase in pro α1(I) and pro α1(III) mRNA after 24 h of IGF-1 treatment, respectively. This effect seems to be specific as the abundance of mRNA for the pro α2(I) chain of type I procollagen or TIMP-II was unchanged. When another 4 strains of dermal fibroblasts were treated with IGF-1, a significant increase (16.94±1.13 vs 10.87±1.79, p<0.01, N=4) in the expression of type I procollagen mRNA was found. This was consistent with a significant increase in collagen production, as measured by hydroxyproline in conditioned medium (2.04±0.3 ng/1000 cells vs 1.35±0.4 ng/1000 cells, p<0.01, N=4). The effects of IGF-1 were temporary, since removal of IGF-1 from media caused a reduction in expression of type I procollagen mRNA to its basal level within 48 h.
Enhanced expression of IGF-1 mRNA in post-burn HSc tissues and the potentially fibrogenic effects of this growth factor on dermal fibroblasts, suggest that it could contribute to the accumulation of type I and type III collagen found in many fibroproliferative disorders such as HSc.
Similar content being viewed by others
References
Scott PG, Ghahary A, Chambers MM, Tredget EE: Biological basis of hypertrophic scarring. In: Advances in Structural Biology, JAI Press Inc, Greenwich, Connecticut, 1994, Vol 3: pp 157–202
Bitterman PB, Henke CA: Fibroproliferative disorders. Chest 99: 81s-84s, 1991
Linares HA, Kischer CW, Dobrkovsky K, Larson DL: The histotypic organization of the hypertrophic scar in humans. J Invest Dermatol 59: 323–331, 1972
Ghahary A, Shen YJ, Gong Y, Scott PG, Tredget EE: Enhanced expression of mRNA for transforming growth factor β1, type 1, type III procollagen in human post-burn hypertrophicscar tissues. J Lab Clin Med 122: 465–473, 1993
Ghahary A, Shen YJ, Scott PG, Tredget EE: Expression of fibronectin mRNA in hypertrophic and normal dermal tissues andin vitro regulation by interferon alpha-2b. Wound Repair and Regeneration 1(3): 166–174, 1993
Ghahary A, Scott PG, Malhotra S, Friedland UA, Harrop AR, Forsyth NL, Tredget EE: Differential expression of type I and type III procollagen mRNA in human hypertrophic burn fibroblasts. Biomed Lett 47: 169–184, 1992
Baird A, Mormede P, Bohlen P: Immunoreactive fibroblast growth factor in cells of peritoneal exudate suggest its identity with macrophage derived growth factor. Biochem Biophys Res Comm 126: 358–364, 1985
Shimokado K, Raines EW, Madtes DK, Barrett TB, Benditt AP, Ross R: A significant part of macrophage-derived growth factor consists of at least two forms of PDGF. Cell 43: 277–286, 1985
Assoian RK, Fleurdelys BE, Stevenson HC, Miller PJ, Madtes DK, Raines EW, Ross R, Sporn MB: Expression and secretion of type B transforming growth factor by activated human macrophage. Proc Natl Acad Sci USA 84: 6020–6024, 1987
Rappolee, DA, Mark D, Banda MJ, Werb Z: Wound macrophages express TGF-α and other growth factorsin vivo: analysis of mRNA phenotyping. Science 241: 708–712, 1988
Wahl LM, Whal SM: Inflammation. In: Cohen, Diegelmann, Lindblad (eds). Wound Healing. W.B. Saunders, Philadelphia, PA, 1992, pp 40–53
McGee G, Davidson JM, Buckley A, Sommer A, Woodword SC, Aquino AM, Barbour R, Dermetriou AA: Recombinant basic fibroblast growth factor accelerates wound healing. J Surg Res 45: 145–153, 1988
Schultz GS, White M, Mitchell R, Brown G, Lynch J, Twadzik DR, Todaro GJ, Epithelial wound healing enhanced by transforming growth factor. Science 235: 350–352, 1987
Sporn MB, Roberts AB, Wakefield LM, de Crombrugghe B: Some recent advances in the chemistry and biology of transforming grwoth factor. J Cell Biol 105: 1039–1045, 1987
Lynch SE, Nixon JC, Colvin RB, Antoniades HN: Role of platelet-derived growth factor in wound healing-synergistic effects with other growth factors. Proc Natl Acad Sci USA 84: 7696–7700, 1987
Rossi P, Karsenty G, Roberts AB, Roche NS, Sporn MB, de Crombrugghe B: A nuclear factor 1 binding site mediates the transcriptional activation of a type I collagen promoter by transforming growth factor-β. Cell 52: 405–414, 1988
Roberts AB, Sporn MB, Assoian RK, Smith JM, Roche S, Wakefield LM, Heine UI, Liotta LA, Falanga V, Kehrl JH, Fauci AS: Transforming growth factor B rapid induction of fibrosis and angiogenesis in vivo and stimulation of collagen formationin vitro. Proc Natl Acad Sci USA 83: 4167–4171, 1986
Pierce GF, Mustoe TA, Lingelbach J, Maskakowsi VR, Griffin GL, Senior RM. Deuel TF: Platelet derived-growth factor and transforming growth factor inducein vivo andin vitro tissue repair activities by unique mechanisms. J Cell Biol 429–440, 1989
Morgan CJ, Pledger WJ: Fibroblast proliferation. In: Cohen, Diegelmann, Lindblad (eds). Wound Healing. W.B. Saunders, Philadelphia, PA, 1992, pp 63–73
Rockwell WB, Cohen IK, Ehrlich HP: Keloids and hypertrophic scars comprehensive review. Plast Reconstr Surg 84: 827–837, 1989
Nakano T, Scott PG: Purification and characterization of a gelatinase produced by fibroblasts from human gingiva. Biochem Cell Biol 64: 387–393, 1986
Uitto J, Booth BA, Polak KI: Collagen biosynthesis by human skin fibroblasts: Isolation and further characterization of type I and type III procollagen synthesized in culture. Biochim Biophys Acta 624: 545–561, 1980
Chirgwin JMt Pryzbyla AE, MacDonald RJ, Rutter WJ: Isolation of biologically active ribonucleic acid from sources enriched in ribonucleases. Biochem 18: 5294–5299, 1979
Thomas PS: Hybridization of denatured RNA and small DNA fragments transferred tonitrocellulose. Proc Natl Acad Sci USA 77: 5201–5205, 1980
Tredget EE, Shen Y, Forsyth N, Smith C, Scott PG, Ghahary A: The effect of interferon alpha-2b on hypertrophic scar fibroblastsin vitro. Wound Repair and Regeneration 1(3): 156–165, 1993
Murphy LJ, Bell GJ, Friesen HG: Tissue distribution of insulin-like growth factor-1 and II mRNA in adult rats. Endocrinol 120: 1279–1282, 1987
Sikbergeld A, Lazar L, Erster B, Keret R, Tepper R, Laron Z: Serum growth hormone binding protein activity in healthy neonates, children, and young adults-correlation with age, height and weight. Clin Endocrinol 31: 295–303, 1989
Spencer EM, Skover G, Hunt TK: Growth factors and other agents of wound healing. In: E. Barbul, E. Pines, M. Caldwell, T.K. Hunt (eds): Biological and Clinical Implication. Liss: NY, 1988, pp 103–116
Gartner MH, Benson JD, Caldwell MD: Insulin-like growth factors I and II expressioninthe healingwound. J Surg Res 52: 389–394, 1992
Hunt TK, Hussain Z: Wound microenvironment. In: Cohen, Diegelmann, Lindblad (eds). Wound Healing. W.B. Saunders, Philadelphia, PA, 1992, pp 274–281
Russel WE, Van-Wyk JJ, Pledger WJ: Proc Natl Acad Sci USA 81: 2388–2392, 1984
Clemmons DR, Van Wyk JJ. Somatomedins: Physiological control and effects on cell proliferation. In: R. Baserga (ed.). Handbook of Experimental Pharmacology. Springer-Verlag, New York, 1981, pp 161–164
Narayanan AS, Meyers DF, Page RC, Welgus HG: Action of mammalian collagenase on the type I trimer collagen. Collagen Rel Res 4: 289–296, 1984
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ghahary, A., Shen, Y.J., Nedelec, B. et al. Enhanced expression of mRNA for insulin-like growth factor-1 in post-burn hypertrophic scar tissue and its fibrogenic role by dermal fibroblasts. Mol Cell Biochem 148, 25–32 (1995). https://doi.org/10.1007/BF00929499
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00929499