Research Article| Volume 17, ISSUE 2, P123-131, June 1998

Fibroblast-migration in a wound model of ascorbic acid-supplemented three-dimensional culture system: the effects of cytokines and malotilate, a new wound healing stimulant, on cell-migration


      To assess the migratory response of fibroblasts in vitro, normal human dermal fibroblasts (NHDF) were cultured in the presence of l-ascorbic acid 2-phosphate to induce a multilayered structure. Round wounds were made by punching, and the migratory response was evaluated by counting the number of migrating cells in the wounded areas. Collagenase activity in the culture-medium was then measured. When the wound model was treated with bFGF, IL-1α or PDGF, the migratory response was facilitated with increased collagenase secretion. In contrast, treatment with TGF-β reduced the migratory response and collagenase secretion. Since the multilayered structure is rich in collagenous matrix, degradation of the matrix by secreted collagenase is probably necessary for the cells to migrate into the wounded areas. Furthermore, malotilate, which is now under development as an agent for wound therapy, facilitated the migratory response of NHDF with increased collagenase secretion in this wound model, suggesting that the wound healing effect of malotilate is in part attributable to stimulated migration of fibroblasts to wounded areas subsequent to extracellular matrix-degradation.


      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Journal of Dermatological Science
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Bigg H.F.
        • Cawston T.E.
        All-trans-retinoic acid interacts synergistically with basic fibroblast growth factor and epidermal growth factor to stimulate the production of tissue inhibitor of metalloproteinases from fibroblasts.
        Arch Biochem Biophys. 1995; 319: 74-83
        • Buckley-Sturrock A.
        • Woodward S.C.
        • Senior R.M.
        • Griffin G.L.
        • Klagsbrun M.
        • Davidson J.M.
        Differential stimulation of collagenase and chemotactic activity in fibroblasts derived from rat wound repair tissue and human skin by growth factors.
        J Cell Physiol. 1989; 138: 70-78
        • McNeil P.L.
        • Muthukrishnan L.
        • Warder E.
        • D'Amore P.A.
        Growth factors are released by mechanically wounded endothelial cells.
        J Cell Biol. 1989; 109: 811-822
        • Ellis I.
        • Grey A.M.
        • Schor A.M.
        • Schor S.L.
        Antagonistic effects of TGF-β1 and MSF on fibroblast migration and hyaluronic acid synthesis. Possible implications for dermal wound healing.
        J Cell Sci. 1992; 102: 447-456
        • Hata R.
        • Senoo H.
        l-ascorbic acid 2-phosphate stimulates collagen accumulation, cell proliferation, and formation of a three-dimensional tissuelike substance by skin fibroblasts.
        J Cell Physiol. 1989; 138: 8-16
        • Ishikawa O.
        • Yokoyama Y.
        • Miyachi Y.
        Disaccharide analysis of dermal fibroblast-derived glycosaminoglycans in the three-dimensional culture.
        J Dermatol Sci. 1994; 8: 203-207
      1. Suzuki H, Ichida F, Takino T, Nagashima H, Hirayama C, Fujisawa K, Furuta S, Monna T, Yamamoto S, Oda T. Therapeutic effects of malotilate on chronic hepatitis and liver cirrhosis: A double blind, controlled multicenter trial. In: Oda T, Tygstrup N, editors, Hepatotrophic Agent Malotilate, Proceedings of a Symposium on Malotilate held at 7th World Congress of Gastroenterology, Stockholm, June 15, 1982. Amsterdam: Excerpta Medica, Prinston, 1983:54–68.

        • Niwano Y.
        • Katoh M.
        • Uchida M.
        • Sugimoto T.
        Acceleration of liver regeneration by malotilate in partially hepatectomized rats.
        Jpn J Pharmacol. 1986; 40: 411-415
        • Niwano Y.
        • Koga H.
        • Sakai A.
        • Kanai K.
        • Hamaguchi H.
        • Uchida M.
        • Tachikawa T.
        Wound healing effect of malotilate in rats.
        Arzneim-Forsch/Drug Res. 1996; 46: 450-455
        • Postlethwaite A.E.
        • Lachman L.B.
        • Mainardi C.L.
        • Kang A.H.
        Interleukin 1 stimulation of collagenase production by cultured fibroblasts.
        J Exp Med. 1983; 157: 801-806
        • Circolo A.
        • Welgus H.G.
        • Pierce G.F.
        • Kramer J.
        • Strunk R.C.
        Differential regulation of the expression of proteinases/antiproteinases in fibroblast. Effects of interleukin-1 and platelet-derived growth factor.
        J Biol Chem. 1991; 266: 12283-12288
        • Overall C.M.
        • Wrana J.L.
        • Sodek J.
        Independent regulation of collagenase, 72-kDa progelatinase, and metalloendoproteinase inhibitor expression in human fibroblasts by transforming growth factor-β.
        J Biol Chem. 1989; 264: 1860-1869
        • Igarashi S.
        • Hatahara T.
        • Nagai Y.
        • Hori H.
        • Sakakibara K.
        • Katoh M.
        • Sakai A.
        • Sugimoto T.
        Anti-fibrotic effect of malotilate on liver fibrosis induced by carbon tetrachloride in rats.
        Jpn J Exp Med. 1986; 56: 235-245
        • Ala-Kokko L.
        • Stenbäck F.
        • Ryhänen L.
        Preventive effect of malotilate on dimethylnitrosamine-induced liver fibrosis in the rat.
        J Lab Clin Med. 1989; 113: 117-183
        • Chan B.P.
        • Chan K.M.
        • Maffulli N.
        • Webb S.
        • Lee K.K.
        Effect of basic fibroblast growth factor. An in vitro study of tendon healing.
        Clin Orthop. 1997; 342: 239-247
        • Dunlevy J.R.
        • Couchman J.R.
        Interleukin-8 induces motile behavior and loss of focal adhesions in primary fibroblasts.
        J Cell Sci. 1995; 108: 311-321
        • Seppä H.
        • Grotendorst G.
        • Seppä S.
        • Schiffmann E.
        • Martin G.R.
        Platelet-derived growth factor is chemotactic for fibroblasts.
        J Cell Biol. 1982; 92: 584-588
        • Porras-Reyes B.H.
        • Blair H.C.
        • Jeffrey J.J.
        • Mustoe T.A.
        Collagenase production at the border of granulation tissue in a healing wound. Macrophage and mesenchymal collagenase production in vivo.
        Connect Tiss Res. 1991; 27: 63-71
        • Moses M.A.
        • Marikovsky M.
        • Harper J.W.
        • Vogt P.
        • Eriksson E.
        • Klagsbrun M.
        • Langer R.
        Temporal study of the activity of matrix metalloproteinases and their endogenous inhibitors during wound healing.
        J Cell Biochem. 1996; 60: 379-386
        • Girard M.T.
        • Matsubara M.
        • Kublin C.
        • Tessier M.J.
        • Cintron C.
        • Fini M.E.
        Stromal fibroblasts synthesize collagenase and stromelysin during long-term tissue remodeling.
        J Cell Sci. 1993; 104: 1001-1011