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Re-epithelialization of Human Oral Keratinocytes in vitro
J.A. Garlick
Department of Oral Biology and Pathology, School of Dental Medicine, State University of New York at Stony Brook, Stony Brook, New York 11794-8702
W.C. Parks
Division of Dermatology at Jewish Hospital, Washington University Medical Center, St. Louis, Missouri 63110
H.G. Welgus
Division of Dermatology at Jewish Hospital, Washington University Medical Center, St. Louis, Missouri 63110
L.B. Taichman
Department of Oral Biology and Pathology, School of Dental Medicine, State University of New York at Stony Brook, Stony Brook, New York 11794-8702, Department of Dermatology, School of Medicine, State University of New York at Stony Brook
Re-epithelialization involves interactions between keratinocytes and the extracellular matrix upon which these cells move. It is hypothesized that keratinocytes are activated when wounded, and the resultant phenotypic change directs re-epithelialization. We have adapted organotypic cultures, in which oral gingival keratinocytes are fully differentiated, to study re-epithelialization following wounding. To elucidate keratinocyte behavior and phenotype during re-epithelialization, we have investigated this process in the presence and absence of the growth factor TGF-β1 and have monitored expression of MMP-1 (Type I collagenase) mRNA by in situ hybridization. In addition, we have followed proliferation and migration of wound keratinocytes by genetically marking these cells with a retroviral vector and by measuring their proliferative index. We found that keratinocytes grown without TGF-β1 were hyperproliferative in response to wounding, and re-epithelialization was complete by 24 h. However, 2.5 ng/mL TGF-β1 induced a transient delay in re-epithelialization, a reduction in proliferation, and fewer clusters of genetically marked cells. Keratinocytes expressed MMP-1 mRNA only when they covered the wounded surface, suggesting that the cells acquire a collagenolytic phenotype during re-epithelializaation and that contact with different ECM components may modulate keratinocyte expression of MMP-1. We conclude that the phenotype of oral keratinocytes is altered during re-epithelialization in vitro and that this process is modulated by TGF-pl. Reepithelialization occurs as keratinocytes are activated to move over the wound bed. Understanding the phenotype of wounded keratinocytes may facilitate treatment of chronic oral wounds and periodontal disease.
Key Words: oral keratinocytes • re-epithelialization • matrix metalloproteinase • transforming growth factor-β • wound healing
REFERENCES
- Ammann AJ, Beck LS, Deguzman L., Hirabayashi SE, Lee WP, McFatridge LL, et al.(1993). Transforming growth factor-β, effect on soft tissue repair. Ann NY Acad Sci 593:124-134. Barnard JA, Lyons RM, Moses HL (1990). The cell biology of transforming growth factor β. Biochim Biophys Acta 1032:79-87.[Medline]
[Order article via Infotrieve]
- Beck LS, Chen TL, Hirabayashi SE, Deguzman L., Lee WP, McFatridge LL, et al.(1990). Accelerated healing of ulcer wounds in the rabbit ear by recombinant human transforming growth factor-β1. Growth Factors 2:273-282. Bell E., Ehrlich HP, Buttle DJ, Nakatsuji T. (1981). Living tissue formed in vitro and accepted as skin-equivalent tissue of full thickness. Science 211:1052-1054.[Abstract/Free Full Text]
- Chen TL, Bates RL, Xu Y., Ammann AJ, Beck LS ( 1992). Human recombinant transforming growth factor-β1 modulation of biochemical and cellular events in healing of ulcer wounds. J Invest Dermatol 98:428-435.[Medline]
[Order article via Infotrieve]
- Cromack DT, Pierce GF, Mustoe TA (1991). TGF-β and PDGF mediated tissue repair: Identifying mechanisms of action using impaired and normal models of wound healing. In: Clinical and experimental approaches to dermal and epidermal repair: Normal and chronic wounds. New York: Wiley-Liss Inc., pp. 359-373.
- Donoff RB, McLennan JE, Grillo HC (1971). Preparation and properties of collagenases from epithelium and mesenchyme of healing mammalian wounds. Biochim Biophys Acta 227:639-653.[Medline]
[Order article via Infotrieve]
- Fini ME, Girard MT, Matsubara M. (1992). Collagenolytic/gelatinolytic enzymes in corneal wound healing. Acta Ophthalmol Suppl 202:26-33.
- Gailit J., Welch MP, Clark Raf (1994). TGF-β stimulates expression of keratinocyte integrins during re-epithelialization of cutaneous wounds. J Invest Dermatol 103:221-227.[CrossRef][Medline]
[Order article via Infotrieve]
- Garlick JA, Taichman LB (1993). The fate of genetically marked human oral keratinocytes in vitro. Arch Oral Biol 38:903-910.[Medline]
[Order article via Infotrieve]
- Garlick JA, Taichman LB (1994a). Fate of human keratinocytes
- during reepithelialization in an organotypic culture model. Lab Invest 70:916-924.
- Garlick JA, Taichman LB (1994b). Effect of TGF-β1 on re-epithelialization of human keratinocytes in vitro: An organotypic model. J Invest Dermatol 103:554-559.[CrossRef][Medline]
[Order article via Infotrieve]
- Garlick JA, Katz AB, Fenjves ES, Taichman LB (1991). Retrovirus-mediated transduction of cultured epidermal keratinocytes. J Invest Dermatol 97:824-829.[CrossRef][Medline]
[Order article via Infotrieve]
- Gipson IK, Spurr-Michaud S., Tisdale A., Elwell J., Stepp MA (1993). Redistribution of the hemidesmosome components alpha6 beta4 integrin and bullous pemphigoid antigens during epithelial wound healing. Exp Cell Res 207:86-98. Goldberg GI, Wilhelm SM, Kronberger A., Bauer EA, Grant GA, Eisen AZ (1986). Human fibroblast collagenase. Complete primary structure and homology to an oncogene transformation-induced rat protein. J Biol Chem 261:6600-6605.[Abstract/Free Full Text]
- Grillo HC, Gross J. (1967). Collagenolytic activity during mammalian wound repair. Dev Biol 15:300-317.[CrossRef][Medline]
[Order article via Infotrieve]
- Grinnell F. (1992). Wound repair, keratinocyte activation and integrin modulation. J Cell Sci 101:1-5.[Free Full Text]
- Jaffee EM, Dranoff G., Cohen LK, Hauda KM, Clift S., Marshall FF, et al.(1993). High efficiency gene transfer into primary human tumor explants without cell selection. Cancer Res 53:2221-2226.[Abstract/Free Full Text]
- Jones SC, Curtsinger LJ, Whalen JD, Pietsch JD, Ackerman D., Brown GL, et al.(1991). Effect of topical recombinant TGF-β on healing of partial thickness injuries. J Surg Res 51:344-352. Kiritsy CP, Lynch AB, Lynch SE (1993). Role of growth factors in cutaneous wound healing: A review. Crit Rev Oral Biol Med 4:729-760.[Abstract/Free Full Text]
- Ksander GA, Ogawa Y., Chu GH, McMullin H., Rosenblatt JS, McPherson JM (1990). Exogenous transforming growth factor-beta-2 enhances connective tissue formation and wound strength in guinea pig dermal wound healing by secondary intent. Ann Surg 211:288-294.[Medline]
[Order article via Infotrieve]
- Larjava H., Salo T., Haapasalmi K., Kramer RH, Heino J. (1993). Expression of integrins and basement membrane components by wound keratinocytes. J Clin Invest 92:1425-1435.[Medline]
[Order article via Infotrieve]
- Levine JH, Moses HL, Gold LI, Nanney LB (1993). Spatial and temporal patterns of immunoreactive transforming growth factor β1, β2, and β3 during excisional wound repair. Am J Pathol 143:368-380.[Abstract]
- Lin H-Y., Wells BR, Taylor RE, Birkedal-Hansen H. (1987). Degradation of type I collagen by rat mucosal keratinocytes: Evidence for secretion of a specific epithelial collagenase. J Biol Chem 262:6823-6831.[Abstract/Free Full Text]
- Mackenzie IC, Dabelsteen E., Zimmermann K. (1977). The relationship between expression of epithelial B-like blood group antigen, cell movement and cell proliferation. Acta Path Microbiol Scand Sect A 85:49-56.
- Makela M., Salo T., Uitto V-J., Larjava H. (1994). Matrix metalloproteinases (MMP-2 and MMP-9) of the oral cavity: Cellular origin and relationship to periodontal status. J Dent Res 73:1397-1406.
- Matrisian LM (1990). Metalloproteinases and their inhibitors in matrix remodeling. Trends Genet 6:121-125.[CrossRef][Medline]
[Order article via Infotrieve]
- Nickoloff BJ, Mitra RS, Riser BL, Dixit VM, Varani J. (1988). Modulation of keratinocyte motility: correlation with production of extracellular matrix molecules in response to growth promoting and antiproliferative factors. Am J Pathol
- :543-551.
- Parenteau NL, Nolte CM, Bilbo P., Rosenberg M., Wilkins LM, Johnson EW, et al. (1991). Epidermis generated in vitro: practical considerations and applications. J Cell Biochem 45:245-251. Parenteau NL (1994). Skin equivalents. In: The keratinocyte handbook. Vol. II. Leigh IM, Watt FW, editors. Cambridge: Cambridge University Press, pp. 45-56.
- Prosser IW, Stenmark KR, Suthar M., Crouch EC, Mecham RP, Parks WC (1989). Regional heterogeneity of elastin and collagen gene expression in intralobar arteries in response to hypoxic pulmonary hypertension as demonstrated by in situ hybridization. Am J Pathol 135:1073-1088.[Abstract]
- Quaglino D Jr, Nanney LB, Ditesheim JA, Davidson JM (1991). Transforming growth factor-beta stimulates wound healing and modulates extracellular matrix gene expression in pig skin: incisional wound model. J Invest Dermatol 97:34-42. Rheinwald JG, Green H. (1975). Serial cultivation of strains of human epidermal keratinocytes; the formation of keratinizing colonies from single cells. Cell 6:331-343. Saarialho-Kere UK, Chang ES, Welgus HG, Parks WC (1992). Distinct localization of collagenase and TIMP expression in wound healing associated with ulcerative pyogenic granuloma. J Clin Invest 90:1952-1957.[Medline]
[Order article via Infotrieve]
- Saarialho-Kere UK, Kovacs SO, Pentland AP, Olerud JE, Welgus HG, Parks WC (1993a). Cell-matrix interactions modulate interstitial collagenase expression by human keratinocytes actively involved in wound healing. J Clin Invest 92:2858-2866.[Medline]
[Order article via Infotrieve]
- Saarialho-Kere UK, Welgus HG, Parks WC (1993b). Distinct mechanisms regulate interstitial collagenase and 92 kDa gelatinase expression in human monocytic-like cells exposed to bacterial endotoxin. J Biol Chem 268:17354-17361. Saarialho-Kere UK, Pentland AP, Birkedal-Hansen H, Parks WC, Welgus HG (1994). Distinct populations of basal keratinocytes express stromelysin-1 and stromelysin-2 in chronic wounds. J Clin Invest 94:79-88.
- Salo T., Makela M., Kylmaniemi M., Autio-Harmainen H., Larjava H. (1994). Expression of matrix metalloproteinase-2 and -9 during early human wound healing. Lab Invest 70:176-182. Sciubba JJ (1977). Regeneration of the basal lamina complex during epithelial wound healing. J Periodont Res 12:204-217.[CrossRef][Medline]
[Order article via Infotrieve]
- Uitto V-J., Larjava H. (1991). Extracellular matrix molecules and their receptors: An overview with special emphasis on periodontal tissues. Crit Rev Oral Biol Med 2:323-354. Winter GD (1972). Epidermal regeneration studied in the domestic pig. In: Epidermal wound healing. Maibach HI, Rovee DT, editors. Chicago: Year Book Publishing, pp. 71-112.
- Woodley DT, O'Keefe EJ, Prunieras M. (1985). Cutaneous wound healing: A model for cell-matrix interactions. J Am Acad Dermatol 12:420-433.[Medline]
[Order article via Infotrieve]
- Wu Y-J., Parker LM, Binder NE, Beckett MA, Sinard JH, Griffiths CT, Reinwald JG (1982). The mesothelial keratins: a new family of cytoskeletal proteins identified in cultured mesothelial cells nonkeratinizing epithelia. Cell 31:693-703.[CrossRef][Medline]
[Order article via Infotrieve]
Journal of Dental Research, Vol. 75, No. 3,
912-918 (1996)
DOI: 10.1177/00220345960750030801
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