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Morphometric Studies of Collagen and Fibrin Lattices Contracted by Human Gingival Fibroblasts; Comparison with Dermal Fibroblasts

Centre d'Etudes des Biomateriaux et Interfaces, EA 2068, IFR 53 Biomolecules, UFR d'Odontologie, 2, rue du General Koenig, 51100 Reims, France

W. Hornebeck

Laboratoire de Biochimie et Biologie Moleculaire, UPRESA 6021 CNRS, IFR 53 Biomolecules, Faculte de Medecine, 51, rue Cognacq-Jay, 51095 Reims France

G. Godeau

Laboratoire de Biochimie, Faculte de Chirurgie Dentaire, Universite Rene Descartes, 1, rue Maurice Arnoux, 92120 Montrouge, France

B. Pellat

Laboratoire de Biochimie, Faculte de Chirurgie Dentaire, Universite Rene Descartes, 1, rue Maurice Arnoux, 92120 Montrouge, France

P. Gillery

Laboratoire de Biochimie et Biologie Moleculaire, UPRESA 6021 CNRS, IFR 53 Biomolecules, Faculte de Medecine, 51, rue Cognacq-Jay, 51095 Reims France

EX Maquart

Laboratoire de Biochimie et Biologie Moleculaire, UPRESA 6021 CNRS, IFR 53 Biomolecules, Faculte de Medecine, 51, rue Cognacq-Jay, 51095 Reims France

D. Laurent-Maquin

Centre d'Etudes des Biomateriaux et Interfaces, EA 2068, IFR 53 Biomolecules, UFR d'Odontologie, 2, rue du General Koenig, 51100 Reims, France

Cell shape variations and substratum re-organization during contraction of floating collagen and fibrin lattices seeded with human gingival fibroblasts were determined by computerized image analysis of light and scanning electron microscopic images. Data were compared with those obtained with lattices populated with human dermal fibroblasts. The extent of collagen lattice contraction was similar with both cell types, resulting in a two-fold decrease in the area fractions occupied by collagen fibers. Fibroblasts exhibited a rounded shape (form factors equal to 0.8 and 0.7 for gingival and dermal cells, respectively) at day 1 of culture; they possessed a more elongated appearance (with form factors equal to 0.3 and 0.15 for gingival and dermal cells, respectively) at day 7. Continuous (gingival) and discontinuous (dermal) layers of cells were evidenced at the cortex of lattices. Contractions were associated with a significant reduction of the diameters of collagen fibers. Re-organization of substratum, as analyzed by the "Rose of Directions" technique, was evidenced only at the vicinity of filipodia where fibers ran parallel to these protrusions. Several lvsed matrix cavities were observed when fibrin lattices were populated with gingival but not dermal fibroblasts at day 5 of culture. Although cells in fibrin lattices exhibited morphometric parameters comparable with those in collagen lattices, no fibroblast layers could be demonstrated at gel peripheries. Fibrin matrices consisted of an isotropic network of entangled fibrin filaments from the start of culture, and only a slight reduction of the diameters of fibrin fibers could be evidenced in dermal fibroblast-populated lattices. Fibrinolysis at the vicinity of gingival fibroblasts led to an entire re-organization of substratum toward the formation of larger fibers. The differential behavior of gingival vs. dermal fibroblasts inside fibrin but not collagen matrices could therefore partly explain the increased rate of remodeling of gingiva as compared with dermis.

Key Words: fibroblasts • collagen lattices • fibrin lattices • gingiva • skin

REFERENCES

• Abercrombie M., Flint MH, James DW (1956). Wound contraction in relation to collagen formation in scorbutic guinea-pigs. J Embryol Exp Morphol 4:167-175.[Medline] [Order article via Infotrieve]
• Allen TD, Schor SL, Schor AM (1984). An ultrastructural review of collagen gels, a model system for cell-matrix, cell-basement membrane and cell-cell interactions. Scan Electron Microsc 1:375-390.
• Anderson TF (1966). Electron microscopy of microorganisms. In: Physical techniques in biological research. 2nd ed. Vol. IIIA. Pollister AW, editor. New York: Academic Press, pp. 319-387.
• Andujar MB, Melin M., Guerret S., Grimaud JA (1992). Cell migration influences collagen gel contraction. J Submicr Cytol Path 24:145-154.
• Bell E., Ivarsson B., Merrill C. (1979). Production of a tissue-like structure by contraction of collagen lattices by human fibroblasts of different proliferative potential in vitro. Proc Natl Acad Sci USA 76:1274-1278.[Abstract/Free Full Text]
• Bellows CG, Melcher AH, Aubin JE (1981). Contraction and organization of collagen gels by cells cultured from periodontal ligament, gingiva, and bone suggest functional differences between cell types. J Cell Sci 50:299-314.[Abstract]
• Bellows CG, Melcher AH, Bhargava U., Aubin JE (1982). Fibroblasts contracting three-dimensional collagen gels exhibit ultrastructure consistent with either contraction or protein secretion. J Ultrastruct Res 78:178-192.[CrossRef][Medline] [Order article via Infotrieve]
• Brown LF, Lanir N., McDonagh J., Tognazzi K., Dvorak AM, Dvorak HF (1993). Fibroblast migration in fibrin gel matrices. Am J Pathol 142:273-283.[Abstract]
• Buttle DJ, Ehrlich HP (1983). Comparative studies of collagen lattice contraction utilizing a normal and a transformed cell line. J Cell Physiol 116:159-166.[Medline] [Order article via Infotrieve]
• Ciano PS, Colvin RB, Dvorak AM, McDonagh J., Dvorak HF (1986). Macrophage migration in fibrin gel matrices. Lab Invest 54:62-70.[Medline] [Order article via Infotrieve]
• Clark Raf (1991). Cutaneous wound repair. In: Physiology, biochemistry, and molecular biology of the skin. 2nd ed. Vol. 1. Goldsmith LA, editor. New York: Oxford University Press, pp. 576-601.
• Curatolo L., Balconi G., Borgia R., Morasca L., Donati MB (1980). Fibrin clot retractile activity of mouse fibroblasts during growth and aging. In Vitro 16:731-737.[Medline] [Order article via Infotrieve]
• Docherty R., Forrester JV, Lackie JM, Gregory DW (1989). Glycosaminoglycans facilitate the movement of fibroblasts through three-dimensional collagen matrices. J Cell Sci 92:263-270.[Abstract/Free Full Text]
• Dodd Njf, Schor SL, Rushton G. (1982). The effects of a collagenous extracellular matrix on fibroblast membrane organization. An ESR spin label study. Exp Cell Res 141:421-431.[Medline] [Order article via Infotrieve]
• Ehrlich H., Rajaratman JB (1990). Cell locomotion forces versus cell contraction forces for collagen lattice contraction: An in vitro model of wound contraction. Tissue Cell 22:407-417.[CrossRef][Medline] [Order article via Infotrieve]
• Emonard H., Peyrol S., Grimaud JA (1991). Tissue hyperfibrils are degraded forms of collagen fibrils. An ultrastructural study employing enzymatic treatments. J Submicrosc Cytol Pathol 23:427-430.[Medline] [Order article via Infotrieve]
• Farsi Jma, Aubin JE (1984). Microfilament rearrangements during fibroblast-induced contraction of three-dimensional hydrated collagen gels. Cell Motility 4:29-40.[CrossRef][Medline] [Order article via Infotrieve]
• Fiszer-Szafarz B., Szafarz D., Guevara de Murillo A. (1981). A general, fast, and sensitive micromethod for DNA determination: application to rat and mouse liver, rat hepatoma, human leucocytes, chicken fibroblasts, and yeast cells. Anal Biochem 110:165-170.[CrossRef][Medline] [Order article via Infotrieve]
• Flook A. (1987). The quantitative measurement of partial shape. Acta Stereol 6:1009-1021.
• Gabbiani G. (1994). Modulation of fibroblastic cytoskeletal features during wound healing and fibrosis. Path Res Pract 190:851-853.
• Gabbiani G., Hirschel BJ, Ryan GB, Statkov PR, Majno G. (1972). Granulation tissue as a contractile organ: a study of structure and function. J Exp Med 135:719-734.[Abstract]
• Germain L., Jean A., Auger FA, Garrel DR (1994). Human wound healing fibroblasts have greater contractile properties than dermal fibroblasts. J Surg Res 57:268-273.[CrossRef][Medline] [Order article via Infotrieve]
• Gillery P., Maquart FX, Borel JP (1986). Fibronectin-dependence of the contraction of collagen lattices by human skin fibroblasts. Exp Cell Res 167:29-37.[CrossRef][Medline] [Order article via Infotrieve]
• Gillery P., Bellon G., Coustry F., Borel JP (1989). Culture of fibroblasts in fibrin lattices: Models for the study of metabolic activities of the cells in physiological conditions. J Cell Physiol 140:483-490.[Medline] [Order article via Infotrieve]
• Godeau G., Gonnord G., Jolivet O., Schoevaert D., Pompidou A., Lagrue G., et al. (1986). A selective histochemical method for the quantitative estimation of elastic fibers by computerized morphometric analysis. Effect of colchicin treatment. Anal Quant Cytol Histol 8:321-325.[Medline] [Order article via Infotrieve]
• Grinnell F. (1994). Fibroblasts, myofibroblasts and wound contraction. J Cell Biol 124:401-404.[Free Full Text]
• Grinnell F., Lamke CR (1984). Reorganization of hydrated collagen lattices by human skin fibroblasts. J Cell Sci 66:51-63.[Abstract]
• Grinnell F., Nakagawa S., Ho CH (1989). The collagen recognition sequence for fibroblasts depends on collagen topography. Exp Cell Res 182:668-672.[CrossRef][Medline] [Order article via Infotrieve]
• Hassell TM (1993). Tissues and cells of the periodontium. Periodontology 2000 3:9-38.
• Katagiri Y., Hiroyama T., Akamatsu N., Suzuki H., Yamazaki H., Tanoue K. (1995). Involvement of vβ3 integrin in mediating fibrin gel retraction. J Biol Chem 270:1785-1790.[Abstract/Free Full Text]
• Kleinman HK, Luckenbill-Edds L., Cannon FW, Sephel GC (1987). Use of extracellular matrix components for cell culture. Anal Biochem 166:1-13.[CrossRef][Medline] [Order article via Infotrieve]
• Knox P., Crooks S., Scaife MC, Patel S. (1987). Role of plasminogen, plasmin and plasminogen activators in the migration of fibroblasts into plasma clots. J Cell Physiol 132:501-508.[CrossRef][Medline] [Order article via Infotrieve]
• Langholz O., Rockel D., Mauch C., Kozlowska E., Bank H., Krieg T., et al. (1995). Collagen and collagenase gene expression in three-dimensional collagen lattices are differentially regulated by 1β1 and 2β1 integrins. J Cell Biol 131:1903-1915.[Abstract/Free Full Text]
• Lorimier S., Gillery P., Hornebeck W., Chastang F., Laurent-Maquin D., Bouthors S., et al. (1996). Tissue origin and extracellular matrix control neutral proteinase activity in human fibroblast three-dimensional cultures. J Cell Physiol 168:188-198.[CrossRef][Medline] [Order article via Infotrieve]
• Majno G., Gabbiani G., Hirschel BJ, Ryan GB, Stakov PR (1971). Contraction of granulation tissue in vitro: similarity to smooth muscle. Science 173:548-550.[Abstract/Free Full Text]
• Mauch C., Adelmann-Grill B., Hatamochi A., Krieg T. (1989). Collagenase gene expression in fibroblasts is regulated by a three-dimensional contact with collagen. FEBS Lett 250:301-305.[CrossRef][Medline] [Order article via Infotrieve]
• Meyer F. (1978). Contrast feature extraction. In: Practical metallography. Spec iss. Chermant JL, editor. Stuttgart: Riederer Verlag, pp. 374-380.
• Mirshahi M., Azzarone B., Soria J., Mirshahi F., Soria C. (1991). The role of fibroblasts in organization and degradation of a fibrin clot. J Lab Clin Med 117:274-281.[Medline] [Order article via Infotrieve]
• Murphy KG, Daniel JC (1987). Human periodontal ligament in vitro: Cell culture passage effect on collagen gel contraction. J Periodont Res 22:342-347.[CrossRef][Medline] [Order article via Infotrieve]
• Niewiarowski S., Regoeczi E., Mustard JF (1972). Adhesion of fibroblasts to polymerizing fibrin and retraction of fibrin induced by fibroblasts. Proc Soc Exp Biol Med 140:199-204.[CrossRef][Medline] [Order article via Infotrieve]
• Nishiyama J., Tominaga N., Nakajima K., Hayashi T. (1988). Quantitative evaluation of the factors affecting the process of fibroblast-mediated collagen gel contraction by separating the process in 3 phases. Collagen Rel Res 8:259-273.
• Nomine JP, Vincent L., Meyer F., Serra J., Escande JP (1987). Cutaneous aging and mathematical morphology. Acta Stereol 6:895-900.
• Nusgens B., Merrill C., Lapiere C., Bell E. ( 1984). Collagen biosynthesis by cells in a tissue equivalent matrix in vitro. Collagen Rel Res 4:351-363.
• Odland GF (1991). Structure of the skin. In: Physiology, biochemistry, and molecular biology of the skin. 2nd ed. Vol. 1. Goldsmith LA, editor. New York: Oxford University Press, pp. 3-62.
• Sarber R., Hull B., Merrill C., Soranno T., Bell E. ( 1981). Regulation of proliferation of fibroblasts of low and high population doubling levels grown in collagen lattices. Mech Ageing Dev 17:107-117.[CrossRef][Medline] [Order article via Infotrieve]
• Schmitt-Graff A., Desmouliere A., Gabbiani G. (1994). Heterogeneity of myofibroblast phenotypic features: an example of fibroblastic cell plasticity. Virchows Arch 425:3-24.[Medline] [Order article via Infotrieve]
• Serra J. (1982). Image analysis and mathematical morphology. London: Academic Press, pp. 283-286.
• Sharaf MN, Page RC ( 1988). Contraction of collagen gels by fibroblasts cultured from normal and inflamed gingivae. Dtsch Zahnärztl 43:693-695.
• Stopak D., Harris AK (1982). Connective tissue morphogenesis by fibroblast traction. 1. Tissue culture observations. Dev Biol 90:383-398.
• Tomasek JJ, Hay ED (1984). Analysis of the role of microfilaments and microtubules in acquisition of bipolarity and elongation of fibroblasts in hydrated collagen gels. J Cell Biol 99:536-549.[Abstract/Free Full Text]
• Tomasek JJ, Hay ED, Fujiwara K. (1982). Collagen modulates cell shape and cytoskeleton of embryonic corneal and fibroma fibroblasts: distribution of actin, alpha-actinin, and myosin. Dev Biol 92:107-122.[CrossRef][Medline] [Order article via Infotrieve]
• Tuan TL, Grinnell F. (1989). Fibronectin and fibrinolysis are not required for fibrin gel contraction by human skin fibroblasts. J Cell Physiol 140:577-583.[CrossRef][Medline] [Order article via Infotrieve]
• Wikesjo UM, Nilveus RE, Selvig KA (1992). Significance of early healing events on periodontal repair: a review. J Periodontol 63:158-165.[Medline] [Order article via Infotrieve]
• Yamamoto N. (1989). Effect of dimethyl sulfoxide on collagen gel contraction by fibroblasts. Biomed Res 10:83-94.
• Yamato M., Yamamoto K., Hayashi T. (1993). Age-related changes in collagen gel contraction by cultured human lung fibroblasts resulting in cross-over of contraction curves between young and aged cells. Mech Ageing Dev 67:149-158.[Medline] [Order article via Infotrieve]
• Yamato M., Adachi E., Yamamoto K., Hayashi T. (1995). Condensation of collagen fibrils to the direct vicinity of fibroblasts as a cause of gel contraction. J Biochem 117:940-946.[Abstract/Free Full Text]

Journal of Dental Research, Vol. 77, No. 9, 1717-1729 (1998)
DOI: 10.1177/00220345980770090801


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