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Adhesion and spreading of corneal endothelial cells on collagens type I and IV in vitro: a model to study mechanisms of endothelial repair

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Research in Experimental Medicine

Summary

In vitro assays using endothelial cells (EC, bovine corneal) were performed to study adhesion and spreading on collagen types I and IV. Adhesion was quantitatively analyzed by counting the EC under a light microscope. Spreading was determined by measuring cell area using a scanning electron microscope (SEM). Collagen types I, IV, and IV-F, a mixture of 70, 120, and 140 KD fragments of type IV, all promoted EC adhesion, Types IV and IV-F showed evidence of giving a more marked adhesion than type I. A study of cell area, carried out under identical conditions, such as those in the adhesion assay, showed that types I and IV-F, but not type IV, promoted cell spreading. This provides evidence that cell adhesion and spreading are indeed separate biological phenomena. Furthermore, the ability of fragments of type IV collagen to promote both cell adhesion and spreading may represent an inherent repair mechanism in damaged endothelium.

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References

  1. Berliner JA (1981) Regulation of endothelial cell DNA synthesis and adherence. In Vitro 17:985–992

    Article  CAS  Google Scholar 

  2. Burridge K, Fath K, Kelly T, Nuckolls G, Turner C (1988) Focal adhesions: transmebrane junctions between the extracellular matrix and the cytoskeleton. Annu Rev Cell Biol 4:487–525

    Article  PubMed  CAS  Google Scholar 

  3. Curtis ASG, McMurray H (1986) Conditions for fibroblast adhesion without fibronectin. J Cell Sci 86:25–33

    PubMed  CAS  Google Scholar 

  4. Dieringer H (1982) Untersuchungen zur molekularen und makromolekularen Struktur von Typ IV Kollagen aus menschlicher Plazenta. Ph. D. Thesis, Faculty of Science, Ludwig-Maximilians-University, Munich

    Google Scholar 

  5. Dieringer H, Hollister DW, Glanville RW, Sakai LY, Kühn K (1985) Structural studies of human basement-membrane collagen with the use of a monoclonal antibody. Biochem J 227:217–222

    PubMed  CAS  Google Scholar 

  6. Eldor A, Vlodavsky I, Hy-Am E, Atzmon R, Weksler BB, Raz A, Fuks Z (1983) Cultured endothelial cells increase their capacity to synthesize prostacyclin following the formation of a contact inhibited cell monolayer. J Cell Physiol 114:179–183

    Article  PubMed  CAS  Google Scholar 

  7. Fritz H, Jochum M, Geiger R, Duswald KH, Dittmer H, Kortmann H, Neumann S, Lang H (1986) Granulocyte proteinases as mediators of unspecific proteolysis in inflammation: a review. Folia Histochem Cytobiol 24:99–115

    PubMed  CAS  Google Scholar 

  8. Gold LI, Pearlstein E (1980) Fibronectin-collagen binding and requirement during cellular adhesion. Biochem J 186:551–559

    PubMed  CAS  Google Scholar 

  9. Gospodarowicz D, Mescher AR, Birdwell CR (1977) Stimulation of corneal endothelial cell proliferation in vitro by fibroblast and epidermal growth factors. Exp Eye Res 25:75–89

    Article  PubMed  CAS  Google Scholar 

  10. Jaffe EA, Mosher DF (1978) Synthesis of fibronectin by cultured human endothelial cells. J Exp Med 148:1779–1791

    Article  Google Scholar 

  11. Kühn K, Wiedemann H, Timpl R, Risteli J, Dieringer H, Voss T, Glanville RW (1981) Macromolecular structure of basement membrane collagens. Identification of 7S collagen as a crosslinking domain of type IV collagen. FEBS Lett 125:123–128

    Article  PubMed  Google Scholar 

  12. Lapis K, Paku S, Liotta LA (1988) Endothelialization of embolized tumor cells during metastasis formation. Clin Exp Metastasis 6:73–89

    Article  PubMed  CAS  Google Scholar 

  13. Lasser A (1983) The mononuclear phagocytic system: a review. Hum Pathol 14:108–126

    Article  PubMed  CAS  Google Scholar 

  14. Macarak EJ, Howard PS (1983) Adhesion of endothelial cells to extracellular matrix proteins. J Cell Physiol 116:76–86

    Article  PubMed  CAS  Google Scholar 

  15. Pierschbacher MD, Ruoslahti E (1984) Cell attachment activity of fibronectin can be duplicated by small synthetic fragments of the molecule. Nature 309:30–33

    Article  PubMed  CAS  Google Scholar 

  16. Ross R, Raggiotto A, Bowen-Pope D, Raines E (1984) The role of endothelial injury and platelet and macrophage interactions in atherosclerosis. Circulation 70 [Suppl III]:77–82

    Google Scholar 

  17. Ryan GB, Majno G (1977) Acute inflammation: a review. Am J Pathol 86:183–276

    PubMed  CAS  Google Scholar 

  18. Sage H, Pritzl P, Bornstein P (1981) Secretory phenotypes of endothelial cells in culture: comparison of aortic, venous, capillary, and corneal endothelium. Arteriosclerosis 1:427–442

    Article  PubMed  CAS  Google Scholar 

  19. Timpl R (1989) Structure and biological activity of basement membrane proteins. Eur J Biochem 180:487–502

    Article  PubMed  CAS  Google Scholar 

  20. Tryggvason K, Höyhtyä M, Salo T (1987) Proteolytic degradation of extracellular matrix in tumor invasion. Biochim Biophys Acta 907:191–217

    PubMed  CAS  Google Scholar 

  21. Vlodavsky I, Folkman J, Sullivan R, Fridman R, Ishai-Michaeli R, Sassa J, Klagsbrun M (1987) Endothelial cell-derived basic fibroblast growth factor: synthesis and deposition into subendothelial extracellular matrix. Proc Natl Acad Sci USA 84:2292–2296

    Article  PubMed  CAS  Google Scholar 

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Rixen, H., Kirkpatrick, C.J., Axer, T. et al. Adhesion and spreading of corneal endothelial cells on collagens type I and IV in vitro: a model to study mechanisms of endothelial repair. Res. Exp. Med. 190, 203–211 (1990). https://doi.org/10.1007/PL00020023

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  • DOI: https://doi.org/10.1007/PL00020023

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