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Indication of fibroblast apoptosis during the maturation of disc-shaped mechanically stressed collagen lattices

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Abstract

Human dermal fibroblasts seeded in tense disc-shaped collagen gels organized gradually into a tissue composed of three distinct superimposed layers of cells: a basal layer of aligned myofibroblasts, a middle layer of unoriented fibroblasts, and an upper layer of myofibroblasts oriented orthogonally to the basal myofibroblasts. Confocal observation of α-smooth muscle actin (α-SMA) immunolabelling as a marker of myofibroblasts showed that the upper myofibroblasts disappeared during maturation of the lattices. Observation of morphological modifications of cells, typical chromatin condensation identified by Hoechst 33258 staining, and detection of ssDNA after formamide-induced denaturation suggested the involvement of apoptosis in myofibroblast disappearance. It is suggested that the model of disc-shaped stressed collagen lattice is thus able to mimic conditions in wound repair: on the one hand, wound contraction during which fibroblasts undergo mechanical stress and, on the other, apoptotic disappearance of cells at the end of tissue retraction.

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References

  1. Adams LW, Priestley GC (1988) Contraction of collagen lattices by skin fibroblasts: drug-induced changes. Arch Dermatol Res 280:114–118

    CAS  PubMed  Google Scholar 

  2. Bard J, Hay E (1975) The behavior of fibroblasts from the developing avian cornea: morphology and movement in situ and in vitro. J Cell Biol 67:400–418

    CAS  PubMed  Google Scholar 

  3. Baschong W, Sütterling R, Aebi U (1997) Punch wounded, fibroblast populated collagen matrices: a novel approach for studying cytoskeletal changes in three dimensions by confocal laser scanning microscopy. Eur J Cell Biol 72:189–201

    CAS  PubMed  Google Scholar 

  4. Bell E, Ivarson B, Merill 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 U S A 76:1274–1278

    CAS  PubMed  Google Scholar 

  5. Bride J, Lucarz-Bietry A, Viennet C, Humbert Ph (2002) Organisation spatiale des lattices de collagène tendus observés au microscope confocal. In: Actualités en ingénierie cutanée, vol 2. Eska, Paris, pp 229–240

  6. Chamson A, Sudre F, Le Guen C, Lee J, Rattner A, Frey J (1997) Morphological alteration of fibroblasts mechanically stressed in a collagen lattice. Arch Dermatol Res 289:596–599

    CAS  PubMed  Google Scholar 

  7. Brown RA, Prajapati R, MacGrouther DA, Yannas IV, Eastwood M (1998) Tensional homeostasis in dermal fibroblasts: mechanical responses to mechanical loading in three-dimensional substrates. J Cell Physiol 175:323–332

    CAS  PubMed  Google Scholar 

  8. Coulomb B, Dubertret L, Bell E, Touraine R (1984) The contractility of fibroblasts in a collagen lattice is reduced by corticosteroids. J Invest Dermatol 82:341–344

    CAS  PubMed  Google Scholar 

  9. Darby I, Skalli O, Gabbiani G (1990) Alpha-smooth muscle actin is transiently expressed by myofibroblasts during experimental wound healing. Lab Invest 63:21–29

    CAS  PubMed  Google Scholar 

  10. Darby I, Bisucci T, Hewitson T, MacLellan D (1997) Apoptosis is increased in a model of diabetes-impaired wound healing in genetically diabetic mice. Int J Biochem Cell Biol 29:191–200

    CAS  PubMed  Google Scholar 

  11. Delvoye P, Wiliquet P, Levêque JL, Nusgens BV, Lapière CM (1991) Measurement of mechanical forces generated by skin fibroblasts embedded in a three-dimensional collagen gel. J Invest Dermatol 97:898–902

    CAS  PubMed  Google Scholar 

  12. Desmoulière A, Redard M, Darby I, Gabbiani G (1995) Apoptosis mediates the decrease in cellularity during the transition between granulation tissue and scar. Am J Pathol 146:156–166

    Google Scholar 

  13. Eastwood M, MacGrouther DA, Brown RA (1994) A culture force monitor for measurement of contraction forces generated in human dermal fibroblast culture: evidence for cell-matrix mechanical signalling. Biochim Biophys Acta 1201:186–192

    CAS  PubMed  Google Scholar 

  14. Eastwood M, Mudera VC, MacGrouther DA, Brown RA (1998) Effect of precise mechanical loading on fibroblast populated collagen lattices: morphological changes. Cell Motil Cytoskeleton 40:13–21

    CAS  PubMed  Google Scholar 

  15. Ehrlich HP, Rajaratnam 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

    CAS  PubMed  Google Scholar 

  16. Elsdale T, Bard J (1972) Cellular interaction in mass cultures of human diploid fibroblasts. Nature 236:152–155

    CAS  PubMed  Google Scholar 

  17. Elsdale T, Foley R (1969) Morphogenetic aspects of multilayering in Petri dish cultures of human fetal lung fibroblasts. J Cell Biol 41:298–311

    CAS  PubMed  Google Scholar 

  18. Fluck J, Querfeld C, Cremer A, Niland S, Krieg T, Sollberg S (1998) Normal human primary fibroblasts undergo apoptosis in three-dimensional contractile collagen gels. J Invest Dermatol 110:153–157

    Google Scholar 

  19. Frankfurt OS, Krishan A (2001) Identification of apoptotic cells by formamide-induced DNA denaturation in condensed chromatin. J Histochem Cytochem 49:369–378

    CAS  PubMed  Google Scholar 

  20. Grinnell F (1994) Fibroblasts, myofibroblasts and wound contraction. J Cell Biol 124:401–404

    CAS  PubMed  Google Scholar 

  21. Grinnell F, Zhu M, Carlson MA, Abrams JM (1999) Release of mechanical tension triggers apoptosis of human fibroblasts in a model of regressing granulation tissue. Exp Cell Res 248:608–619

    Article  CAS  PubMed  Google Scholar 

  22. Hinz B, Mastrangelo D, Iselin CE, Chaponnier C, Gabbiani G (2001) Mechanical tension controls granulation tissue contractile activity and myofibroblast differentiation. Am J Pathol 159:1009–1020

    CAS  PubMed  Google Scholar 

  23. Ishikawa O, Kondo A, Okada K, Miyachi Y, Furumura M (1997) Morphological and biochemical analyses on fibroblasts and self-produced collagens in a novel three-dimensional culture. Br J Dermatol 136:6–11

    CAS  PubMed  Google Scholar 

  24. Kischer CW (1992) The microvessels in hypertrophic scars, keloids and related lesions: a review. J Submicrosc Cytol Pathol 24:281–296

    CAS  PubMed  Google Scholar 

  25. Kolodney MS, Wysolmerski RB (1992) Isometric contraction by fibroblasts and endothelial cells in tissue culture: a quantitative study. J Cell Biol 117:73–82

    CAS  PubMed  Google Scholar 

  26. Lambert CA, Soudant EP, Nusgens BV, Lapiere CM (1992) Pretranslational regulation of extracellular matrix macromolecules and collagenase expression in fibroblasts by mechanical forces. Lab Invest 66:444–451

    CAS  PubMed  Google Scholar 

  27. Lee J, Rattner A, Chepda T, Frey J, Chamson A (2002) Production of matrix metalloproteinase 2 in fibroblast reaction to mechanical stress in a collagen gel. Arch Dermatol Res 294:405–410

    PubMed  Google Scholar 

  28. Lopez-Valle CA, Auger FA, Rompré P, Bouvard V, Germain L (1992) Peripheral anchorage of dermal equivalents. Br J Dermatol 127:365–371

    CAS  PubMed  Google Scholar 

  29. Lucarz-Biétry A, Chapuis JF, Agache P, Humbert P (1995) A histological and ultrastructural comparative study of retracted and tense collagen lattices. Eur J Dermatol 5:524–530

    Google Scholar 

  30. Nagata M, Takenaka H, Shibagaki R, Kishimoto S (1999) Apoptosis and P53 protein expression increase in the process of burn wound healing in guinea-pig skin. Br J Dermatol 140:829–836

    CAS  PubMed  Google Scholar 

  31. Nakagawa S, Pawclek P, Grinnell F (1989) Extracellular matrix organization modulates fibroblast growth and growth factor responsiveness. Exp Cell Res 182:572–582

    CAS  PubMed  Google Scholar 

  32. Niland S, Cremer A, Fluck J, Eble A, Krieg T, Sollberg S (2001) Contraction dependent apoptosis of normal dermal fibroblasts. J Invest Dermatol 116:686–692

    Google Scholar 

  33. Nishiyama T, Tsunenaga M, Nakayama E, Adachi E, Hayashi T (1989) Growth rate of human fibroblasts is repressed by the culture within reconstituted collagen matrix but not by the culture on the matrix. Matrix 9:193–199

    CAS  PubMed  Google Scholar 

  34. Novotny GEK, Gnoth C (1971) Variability of fibroblast morphology in vivo: a silver impregnation study on human digital dermis and subcutis. J Anat 177:195–207

    Google Scholar 

  35. Olerud JE, Chiu DS, Usui ML, Gibran NS, Ansel JC (1998) Protein gene product 9.5 is expressed by fibroblasts in human cutaneous wounds. J Invest Dermatol 111:565–572

    CAS  PubMed  Google Scholar 

  36. Porter RA, Brown RA, Eastwood MA, Occleston NL, Khaw PT (1998) Ultrastructural changes during contraction of collagen lattices by ocular fibroblasts. Wound Repair Regen 6:157–166

    CAS  PubMed  Google Scholar 

  37. Ryan BR, Cliff WJ, Gabbiani G, Irle C, Montandon D, Statkov PR, Majno G (1974) Myofibroblasts in human granulation tissue. Hum Pathol 5:55–67

    CAS  PubMed  Google Scholar 

  38. Sarber R, Hull B, Merrill C, Sorano 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

    CAS  PubMed  Google Scholar 

  39. Skalli O, Gabbiani G (1988) The biology of the myofibroblast relationships to wound contraction and fibrocontractive diseases. In: Clark RAF, Henson PM (eds) The molecular and cellular biology of wound repair. Plenum Publishing, New York, pp 373–402

  40. Sperandio S, de Belle I, Bredesen DE (2000) An alternative non apoptotic form of programmed cell death. Proc Natl Acad Sci U S A 97:14376–14381

    CAS  PubMed  Google Scholar 

  41. Tomasek JJ, Haaksma CJ, Eddy RJ, Vaughan MB (1992) Fibroblast contraction occurs on release of tension in attached collagen lattices: dependency on an organized actin cytoskeleton and serum. Anat Rec 359:68

    Google Scholar 

  42. Willingham MC (1999) Cytochemical methods for the detection of apoptosis. J Histochem Cytochem 47:1101–1109

    CAS  PubMed  Google Scholar 

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Acknowledgements

The authors thank the “Centre de Microscopie Electronique” of Besançon (France), Isabelle Bruey and Marie-Pierre Decour for their assistance in the organization of the manuscript.

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Authors and Affiliations

  1. Laboratoire d’Ingénierie et de Biologie Cutanées, UFR Médecine-Pharmacie, 25030, Besançon Cedex, France

    Jacqueline Bride, Celine Viennet, Annie Lucarz-Bietry & Philippe Humbert

  2. Service de Dermatologie 1, Hôpital Saint-Jacques - 2, Place Saint-Jacques, 25030, Besançon Cedex, France

    Philippe Humbert

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  1. Jacqueline Bride
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  2. Celine Viennet
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  3. Annie Lucarz-Bietry
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  4. Philippe Humbert
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Correspondence to Philippe Humbert.

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Bride, J., Viennet, C., Lucarz-Bietry, A. et al. Indication of fibroblast apoptosis during the maturation of disc-shaped mechanically stressed collagen lattices. Arch Dermatol Res 295, 312–317 (2004). https://doi.org/10.1007/s00403-003-0438-z

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  • DOI: https://doi.org/10.1007/s00403-003-0438-z

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