Abstract
The fibroblast-populated collagen lattice (FPCL) was intended to act as the dermal component for “skin-equivalent” or artificial skin developed for skin grafting burn patients. The “skin-equivalent” was clinically unsuccessful as a skin graft, but today it is successfully used as a dressing for the management of chronic wounds. The FPCL has, however, become an instrument for investigating cell–connective tissue interactions within a three-dimensional matrix. Through the capacity of cell compaction of collagen fibrils, the FPCL undergoes a reduction in volume referred to as lattice contraction. Lattice contraction proceeds by cell-generated forces that reduce the water mass between collagen fibers, generating a closer relationship between collagen fibers. The compaction of collagen fibers is responsible for the reduction in the FPCL volume. Cell-generated forces through the linkage of collagen fibers with fibroblast’s cytoskeletal actin-rich microfilament structures are responsible for the completion of the collagen matrix compaction. The type of culture dish used to cast FPCL as well as the cell number will dictate the mechanism for compacting collagen matrices. Fibroblasts, at moderate density, cast as an FPCL within a petri dish and released from the surface of the dish soon after casting compact collagen fibers through cell tractional forces. Fibroblasts at moderate density cast as an FPCL within a tissue culture dish and not released for 4 days upon release show rapid lattice contraction through a mechanism of cell contraction forces. Fibroblasts at high density cast in an FPCL within a petri dish, released from the surface of the dish soon after casting, compact a collagen lattice very rapidly through forces related to cell elongation. The advantage of the FPCL contraction model is the study of cells in the three-dimensional environment, which is similar to the environment from which these cells were isolated. In this chapter methods are described for manufacturing collagen lattices, which assess the three forces involved in compacting and/or organizing collagen fibrils into thicker collagen fibers. The clinical relevance of the FPCL contraction model is related to advancing our understanding of wound contraction and scar contracture.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
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
Bell E, Parenteau N, Gay R, Nolte C, Kemp P, Bilbo P, Ekstein B, Johnson E (1991) The living skin equivalent: its manufacture, its organotypic properties and its responses to irritants. Toxicol In Vitro 5:591–596
Clark RA, Folkvord JM, Hart CE, Murray MJ, McPherson JM (1989) Platelet isoforms of platelet-derived growth factor stimulate fibroblasts to contract collagen matrices. J Clin Invest 84:1036–1040
Brem H, Balledux J, Bloom T, Kerstein MD, Hollier L (2000) Healing of diabetic foot ulcers and pressure ulcers with human skin equivalent: a new paradigm in wound healing. Arch Surg 135:627–634
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
Ehrlich HP, Rittenberg T (2000) Differences in the mechanism for high versus moderate density fibroblast populated collagen lattice contraction. J Cell Physiol 185:432–439
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 232:359–368
Gabbiani G, Ryan GB, Majno G (1971) Presence of modified fibroblasts in granulation tissue and their possible role in wound contraction. Experientia 27:549–555
Ehrlich HP, Rajaratnam JBM (1990) Cell locomotion forces versus cell contraction forces for collagen lattice contraction: an in vitro model of wound contraction. Tissue Cell 22:407–417
Vaughan MB, Howard EW, Tomasek JJ (2000) Transforming growth factor-beta1 promotes the morphological and functional differentiation of the myofibroblast. Exp Cell Res 257:180–189
Ehrlich HP, Griswold TR, Rajaratnam JBM (1986) Studies of vascular smooth muscle cells and dermal fibroblasts in collagen matrices: effects of heparin. Exp Cell Res 164:154–163
Souren JM, Ponec M, van Wijk R (1989) Contraction of collagen by human fibroblasts and keratinocytes. In Vitro Cell Dev Biol 25:1039–1045
Borland K, Ehrlich HP, Muffly K, Dills WL Jr, Halls PF (1986) The interaction of rat Sertoli cells with a collagen lattice in vitro. In Vitro Cell Dev Biol 22:661–669
Friedl P, Maaser K, Klein CE, Niggemann B, Krohne G, Zänker KS (1997) Migration of highly aggressive MV3 melanoma cells in 3-dimensional collagen lattices results in local matrix reorganization and shedding of alpha2 and beta1 integrins and CD44. Cancer Res 57:2061–2070
Lackie JM, Brown AF (1983) Adhesion and the locomotion of neutrophils on surfaces and in matrices. Agents Actions Suppl 12:73–90
Moyer KE, Saggers GC, Ehrlich HP (2004) Mast cells promote fibroblast populated collagen lattice contraction through gap junction intercellular communication. Wound Repair Regen 12:269–275
Greco RM, Ehrlich HP (1992) Differences in cell division and thymidine incorporation with rat and primate fibroblasts in collagen lattices. Tissue Cell 24:843–851
Ehrlich HP (1988) The modulation of contraction of fibroblast populated collagen lattices by types I, II, and III collagen. Tissue Cell 20:47–50
Ehrlich HP (1988) Wound closure: evidence of cooperation between fibroblasts and collagen matrix in scar contracture. Eye 2:149–157
Montesano R, Orci L (1988) Transforming growth factor beta stimulates collagen-matrix contraction by fibroblasts: implications for wound healing. Proc Natl Acad Sci USA 85:4894–4897
Hinz B, Celetta G, Tomasek JJ, Gabbiani G, Chaponnier C (2001) Alpha-smooth muscle actin expression up regulates fibroblast contractile activity. Mol Biol Cell 12:2730–2741
Seppä H, Grotendorst G, Seppä S, Schiffmann E, Martin GR (1982) Platelet-derived growth factor in chemotactic for fibroblasts. J Cell Biol 92:584–588
Bowman N, Donahue H, Ehrlich HP (1998) Gap junctional intercellular communication contribution to the contraction of rat osteoblast populated collagen lattices. J Bone Miner Res 13:1700–1706
Rittenberg T, Ehrlich HP (1992) Free fatty acids and dialyzed serum alterations of fibroblast populated collagen lattice contraction. Tissue Cell 24:243–251
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media New York
About this protocol
Cite this protocol
Ehrlich, H.P., Moyer, K.E. (2013). Cell-Populated Collagen Lattice Contraction Model for the Investigation of Fibroblast Collagen Interactions. In: Gourdie, R., Myers, T. (eds) Wound Regeneration and Repair. Methods in Molecular Biology, vol 1037. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-505-7_3
Download citation
DOI: https://doi.org/10.1007/978-1-62703-505-7_3
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-504-0
Online ISBN: 978-1-62703-505-7
eBook Packages: Springer Protocols