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1 March 2006 COMPLEMENTARY ROLES OF MICROTUBULES AND MICROFILAMENTS IN THE LUNG FIBROBLAST-MEDIATED CONTRACTION OF COLLAGEN GELS: DYNAMICS AND THE INFLUENCE OF CELL DENSITY
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Abstract
Fibroblasts are important cellular components in wound healing, scar formation, and fibrotic disorders; and the fibroblast-populated collagen-gel (FPCG) model allows examination of fibroblast behavior in an in vitro three-dimensional environment similar to that in vivo. Contraction of free-floating FPCGs depends on an active and dynamic cytoskeleton, and the contraction dynamics are highly influenced by cell density. We investigated mechanistic differences between high– and low–cell density FPCG contraction by evaluating contraction dynamics in detail, using specific cytoskeletal disruptors. Collagen gels were seeded with human lung fibroblasts at either high (HD) or low (LD) density, and incubated with or without cytoskeletal disruptors colchicine (microtubules) or cytochalasin D (microfilaments). Gel area was measured daily. FPCG contraction curves were essentially sigmoidal, featuring an initial period of no contraction (lag phase), followed by a period of rapid contraction (log phase). Contraction curves of HD-FPCGs were distinct from those of LD-FPCGs. For example, HD-FPCGs had a negligible lag phase (compared with 3 d for LD-FPCGs) and exhibited a higher rate of log-phase contraction. Both colchicine and cytochalasin dose-dependently inhibited contraction but specifically affected different phases of contraction in HD- and LD-FPCGs; and colchicine inhibited LD-FPCGs much more than HD-FPCGs. The data indicate that LD- and HD-FPCGs contract through different primary mechanisms. Microtubules and microfilaments are both complementarily and dynamically involved in the contraction of FPCGs, and cell density influences primary cytoskeletal mechanisms. These results provide valuable information about fibroblast behavior in healing and fibrosis, and may suggest novel treatment options.
ROBERT A. REDDEN and EDWARD J. DOOLIN "COMPLEMENTARY ROLES OF MICROTUBULES AND MICROFILAMENTS IN THE LUNG FIBROBLAST-MEDIATED CONTRACTION OF COLLAGEN GELS: DYNAMICS AND THE INFLUENCE OF CELL DENSITY," In Vitro Cellular & Developmental Biology - Animal 42(3), (1 March 2006). https://doi.org/10.1290/0509065.1
Received: 29 September 2005; Accepted: 16 January 2006; Published: 1 March 2006
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