Attachment kinetics, proliferation rates and vinculin assembly of bovine osteoblasts cultured on different pre-coated artificial substrates

Primary bovine osteoblasts were used to study in-vitro effects of attachment on vinculin assembly in cells cultured on various artificial substrates. Materials coated with fibronectin and bovine serum albumin (BSA) as well as untreated materials (tissue culture polystyrene and Aclar foils) were chosen to investigate substrate-dependent proliferation during the first 3 days of culture. Proliferation was highest on fibronectin-coated substrates, followed by BSA-coated and untreated substrates. During the first 24 h of cultivation, cell attachment kinetics revealed no significant difference between the various substrates. After 24 h detachment rates obtained by calcium depletion with ethylenediaminetetraacetic acid were highest on uncoated materials, followed by BSA- and fibronectin-coated substrates. Phase contrast microscopy revealed typical osteoblast morphology after cell adhesion for 24 h. The dynamic attachment process was concomitant with the reassembly of vinculin into streak-like focal contacts clustered on the ventral side of cells. The kinetics of vinculin reassembly were independent of the underlying coating. Thus, fibronectin coating of artificial substrates increased the attachment strength and proliferation rate of osteoblasts. While the reassembly of vinculin in focal contacts seems to be a prerequisite of osteoblast attachment in vitro, it does not seem to have profound effects on the subsequent cell behaviour on artificial substrates. © Chapman & Hall

[1]  Y. Missirlis,et al.  Modern aspects of protein adsorption on biomaterials , 1991 .

[2]  E. Ruoslahti,et al.  Superfibronectin is a functionally distinct form of fibronectin , 1994, Nature.

[3]  Richard O. Hynes,et al.  Integrins: A family of cell surface receptors , 1987, Cell.

[4]  B. Geiger,et al.  Microfilament-organizing centers in areas of cell contact: cytoskeletal interactions during cell attachment and locomotion , 1984, The Journal of cell biology.

[5]  R. Ezzell,et al.  Expression of chicken vinculin complements the adhesion-defective phenotype of a mutant mouse F9 embryonal carcinoma cell , 1993, The Journal of cell biology.

[6]  J. Feramisco,et al.  Microinjection and localization of a 130K protein in living fibroblasts: a relationship to actin and fibronectin , 1980, Cell.

[7]  T. Hunter,et al.  Integrin-mediated signal transduction linked to Ras pathway by GRB2 binding to focal adhesion kinase , 1994, Nature.

[8]  B. Geiger A 130K protein from chicken gizzard: Its localization at the termini of microfilament bundles in cultured chicken cells , 1979, Cell.

[9]  J. Meigs,et al.  Reorganization of alpha-actinin and vinculin induced by a phorbol ester in living cells , 1986, The Journal of cell biology.

[10]  M. Bissell,et al.  The Influence of Extracellular Matrix on Gene Expression: Is Structure the Message? , 1987, Journal of Cell Science.

[11]  E. Ruoslahti,et al.  Motility of fibronectin receptor-deficient cells on fibronectin and vitronectin: collaborative interactions among integrins [published erratum appears in J Cell Biol 1992 Jul;118(1):217] , 1992, The Journal of cell biology.

[12]  D. Puleo,et al.  Osteoblasts on hydroxyapatite, alumina and bone surfaces in vitro: morphology during the first 2 h of attachment. , 1992, Biomaterials.

[13]  D. Jones,et al.  Biochemical signal transduction of mechanical strain in osteoblast-like cells. , 1991, Biomaterials.

[14]  W. T. Chen,et al.  Immunoelectron microscopic studies of the sites of cell-substratum and cell-cell contacts in cultured fibroblasts , 1982, The Journal of cell biology.

[15]  D. Puleo,et al.  Mechanisms of fibronectin-mediated attachment of osteoblasts to substrates in vitro. , 1992, Bone and mineral.

[16]  D. Puleo,et al.  RGDS tetrapeptide binds to osteoblasts and inhibits fibronectin-mediated adhesion. , 1991, Bone.

[17]  Frank P. Luyten,et al.  Differentiation of canalicular cell processes in bone cells by basement membrane matrix components: Regulation by discrete domains of laminin , 1990, Cell.

[18]  D. Puleo,et al.  Formation of focal contacts by osteoblasts cultured on orthopedic biomaterials. , 1992, Journal of biomedical materials research.

[19]  E Ruoslahti,et al.  New perspectives in cell adhesion: RGD and integrins. , 1987, Science.