title: Attachment of human bone cells to tissue culture polystyrene and to unmodified polystyrene: the effect of surface chemistry upon initial cell attachment.
abstract: Cell culture studies have often been used in the determination of the suitability of biomaterials as surfaces for the attachment and growth of cells. For such studies of surfaces for potential use in bone implants, cells derived from bone may be maintained in culture on tissue culture polystyrene (TCPS). We have determined the contribution that serum fibronectin (FN) or vitronectin (VN) make to the attachment and spreading of cells cultured from explanted human bone (bone-derived cells) during the first 90 min following seeding on culture surfaces. The attachment of bone-derived cells to TCPS was simulated two-fold by the addition of 10% (v/v) fetal bovine serum (FBS) to the seeding culture medium. The roles of FN and VN were determined by selective removal of the FN or VN from the FBS prior to addition to the culture medium. FBS from which the VN had been removed did not have this stimulatory activity. In contrast, the attachment of bone-derived cells onto TCPS from medium containing FN-depleted serum (which contained VN) was the same as when intact FBS was used. There was incomplete attachment of bone-derived cells (27% of cells) when seeded in medium containing FBS depleted of both VN and FN. Our results show that for human bone-derived cells, the attachment onto TCPS of cells planted in medium containing FBS during the first 90 min of culture is principally as a result of adsorption onto the surface of serum VN. As unmodified polystyrene (PS) has also been used previously as a model biomaterial surface, PS was compared to TCPS for attachment of the bone-derived cells. Attachment of bone-derived cells to TCPS was twice that onto PS, both when the medium was serum-free and when it contained FBS. Bone-derived cells attached to TCPS or PS onto which purified VN or FN had been precoated, with VN adsorbed onto PS being as effective as was VN adsorbed onto TCPS. With FN, there was an effect of the polystyrene surface chemistry which was evident in that suboptimal concentrations of FN had a slightly higher potency when adsorbed onto TCPS than did the same concentrations of FN coated onto PS. When preadsorbed onto TCPS, the potency of FN for attachment of bone-derived cells was at least equal to that of VN.
authors.affiliation: CSIRO Division of Biomolecular Engineering, Sydney Laboratories, North Ryde, NSW, Australia.
authors.forename: J G
authors.initials: JG
authors.lastname: Steele
authors.forename: C
authors.initials: C
authors.lastname: McFarland
authors.forename: B A
authors.initials: BA
authors.lastname: Dalton
authors.forename: G
authors.initials: G
authors.lastname: Johnson
authors.forename: M D
authors.initials: MD
authors.lastname: Evans
authors.forename: C R
authors.initials: CR
authors.lastname: Howlett
authors.forename: P A
authors.initials: PA
authors.lastname: Underwood
mesh_terms: D001842:Bone and Bones / Q000166:cytology*; D002448:Cell Adhesion; D002478:Cells, Cultured; D002648:Child; D046508:Culture Techniques / Q000295:instrumentation*; D005353:Fibronectins; D006023:Glycoproteins; D006801:Humans; D011137:Polystyrenes / Q000737:chemistry*; D013499:Surface Properties; D019096:Vitronectin
chemical_list: D005353:Fibronectins; D006023:Glycoproteins; D011137:Polystyrenes; D019096:Vitronectin