Steele J G, McFarland C, Dalton B A, Johnson G, Evans M D, Howlett C R, Underwood P A
CSIRO Division of Biomolecular Engineering, Sydney Laboratories, North Ryde, NSW, Australia.
J Biomater Sci Polym Ed. 1993;5(3):245-57. doi: 10.1163/156856293x00339.
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.
细胞培养研究常常被用于确定生物材料作为细胞附着和生长表面的适用性。对于此类潜在用于骨植入物的表面研究,源自骨的细胞可以在组织培养聚苯乙烯(TCPS)上进行培养。我们已经确定了血清纤连蛋白(FN)或玻连蛋白(VN)在接种到培养表面后的最初90分钟内,对从人骨外植体培养的细胞(骨源性细胞)的附着和铺展所起的作用。通过向接种培养基中添加10%(v/v)胎牛血清(FBS),骨源性细胞对TCPS的附着被模拟了两倍。FN和VN的作用是通过在添加到培养基之前从FBS中选择性去除FN或VN来确定的。去除了VN的FBS没有这种刺激活性。相反,骨源性细胞从含有耗尽FN的血清(其中含有VN)的培养基附着到TCPS上的情况与使用完整FBS时相同。当接种在同时耗尽VN和FN的FBS的培养基中时,骨源性细胞存在不完全附着(27%的细胞)。我们的结果表明,对于人骨源性细胞,在培养的最初90分钟内,接种在含有FBS的培养基中的细胞附着到TCPS上主要是由于吸附到血清VN的表面。由于未改性的聚苯乙烯(PS)先前也被用作模型生物材料表面,因此将PS与TCPS进行了比较,以观察骨源性细胞的附着情况。无论培养基是无血清的还是含有FBS的,骨源性细胞对TCPS的附着都是对PS的两倍。骨源性细胞附着到预先包被有纯化VN或FN的TCPS或PS上,吸附到PS上的VN与吸附到TCPS上的VN效果相同。对于FN,聚苯乙烯表面化学有影响,这表现为次优浓度的FN吸附到TCPS上时比相同浓度的FN包被到PS上时具有略高的效力。当预先吸附到TCPS上时,FN对骨源性细胞附着的效力至少与VN相等。