Karlsson M, Pålsgård E, Wilshaw P R, Di Silvio L
Center for Surface Biotechnology, BMC, Husarg. 3, D7:3, Box 577, 751 23 Uppsala, Sweden.
Biomaterials. 2003 Aug;24(18):3039-46. doi: 10.1016/s0142-9612(03)00146-7.
In the present study we have used a characterised primary human cell culture model to investigate cellular interactions with nano-porous alumina. This material, prepared by anodisation, is being developed as a coating on titanium alloy implants. The structure of the alumina, as determined by X-ray diffraction and transmission electron microscopy, was amorphous. When studying cell/material interactions we used both biochemical and morphological parameters. Cell viability, proliferation and phenotype were assessed by measurement of redox reactions in the cells, cellular DNA, tritiated thymidine ([3H]-TdR) incorporation and alkaline phosphatase (ALP) production. Results showed a normal osteoblastic growth pattern with increasing cell numbers during the first 2 weeks. A peak in cell proliferation was seen on day 3, after which cell growth decreased, followed by an increase in ALP production, thus indicating that the osteoblastic phenotype was retained on the alumina. Cell adhesion was observed, the osteoblast-like cells having a flattened morphology with filipodia attached to the pores of the material. SDS-PAGE and western blot measurements showed that the nano-porous alumina was able to adsorb fibronectin. Trace amounts of aluminium ions were measured in the surrounding medium, but no adverse effect on cell activity was observed.
在本研究中，我们使用了一种经过表征的原代人细胞培养模型来研究细胞与纳米多孔氧化铝之间的相互作用。这种通过阳极氧化制备的材料正被开发用作钛合金植入物的涂层。通过X射线衍射和透射电子显微镜确定，氧化铝的结构为非晶态。在研究细胞/材料相互作用时，我们使用了生化和形态学参数。通过测量细胞中的氧化还原反应、细胞DNA、氚标记胸腺嘧啶核苷（[3H]-TdR）掺入量和碱性磷酸酶（ALP）产量来评估细胞活力、增殖和表型。结果显示，在前两周细胞数量增加，呈现出正常的成骨细胞生长模式。在第3天观察到细胞增殖高峰，之后细胞生长下降，随后ALP产量增加，这表明成骨细胞表型在氧化铝上得以保留。观察到细胞黏附现象，成骨样细胞形态扁平，有丝状伪足附着在材料的孔隙上。SDS-PAGE和蛋白质印迹测量表明，纳米多孔氧化铝能够吸附纤连蛋白。在周围培养基中检测到微量铝离子，但未观察到对细胞活性有不利影响。
