In vitro study of monocyte viability during the initial adhesion to albumin- and fibrinogen-coated surfaces.
In vitro study of monocyte viability during the initial adhesion to albumin- and fibrinogen-coated surfaces.
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DOI：
10.1016/s0142-9612(00)00246-5
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发表时间：
2001-04
期刊：
影响因子：
14
通讯作者：
M. Werthén;A. Sellborn;M. Källtorp;H. Elwing;P. Thomsen
中科院分区：
文献类型：
--
作者：
M. Werthén;A. Sellborn;M. Källtorp;H. Elwing;P. Thomsen
表面粘附的单核细胞和巨噬细胞在生物材料的炎症反应中起着重要作用。在本研究中的粘附，生存能力和凋亡的变化，在材料表面粘附单核细胞在体外细胞表面相互作用的第一个小时进行了研究，使用组织培养聚苯乙烯表面包被人白蛋白和纤维蛋白原。采用两步梯度离心法富集人外周血单核细胞，并将其重悬于含10%胎牛血清的RPMI中（1×106/ml）。将细胞加入到用人纤维蛋白原或白蛋白包被的聚苯乙烯表面上，并在37°C（5%C02，100%湿度）下孵育30分钟、1、2、3和24小时。分别使用膜联蛋白-V-荧光素和碘化丙啶染色对粘附细胞进行早期凋亡变化（暴露的磷脂酰丝氨酸）和细胞死亡染色。在纤维蛋白原包被的表面上观察到双相粘附，在30 min和24 h后具有最高粘附细胞数，而1- 3 h后细胞数显著减少。白蛋白上的贴壁细胞数在所有短时间孵育后均相对较低，但在24 h后达到较高水平;白蛋白和纤维蛋白原上的贴壁死细胞数在孵育1 h后均达到最高水平（分别为约30%和约15%）。在24小时培养物中，两个表面上的贴壁细胞的活力都很高（95-100%）。细胞-材料表面相互作用30分钟后，仅在白蛋白包被的表面上清楚地观察到早期凋亡变化染色阳性的活细胞。因此，细胞死亡，包括凋亡性死亡，似乎在单核细胞和外源性表面之间的初始相互作用过程中发挥重要作用。
Surface adherent monocytes and macrophages play a central role in the inflammatory response to biomaterials. In the present study the adhesion, viability and apoptotic changes in material surface adherent monocytes during the first hours of cell-surface interactions in vitro were studied, using tissue culture polystyrene surfaces coated with human albumin and fibrinogen. Human peripheral blood monocytes were enriched by a two-step gradient centrifugation and resuspended (1×106/ml) in RPMI with 10% fetal bovine serum. The cells were added to polystyrene surfaces coated with human fibrinogen or albumin and incubated in 37°C (5% CO2, 100% humidity) for 30min, 1, 2, 3 and 24h. The adherent cells were stained for early apoptotic changes (exposed phosphatidylserine) and cell death using Annexin-V-fluorescein and propidium iodide staining, respectively. A bi-phasic adhesion was observed on the fibrinogen coated surface, having the highest number of adherent cells after 30min and 24h, while the cell number was markedly reduced after 1–3h. The number of adherent cells on albumin was relatively low after all short time incubations but had reached a high level after 24h.The number of adherent dead cells was highest after 1h on both albumin (∼30%) and fibrinogen (∼15%). In the 24h cultures, the viability of adherent cells was high on both surfaces (95–100%). Viable cells staining positive for early apoptotic changes could only be clearly observed on the albumin coated surface, after 30min of cell–material surface interaction. Cell death, including apoptotic death, thus seems to play an important role during the initial interactions between monocytes and a foreign surface.