Institute of Advanced Biomedical Engineering and Science and Global Center of Excellence (G-COE) Program, Tokyo Women's Medical University, 8-1 Kawadacho, Shinjuku-ku, Tokyo 162-8666, Japan.
Institute of Advanced Biomedical Engineering and Science and Global Center of Excellence (G-COE) Program, Tokyo Women's Medical University, 8-1 Kawadacho, Shinjuku-ku, Tokyo 162-8666, Japan.
Acta Biomater. 2014 Aug;10(8):3398-408. doi: 10.1016/j.actbio.2014.03.024. Epub 2014 Mar 27.
A double polymeric nanolayer consisting of poly(N-isopropylacrylamide) (PIPAAm) and hydrophilic polyacrylamide (PAAm) was deposited on tissue culture polystyrene (TCPS) surfaces using electron beam irradiation to form a new temperature-responsive cell culture surface in which the basal hydrophilic PAAm component in the double polymeric layer promotes the hydration of the upper PIPAAm layer and induces rapid cell detachment compared to a conventional temperature-responsive cell culture surface, PIPAAm-grafted TCPS (PIPAAm-TCPS). Take-off angle-dependent X-ray photoelectron spectroscopy spectral analysis demonstrated that the grafted PIPAAm and PAAm components were located in the upper and basal regions of the double polymeric layer, respectively, suggesting that the double polymeric layer forms an inter-penetrating-network-like structure with PAAm at the basal portion of the PIPAAm grafted chains. The wettability of the temperature-responsive cell culture surfaces with the double polymeric layer tended to be more hydrophilic, with an increase in the basal PAAm graft density at a constant PIPAAm graft density. However, when the graft densities of the upper PIPAAm and basal PAAm were optimized, the resulting temperature-responsive cell culture surface with the double polymeric layer exhibited rapid cell detachment while maintaining cell adhesive character comparable to that of PIPAAm-TCPS. The cell adhesive character was altered from cell-adhesive to cell-repellent with increasing PAAm or PIPAAm graft density. The cell adhesive character of the temperature-responsive cell culture surfaces was relatively consistent with their contact angles. These results strongly suggest that the basal PAAm surface properties affect the degree of hydration and dehydration of the subsequently grafted PIPAAm. In addition, the roles of the hydrophilic component in accelerating cell detachment are further discussed in terms of the mobility of the grafted PIPAAm chains. Applications of this insight might be useful for designing temperature-responsive cell culture surfaces for achieving efficient cell culture and quick target cell detachment.
一种由聚（N-异丙基丙烯酰胺）（PIPAAm）和亲水性聚丙烯酰胺（PAAm）组成的双层聚合纳米层通过电子束辐照沉积在组织培养聚苯乙烯（TCPS）表面上，形成一种新的温度响应细胞培养表面，其中双层聚合层中的基本亲水性 PAAm 成分促进了上层 PIPAAm 层的水合作用，并诱导细胞快速脱离，与传统的温度响应细胞培养表面，即 PIPAAm 接枝 TCPS（PIPAAm-TCPS）相比。起飞角依赖的 X 射线光电子能谱光谱分析表明，接枝的 PIPAAm 和 PAAm 组分分别位于双层聚合层的上层和基底区域，表明双层聚合层与 PIPAAm 接枝链的基底部分形成互穿网络状结构。具有双层聚合层的温度响应细胞培养表面的润湿性趋于更亲水，在恒定的 PIPAAm 接枝密度下，基底 PAAm 接枝密度增加。然而，当优化上层 PIPAAm 和基底 PAAm 的接枝密度时，具有双层聚合层的所得温度响应细胞培养表面在保持与 PIPAAm-TCPS 相当的细胞粘附特性的同时表现出快速的细胞脱离。随着 PAAm 或 PIPAAm 接枝密度的增加，细胞粘附特性从细胞粘附变为细胞排斥。温度响应细胞培养表面的细胞粘附特性与其接触角相对一致。这些结果强烈表明基底 PAAm 表面特性影响随后接枝的 PIPAAm 的水合和去水合程度。此外，还进一步讨论了亲水性成分在加速细胞脱离方面的作用，这是基于接枝的 PIPAAm 链的迁移率。这一见解的应用可能有助于设计用于实现高效细胞培养和快速目标细胞脱离的温度响应细胞培养表面。
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