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不同粗糙度的聚苯乙烯和聚醚酰亚胺细胞培养插入物的免疫评估。

Immunological evaluation of polystyrene and poly(ether imide) cell culture inserts with different roughness.

机构信息

Center for Biomaterial Development and Berlin-Brandenburg Centre for Regenerative Therapies, Institute of Polymer Research, Helmholtz-Zentrum Geesthacht, Teltow, Germany.

出版信息

Clin Hemorheol Microcirc. 2012;52(2-4):375-89. doi: 10.3233/CH-2012-1612.

Abstract

For the successful clinical and biological application of polymers, their interaction with cells, tissues, and body fluids has to be well characterized. In order to investigate how the physical, chemical, and mechanical properties of candidate biomaterials influence cell behaviours, the testing sample is usually placed in commercially available cell culture plates. Thus, not only the testing sample itself but also the culture dish material might influence the cell behaviour. Therefore, an insert system was created to exclude this influence and allow investigations of the testing material solely. In this study micropatterned inserts prepared from polystyrene (PS) as well as from poly(ether imide) (PEI) with three different roughness levels of i) Rq = 0.29 μm (PS) and 0.23 μm (PEI); ii) Rq = 3.47 μm (PS) and 3.92 μm (PEI); and iii) Rq = 22.16 μm [corrected] (PS) and 22.65 μm (PEI) were explored with regard of their immuno-compatibility including the determination of potential contaminations with endotoxins or other microbial products. The endotoxin levels of the inserts were determined to be less than 0.07 EU/mL, which is well below the U.S. Food and Drug Administration limit of 0.5 EU/mL and the survival of murine macrophages cultured in the inserts was not impaired. Activation of early immune mechanisms such as complement activation and the generation of reactive oxygen species could not be observed. All tested materials had no influence on the cytokine secretion from cells of whole human blood. The investigated inserts were immuno-compatible and apparently free of contaminations with microbial products. The roughness of the inserts had no stimulatory or inhibitory effect on early immune mechanisms. Conclusively, the 24-well plate insert systems introduced in this study allow investigating the interactions of tailored surface properties such as roughness with many other cell types, without the disadvantage of the standard commercially available culture vessels influencing the biomaterial testing.

摘要

为了成功将聚合物应用于临床和生物领域,必须充分了解其与细胞、组织和体液的相互作用。为了研究候选生物材料的物理、化学和机械性能如何影响细胞行为,通常将测试样品放置在市售的细胞培养板中。因此,不仅测试样品本身,而且培养皿材料也可能影响细胞行为。因此,创建了插入系统以排除这种影响,并仅允许对测试材料进行研究。在这项研究中,探索了由聚苯乙烯(PS)和聚醚酰亚胺(PEI)制成的具有三种不同粗糙度水平的微图案化插入物,粗糙度水平分别为 i)Rq = 0.29 μm(PS)和 0.23 μm(PEI);ii)Rq = 3.47 μm(PS)和 3.92 μm(PEI);和 iii)Rq = 22.16 μm [更正](PS)和 22.65 μm(PEI)。研究了它们的免疫相容性,包括确定内毒素或其他微生物产物的潜在污染。插入物的内毒素水平被确定为低于 0.07 EU/mL,远低于美国食品和药物管理局规定的 0.5 EU/mL 限值,并且在插入物中培养的鼠巨噬细胞的存活率没有受到损害。无法观察到早期免疫机制的激活,如补体激活和活性氧的产生。所有测试材料对全人血细胞的细胞因子分泌均无影响。所研究的插入物具有免疫相容性,显然没有微生物产物的污染。插入物的粗糙度对早期免疫机制没有刺激或抑制作用。总之,本研究中引入的 24 孔板插入系统允许研究诸如粗糙度等定制表面特性与许多其他细胞类型的相互作用,而不会受到标准市售培养容器影响生物材料测试的缺点。

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