Inverse inkjet printed gold micro electrodes for the structured deposition of epithelial cells and fibrin

Rolf Zehbe , Ulrich Gross , Helmut Schubert
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引用次数: 7

Abstract

The micro structured deposition of vital cells is an important challenge in tissue engineering, biosensor technology, and in all research dealing with cell–cell and cell–substrate contacts. Hence, an inkjet printing technology has been developed to manufacture Au-based micro electrodes by sputter coating inversely printed polyester-foils. These electrodes feature minimal structure sizes of 35 μm and consist of an anode and a cathode part. They were used with fibrinogenic epithelial cell suspensions to deposit human keratinocytes (HaCaT), mouse fibroblasts (L-929) and the protein fibrin by applying DC voltage. Subsequently cells were electrophoretically attracted to the anode, following exactly its shape, while the insoluble fibrin was simultaneously precipitated due to the electrically mediated polymerization of the soluble fibrinogen molecule. Furthermore, it was demonstrated that this technique is suitable to co-deposit both cell types in a layered fashion.

The lower voltage boundary for successful deposition was set at approximately 0.8 V needed for the conversion of fibrinogen into fibrin, while the upper voltage boundary was set at approximately 1.85 V, when commencing electrolysis inhibited the deposition of vital cells.

Subsequent to the anodic cell-fibrin deposition, cells were cultivated for up to 4 days and then characterized by FDA + EB staining, methyl violet staining, MNF staining and SEM. The conversion from fibrinogen into fibrin was studied using ATR/FTIR.

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用于上皮细胞和纤维蛋白结构沉积的逆喷墨印刷金微电极
重要细胞的微结构沉积是组织工程、生物传感器技术以及所有涉及细胞-细胞和细胞-基质接触的研究中的一个重要挑战。因此,本文发展了一种喷墨印刷技术,通过溅射涂覆反印刷聚酯箔来制造金基微电极。这些电极的最小结构尺寸为35 μm,由阳极和阴极部分组成。将它们与纤维原性上皮细胞混悬液一起使用,施加直流电压沉积人角质形成细胞(HaCaT)、小鼠成纤维细胞(L-929)和纤维蛋白。随后,细胞被电泳吸引到阳极上,完全遵循其形状,而不溶性纤维蛋白同时由于可溶性纤维蛋白原分子的电介导聚合而沉淀。此外,还证明了该技术适用于以分层方式共同沉积两种细胞类型。成功沉积的低电压边界设置在纤维蛋白原转化为纤维蛋白所需的约0.8 V,而高电压边界设置在约1.85 V,此时开始电解会抑制重要细胞的沉积。阳极细胞-纤维蛋白沉积后,细胞培养4天,然后用FDA + EB染色、甲基紫染色、MNF染色和SEM进行表征。利用ATR/FTIR研究了纤维蛋白原向纤维蛋白的转化。
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