Production of Highly Ordered Nanoporous Alumina and its Application in Cell Cultivation
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Production of Highly Ordered Nanoporous Alumina and its Application in Cell Cultivation Andreas Hoess1, Andrea Staeudte1, Annika Thormann1, Martin Steinhart2, and Andreas Heilmann1 1 Biological Materials and Interfaces, Fraunhofer Institute for Mechanics of Materials, WalterHülse-Strasse 1, Halle, 06120, Germany 2 Experimental Department II, Max Planck Institute of Microstructure Physics, Weinberg 2, Halle, 06120, Germany ABSTRACT Nanoporous alumina membranes were utilized as cell culture substrates. Two different anodization processes were applied in order to obtain highly ordered, self-supporting nanoporous alumina membranes. The resulting membranes exhibit uniform nanopores with constant diameters having aspect ratios (pore depth:pore diameter) larger than 1000. Tentative experiments evaluating the cellular interaction of a hepatoma cell line (HepG2) with the nanoporous alumina membranes will be described. Furthermore, cell proliferation as well as the adhesion behavior of the cells on the porous substrates was investigated. Scanning electron microscopy (SEM) and focused ion beam (FIB) technology were applied to examine the cell morphology. The experiments revealed that the nanoporous membranes can be used as cell culture substrates and that they have no adverse effects on cell proliferation and cell viability. Besides, cells on membranes with pore diameters larger than 200 nm developed small cell extensions (filopodia) which penetrated into the nanopores. INTRODUCTION Electrochemical oxidation of aluminum in acidic electrolytes under constant potential conditions results in the formation of a nanoporous aluminum oxide (alumina) layer on its surface [1-3]. By applying specific techniques in the end of the production process selfsupporting nanoporous alumina membranes can be obtained. They are characterized by regularly distributed, parallel pores which are aligned perpendicular to the membrane surface. By variations in the production process (e.g. applied voltage, used electrolyte, duration of the anodization) the membrane properties, such as pore diameter or membrane thickness, can be influenced. The unique structure of these membranes is highly advantageous for the production of nanostructured device components. However, its application in biomedical fields like drug delivery, tissue engineering or cell cultivation appears also to be promising. The supply of nutrients or molecules to both sides of cells grown on the membranes can be controlled by adjusting membrane properties like pore diameter, porosity and membrane thickness. Such a configuration is associated with fundamental advantages for the cultivation of cells with polar phenotype, e.g. liver cells (hepatocytes), or for the set-up of co-cultures of different cell types on the opposite surfaces of the membrane. Currently nanoporous alumina is proposed as a useful coating of bone implants in order to improve cell ingrowth and implant stability [4-6]. The studies revealed that nanoporous alumina membranes have no detrimental effects on cell v
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