Investigation of Nanofibrillar Influence on Cell-Cell Interactions of Astrocytes by Epi-fluorescence and Atomic Force Mi
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Investigation of Nanofibrillar Influence on Cell-Cell Interactions of Astrocytes by Epi-fluorescence and Atomic Force Microscopies Volkan M. Tiryaki1, Virginia M. Ayres1, Adeel A. Khan2, Dexter A. Flowers3, Ijaz Ahmed4, Roberto Delgado-Rivera5, Sally Meiners6 1 College of Engineering, Michigan State University, East Lansing MI 48824 2 College of Engineering, Western Michigan University, Kalamazoo, MI 49008 3 College of Engineering, Wayne State University, Detroit, MI 48202 4 College of Engineering, Rutgers, State University of New Jersey, Piscataway, NJ 08854 5 Department of Chemistry and Chemical Biology, Rutgers, State University of New Jersey, Piscataway, NJ 08854 6 117 Bedford Court, Piscataway, NJ, 08854 ABSTRACT Long distance intercellular communication between astrocytes on nanofibrillar and planar surfaces was investigated by epi-fluorescence microscopy and atomic force microscopy. We found that astrocytes on nanofibrillar surfaces and astrocytes on planar surfaces diverged in apparent cell-cell contact structures. Astrocytes on nanofibrillar surfaces exhibited a “single cellular process” response, while astrocytes on planar surfaces exhibited a filopodial network response. The possibility that astrocytes can sense their geometrical environment and form different cell-to-cell contacts on nanofibrillar versus planar surfaces, with activation of different signaling pathways, is discussed. INTRODUCTION Cell-cell interactions via direct connective bridges over distances that are much larger than traditional cell membrane-to-cell membrane junction distances are currently of great interest. These interactions are potentially overlooked vectors for the exchange of genetic information or for viral transmission [1]. In the present work, epi-fluorescence microscopy and atomic force microscopy (AFM) were used to investigate the possible long distance (> 50 µm) cell-cell interactions of astrocytes in detail. Astrocytes are key cellular components of the central nervous system that function as cellular intermediaries between capillaries and neurons and also provide a mechanical support environment [2]. In a wound-healing situation, astrocytes form a continuous layer that seals off the wound site. Intercellular communication, initiated prior to cell body contact, may therefore play a role in wound healing. Cultured astrocytes have been shown by our group to support enhanced neurite outgrowth in co-culture with cerebellar granule neurons when they are plated on synthetic polyamide nanofibrillar surfaces compared with planar plastic (ACLAR) surfaces [3]. This may indicate a more physiologically relevant function for astrocytes cultured on nanofibrillar versus planar surfaces, as a critical role of astrocytes in vivo is to support neuronal outgrowth during development and following injury. Nanofibrillar meshworks
in turn structurally resemble the basement membrane that separates astrocytes from capillary endothelial cells in the brain and the spinal cord. In the present work, we therefore explored the hypothesis that astro
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