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S. Kanayama, T. Yoshimatsu, S. Garty, B. Kim, B. D. Ratner, R. O. Wong, T. T. Shen; Imaging the Dynamics of Corneal Epithelial Cell-Polymer Integration Using Multiphoton Microscopy. Invest. Ophthalmol. Vis. Sci. 2010;51(13):1156.
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To determine the feasibility of using live-imaging approaches to visualize and monitor cell-polymer interactions in a corneal epithelial cell-polymer model. Cellular integration with polymeric constructs is an essential component in developing artificial organs such as artificial cornea. Current methods, such as histology and scanning electron microscopy (SEM) are currently the standard procedures for detailed examination of cell-polymer interactions. However, limited information can be derived from "static images" acquired at fixed time points after tissue fixation. Live-cell imaging studies are necessary for assessing how different polymeric structures affect cell dynamics and eventual integration.
A cell-polymer culture model was established to investigate the cellular interactions with a porous polymer: SV40-transfected corneal epithelial cells were seeded and cultured with a sphere-templated polymer material developed for artificial cornea. High resolution SEM was used to determine when after plating (6,12, 24 hours), cells were present within the porous structure. For time-lapse multiphoton imaging, cells were incubated with Calcein AM, then seeded onto the polymer and were acquired at 15 minute intervals over several hours.
Three-dimensional reconstruction of the images revealed the movement of the corneal epithelial cells across and within the polymer, changes in cell morphology, and process extension during integration with the substrate.
Cell-polymer interactions can thus be characterized using time-lapse multiphoton microscopy. This approach could offer new insights for better understanding of the cell polymer interaction process that would guide future designs of polymers before costly in vivo studies are conducted.
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