Three-dimensional (3D) cell cultures have many advantages over two-dimensional cultures. However, seeding cells in 3D scaffolds such as nonwoven fibrous polyethylene terephthalate (PET) matrices has been a challenge task in tissue engineering and cell culture bioprocessing. In this study, a centrifugal seeding method was investigated to improve the cell seeding efficiency in PET matrices with two different porosities (93% and 88%). Both the centrifugal force and centrifugation time were found to affect the seeding efficiency. With an appropriate centrifugation speed, a high 80-90% cell seeding efficiency was achieved and the time to reach this high seeding efficiency was less than 5 min. The seeding efficiency was similar for matrices with different porosities, although the optimal seeding time was significantly shorter for the low-porosity scaffold. Post seeding cell viability was demonstrated by culturing colon cancer cells seeded in PET matrices for over 5 days. The centrifugal seeding method developed in this work can be used to efficiently and uniformly seed small fibrous scaffolds for applications in 3D cell-based assays for high-throughput screening.
Abstract:Tissue engineering is an emerging field in biomedicine, holding enormous promise for regenerative medicine. Scaffolds, within which cells proliferate, are a controlling factor in tissue engineering applications. Upon fabrication, tissue scaffolds must undergo appropriate sterilization to eliminate contaminants. Current sterilization methods are either costly, time consuming, or ineffective. In this study, a quaternary salt, benzalkonium chloride (BAC), was used as a chemical agent for sterilization of nonwoven polyethylene terephthalate (PET) fibers and polylactic acid nanofibers. Treating the PET scaffolds with 0.1% (w/v) BAC for only 2 minutes was effective to eliminate bacterial contaminants in the fibrous matrices. In addition, astrocyte cells were successfully cultured in the PET scaffolds following BAC sterilization, demonstrating the suitability of BAC as a sterilization agent. This chemical sterilization method is also mild and nonabrasive to nanostructured materials such as electrospun polylactic acid nanofibers.
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