(430t) Novel Approaches of Scaffold Sterilization and Cell Seeding for Tissue Engineering Application

Sterilization and seeding are the preliminary steps dictating the performance of the tissue engineering scaffolds and constructs. Although several sterilization methods are available for tissue engineering scaffolds, most of them are ineffective, inconvenient or detrimental to the cells. In this study, a quartenary ammonium salt, Benzalkonium chloride (BAC), was assessed for its sterilization activity. The activity was tested against a range of microorganism stains, such as yeast, Bacillus and Streptococcus. Assessment using 0.1% BAC demonstrated an activity of higher than 99.99% in less than one minute treatment time. In addition, results showed that BAC sterilization followed by proper washing after the sterilization was not detrimental to astrocyte cells. This innocuous yet highly efficient chemical sterilization method is very suitable for sterilization of susceptible nanofibrous materials. Seeding protocols for tissue engineering can vary from static seeding to filtration seeding to centrifugation seeding. Static seeding, which is the commonly used seeding protocol, is very inefficient with a lower than 60% seeding efficiency. Besides, static seeding results in non-uniformity cell distribution depending on the volume of the cell suspension added to the scaffolds. Although filtration seeding can achieve higher seeding efficiency and more uniform cell distribution, it is very time consuming. With our developed centrifugation seeding, we achieved up to 85% seeding efficiency in less than three minutes seeding time. In conclusion, we demonstrated novel protocols for scaffold sterilization and cell seeding for tissue engineering application.