Effects of sterilization methods on gelatin methacryloyl hydrogel properties and macrophage gene expressionin vitro

To assure the long-term safety and functional performance after implantation, it is of critical importance to completely sterilize a biomaterial implant. Ineffective sterilization can cause severe inflammation and infection at the implant site, leading to detrimental events of morbidity and even mortality. Macrophages are pivotal players in the inflammatory and foreign body response after implanting a biomaterial in the body. However, the relationship between the sterilization procedure and macrophage response has not been established. In this study, three commonly used sterilization methods, including autoclaving, ethylene oxide gas and ethanol treatment, were used to sterilize a gelatin methacryloyl hydrogel. The impacts of different sterilization methods on the structure and physical properties of the hydrogel were compared. Macrophage responses to the sterilized hydrogel were analyzed based on their morphology, viability andin vitrogene expression. It was found that the sterilization methods only marginally altered the hydrogel morphology, swelling behavior and elastic modulus, but significantly impacted macrophage gene expression within 48 h and over 7 din vitro. Therefore, when selecting sterilization methods for GelMA hydrogel, not only the sterility and hydrogel properties, such as material destruction and degradation caused by temperature and moisture, should be taken into consideration, but also the cellular responses to the sterilized material which could be substantially different.