Effects of Autoclaving, EtOH, and UV Sterilization on the Chemical, Mechanical, Printability, and Biocompatibility Characteristics of Alginate.

Sterilization is a necessary step during the processing of biomaterials, but it can affect the materials' functional characteristics. This study characterizes the effects of three commonly used sterilization processes-autoclaving (heat-based), ethanol (EtOH; chemical-based), and ultraviolet (UV; radiation-based)-on the chemical, mechanical, printability, and biocompatibility properties of alginate, a widely used biopolymer for drug delivery, tissue engineering, and other biomedical applications. Sterility assessment tests showed that autoclaving was effective against Gram-positive and Gram-negative bacteria at loads up to 108 CFU/mL, while EtOH was the least effective. Nuclear magnetic-resonance spectroscopy showed that the sterilization processes did not affect the monomeric content in the alginate solutions. The differences in compressive stiffness of the three sterilized hydrogels were also not significant. However, autoclaving significantly reduced the molecular weight and polydispersity index, as determined via gel permeation chromatography, as well as the dynamic viscosity of alginate. Printability analyses showed that the sterilization process as well as the extrusion pressure and speed affected the number of discontinuities and spreading ratio in printed and cross-linked strands. Finally, human adipose-derived stem cells demonstrated over 90% viability in all sterilized hydrogels over 7 days, but the differences in cellular metabolic activity in the three groups were significant. Taken together, the autoclaving process, while demonstrating broad spectrum sterility effectiveness, also resulted in most notable changes in alginate's key properties. In addition to the specific results with the three sterilization processes and alginate, this study serves as a roadmap to characterize the interrelationships between sterilization processes, fundamental chemical properties, and resulting functional characteristics and processability of hydrogels.