In-Hydrogel Cell-Free Protein Expression System as Biocompatible and Implantable Biomaterial

Area of Science:

• Biomaterials Science
• Synthetic Biology
• Biomedical Engineering

Background:

• Therapeutic protein delivery faces challenges with centralized manufacturing, limiting point-of-care applications.
• Biomaterials offer potential for localized and dynamic protein production and release.

Purpose of the Study:

• To investigate confined cell-free protein synthesis within hydrogels for point-of-care biomaterial applications.
• To develop functional biomaterials capable of in situ protein production and delivery.

Main Methods:

• Entrapping cell-free extracts (CFEs) and plasmid DNA encoding super folded green fluorescence protein (sGFP) into hydrogels.
• Evaluating agarose hydrogels for protein synthesis, diffusion, and freeze-drying (FD) stability.
• Assessing in vitro biocompatibility and in vivo implantation in a preclinical mouse model.

Main Results:

• Agarose hydrogels efficiently confined cell-free protein synthesis and allowed sGFP diffusion.
• Freeze-dried agarose hydrogels maintained protein synthesis and diffusion capabilities.
• FD-agarose hydrogels demonstrated in vitro biocompatibility and successful in vivo implantation, with immune responses influenced by material properties.

Conclusions:

• Confined cell-free protein synthesis in hydrogels is a viable strategy for dynamic protein production and delivery.
• Freeze-dried hydrogels offer a promising platform for point-of-care biomaterial assembly and therapeutic protein approaches.
• This technology enables in vivo protein synthesis and targeted delivery, advancing protein therapeutics.