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J Biomater Appl. 2013 Sep;28(3):395-406. doi: 10.1177/0885328212454555. Epub 2012 Jul 24.

Protein polymer hydrogels: effects of endotoxin on biocompatibility.

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  • 1Department of Chemical and Biological Engineering, Northwestern University, Evanston, IL, USA.


Protein polymer-based hydrogels have shown potential for tissue engineering applications, but require biocompatibility testing for in vivo use. Enzymatically crosslinked protein polymer-based hydrogels were tested in vitro and in vivo to evaluate their biocompatibility. Endotoxins present in the hydrogel were removed by Trition X-114 phase separation. The reduction of endotoxins decreased TNF-α production by a macrophage cell line in vitro; however, significant inflammatory response was still present compared to collagen control gels. A branched PEG molecule and dexamethasone were added to the hydrogel to reduce the response. In vitro testing showed a decrease in the TNF-α levels with the addition of dexamethasone. In vivo implantations into the epididymal fat pad of C57/BL6 mice, however, indicated a decreased inflammatory mediated immune response with a hydrogel treated with both PEGylation and endotoxin reduction. This study demonstrates the importance of endotoxin testing and removal in determining the biocompatibility of biomaterials.


Hydrogel; biocompatibility; dexamethasone; endotoxins; polyethylene glycol

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