Biocompatibility study of a biological tissue fixed with a naturally occurring crosslinking reagent

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Abstract
A recognized disadvantage of the currently available chemical reagents used to fix bioprostheses is the potential toxic effects a recipient may be exposed to from residues. It is therefore desirable to provide a crosslinking reagent that is of low cytotoxicity and can form stable and biocompatible crosslinked products. To achieve this goal, a naturally occurring crosslinking reagent—genipin—was used by our group to fix biological tissues. Genipin can be obtained from its parent compound geniposide, which can be isolated from the fruits of Gardenia jasminoides ELLIS. In our previous feasibility study, it was found that the cytotoxicity of genipin is significantly lower than both glutaraldehyde and an epoxy compound. Additionally, it was shown that genipin can form stable crosslinked products. The present study further investigates the biocompatibility of a genipin‐fixed porcine pericardium implanted subcutaneously in a growing rat model. The fresh, glutaraldehyde‐, and epoxy‐fixed counterparts were used as controls. It was noted that the inflammatory reaction of the genipin‐fixed tissue was significantly less than its glutaraldehyde‐ and epoxy‐fixed counterparts. Also, the genipin‐fixed tissue has tensile strength and resistance against in vivo degradation comparable to the glutaraldehyde‐fixed tissue. Additionally, the calcium content of the genipin‐fixed tissue measured throughout the entire course of the study was minimal. Nevertheless, further study in calcification for the genipin‐fixed tissue should be conducted in a blood‐contact environment. The results obtained in this subcutaneous study indicate that genipin is a promising crosslinking reagent for biological tissue fixation. However, further durability testing in vitro and in vivo are needed to determine the relative functional merits of this new crosslinker. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 42, 568–576, 1998.