The present invention provides methods for treating tissue to inhibit
post-implant calcification of a biological tissue. In one method of this
invention, a tissue is immersed in or otherwise contacted with a
pretreated glutaraldehyde solution, i.e., a heat-treated or pH-adjusted
glutaraldehyde solution. The tissue may be partially fixed with
glutaraldehyde prior to, after, or concurrently with the step of
contacting the tissue with the pretreated gluteraldehyde. Contact with
the pretreated gluteraldehyde produces free amine groups on the tissue,
which are subsequently blocked by contacting the crosslinked tissue with
a blocking agent. In another embodiment, a tissue is contacted with
either a non-pretreated glutaraldehyde or a pH-adjusted glutaraldehyde
solution for a period of time sufficient to crosslink the tissue. The
crosslinked tissue is then treated with a reducing agent that reduces
aldehyde and carboxylic acid groups on the fixed tissue.