As an ongoing effort to elucidate the mechanisms involved in bioprosthetic heart valve (BHV) calcification, the role of C-reactive protein (CRP) in the tissue calcification process was investigated. The profile of calcium-associated proteins (CAP) on glutaraldehyde-preserved (0.6%) porcine aortic wall, which were subcutaneously implanted in rats for up to 8 weeks, showed a temporal appearance pattern. The total extracted amount of proteins from the control tissues implanted for 8 weeks was significantly greater than that from ethanol-treated tissues (1.78 ± 0.2 vs. 1.27 ± 0.18 µg/mg), indicating that the binding affinity of CAP for BHV pretreated with an anticalcification agent was significantly decreased (p < 0.05). The dye Stains-All method showed that the dark-blue colored bands, representing high calcium binding and phosphorylated proteins, were stained from the extract of the control BHV at the molecular weight varying from 4 to 250 kDa, but rarely seen in the extract of BHV pretreated with ethanol. One of those proteins was exclusively immunoreactive with CRP antibody, while there was no immunoreaction in less calcified tissues. When aortic wall was exposed to an excess amount of CRP in an in vitro simulating model, the calcification rate of aortic wall increased as the concentration of CRP increased. The results of this work clearly revealed that CRP has indirect vascular effects, leading to an increased rate of aortic wall calcification.