Calcification is one of the key factors for short durability of glutaraldehyde-crosslinked bioprosthetic heart valves. We have shown previously that procyanidins (PC)-crosslinked heart valve matrix has low cytotoxicity, perfect mechanical properties, and ideal stability. The aim of this study is to investigate the anticalcification effect of procyanidins and its crosslinked heart valves. Porcine aortic heart valve leaflets were decellularized and crosslinked with PC solution. The inhibition of calcification on PC-crosslinked heart valves was evaluated by soaking valves in simulated body fluid. The anticalcification effect of PC on valvular related cells was evaluated by culturing cells in the presence of PC. The results showed that minerals deposited on non- and glutaraldehyde-crosslinked valvular matrix in simulated body fluid, and PC could inhibit valve matrix mineralization in a dose-dependant manner. In addition, PC inhibited osteodifferentiation and calcification of valvular related cells by suppressing alkaline phosphatase activity and mineral deposition of cells. In conclusion, procyanidins can inhibit calcification of valvular matrix effectively by blocking mineral deposition and suppressing alkaline phosphatase activity and calcification of valvular related cells. Therefore, procyanindins-crosslinked heart valve matrix may be a potential candidate as new bioprosthetic heart valve implants.