This study sought to assess the effect of SspA on the formation of Staphylococcus aureus biofilm extending over the surfaces of Cardiovascular Biomaterial Dacron. SspA was extracted from the surface of staphylococcus aureus biofilm, purified, and then used to influence the adhesion of Staphylococcus aureus and the formation of Staphylococcus aureus biofilm on Dacron biomaterial surfaces. The formation of the Staphylococcus aureus biofilm on cardiovascular biomaterial Dacron surfaces under gradient SspA concentrations was evaluated by confocal laser microscopy. The result revealed that SspA inhibited the formation of Staphylococcus aureus biofilms on cardiovascular biomaterials surfaces effectively, and it was dose dependent. This study indicates that SspA is effective for preventing biomaterial centered infection and this method is conducive to clinical applications.