In the present study, three BMP-2 (bone morphogenetic protein-2) micropatterns were produced on polystyrene (PS) surface by microcontact printing in order to investigate the effect of protein micropattern on biomaterials surface on human chondrocytes behavior. The excellent BMP-2 micropatterns from fluorescent observation were successfully obtained. Cell adhesion and immunohistochemical assay were utilized to explore cell behavior. The results indicated that the protein micropatterns have significant influence on cell adhesion, spread, alignment and protein expression. The cells preferentially adhered on protein micropattern zones. The cells tend to spread towards micropattern direction, leading to better spread. The spread cells could excreted more type II and VI collagen than that of non-spread or poor-spread cells. The results of this paper suggested that extracellular matrix (ECM) protein micropatterns can effectively regulate human chondrocytes behavior, which may be helpful for development of new cartilage tissue.