Aim To compare the adhesion and proliferation of late endothelial progenitor cells (EPC) isolated and expanded from rabbit peripheral blood cultured with nanofibers and alignate nanofibers synthetic poly-L-lactic acid (PLLA) scaffold in vitro for applications in vascular tissue engineering,thus to optimize PLLA scaffold materials. Methods Nanofibrous scaffolds of PLLA by electrospinning were modified by hypothermy plasma and type I collagen was coated onto the materials physically. In vitro,late EPC were cultured on the PLLA scaffold. Furthermore,cell incorporation of DiI-labelled ac-LDL and binding of FITC-UEA-I was determined by fluorescent microscopy. Adherency and proliferation of EPC in two groups at different time points were surveyed and to observe morphological change of late EPC on PLLA scaffold was observed by contrast phase microscope,fluorescent microscope and scanning electron microscope. Results The nanofibrous scaffolds consists of fibers with diameters ranging from 300 nm to 400 nm,whose porosities were more than 90%. The adherency of late EPC on scaffold were completed in 12 hours. The proliferation of late EPC on alignate nanofibers scaffold was markedly increased compared to control groups in every time (P<0.05). Late EPC grew well with PLLA scaffold,yet confused and disorderly in control groups. Late EPC could attach,extend,proliferate and secrete extracellular matrix following fibrous orientation in alignate nanofibers groups of PLLA,which majority of the fibers were oriented along the longitudinal axis which form a unique aligned topography. Conclusions Late EPC are ideal seeding cells for tissue engineering. Late EPC can adhere well to alignate nanofibers of PLLA and proliferate,keep well on cell morphology. This type of PLLA scallfold is a optimal candidate material for late EPC transplantation in vascular tissue engineering.