Objective To observe the attachment and growth of human periodontal ligament cells (hPDLCs) on electrospun poly (L-lactic acid)/hydroxyapatite (PLLA/HA) fibrous biomaterial (called "biomaterial" for short), and to investigate the feasibility of applying this new material in human periodontal tissue regeneration. Methods PLLA/HA fibrous biomaterial was fabricated by electrospinning technique. the hPDLCs were harvested from healthy premolars extracted for or BR>thodontics, and the cells were identified by immunohistochemical method. The compatibility of the biomaterial was evaluated by the MTT assay method. the hPDLCs were seeded onto the PLA/HA fibrous biomaterial, and then the compound was observed under scanning electron microscope; the hPDLCs transfected by the recombinant adenovirus containing green fluorescent protein was cocultured with the biomaterial and observed under confocal laser scanning microscope. Results The hPDLCs were successfully separated, and immunohistochemistry staining of vimentin protein showed that the cells came from the mesoderm; the MTT assay results showed that the cells propagated on the biomaterial well compared with cultured normally; the scanning electon microscope (SEM) results showed the cells on the biomaterial grew and stretched well; the confocal laser scanning microscope results showed that the cells grew along the fiber of the biomaterial, and presented a certain three dimensional structure. Conclusion The results suggested that electrospun PLLA/HA fibrous biomaterial possesses a three dimensional net-like structure and a good cellular compatibility with hPDLCs; the hPDLCs can adhere and propagate well on it. This experiment can provide some basic theory for the use of electrospun PLLA/HA fibrous biomaterial in the regeneration of human periodontal tissue.