The effects of the γ-irradiatted PLLA on the osteoblasts and apatite formation were investigated in vitro. The PLLA sheet was γ-ray irradiated at the dose of 10, 25 or 50 kGy. The mouse osteoblast-like MC3T3-E1 cells and normal human osteoblast NHOst cells were micromass cultured on the PLLA sheet for 2 weeks, and then the proliferation and differentiation of the cells were determined. The proliferations of MC3T3-E1 and NHOst cells hardly changed with increasing irradiation dose. However, the differentiations of MC3T3-E1 and NHOst cells increased with irradiation dose. On the other hand, the surface of the PLLA sheet after soaking in the medium without the cells was characterized by SEM, EDX, FT-IR and XPS. The hydroxyapatite was formed on the surface of the PLLA sheet after soaking, and the amount of hydroxyapatite increased with irradiation dose. In summary, the γ-irradiated PLLA increased the differentiation of osteoblasts and also increased apatite-forming ability even without the osteoblasts. The osteoblast differentiation was enhanced well in the apatite formation on the surface of PLLA after the γ-irradiation.