〔Abstract〕 Objective To investigate the effect of teriparatide(TPTD) on the generation of MC3 T3-E1 cells towards osteogenic differentiationviathe Wnt3 a/β-catenin pathway in a high-glucose environment. Methods The experiment was divided into five groups: low glucose group, low glucose+TPTD group, high glucose group, high glucose+TPTD group, high glucose+TPTD+Wnt3 a inhibitor G244-LM group. Cell proliferation activity was detected by Calcein-AM and CCK-8 assay, cell mineralized nodule formation was observed by ALP and alizarin red staining, and actin formation was analyzed by immunofluorescence assay. Real-time PCR was performed to detectWnt3 a,β-catenin,Tcf1,OPG and COLⅠ mRNA expression. Results TPTD had no significant effect on the proliferative activity of MC3 T3-E1 cells under high glucose condition.The ALP staining area, protein activity and alizarin red staining area of the cells in the low glucose+TPTD group were higher than those in the other four groups(P<0.05); the high glucose group was lower than the low glucose group(P<0.05); the high glucose+TPTD group was higher than the high glucose group and the high glucose+TPTD+G244-LM group(P<0.05). The cytoskeleton in the low glucose+TPTD group was the clearest; the cytoskeleton was less clear in both the high glucose and high glucose+TPTD+G244-LM groups than in the high glucose+TPTD group. Genes such asWnt3 a,β-catenin,Tcf1,OPG and COLⅠ had the highest mRNA levels in the cells of the low glucose+TPTD group(P<0.05); the mRNA levels of all genes were higher in the low glucose group than thosein the high glucose group(P<0.05); the mRNA levels of all genes in the cells of the high glucose+TPTD group were higher than those in the high glucose group and the high glucose+TPTD+G244-LM group(P<0.05). Conclusion High glucose inhibited osteoblast differentiation, and TPTD promoted osteoblast differentiation in high glucose environment by regulating Wnt3 a/β-catenin pathway.