Abstract: Objective To explore the optimum conditions for preparing poly(lactic-co-glycolic) acid (PLGA) nanoparticles and evaluate the bioactivity of hepatocyte growth factor (HGF)-loaded PLGA nanoparticles. Methods Bovine serum albumin (BSA)-loaded PLGA nanoparticles were prepared using a double emulsion-solvent evaporation method. The preparation process of nanoparticles was optimized by orthogonal test with the particle size, encapsulation efficiency (EE), drug loading (DD), and recovery as the indexes. HGF-loaded nanoparticles were then prepared under the optimized conditions. The EE, DD and release characteristics of BSA-loaded nanoparticles and HGF-loaded nanoparticles were evaluated using a BCA kit and HGF ELISA kit. The bioactivity of HGF-loaded nanoparticles was evaluated using CCK8 proliferation assay. Results The HGF-loaded nanoparticles prepared under the optimized conditions had a uniform size with a mean diameter of 234.4±4.8 nm, an EE of (77.75±3.04)% and a recovery rate of (49.33±9.34)%. The in vitro release curve highlighted an initial burst drug release followed by sustained release from the nanoparticles. HGF-loaded nanoparticles obviously promoted the proliferation of Hacat keratinocytes in vitro. Conclusion HGF-loaded nanoparticles prepared using double emulsion-solvent evaporation method under optimized conditions possesses a high EE with a good sustained drug release profile and a good bioactivity.