Influences of preparation conditions on particle size and DNA-loading efficiency for poly(DL-lactic acid-polyethylene glycol) microspheres entrapping free DNA.

Poly-DL-lactic acid-polyethylene glycol (PELA) with different contents and different molecular weight of polyethylene glycol (PEG) was used as a DNA delivery system. DNA-loaded PELA or poly(DL-lactic acid) (PLA) microspheres were prepared by the emulsion evaporation technique, which was based on the water-in-oil-in-water solvent evaporation method. The purpose of the present work was to investigate the factors influencing particle size and DNA loading efficiency for the PELA microspheres containing free DNA. During the preparation process, different conditions were used and the resulting microspheres were characterized by particle size and DNA loading efficiency. Microspheres prepared by PELA with a PEG (molecular weight: 6000 Da) content of 6-10% obtained the highest loading efficiency and smaller particle size among other PELA copolymer and PLA homopolymer. When the solvent of the oil phase was composed of methylene chloride and ethyl acetate (1:1, v/v), the highest loading efficiency and smaller particle size were also obtained for the PELA microspheres. The presence of the surfactant in oil phase influenced both the particle size and loading efficiency. Increasing the concentration of polymer in oil phase resulted in an increase of particle size and loading efficiency for DNA-loaded PELA microspheres. The addition of a hydrophilic polymer into the internal water phase ameliorated the DNA loading efficiency and reduced the particle size. Significant influences of DNA molecular weight and structure on the particle size and loading efficiency were observed. The volume and concentration of the external water phase also influenced the particle size and loading efficiency.