ZHANG Demeng, LOU Ruyun, WANG Yu, LIU Xiudong, YU Weiting, MA Xiaojun
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With the preparation of Chitosan/siRNA nanoparticles based on electrostatic self-assembly, a nucleic acid dye assay with SYBR Gold was established to determine the condensation rate (CR) of siRNA in nanoparticles, and then the influence of n(N)/n(P) ratio, Mw, DD, pH and ion strength of chitosan solution on siRNA CR, nanoparticle size and zeta-potential was investigated by SYBR Gold assay, dynamic light scattering and laser doppler electrophoresis respectively. The results showed that CR rose from (18.6±4.0)% to (92.9±2.0)% as n(N)/n(P) increased from 0.5 to 2 and stayed stable even when n(N)/n(P) reached 150. When n(N)/n(P) was below 2, no stable nanoparticles were detected and the complex was negative charged, while n(N)/n(P) went from 5 to 150, the size of nanoparticles increased from (179.3±5.7) nm to (235.5±8.3) nm and zeta-potential rose from (17.2±1.3) mV to (23.6±0.9) mV. At n(N)/n(P) 150, CR raised with the increase of DD or with the decrease of Bc, the size of nanoparticles increased with the increase of Mw and decreased with the increase of DD or Bc, the zeta-potential of nanoparticles increased with the increase of pH value or DD but decreased with the decrease of Bc. It demonstrated that n(N)/n(P) ratio is the most important factor determining the size, zeta-potential and CR, and nanoparticles with the diameter ranging from 100-300 nm and zeta-potential ranging from 15-25 mV can be easily obtained via adjusting above parameters.