| Objective: To investigation the determinate factors and levels on procedures of Doxorubicin-polybutylcyanoacrylate nanoparticles (DOX-PBCA-NP) and to optimize preparation procedures and then product the Dox-PBCA-NP entrapped Doxorubicin and plasmid p53,in order to reveal its' possibility in combining gene therapy with targeting therapy.Methods: The influential factors were determined preliminarily by one-step methods. On the base of this attempt, uniform design allowed to find the interaction among all possible factors and its' levels. The Zeta potential of suspension systems was altered and ion-pair reagents were added so as to elevate drug content of nanoparticles. By two-step method, p53DNA-Doxorubcin-polybutylcyanoacrylate nanoparticles was prepared.Results:1. The influential factors of drug content of PBCA-NP included: quantity of DOX in initial system; added volume of alpha-BCA monomer; PH value of media; class and volume of surfacant.2. Multiple factors linear regression equation was established: Y =19.456+0.113X1-1.66X2-6.121X3-0.839X4, F=0.888, R=0.686, R2=0.57, P=0.544, and high content of Doxorubicin of PBCA-NP with fine particle size and shape were producted successfully followed the optimized procedure.The optimized recipe for Doxorubicin-polybutylcyanoacrylate nanoparticles is as follow: DOX lO.Omg, BCA 0.25ml, PH=2.5, 1.5%Dextran70.3. The drug content of PBCA-NP was enhanced by addition of iron-pare reagents and increasing Zeta potential of colloid systems. Compared with controlled groups, enhancement of entrapment rate and drug loading capacity were obtained in group with additional Na2SO4 (P<0.05 ) ,but the similar results couldn't be found in NaCl and Na2CO3 groups (P>0.05 ) .Entrapment rate and drug loading capacity of Doxorubicin in 4.0, 8.0, 12.0mg/ml Na2SO4 groups were higher than 0 mg/ml one (P<0.05) ,the biggest of which was ER=84.26+/- 0.27 ,LD =3.57+7-0.01 when system concentration of Na2SO4 was 12.0mg/ml. while system concentration of Na2SO4 increased to 16.0mg/ml, the drug content declined compaired with 12.0mg/ml group (P<0.05 ) . In the same time ,Zeta potential of colloid systems followed this trend (P<0.05) and its' strength are negatively related to the scale of drug content of DOX-PBCA-NP (rER=0.9671, rLD=0.9125, P<0.05) . The drug content of PBCA-NP was intensified by additional ion-pair reagents too (P<0.05). 4.High drug content nanoparticles, p53DNA-DOX-PBCA-NP wasprepared successfully. ER(%) of p53DNA rised markedly ( ER-49.67+7-1.26%, P<0.05) . but there was no difference for Doxorubicin (ER= 82.17+/ -0.22%,P>0.05 ) .Conclusion: The quantity of Doxorubicin and butylcyanoacrylate polymer, PH value in medium, the varies and volume of the surfactants were key factors on drug content of PBCA-NP. The influence of stirring speed is negligible.Taken the optimized recipe, DOX-PBCA-Np with fine particle size and morphology is available and it's drug content is intensified too. The optimized technique of preparation of nanoparticles entrapped Doxorubicin will facilitate the further study. Both change of Zeta potential and employment of ion-pair reagents can elevate the drug content of NPs. the higher drug content of DOX-PBCA-NP resulted from electrolyte Na2SO4 takes on negative correlation with it's alternation of Zeta potential. The presence of p53- DOX-PBCA-NP provides a novel method for studying combination of gene therapy with chemical therapy.
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