Experiment Study Of Epirubicin Dolybutylcyanoacrylate Nan-oparticles To Treat Transplanted Liver Cancer In Rat

Objective:To investigation the determinate factors and levels on procedures of Epirubicin-polybutylcyanoacrylate nanoparticles (EPI-PBCA-NP) and to optimize preparation procedures and then product the EPI-PBCA-NP entrapped Epirubicin 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, Epirubicin-polybutylcyanoacrylate nanoparticles was prepared.Results:1.The influential factors of drug content of PBCA-NP included:quantity of EPI in initial system;added volume of alpha-BCA monomer; PH value of media;class and volume of surfactant.2.Multiple factors linear regression equation was estabilished: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 Epirubicin of PBCA-NP with fine particle size and shape were producted successfully followed the optimized procedure.The optimized recipe for Epirubicin-polybutylcyanoacrylate nanoparticles is as follow: EPI 10.0mg,BCA 0.25ml,PH=2.5,1.5%Dextean70.3.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 Epirubicin in 4.0,8.0,12.0mg/ml Na2SO4 groups were higher than Omg/ml one(P<0.05), the biggest of which was ER=84.26+/-0.27,LD=3.57+/-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.Omg/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 EPI-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,DNA-EPI-PBCA-NP was prepared successfully.Erence for Epirubicin(ER=82.17+/-0.22%)Conclusion:The quantity of Epirubicin 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, EPI-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 Epirubicin 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 EPI-PBCA-NP resulted from electrolyte NaSO4 takes on negative correlation with it's alternation of Zeta potential. The presence of EPI-PBCA-NP provides a novel method for studying combination of gene therapy with chemical therapy. Aim:To observe the difference of the bio-distribution in the body of the rats of the Epirubicin polybutylcyanoacrylate napariticles(EPI-PBCA-NP) freeze-dry dose for the different diameter of the Napariticles and the solution of the Epirubicin (EPI) rejected by the vein,and study the liver targeting for EPI-PBCA-NP with different diameter of nanoparticles to normal liver,and choose the EPI-PBCA-NP for the best liver targeting.Method:EPI-PBCA-NP with various ranges of diameter of the nanoparticles including (22±6.2)nm,(48±9.2)nm,(101±20.3)nm,(143±23.5)nm,(194±28.4)nm were produced by using the method of emulsior polymerization based on the carrier materials of a-polybutylcyanoacrylate.180 rats from kunmin were divided into 6 groups at random,with 30 rats for each group (EPI-PBCA-NP with diameters of the nanoparticles were divided into 5 groups.In addition free Epirubicin was considered as comparative group) EPI-PVCA-NP freeze-dry dose and EPI solution were injected into the veins of rats by the formulation of 2mg/kg.5 rats in each group were killed 5 15,30min,1,5 and 12 hours respectively after drug was given.EPI concentrations were determined using a high effective liquid chromatography with fluorescence detector technique.when obtaining the samples from liver,kidney,spleen,heart,lung and plasma respectively.Result:The EPI concentrations in the lever of rat in all the napariticles groups is markedly higher as compared to free EPI groups during 12h, with the excepting of the group with (22±6.2)nm(p<0.05).whereas the concentration in the heart muscle was not determined or markedly lower than free EPI group(p<0.05)high concentration in kidney did not keep long time,EPI concentration in the lung in the group of(194±28.45)nm was higher than that of (48±9.2)nm (p<0.05).EPI concentration in the group of (101±18.3)nm in the liver of the rats were remarkably higher than that in the other groups (p<0.05),the second is(143±21.4)nm. EPI concentration in the group of (22±6.2)nm was not determined at all in the liver, kidney, heart,spleen and lung.Conclusions:EPI-PVCA-NP with some different ranges of diameters of the nanoparticles has the better liver targeting,and the characteristic of low releasing,which decrease the medicine distribution in the heart and other tissues.So it's a good medicine-diverting system for treating liver cancers. DM-PVCA-NP with the diameter of 100-150nm has the best liver targeting,and this supplies experimental data for producing na-meter medicine-carrier of polybutylcyanoacrylate for treating liver cancers. Objective:To objective the therapeutic effects against transplanted liver cancer with Epirubicin-polybutylcyanoacrylate nanoparticles via hepatic artery.In presentce of magnectic field, the animal bearing transplanted liver tumor was injected Epirubicin-polybutylcyanoacrylate nanoparticles containing Epirubicin in order to observe efficacy of the nanopartlicles for transplanted liver cancer.Methods:A hundred rat model of transplanted liver was established, After one weeks,they were divided into three group at random. Each group is 20.Abdominal exposure was carried out through a midline abdominal incision. The diameter of tumor was detected and a cannula was inserted into the hepatic artery and fixed up it,EPIinistrated each group(equivalent dose of free Epirubicin 0.5mg/kg).1.Normal Saline given by hepatic artery.2.Free Epirubicin given by hepatic artery.3.Epirubicin-polybutylcyanoacrylate nanoparticles given by hepatic artery.The survival days of the animals which survive time was over 7 days post EPIinistration was recored. After one weeks,Abdominal exposure was carried out through a midline abdominal incision again.Volume of tumor, tumor growth rate,degree of tumor necrosis and increaseing longer surviaval(ILS) were detected and investigated.Results:Before therapy,no statistic difference was found in each group volume of tumor (p>0.05).After therapy, statistic difference was found in each group volume of tumor (p<0.05).Normal Saline treated group,Survived time was 12.7days;Free Epirubicin treated group,Survived time was 18.7days. Epirubicin-polybutylcyanoacrylate nanoparticles treated group Survived time was 32.5days.The survival rate of animals of every group was analyzed with increaseing longer surviaval.It demonstrated statistics difference existed each group.The results showed that In presence of magnetic field, the survival time was prolonged in rats treated with Epirubicin-polybutylcyanoacrylate nanoparticles treated group;Tumor tissue of the animals given magnetic carrier in absence of magnetic field was seen patch necrosis by histology examination one week post EPI inistration. In presence of magnetic field The tumor growth inhibition with rat received therapy of Epirubicin-polybutylcyanoacrylate nanoparticles and Epirubicin was remarkable necrosis more extensive in comparision with rat received therapy of normal Saline and Epirubicin The tumor tissue was replace of fibril tissue and non-structure tissue,survived one week post EPIinistration.Conclusion:The survival rate of the animal given with free Epirubicin was not improved compared to NS treated group,Using Epirubicin- polybutylcyanoacrylate nanoparticles in absence field can increase survival rate of animals,compared to NS treated animals or animals treated with free Epirubicin.Using Epirubicin-polybutylcyanoacrylate nanoparticles in presence field can remarkablely increase survival rate of animals,compared to NS treated animals or animals treated with free Epirubicin (P<0.01).