Preparation Of CRGD-modified Harmine Long Circulating Magnetoliposomes And Study On Its Characteristics

Object:To prepare cRGD-mediated harmine long-circulating superparamagnetic nanoliposomes,investigate their in vitro properties and study on their antitumor activity.Method:First of all,content assaying method was determined.Then Arg-Gly-Asp contained circular peptides?cRGD?as a ligand was conjugated to polyethylene glycol?PEG?to form cRGD-DSPE-PEG through chemical conjugation.And then the conjugation were added into of the membrane of liposomes which entrapped harmine?HM?and magnetic particles?MP?via reverse phase evaporation.The optimized preparation conditions were determined according to the performance of particle size,zeta potential,content of Fe²⁺and entrapment efficiency.Furthermore,the configuration of liposomes was observed as well as the release,magnetic and stability properties measured.Finally,the cell intake of liposomes,the influence on nucleus form and their restraint effect on tumor cells were observed and investigated.On one hand,Hoechst fluorescent staining was conducted for the observation of nucleus form.On the other hand,MTT,Annexin V,Coomassie Blue staining were utilized to investigate the survival or apoptosis rate of HepG2 and U87 tumor cell lines before and after incubating with the liposomes.Results:The established ultraviolet spectrophotometry for content determination performed good linearity.Also,the cRGD-HM-MP liposomes were well prepared with particle size of243?3.81nm,zeta potential of 21.8?0.77mV and Fe²⁺content of 131.93?27.6?g mL^-1.The entrapment efficiency was 63.17?5.8%with drug-liposome ratio of 1:10 and 87.1?6.8%with the ratio of 1:20.Leakage of drug through liposomes was up to 29.08±17.01 percent in 14 days.Magnetoliposomes showed significant delayed release effects compared to drug solution.n the assays of antitumor activity,the cRGD-HM-MP performed stronger cell growth containment in the magnetic field than other groups with p values under 0.05after statistic analyses were done.The survival rate of HepG2 and U87 cells in the high concentration group after 36 hours was 27.02±0.68%and 44.65±2.29%,considerably lower than blank lipoome group,HM solution group,blank magnetic nanoparticle group and lower concentration group without magnetic field,while the apoptosis rate was30.82±0.5%and 36.73±0.44%,dramatically higher.In addition,this group also showed lowest total protein content with a P value of 0.844 after rank-sum test.Conclusion:In this research,the cRGD-HM-MP liposomes were successfully prepared and magnetic nanoparticles were well entrapped.The selected liposomes showed fairy properties in particle size,zeta potential,Fe²⁺content and entrapment efficiency and showed the significant delayed release effect.The cRGD-HM-MP liposomes were confirmed to show strong growth inhibitory effect on both HepG2 and U87 cells,which suggests the potential usage of this product in pharmaceutics.Also this study draw a solidity basis for further researches.