The Study Of Honokiol Nanoparticles-loaded Microbubbles

In recent years,the microbubbles(MBs)as a drug carrier for the delivery of drug or gene,had gradually received wide attention.Lipid shell provided the space for drug,and the inert gas in the center made MBs more stable.After the drug-loaded microbubbles intravenously being injected into the body through the blood circulation,the ultrasound images can monitor the process of microbubbles reaching the target tissue.Being exposed to a certain intensity ultrasound,the microbubbles fractured into pieces in the target tissue.A series of effects of microbubbles fracture,increased the absorption of drug and improved efficacy.In this research,we chose honokiol(HK)as the model drug,the microbubble as a drug carrier.Honokiol nanoparticles(HKNs)loaded microbubbles(HKN-MBs)were prepared.The use of ultrasound-targeted microbubble destruction made the drug release at the target tissue.In vitro and in vivo experiments,we explored the effect of HKN-MBs combined with ultrasound targeted microbubble destruction ontumor.The study provided a new strategie for the construction of a new pharmaceutical dosage form.Drug-loaded microbubbles combined with ultrasound targeted microbubble destruction provided a new way of tumor targeted therapy.Part ? Establishment the method for in vitro analysis ofHKN-MBsHPLC method for the determination of HK was established.The peak time of HK was about 5.1 min with a good shape.The lecithin would not affect the determination of the drug.The concentration range was 0.2-50.0 ?g/mL,which had a good linear relationship,the standard curve equation was A=40.68-3.0458 C,r=0.9999.Recoveries,intra-day precision,inter-day precision were in line with the requirements of methodology.Part ? Preparation of HKN-MBs1.The HKNs were prepared through a desolvation method.2.The microbubbles were prepared by mechanical shaking,which encapsulated the HKNs and perfluorocarbon gas(HKN-MBs).3.After centrifugation,HKN-MBs rose to the upper layer and exhibited homogeneous turbidity when gently shaken.Part ? Investigation HKN-MBs pharmaceutics properties1.HKN-MBs were evidently larger than blank MBs,which indicated that HKNs loading increased the size of the MBs.MBs were uniform size,and had better dispersion degree.2.The mean diameters of blank MBs and HKN-MBs were(470.5±46.21)and(1447.0±182.4)nm,respectively.And the mean diameters of HKNs were(159.3±45.03)nm.3.The EE% and LE% of HKN-MBs were(67.78%±1.38)% and(3.48%±0.20)%,respectively.4.To further investigate the internal composition of the HKN-MBs,the FITC-labeled HKN-MBs were observed using CLSM.This proved that the internal structure of HKN-MBs included both HKNs and fluorocarbon gas.5.In vitro analysis of US-enhanced imaging showed that both saline and HKNs showed no US contrast imaging effects.However,HKN-MBs and blank MBs brought about an enhanced US imaging in 2% agarose gel phantom.Part ? The inhibition of HKN-MBs on Lewis lung tumor1.The tumor weight of saline control group was(934.9±81.2)mg.The tumor weight of experimental group HKNs is(655.0±73.4)mg,and tumor inhibition rate was 32.85%.The tumor weight of experimental group HKN-MBs is(627.8±64.1)mg,and tumor inhibition rate was 29.94%.The tumor weight of experimental group HKN-MB+US is(478.8±41.7)mg,and tumor inhibition rate was 48.78%.The results exhibited that the experimental group HKN-MB+US showed a strong anti-tumor effect.2.Tumor growth of saline control group,was the fastest,while the experimental group HKNs and HKN-MBs relatively slower,and HKN-MB+US group was the slowest.Tumors in the experimental group were significantly smaller than the tumors in the control group.And tumors in the experimental group HKN-MB+US was minimum,it can be seen from the tumor growth curves and tumor size that the experimental group HKN-MB + US had a strong anti-tumor effect.3.In vivo analysis of US-enhanced imaging showed the HKNs and saline groups showed no US images,whereas both HKN-MBs and blank MBs improved US imaging on the tumor sites.4.HE staining experiments showed that a large range of necrosis were observed in the tumors of experimental group HKN-MB+US compared with those in other groups,which proved that the experimental group HKN-MB+US had a strong anti-tumor effect.