Preparation and in Vitro Targeting Experiment of Vincristine-Loaded Ul-trasound Microbubbles

YANG Hong; CHEN Qiuhong; HUANG Juan; GUO Jingyi; WANG Yuyuan; WU Qi

doi:10.3969/j.issn.2095-3097.2024.02.004

Abstract：Objective To prepare vincristine(VCR)-loaded lipid ultrasound(US)microbubbles(VCR-MBs)tar-geting to peripheral nerve myelin cells,and to observe the in vitro targeting experiment.Methods VCR-MBs were prepared by thin film hydration and mechanical oscillation method,and targeted VCR-MBs(VCR-TMBs)was prepared by biotin-avidin bridging method.The morphology,particle size,stability,in vitro development,encapsulation rate and targeting properties of VCR-TMBs were observed.The effect of VCR-TMBs on cell injury and apoptosis was detected by flow cytometry.The rat renal artery was treated by VCR-TMBs combined with US in vitro,and demyelination was observed by HE staining.Results MBs,VCR-MBs and VCR-TMBs were all circular and evenly distributed under light microscope.The particle sizes of MBs,VCR-MBs and VCR-TMBs were(3.22±1.07),(3.48±1.06)and(3.22±0.79)μm,respectively,with no significant differ-ence(P>0.05).Compared with MBs and VCR-MBs,the concentration of VCR-TMBs decreased significantly after3 days of storage(P<0.05).The encapsulation rate and drug loading rate of VCR-TMBs determined by high-performance liquid chro-matography were(82.90±9.98)%and(2.07±0.25)%,respectively.VCR-TMBs was detected by immunofluorescence and could be targeted to accumulate on Schwann cell membrane.Flow cytometry showed that VCR-TMBS +US had better ap-optotic effect than VCR +US and VCR-MBS +US(P<0.05).HE staining showed that VCR-TMBs +US could effectively de-myelinate and denaturate the renal artery nerve in vitro.Conclusion The successful preparation of VCR-TMBs can target Schwann cells,promote cell apoptosis and demyelinate renal artery nerves of rats,which can be a preliminary preparation for the later realization of renal artery nerve dessympathization by VCR-TMBs in vivo.