Preparation And Molecular Imaging Investigation In Breast Cancer Of Targeted Gold Nanoshelled Poly (Lactic-co-glycolic Acid) Ultrasound Contrast Agent Carrying Anti P53 Antibody

Objective:To prepare and characterize of targeted gold nanoshelled poly?D,L-lactide-co-glycolic acid?ultrasound contrast agent?UCAs?carrying anti p53antibody?p53-PLGA@Au NCs?,and we aimed to investigate its targeting ability and ultrasound contrast behavior in vitro and vivo.Methods:In our study,the nanoscale tUCAs were fabricated through filling perfluoropropane?C₃F₈?into PLGA nanocapsules?PLGA NCs?followed by the formation of gold nanoshell on the surface,then conjugated with anti p53 antibody which has high specificity with the p53 protein overexpressing in breast cancer.Scanning electron microscope?SEM?,high-resolution transmission electron microscope?HR-TEM?,dynamic laser scattering?DLS?were used to observe the morphology,structure,the size and zeta potential of the NCs.MTT assays were used to study the cytotoxicity effect of p53-PLGA@Au NCs.LSCM and flow cytometry?FCM?were used to evaluate its targeting ability in vitro qualitatively and quantitatively.And the preliminary evaluation of the ultrasound contrast behavior of p53-PLGA@Au NCs in vitro and in vivo was carried out by using ultrasonic diagnostic instrument.Results:SEM and TEM images revealed the spherical morphology and the surface of the highly dispersed p53-PLGA@Au NPs.Its average diameter was 247.7±108.2 nm,the zeta potential was-34.2±5.8 mV and the polydispersity index was 0.127,implying the solution has uniform particle size distribution and good stability.Compared with 231cell,MCF-7 cells incubated with p53-PLGA@Au NPs show bright green fluorescence signals.MCF-7 cells incubated with p53-PLGA@Au NPs in FCM assay showed remarkably higher fluorescence signals?73±5.21%positively labeling cells?compared with 231 cells incubated with the same NPs?2.21±0.45%positively labeling cells??P<0.001?.Observed from US contrast-enhanced images,the tubes filled with p53-PLGA@Au NPs displayed a strong dotted echo and the length of imaging time met the requirement of the clinic.The breast cancer?MCF-7?xenograft tumor in situ model of nude mouse was established and the green increased enhancement signal was obvious in 22 MHz and 50 MHz frequency respectively.And the time intensity curve data also implied that the imaging effect of p53-PLGA@Au NPs was better than those of other groups.Conclusion:The p53-PLGA@Au NCs was successfully prepared.Moreover,p53-PLGA@Au NCs has good biocompatibility and provide excellent targeted contrast enhancement for ultrasound molecular imaging in vitro and vivo,which may have potential applications in the early diagnosis of breast cancer in the future.