The Study Of Targeted Polymer Contrast Agent Loaded Gold Nanorods And Liquid Fluorocarbon And The Development Of Their Targeting And Dual-modal Imaging In Vitro

PART Ⅰ PREPARATION OF VCAM-1 RECEPTOR-TARGETED DUAL-MODAL PHOTOACOUSTIC/ULTRASOUND NANOSCALE MOLECULAR PROBE WITH GOLD NANORODS AND LIQUID FLUOROCARBON AND THEIR PHYSICAL CHARACTERISTICS AND TARGETED ABILITY IN VITROObjective To prepare vascular cell adhesion molecule-1(VCAM-1)-targeted polymer photoacoustic/ultrasound dual-modal photoacoustic/ultrasound nanoscale molecular probe with gold nanorods and liquid perflurocarbon and explore their physical characteristics targeting ability in vitro.Methods The polymer PLGA nanoparticles(GNPs) loaded gold nanorods and liquid perflurocarbon(Perfluorohexane, PFH)and only PFH nanoparticles(NPs)and only gold nanorods(Au-NPs) were synthesized by double emulsion and the VCAM-1-targeted GNPs were accomplished via carbodiimide method. The appearance、size and zata potential of three kinds of nanoparticles were observed by light microscopy and transmission electron microscope. The coupling efficiency and imaging of GNPs with VCAM-1 were detected. The optical droplet vaporization(ODV)of GNPs, NPs and Au-NPs were motivated by laser apparatus. The VCAM-1-GNPs was irradiated by laser irradiation with different energy and the same wave length and observed by light microscope, and VCAM-1-GNPs, NPs, Au-NPs was sitimulated by energy(200 mj). The basic physical characteristics of VCAM-1-GNPs were observed with light microscope and electron microscopy. The HUVECs was simulated by some different concentration of TNF-α and the product-VCAM-1 was detected by Western Blot, The VCAM-1-GNPs with HUVECs of targeted group which was stimulated by the best concentration of TNF-α and non-targeted one which was not were observed by light micriscope.Results The VCAM-1-GNPs were synthesized successfully. Their average diameter was(615.86±8.60) nm,those nanoparticles were similar. The coupling efficiency of GNPs with VCAM-1 was 99.87 %. VCAM-1-GNPs could be motivated by the energy which was equal or greater than 140 mj, the ODV imaging of VCAM-1-GNPs simulated by laser(energy was 200 mj) was observed and the ODV imaging of Au-NPs and NPs were not obvious. The protein of VCAM-1 expressed clearly which was effected by TNF-α. The differences were statistical significance compared group 10 ng/ml with groups lower concentrations(p<0.05) while was no statistical significance compared with group 100 ng/ml(p>0.05). The VCAM-1-GNPs had excellent coupling efficiency with HUVECs which was stimulated while the other group had poor connection.Conclusion The targeted polymer nanoparticles coated gold nanorods and perflurocarbon was prepared and morpholopy was well. The ODV was obvious. The VCAM-1-GNPs have excellent targeting ability in vitro. Those results have become a solid foundation for the dual-modal targeting imaging in vivo.PART Ⅱ THE TARGETING NANSCALE MOLECULAR CONTRAST AGENTS OF CARRYING NANORODS AND LIQUID FLUOROCARBON AND THEIR DEVELOPMENT IN VITROObjective to study the bimodal(ultrasound /photoacoustic) imaging in vitro of nanoscale contrast agents which capsuled Gold nanorods and liquid fluorocarbon(perfluorhexane, PFH).Methods The targeting polymer PLGA nanoparticles(VCAM-1-GNPs)which contained nanorods and PFH were prepared as previously, as well as Au-NPs which contained only nanorods and W-NPs contained only water; The two-dimensional ultrasound and photoacoustic of different concentrations of VCAM-1-GNPs was observed. Two-dimensional ultrasound and photoacoustic signal graph of VCAM-1-GNPs, A-NPs, NPs were developed before and after near-infrared laser irradiation..Results The signals of two-dimensional ultrasound of VCAM-1-GNPs were hypoechoic before laser irradiated, but their grayscale and contrast intensity significantly increased after laser excitation and their PA signal was obvious; The signals of two-dimensional ultrasound of Au-NPs were hypoechoic before and after laser excitation and their PA signal was obvious; The NPs of two-dimensional ultrasound and contrast mode were hypoechoic before and after laser excitation and their PA signal was not obvious; As the concentration increased,more and more higer of the signals of two-dimensional ultrasound and PA.Conclude The polymer nanoparticles carried gold nanorods and perflurocarbon were prepared. The dual-modal development of VCAM-1-GNPs in vitro was achieved. the VCAM-1-GNPs has potential to become an excellent nanoscale molecular imaging probe and used for internal imaging and therapy.