Construction of novel amphiphilic chitosan-polylactide graft copolymer nanodroplets for contrast enhanced ultrasound tumor imaging.

Abstract

In this experiment, a new amphiphilic chitosan-poly(lactide) graft copolymer was synthesized and characterized by IR, 1H-NMR, XRD, TGA. The obtained chitosan-poly (lactide) graft copolymer was used as the matrix material to prepare nanodroplets (NDs) encapsulating with liquid PFP by double-emulsion and solvent evaporation method. The resulting NDs were characterized by photon correlation spectroscopy and transmission electron microscopy (TEM). The biocompatibility was explored by cytotoxicity assay, cell migration assay and blood biochemistry analysis. The experiments of ultrasonic imaging in vitro and in vivo were carried out with a B-mode clinical ultrasound imaging system. The results of FI-IR and 1H-NMR confirmed the successful grafting reaction of polylactic acid(PLLA) to chitosan with a graft rate of 365%. The average size of the NDs was 101.1 ± 2.7 nm, with the polydispersity index (PDI) of 0.127 ± 0.020, and the zeta potential was -31.8 ± 1.5 mV. From the TEM results, NDs were highly dispersed and had a spherical shape with a distinct capsule structure. The NDs exhibited good stability during storage at 4°C. The NDs solution with different concentrations did not affect cell growth and showed good biocompatibility in cytotoxicity, cell migration and blood biochemistry studies. Under the irradiation of ultrasonic waves, the NDs formed an ultrasonic high signal, which could significantly enhance the ultrasound imaging of tumor tissue in vivo. Taken together, the NDs hold great potential for ultrasound imaging as a nanosized contrast agent.