The nanoscale emulsions were created by the injection of MNC-laden organic solvent phase into an aqueous continuous phase containing carboxyl polysorbate 80 under ultrasonication and vigorous stirring.
The interface of emulsions with continuum was stabilized by carboxyl polysorbate 80 and MNC within nanoemulsions and was enveloped by carboxyl polysorbate 80 during a solvent evaporation . As described in the experimental section, Apt was conjugated with carboxylated MNC to prepare Apt-MNC for molecular MR imaging of VEGFR2. The selleck morphology of Apt-MNC was observed by TEM. Uniformity and spherical shape of MNC from Apt-MNC were observed; the average diameter of MNC was 11.7 ± 1.0 nm and clustering of MNC was not observed (Figure 2b). The hydrodynamic BMN 673 in vivo diameter of Apt-MNC (34.0 ± 5.8 nm) was slightly increased https://www.selleckchem.com/products/lcz696.html compared with that of carboxylated MNC (31.5 ± 2.2 nm) due to Apt conjugation (Figure 2c). Carboxylated MNC possessed negative surface charge due to the negatively charged surface carboxylate in an aqueous phase. Apt-MNC showed a slightly changed surface charge of −17.0 ± 0.5 mV after Apt conjugation (Figure 2c). These data indicate that Apt was successfully conjugated with carboxylated MNC and Apt-MNC was well dispersed in an aqueous phase, with its monodispersity due to the presence of modified polysorbate 80 molecules. Additionally, negatively charged Apt-MNC surface repulsed nonspecific binding on negatively charged cell surface, increasing
the aptamer-mediated specific binding on VEGFR2 . Thus, the characteristics of Apt-MNC were suitable for a potential MR imaging selleck compound probe to detect the biomarker.
The prepared Apt-MNC exhibited a superparamagnetic property without magnetic hysteresis at zero magnetic field, and the saturation magnetization value was 98.8 emu g−1 Fe at 1.5 T. These magnetic properties were highly acceptable as a sensitive MR imaging contrast agent (Figure 3a). The T2-weighted MR imaging of Apt-MNC solution at various Fe concentrations was obtained to evaluate the capability of imaging contrast effect. The increase of Fe concentration accelerates transverse relaxation to shorten the T2 relaxation time (T2), resulting in a decreased signal intensity with dark contrast. The T2 relaxation rate (R2 = 1/T2, s−1) was plotted versus Fe concentration (mM) to determine the relaxivity coefficient (r2) as 214.5 s−1 mM−1, which is higher than that of commercial MR imaging contrast agents (ferumoxide, 190.5 s−1 mM−1) (Figure 3b) . Figure 3 Properties of Apt-MNC as a contrast agent. (a) Magnetic hysteresis loop of Apt-MNC. (b) T2-weighted images and relaxivity coefficient (r2) of Apt-MNC. To assess the biocompatibility of Apt-MNC, we investigated the in vitro cytotoxicity of carboxylated MNC and Apt-MNC in U87MG cells by monitoring the effects on cell viability and proliferation. Cell viabilities were examined after incubation with various concentrations of carboxylated MNC and Apt-MNC for 24 h.