Abstract

Superparamagnetic iron oxide nanoparticles (SPIO) are regarded as advanced tools in biomedical science in disease diagnostics and therapies. We report the synthesis of antibody-conjugated nanoparticles (Ab-SPIO) that combine MRI behavior of nanoparticles with the selective and specific targeting of cellular proteins. Our novel Ab-SPIO will serve as a MRI biomarker for preventive PAH diseases. It is known that adenosine 1-type receptors (A1R) are involved in several cardiovascular diseases and offer promising therapeutic potential. In our study, we developed new A1R antibodies (Ab) conjugated SPIO nanocarriers as a specific Ab-MRI contrast agent. Spherical magnetite nanoparticles with a hydrodynamic diameter of 145 nm and particle size distribution of 15 - 60 nm were obtained. Surface of SPIO nanoparticles was stabilized by biocompatible polymer carboxymethyl cellulose (CMC) and precisely characterized in stability by measuring of zeta potential (- 43 mV). Strong magnetic response with a saturation magnetization of 75 Am2/kg confirmed appropriate magnetic behavior for MRI application. Oriented immobilization of antibody (Adenosine A1-R Antibody (H-40)) on free carboxyl groups of CMC provides active targeting of adenosine receptors. We conducted microscopic evaluation of the Ab-SPIO probe in VVEC cells, localization (plasma membrane vs. intracellular), and we determined the effects of hypoxia on A1R expression, compared A1R expression in VVEC-Hyp vs. VVEC-Co, and determined the effects of acute hypoxia on A1R expression in VVEC-Hyp vs. VVEC-Co. The experiments are proposed for MRI imaging of VV in control and hypoxic animals. Functionalized A1R-Ab-SPIO complexes will be utilized as a specific MRI biomarker in early disease diagnostics.