Study of therapeutic targeting of Tumor Necrosis Factor-alpha(TNF-α) in Lymphocytes and Macrophages by using nanoparticles for limiting Coronary Artery Disease (CAD)*

R. Shabbir, A. Raza, A. Liaquat, S. Shah, M. Hamza, M. Fayyaz, Z. Hussain, N.M. Butt
Preston University Kohat,,
Pakistan

Keywords: coronary artery disease ,TNF-alpha, inflammation, nanomedicine

Summary:

Cardiovascular disease (CVD) affects a significant number of people around the world which results in high mortality rates. A multitude of environmental and genetic factors contribute to the pathogenesis of CVD. In addition to the current treatment strategies using the modern concepts of Nanotechnology, now attention is focused on the identification and targeting of novel CVD risk factors; one such factor is an inflammatory cytokine Tumor Necrosis Factor-alpha (TNF-alpha). This cytokine is implicated in coronary atherosclerosis and cardiac disease (cardiomyopathy). Cardiologists consider TNF as an inflammatory as well as a prognostic factor of CAD complications. From this aspect it could be that the TNF neutralization results in limiting CAD among patients. Use of conventional drugs has addressed CAD problem to some extent but improvement in the treatment strategies in the emerging area of nanomedicine is demanding to deal with the rising number of CAD cases. The study aimed to investigate the effect of nanoparticles on TNFα level to formulate and investigate the probable Nano-drugs with greater efficacies, improved stabilization and lower toxicities in the Coronary Artery Disease (CAD) patients. Hence, the study comprises of three aspects i.e. chemical synthesis & characterization of nanoparticles, their cytotoxicity (hemolysis of RBC’s, cell viability) and exposure of the nanoparticles to lymphocytes and macrophage in order to reduce TNF-alpha level. For this purpose, Au, ZnO and MgO nanoparticles were synthesized and characterized by using UV-Spectroscopy, X-ray diffraction (XRD) and Scanning Electron Microscopy (SEM). First, hemolysis assay and MTT assay was conducted to determine the effect of different concentrations of ZnO, MgO and Au nanoparticles on red blood cells,lymphocytes and macrophages. After that, clinical blood samples of CAD patients were collected to culture lymphocytes and macrophages. Mature lymphocytes and macrophages were exposed to different concentrations of 5, 10, 25, 50 and 100µg/ml of nanoparticles. A significant reduction in TNF level was observed in all groups when compared with control group as well as plasma isolated from patient's blood. A two fold decrease in TNF alpha level was observed at 100µg/ml followed by 50, 25, 5µg/ml in both lymphocytes and macrophages. To our knowledge so far no study on this important aspect of the effect of nanoparticles on TNF-alpha of triple-vessel coronary artery disease has been made. Further work on a larger number of CAD patients is in progress.