The formation of a cell-free layer (CFL) results from the axial migration of red blood cells (RBC) towards the center of the flow due to the combination of shear induced and compression and tension forces on the cells. This can be enhanced by RBC aggregation which is dominantly present in athletic species. While the formation of CFL in the microcirculatory vessels has been known to play an important role in modulating blood flow resistance, it is also capable of modulating the wall shear stress (WSS) acting on the endothelial cells. This in turn leads to the whole cascade of vasoregulatory mechanism which involves the synthesis and diffusion of nitric oxide (NO), particularly in the arteriolar networks. As such, this project aims to elucidate the influence of CFL variations on the diffusion of NO and oxygen in the microcirculation. In addition, the effect of RBC aggregation on the dynamics of CFL formation was investigated to simulate the potential microvascular load in the arterioles under pathological blood flow conditions. In order to achieve these aims, an integrative approach was adopted to include both experimental work and numerical modelling in this project, to study the dynamics of NO diffusion in an arteriole. The investigation of hemodynamics was also extended to the network level using a laser speckle contrast imaging system, which provides a more accurate representation of the perfusion state in the tissues.