Dispersed particles traveling at a high throughput in microchannels laterally migrate and focus into a streamline at each channel face. The focusing attractors within the cross-section are determined by the balance between the lift forces. However, particles in close proximity (e.g. due to high concentration and abrupt particle contact) suffer a breakdown of distinct focusing due to excessive hydrodynamic interaction. Here, I present numerical investigations into the effects of the strong hydrodynamic interaction on the inertial focusing. The direct numerical simulation is used to calculate the focusing/defocusing of particles, specifically since the particle-induced disturbance flows vary at the particle scale and hence affect the individual particle motion. The simulated defocusing of many-body systems prefer finite interparticle separation, in contrast with sedimentation of two mobile particles, whereby the trailing particle catches up with the leading particle due to reduced drag in its wake. I numerically demonstrate that the finite separation between nearest neighbors is a consequence of hydrodynamic repulsive motion unique to wall-bound shear flows. The author further presents direct demonstrations of the effects of the strong hydrodynamic interaction on the inertial focusing in an experimentally unachievable manner. The excessive hydrodynamic interaction drastically dissipates the near-wall focusing attractors and thus causes irreversible defocusing by breaking the balance between the lift forces. Unexpectedly, I also find that moderate hydrodynamic interaction can alter focusing speed on specific conditions, suggesting that an optimum concentration may significantly boost the inertial focusing in microfluidic-based applications. (C) 2020 The Society of Powder Technology Japan. Published by Elsevier B.V. and The Society of Powder Technology Japan. All rights reserved.