Neuraminidase-Mediated Desialylation Modulates Red Blood Cell Aggregation

The glycocalyx of red blood cells (RBCs), a negatively charged surface layer rich in sialic acid residues, plays a crucial role in modulating RBC aggregation. In pathological conditions such as diabetes and sepsis, glycocalyx degradation is often observed along with abnormal RBC aggregation. However, the mechanistic relationship between these phenomena remains poorly defined. In this study, we investigate the effects of enzymatic glycocalyx degradation on RBC aggregation under physiologically relevant flow conditions. Using neuraminidase from Clostridium perfringens (C. welchii) at varying concentrations, we selectively removed sialic acid residues from the RBC glycocalyx, simulating different levels of desialylation observed in health and disease. Confocal microscopy confirmed the dose-dependent depletion of membrane sialic acid, while microfluidic experiments revealed a significant increase in both the size and stability of the RBC aggregates after enzymatic treatment. Our findings suggest that glycocalyx integrity is a crucial biophysical determinant of RBC aggregation, likely influencing both electrostatic repulsion and hydrodynamic forces. This study provides new insights into how the enzymatic modification of the glycocalyx contributes to pathological hemorheology and may inform future strategies for the diagnosis or treatment of vascular diseases.