Microrheology of Mucin-Albumin Assembly Using Diffusing Wave Spectroscopy

Mucus plays an important role in the protection of the epithelial cells from various pathogens and low pH environments besides helping in the absorption of nutrients. Alteration of the rheology of the mucus layer leads to various disease conditions such as cystic fibrosis, Crohn's disease, and gastric ulcers, among others. Importantly, mucus consists of various mucins along with proteins such as immunoglobulin, lysozyme, and albumin. In the present study, we explore the effect of pH on the interactions between bovine serum albumin (BSA) and porcine gastric mucins using diffusing wave spectroscopy (DWS). The study unveils that BSA actively binds with mucin to form mucin-BSA complexes, which is largely driven by electrostatic interactions. Interestingly, such physical interactions significantly alter the microrheology of these biomaterials, which is indicated by a reduction in the diffusivity of tracer particles in DWS. An array of DWS experiments suggests that the interaction between mucin and BSA is the highest at pH 7.4 and the least at pH 3. Further analyses using atomic force microscopy showed the formation of a compact cross-linked colloidal network of mucin-BSA complexes at pH 7.4, which is the main reason for the reduction in the diffusivity of the tracer particles in DWS. Furthermore, the circular dichroism analysis revealed that the secondary structures of mucin-BSA complexes are markedly different from those of only mucin at pH 7.4. Importantly, such a difference has not been observed at pH 3, which confirms that largely electrostatic interactions drive the formation of mucin-BSA complexes at neutral pH. In such a scenario, the presence of Ca²⁺ ions is also found to facilitate bridging between BSA molecules, which is also reflected in the microrheology of the suspension of BSA-mucin complexes.