Tepho, N. | bioRxiv Science

bioRxiv2025

High-throughput Mucus Microrheology for Phenotyping and Disease Modeling

Mucus mechanics regulate barrier, clearance, and transport functions across epithelial tissues, yet quantitative rheology remains difficult to scale because available assays require large volumes, specialized equipment, or extensive manual analysis. Here, a high-throughput microrheology workflow based on Differential Dynamic Microscopy is established for frequency-dependent measurements from 3-10 uL mucus samples and intact mucosal surfaces using standard epifluorescence microscopy. The workflow combines preloaded tracer chambers, controlled sample handling, short video acquisition, and automated analysis to reduce operator intervention while preserving sensitivity to mucus relaxation on 1--20 s time scales. Validation against shear rheometry and particle tracking in reconstituted mucin gels shows robust recovery of viscoelastic trends across mucus-like material states. Applications in human airway epithelial air--liquid interface cultures resolve culture-medium, age, donor, COPD, cigarette-smoke, cystic-fibrosis treatment, and IL-13-associated differences, including spatial heterogeneity in situ. Measurements of clinical cervical mucus further demonstrate parallel testing of scarce specimens and sensitivity to osmotic swelling. This platform provides a scalable route to quantitative mucus phenotyping for disease modeling, therapeutic testing, and future precision-medicine studies.

bioengineering↗

bioRxiv2023

Structure-function relationships of mucociliary clearance in the human airways

Mucociliary clearance is a vital defense mechanism of the human airways, protecting against harmful particles and infections. When this process fails, it contributes to respiratory diseases like chronic obstructive pulmonary disease (COPD) and asthma. While advances in single-cell transcriptomics have revealed the complexity of airway composition, much of what we know about how airway structure impacts clearance relies on animal studies. This limits our ability to create accurate human-based models of airway diseases. Here we show that the airways in female rats and in humans exhibit species-specific differences in the distribution of ciliated and secretory cells as well as in ciliary beat, resulting in significantly higher clearance effectiveness in humans. We further reveal that standard lab-grown cultures exhibit lower clearance effectiveness compared to human airways, and we identify the underlying structural differences. By combining diverse experiments and physics-based modeling, we establish universal benchmarks to assess human airway function, interpret preclinical models, and better understand disease-specific impairments in mucociliary clearance.

biophysics↗