Myofibroblasts slow down defect recombination dynamics in mixed cell monolayers

TL;DR

This study investigates how myofibroblasts, which are less mobile, influence defect dynamics and cellular organization in mixed cell monolayers, revealing their role in slowing defect recombination and localizing at specific topological defects.

Contribution

It demonstrates that myofibroblasts slow defect relaxation and preferentially localize at -1/2 defects, affecting collective cell behavior in mixed monolayers.

Findings

Increasing myofibroblast fraction increases disorder and slows defect recombination.

Myofibroblasts localize at -1/2 defects, fibroblasts at +1/2 defects.

Myofibroblast localization at defects impedes defect mobility and alters mechanotransduction.

Abstract

Cellular organization and mechanotransduction pathways are crucial regulators of tissue morphogenesis, whereas their dysregulation contributes to pathologies. Overactive myofibroblasts are key drivers of fibrosis, yet how their presence alters collective cellular ordering remains unclear. Owing to steric interactions, elongated cells exhibit local order. Topological defects, where alignment is disrupted, have been postulated to serve as mechanical centers. In this study, we examine how incorporating slower moving myofibroblast phenotype impacts defect relaxation in monolayers consisting of co-cultured fibroblasts and myofibroblasts. In this system, myofibroblasts act as the less active component. Increasing their fraction increases disorder strength and slows defect recombination. On microgrooved surfaces, higher myofibroblast concentrations lead to worse alignment, suggesting single-cell mechanosensing and cell-cell interactions act jointly. Furthermore, we found that myofibroblasts preferentially localize at negatively charged -1/2 defects, whereas fibroblasts localize at +1/2 defects. Consequently, the slowdown of recombination dynamics can be partially attributed to myofibroblasts' preferential association with the less mobile -1/2 defects, increasing local friction and impeding defect mobility. This localization may also reduce compressive stress on myofibroblasts, as indicated by immunofluorescence of a downstream mechanotransducer. This work provides insights into possible connections between topological defects and cell motility in mixed phenotype monolayers.