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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 centres. 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\u2013cell interactions act jointly. Furthermore, we found that myofibroblasts preferentially localize at negatively charged -12 defects, whereas fibroblasts localize at +12 defects. Consequently, the slowdown of recombination dynamics can be partially attributed to myofibroblasts\u2019 preferential association with the less mobile -12 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.<\/jats:p>","DOI":"10.1098\/rsif.2026.0007","type":"journal-article","created":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T07:08:57Z","timestamp":1785913737000},"source":"Crossref","is-referenced-by-count":0,"title":["Myofibroblasts slow down defect recombination dynamics in mixed cell monolayers"],"prefix":"10.1098","volume":"23","author":[{"given":"Zhaofei","family":"Zheng","sequence":"first","affiliation":[{"name":"Department of Mechanical Engineering, Yale University 1 , New Haven, CT, USA"},{"name":"Department of Physics, Carnegie Mellon University 2 , Pittsburgh, PA, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuxin","family":"Luo","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering, Yale University 1 , New Haven, CT, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering, Yale University 1 , New Haven, CT, USA"},{"name":"Department of Radiation Oncology, University of Maryland Medical Center 3 , Baltimore, MD, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9627-8722","authenticated-orcid":false,"given":"Yimin","family":"Luo","sequence":"additional","affiliation":[{"name":"Department of Mechanical Engineering, Yale University 1 , New Haven, CT, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"175","published-online":{"date-parts":[[2026,8,5]]},"reference":[{"key":"2026080503084364800_bib1","doi-asserted-by":"crossref","first-page":"541","DOI":"10.1080\/0267829031000097457","article-title":"Liquid crystals in biology I. 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