{"doi":"10.1101/2025.04.22.650031","title":"Aligned collagen fibers drive distinct traction force signatures to regulate contact guidance","abstract":"Abstract Cellular forces on isotropically deposited extracellular matrix (ECM) have been measured extensively. However, in vivo , cells exert traction force on collagen fiber networks within ECM. Often times collagen fibers are aligned as in cancer, fibrosis and during wound healing. How forces are transmitted on aligned collagen fibers and how the cytoskeleton regulates this is unknown. Here, we develop a dual-traction force microscopy (d-TFM) approach that includes collagen fibers attached to flexible substrates with fiduciary markers on both the collagen fibers and underlying flexible substrates. This allows for the measurement of traction forces on collagen fibers in the plane of the cell, ensuring a physiologically relevant environment and accurately quantifying traction force. We find that the elastic modulus of the substrate determines the steady-state traction stress exerted by spreading cells on aligned collagen fibers, but does not affect traction force kinetics. Furthermore, collagen fiber networks on the same elastic modulus as isotropically adsorbed collagen result in higher traction stresses. Formins and Arp2/3 modulate traction stress differently, where formins affect traction stress magnitude, while Arp2/3 affects traction stress kinetics. Interestingly, we found that there is a positive correlation between traction force, migration speed and directionality on aligned collagen fibers. However, traction force does not seem to correlate with speed and directionality across distinct collagen organizational structures and across cell lines. These findings underscore the complex interplay between the mechanics of collagen fiber networks, cytoskeletal regulators and cellular traction forces, providing insights into how cells navigate complex fiber networks during migration. Graphical Abstract","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2025,"id":557661,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9599,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1349242,"name":"Azarnoosh Foroozandehfar","orcid":"0009-0003-3755-9510","position":1,"is_corresponding":false},{"id":1422578,"name":"Fred Rogers Namanda","orcid":null,"position":2,"is_corresponding":false},{"id":1313410,"name":"I Schneider","orcid":"0000-0002-4414-3842","position":3,"is_corresponding":false},{"id":1457788,"name":"Gopal Niraula","orcid":"0000-0002-8776-7794","position":0,"is_corresponding":true}],"reference_count":69,"raw_metadata":null,"created_at":"2026-07-19T02:55:21.727661Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}