Work

Collagen V Presented at the Cell-Substrate Interface Mimics Substrate Stiffening to Enhance Fibroblast Traction, Adhesion Turnover, and Persistent Migration

Year: 2026

Type: preprint

Abstract: Collagen type V (Col-V) is a minor fibrillar collagen that regulates collagen fibril assembly, and its haploinsufficiency causes classical Ehlers-Danlos syndrome, in which wound healing is delayed. Whether Col-V also changes how fibroblasts adhere to, pull on, and migrate across a collagen-presenting surface has not been tested directly. Here, we covalently coupled Col-I or Col-V to soft (2.7 kPa) silicone substrates and quantified the mechanical response of NIH/3T3 fibroblasts. On Col-V, cells generated about 1.5-fold higher traction, spread more, and formed smaller focal adhesions that each carried more force. Adhesions on Col-V assembled and disassembled faster. Single fibroblasts on Col-V migrated more slowly but more persistently, and fibroblast monolayers closed a 500 µm gap faster. Raising substrate stiffness to 13 kPa reproduced this phenotype on Col-I, including higher traction, larger spread area, and smaller focal adhesions, indicating that Col-V presentation produces an adhesion state that otherwise requires a stiffer substrate. In separate reconstituted collagen hydrogels, co-assembly with 10 to 20% Col-V yielded thinner, straighter fibrils, and 20% Col-V increased the indentation modulus of the gel. These results identify the collagen type presented at the cell-material interface as a design parameter that can partly substitute for bulk stiffness in controlling fibroblast traction and migration, and they suggest Col-V as a tunable component of collagen-based biomaterials. Statement of significance: Substrate stiffness is a central design parameter for cell-instructive biomaterials, but it is difficult to change without also changing other material properties. We show that the type of collagen presented at the cell-material interface can stand in for a roughly fivefold increase in substrate stiffness. On soft substrates coupled with collagen V, a minor fibrillar collagen, fibroblasts generated traction, spreading, and compact focal adhesions comparable to those on a stiffer collagen I substrate, turned over adhesions faster, migrated more persistently, and closed wounds faster. Collagen V refined fibril architecture and stiffened reconstituted collagen hydrogels. These findings point to collagen V as a tunable component for collagen-based coatings and scaffolds intended to direct fibroblast migration, including wound repair.

Source: bioRxiv (Cold Spring Harbor Laboratory)

DOI: 10.64898/2026.10.21.760760