Collagen V Presented at the Cell-Substrate Interface Mimics Substrate Stiffening to Enhance Fibroblast Traction, Adhesion Turnover, and Persistent Migration
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.
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BibTeX
@misc{royer2026collagen,
title = {{Collagen V Presented at the Cell-Substrate Interface Mimics Substrate Stiffening to Enhance Fibroblast Traction, Adhesion Turnover, and Persistent Migration}},
author = {Shaina P. Royer and Himja Tiwari and David Cho and Sesha Sai Ramineni and Majid Rostami and Mohanish Chandurkar and Alexander X. Cartagena-Rivera and Sangyoon J. Han},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.21.760760},
url = {https://www.biorxiv.org/content/10.64898/2026.10.21.760760v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - Collagen V Presented at the Cell-Substrate Interface Mimics Substrate Stiffening to Enhance Fibroblast Traction, Adhesion Turnover, and Persistent Migration
AU - Shaina P. Royer
AU - Himja Tiwari
AU - David Cho
AU - Sesha Sai Ramineni
AU - Majid Rostami
AU - Mohanish Chandurkar
AU - Alexander X. Cartagena-Rivera
AU - Sangyoon J. Han
PY - 2026
DA - 2026/10/21
PB - bioRxiv
DO - 10.64898/2026.10.21.760760
UR - https://www.biorxiv.org/content/10.64898/2026.10.21.760760v1
AB - 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.
SN - 2692-8205
ER -