Fiber alignment alone is insufficient to predict cellular contact guidance responses in engineered 3D collagen hydrogels
- 1Rochester Institute of Technology
Abstract
In breast cancer, collagen fiber alignment and the second harmonic generation forward-to-backward ratio (F/B), an optical measurement sensitive to fibril-scale organization within collagen fibers, have each been associated with metastatic outcome. Although both are measured at the tumor-stromal interface, in vitro studies have largely examined them separately, because experimental systems have not allowed the two to be varied independently. To address this technical gap, we used a microfluidic biofabrication approach to generate paired aligned and unaligned regions within 3D collagen hydrogels via extensional strain and varied measured F/B across gels by changing the pre-gel pH. The resulting library covered alignment values reported in breast tumors and F/B values associated with poorer metastatic outcome in invasive ductal carcinoma at matched matrix stiffness, pore diameter, and fiber fraction. In MDA-MB-231 cells, alignment biased migration along the fiber axis without changing migration speed, as expected, but the magnitude of the contact guidance response varied with measured F/B. In contrast, MCF-7 cells showed little overall contact guidance, yet migrated ~30% faster in high F/B than low F/B gels in both aligned and unaligned regions. Together, these results show that cells migrated differently in matrices with similar fiber alignment. Thus, including measured F/B may provide additional information to help anticipate how cells migrate in collagen environments that appear similar based on alignment alone.
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BibTeX
@misc{esmaili2026fiber,
title = {{Fiber alignment alone is insufficient to predict cellular contact guidance responses in engineered 3D collagen hydrogels}},
author = {P. Esmaili and T. M. Elias and I. M. Joshi and R. A. Simon and D. De Silva and S. Farajollahi and O. M. Harris and E. B. Brown and V. V. Abhyankar},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.29.755189},
url = {https://www.biorxiv.org/content/10.64898/2026.09.29.755189v1},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - Fiber alignment alone is insufficient to predict cellular contact guidance responses in engineered 3D collagen hydrogels
AU - P. Esmaili
AU - T. M. Elias
AU - I. M. Joshi
AU - R. A. Simon
AU - D. De Silva
AU - S. Farajollahi
AU - O. M. Harris
AU - E. B. Brown
AU - V. V. Abhyankar
PY - 2026
DA - 2026/09/29
PB - bioRxiv
DO - 10.64898/2026.09.29.755189
UR - https://www.biorxiv.org/content/10.64898/2026.09.29.755189v1
AB - In breast cancer, collagen fiber alignment and the second harmonic generation forward-to-backward ratio (F/B), an optical measurement sensitive to fibril-scale organization within collagen fibers, have each been associated with metastatic outcome. Although both are measured at the tumor-stromal interface, in vitro studies have largely examined them separately, because experimental systems have not allowed the two to be varied independently. To address this technical gap, we used a microfluidic biofabrication approach to generate paired aligned and unaligned regions within 3D collagen hydrogels via extensional strain and varied measured F/B across gels by changing the pre-gel pH. The resulting library covered alignment values reported in breast tumors and F/B values associated with poorer metastatic outcome in invasive ductal carcinoma at matched matrix stiffness, pore diameter, and fiber fraction. In MDA-MB-231 cells, alignment biased migration along the fiber axis without changing migration speed, as expected, but the magnitude of the contact guidance response varied with measured F/B. In contrast, MCF-7 cells showed little overall contact guidance, yet migrated ~30% faster in high F/B than low F/B gels in both aligned and unaligned regions. Together, these results show that cells migrated differently in matrices with similar fiber alignment. Thus, including measured F/B may provide additional information to help anticipate how cells migrate in collagen environments that appear similar based on alignment alone.
ER -