Hydrogel dimensionality directs mechanics-dependent nascent matrix deposition by tendon fibroblasts
Abstract
Tendon fibroblasts continuously synthesize and remodel extracellular matrix (ECM) in response to their surrounding mechanical environment in vivo and in vitro, yet how dimensionality regulates the deposition of newly synthesized matrix remains poorly understood. Here, we investigated nascent matrix deposition by primary mouse tail tendon fibroblasts cultured on flat two-dimensional (2D) or encapsulated within three-dimensional (3D) norbornene-modified hyaluronic acid hydrogels with tunable mechanical properties. Newly synthesized matrix was visualized using metabolic labeling with azido-galactosamine (GalNAz), which labels nascent glycoproteins deposited during culture. In 2D, increasing hydrogel elastic modulus from 10 to 25 kPa enhanced nascent matrix deposition. This response was accompanied by increased cell spreading and YAP nuclear localization, indicating increased cellular mechanosensing. In contrast, tendon fibroblasts encapsulated within enzymatically degradable 3D hydrogels showed progressive matrix deposition over time but deposited less nascent matrix in stiffer (~50 kPa) hydrogels compared with softer 5 and 10 kPa hydrogels. YAP nuclear localization was increased in softer hydrogels compared with 50 kPa hydrogels. Bulk RNA sequencing showed increased expression of tendon matrisome-associated genes in softer 3D hydrogels, including increased tenascin C and collagen type I expression. These findings demonstrate that hydrogel dimensionality determines the relationship between matrix mechanics and nascent matrix deposition. This work establishes a framework for studying how engineered microenvironments regulate the matrix-producing behavior of tendon fibroblasts and provides insight into the design of biomaterials for investigating tendon biology and matrix remodeling.
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BibTeX
@misc{liu2026hydrogel,
title = {{Hydrogel dimensionality directs mechanics-dependent nascent matrix deposition by tendon fibroblasts}},
author = {Joshua Yu-Chung Liu and Ziqian Yang and Stephanie S. Steltzer and Mingyue Fan and Evelyn S. Navarro Salazar and Jiahao Liang and Megan L. Killian and Claudia Loebel},
howpublished = {bioRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.23.761444},
url = {https://www.biorxiv.org/content/10.64898/2026.10.23.761444v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Hydrogel dimensionality directs mechanics-dependent nascent matrix deposition by tendon fibroblasts
AU - Joshua Yu-Chung Liu
AU - Ziqian Yang
AU - Stephanie S. Steltzer
AU - Mingyue Fan
AU - Evelyn S. Navarro Salazar
AU - Jiahao Liang
AU - Megan L. Killian
AU - Claudia Loebel
PY - 2026
DA - 2026/10/23
PB - bioRxiv
DO - 10.64898/2026.10.23.761444
UR - https://www.biorxiv.org/content/10.64898/2026.10.23.761444v1
AB - Tendon fibroblasts continuously synthesize and remodel extracellular matrix (ECM) in response to their surrounding mechanical environment in vivo and in vitro, yet how dimensionality regulates the deposition of newly synthesized matrix remains poorly understood. Here, we investigated nascent matrix deposition by primary mouse tail tendon fibroblasts cultured on flat two-dimensional (2D) or encapsulated within three-dimensional (3D) norbornene-modified hyaluronic acid hydrogels with tunable mechanical properties. Newly synthesized matrix was visualized using metabolic labeling with azido-galactosamine (GalNAz), which labels nascent glycoproteins deposited during culture. In 2D, increasing hydrogel elastic modulus from 10 to 25 kPa enhanced nascent matrix deposition. This response was accompanied by increased cell spreading and YAP nuclear localization, indicating increased cellular mechanosensing. In contrast, tendon fibroblasts encapsulated within enzymatically degradable 3D hydrogels showed progressive matrix deposition over time but deposited less nascent matrix in stiffer (~50 kPa) hydrogels compared with softer 5 and 10 kPa hydrogels. YAP nuclear localization was increased in softer hydrogels compared with 50 kPa hydrogels. Bulk RNA sequencing showed increased expression of tendon matrisome-associated genes in softer 3D hydrogels, including increased tenascin C and collagen type I expression. These findings demonstrate that hydrogel dimensionality determines the relationship between matrix mechanics and nascent matrix deposition. This work establishes a framework for studying how engineered microenvironments regulate the matrix-producing behavior of tendon fibroblasts and provides insight into the design of biomaterials for investigating tendon biology and matrix remodeling.
SN - 2692-8205
ER -