Characterizing the Impact of Fabrication Methods on Mechanically Tunable Gelatin Hydrogels for Cardiac Fibrosis Studies

The mechanical properties of the extracellular matrix critically influence cell behavior in both physiological and pathophysiological states, including cardiac fibrosis. In vitro models have played a critical role in assessing biological mechanisms. In this study, we characterized mechanically tunab...

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Main Authors: Jordyn Folh, Phan Linh Dan Tran, Renita E. Horton
Format: Article
Language:English
Published: MDPI AG 2025-07-01
Series:Bioengineering
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Online Access:https://www.mdpi.com/2306-5354/12/7/759
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author Jordyn Folh
Phan Linh Dan Tran
Renita E. Horton
author_facet Jordyn Folh
Phan Linh Dan Tran
Renita E. Horton
author_sort Jordyn Folh
collection DOAJ
description The mechanical properties of the extracellular matrix critically influence cell behavior in both physiological and pathophysiological states, including cardiac fibrosis. In vitro models have played a critical role in assessing biological mechanisms. In this study, we characterized mechanically tunable enzymatically crosslinked gelatin-microbial transglutaminase (mTG) hydrogels for modeling cardiovascular diseases. Gelatin hydrogels were fabricated via direct mixing or immersion crosslinking methods. Hydrogel formulations were assessed using the Piuma nanoindenter and Instron systems. This study investigates the effects of fabrication methods, UV ozone (UVO) sterilization, crosslinking methods, and incubation media on hydrogel stiffness. Further, this study examined the response of murine cardiac fibroblasts to hydrogel stiffness. The hydrogels exhibited modulus ranges relevant to both healthy and fibrotic cardiac tissues. UVO exposure led to slight decreases in hydrogel modulus, while the fabrication method had a significant impact on the modulus. Hydrogels incubated in phosphate buffered saline (PBS) were stiffer than those incubated in Medium 199 (M199), which correlated with lower pH in PBS. Fibroblasts cultured on stiffer hydrogels display enhanced smooth muscle actin (SMA) expression, suggesting sensitivity to material stiffness. These findings highlight how fabrication parameters influence the modulus of gelatin-mTG hydrogels for cardiac tissue models.
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spelling doaj-art-3f3fccf881704588a080ee8549f0e5ad2025-08-20T03:36:13ZengMDPI AGBioengineering2306-53542025-07-0112775910.3390/bioengineering12070759Characterizing the Impact of Fabrication Methods on Mechanically Tunable Gelatin Hydrogels for Cardiac Fibrosis StudiesJordyn Folh0Phan Linh Dan Tran1Renita E. Horton2Cardiovascular Tissue Engineering Laboratory, Biomedical Engineering Department, Cullen College of Engineering, University of Houston, Houston, TX 77204, USACardiovascular Tissue Engineering Laboratory, Biomedical Engineering Department, Cullen College of Engineering, University of Houston, Houston, TX 77204, USACardiovascular Tissue Engineering Laboratory, Biomedical Engineering Department, Cullen College of Engineering, University of Houston, Houston, TX 77204, USAThe mechanical properties of the extracellular matrix critically influence cell behavior in both physiological and pathophysiological states, including cardiac fibrosis. In vitro models have played a critical role in assessing biological mechanisms. In this study, we characterized mechanically tunable enzymatically crosslinked gelatin-microbial transglutaminase (mTG) hydrogels for modeling cardiovascular diseases. Gelatin hydrogels were fabricated via direct mixing or immersion crosslinking methods. Hydrogel formulations were assessed using the Piuma nanoindenter and Instron systems. This study investigates the effects of fabrication methods, UV ozone (UVO) sterilization, crosslinking methods, and incubation media on hydrogel stiffness. Further, this study examined the response of murine cardiac fibroblasts to hydrogel stiffness. The hydrogels exhibited modulus ranges relevant to both healthy and fibrotic cardiac tissues. UVO exposure led to slight decreases in hydrogel modulus, while the fabrication method had a significant impact on the modulus. Hydrogels incubated in phosphate buffered saline (PBS) were stiffer than those incubated in Medium 199 (M199), which correlated with lower pH in PBS. Fibroblasts cultured on stiffer hydrogels display enhanced smooth muscle actin (SMA) expression, suggesting sensitivity to material stiffness. These findings highlight how fabrication parameters influence the modulus of gelatin-mTG hydrogels for cardiac tissue models.https://www.mdpi.com/2306-5354/12/7/759hydrogelmodulusmechanotransductionfibroblastsfibrosis
spellingShingle Jordyn Folh
Phan Linh Dan Tran
Renita E. Horton
Characterizing the Impact of Fabrication Methods on Mechanically Tunable Gelatin Hydrogels for Cardiac Fibrosis Studies
Bioengineering
hydrogel
modulus
mechanotransduction
fibroblasts
fibrosis
title Characterizing the Impact of Fabrication Methods on Mechanically Tunable Gelatin Hydrogels for Cardiac Fibrosis Studies
title_full Characterizing the Impact of Fabrication Methods on Mechanically Tunable Gelatin Hydrogels for Cardiac Fibrosis Studies
title_fullStr Characterizing the Impact of Fabrication Methods on Mechanically Tunable Gelatin Hydrogels for Cardiac Fibrosis Studies
title_full_unstemmed Characterizing the Impact of Fabrication Methods on Mechanically Tunable Gelatin Hydrogels for Cardiac Fibrosis Studies
title_short Characterizing the Impact of Fabrication Methods on Mechanically Tunable Gelatin Hydrogels for Cardiac Fibrosis Studies
title_sort characterizing the impact of fabrication methods on mechanically tunable gelatin hydrogels for cardiac fibrosis studies
topic hydrogel
modulus
mechanotransduction
fibroblasts
fibrosis
url https://www.mdpi.com/2306-5354/12/7/759
work_keys_str_mv AT jordynfolh characterizingtheimpactoffabricationmethodsonmechanicallytunablegelatinhydrogelsforcardiacfibrosisstudies
AT phanlinhdantran characterizingtheimpactoffabricationmethodsonmechanicallytunablegelatinhydrogelsforcardiacfibrosisstudies
AT renitaehorton characterizingtheimpactoffabricationmethodsonmechanicallytunablegelatinhydrogelsforcardiacfibrosisstudies