View in EDS

TNF-α-induced contractile dysfunction in three-dimensional engineered muscle.

Saved in:

Bibliographic Details
Title:	TNF-α-induced contractile dysfunction in three-dimensional engineered muscle.
Authors:	Tamura, Yukinori¹ (AUTHOR) ytamura@nutr.kobegakuin.ac.jp, Ishizaka, Junpei¹ (AUTHOR), Yokoyama, Sho² (AUTHOR), Ochi, Ayune¹ (AUTHOR), Kishishita, Kota² (AUTHOR), Nakajima, Ryo² (AUTHOR), Sakai, Maho³ (AUTHOR), Zeng, Ying⁴ (AUTHOR), Okugawa, Airi⁴ (AUTHOR), Higuchi, Ryosuke³ (AUTHOR), Fujisato, Toshia³ (AUTHOR), Mizutani, Ken-ichi⁴ (AUTHOR), Nakamura, Tomohiro⁵ (AUTHOR)
Source:	Journal of Bioscience & Bioengineering. Apr2026, Vol. 141 Issue 4, p300-307. 8p.
Subjects:	Tumor necrosis factors, Muscle contraction, Tissue engineering, Cytokines, RNA sequencing, Muscular atrophy, Intracellular calcium, Skeletal muscle physiology
Abstract:	Three-dimensional engineered muscle (3D-EM) provides a physiologically relevant model for examining skeletal muscle function. Tumor necrosis factor-α (TNF-α), a pro-inflammatory cytokine elevated in chronic conditions such as sarcopenia and cachexia, has been linked to muscle weakness. However, the mechanism underlying this effect remains unclear. In this study, we used a 3D-EM system to evaluate the direct impact of TNF-α on muscle contractile force. 3D-EM was produced by seeding C2C12 myoblasts with type I collagen on a culture device, followed by 15 days of differentiation. Constructs were then treated with TNF-α for 48 or 72 h, and contractile output was measured during electrical pulse stimulation. Immunohistochemical analysis and RNA sequencing (RNA-seq) with subsequent enrichment analysis were conducted to assess tissue structure and transcriptomic changes. After 48 h, TNF-α reduced contractile force by 60 %, and after 72 h, by 90 % relative to controls. Immunohistochemistry showed myotube atrophy accompanied by loss of fast-twitch fibers. RNA-seq combined with Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses indicated suppression of extracellular matrix, sarcomere organization, and calcium signaling pathways. These results suggest that TNF-α reduced force generation in 3D-EM by impairing extracellular matrix integrity, sarcomeric structure, and calcium-dependent contraction mechanisms, with preferential effects on fast-twitch fibers. Overall, this study offers mechanistic insight into the basis of sarcopenia and demonstrates the utility of 3D-EM as a model of cytokine-induced muscle weakness. • TNF-α caused a time-dependent decline in contractile force of 3D-engineered muscle. • Preferential atrophy of fast-twitch myotubes was induced by TNF-α. • RNA-seq revealed downregulation of ECM, sarcomeric, and calcium-handling genes. • Classical proteolytic pathways were not robustly activated by TNF-α. • 3D-EM provides a physiologically relevant model for inflammatory muscle weakness. [ABSTRACT FROM AUTHOR]
	Copyright of Journal of Bioscience & Bioengineering is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
Database:	Engineering Source

Description
Abstract:	Three-dimensional engineered muscle (3D-EM) provides a physiologically relevant model for examining skeletal muscle function. Tumor necrosis factor-α (TNF-α), a pro-inflammatory cytokine elevated in chronic conditions such as sarcopenia and cachexia, has been linked to muscle weakness. However, the mechanism underlying this effect remains unclear. In this study, we used a 3D-EM system to evaluate the direct impact of TNF-α on muscle contractile force. 3D-EM was produced by seeding C2C12 myoblasts with type I collagen on a culture device, followed by 15 days of differentiation. Constructs were then treated with TNF-α for 48 or 72 h, and contractile output was measured during electrical pulse stimulation. Immunohistochemical analysis and RNA sequencing (RNA-seq) with subsequent enrichment analysis were conducted to assess tissue structure and transcriptomic changes. After 48 h, TNF-α reduced contractile force by 60 %, and after 72 h, by 90 % relative to controls. Immunohistochemistry showed myotube atrophy accompanied by loss of fast-twitch fibers. RNA-seq combined with Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses indicated suppression of extracellular matrix, sarcomere organization, and calcium signaling pathways. These results suggest that TNF-α reduced force generation in 3D-EM by impairing extracellular matrix integrity, sarcomeric structure, and calcium-dependent contraction mechanisms, with preferential effects on fast-twitch fibers. Overall, this study offers mechanistic insight into the basis of sarcopenia and demonstrates the utility of 3D-EM as a model of cytokine-induced muscle weakness. • TNF-α caused a time-dependent decline in contractile force of 3D-engineered muscle. • Preferential atrophy of fast-twitch myotubes was induced by TNF-α. • RNA-seq revealed downregulation of ECM, sarcomeric, and calcium-handling genes. • Classical proteolytic pathways were not robustly activated by TNF-α. • 3D-EM provides a physiologically relevant model for inflammatory muscle weakness. [ABSTRACT FROM AUTHOR]
ISSN:	13891723
DOI:	10.1016/j.jbiosc.2025.12.008