Controllable assembly of skeletal muscle-like bundles through 3D bioprinting

Abstract 3D printing is an effective technology for recreating skeletal muscle tissue in vitro. To achieve clinical skeletal muscle injury repair, relatively large volumes of highly aligned skeletal muscle cells are required; obtaining these is still a challenge. It is currently unclear how individual skeletal muscle cells and their neighbouring components co-ordinate to establish anisotropic architectures in highly homogeneous orientations. Here, we demonstrated a 3D printing strategy followed by sequential culture processes to engineer skeletal muscle tissue. The effects of confined printing on the skeletal muscle during maturation, which impacted the myotube alignment, myogenic gene expression, and mechanical forces, were observed. Our findings demonstrate the dynamic changes of skeletal muscle tissue during in vitro 3D construction and reveal the role of physical factors in the orientation and maturity of muscle fibres.

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THE ROLE OF SERUM PROTEINS IN THE DIFFERENTIATION OF SKELETAL MUSCLE TISSUE IN VITRO

Molecular and Cellular Aspects of Muscle Function ◽

10.1016/b978-0-08-026817-0.50014-7 ◽

1981 ◽

pp. 77-78

Author(s):

M. Tolar ◽

J. Michl ◽

I. Hájek ◽

M. Klicpera

Keyword(s):

Skeletal Muscle ◽

Muscle Tissue ◽

Serum Proteins ◽

Skeletal Muscle Tissue

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1-Deoxysphingolipids, Early Predictors of Type 2 Diabetes, Compromise the Functionality of Skeletal Myoblasts

Frontiers in Endocrinology ◽

10.3389/fendo.2021.772925 ◽

2021 ◽

Vol 12 ◽

Author(s):

Duyen Tran ◽

Stephen Myers ◽

Courtney McGowan ◽

Darren Henstridge ◽

Rajaraman Eri ◽

...

Keyword(s):

Type 2 Diabetes ◽

Skeletal Muscle ◽

Glucose Uptake ◽

Muscle Cells ◽

Skeletal Muscle Cells ◽

Muscle Dysfunction ◽

Dependent Manner

Metabolic dysfunction, dysregulated differentiation, and atrophy of skeletal muscle occur as part of a cluster of abnormalities associated with the development of Type 2 diabetes mellitus (T2DM). Recent interest has turned to the attention of the role of 1-deoxysphingolipids (1-DSL), atypical class of sphingolipids which are found significantly elevated in patients diagnosed with T2DM but also in the asymptomatic population who later develop T2DM. In vitro studies demonstrated that 1-DSL have cytotoxic properties and compromise the secretion of insulin from pancreatic beta cells. However, the role of 1-DSL on the functionality of skeletal muscle cells in the pathophysiology of T2DM still remains unclear. This study aimed to investigate whether 1-DSL are cytotoxic and disrupt the cellular processes of skeletal muscle precursors (myoblasts) and differentiated cells (myotubes) by performing a battery of in vitro assays including cell viability adenosine triphosphate assay, migration assay, myoblast fusion assay, glucose uptake assay, and immunocytochemistry. Our results demonstrated that 1-DSL significantly reduced the viability of myoblasts in a concentration and time-dependent manner, and induced apoptosis as well as cellular necrosis. Importantly, myoblasts were more sensitive to the cytotoxic effects induced by 1-DSL rather than by saturated fatty acids, such as palmitate, which are critical mediators of skeletal muscle dysfunction in T2DM. Additionally, 1-DSL significantly reduced the migration ability of myoblasts and the differentiation process of myoblasts into myotubes. 1-DSL also triggered autophagy in myoblasts and significantly reduced insulin-stimulated glucose uptake in myotubes. These findings demonstrate that 1-DSL directly compromise the functionality of skeletal muscle cells and suggest that increased levels of 1-DSL observed during the development of T2DM are likely to contribute to the pathophysiology of muscle dysfunction detected in this disease.

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Role of laminin and fibronectin in selecting myogenic versus fibrogenic cells from skeletal muscle cells in vitro

Developmental Biology ◽

10.1016/0012-1606(86)90331-3 ◽

1986 ◽

Vol 117 (2) ◽

pp. 628-635 ◽

Cited By ~ 83

Author(s):

Uwe Kühl ◽

Mücella Öcalan ◽

Rupert Timpl ◽

Klaus von der Mark

Keyword(s):

Skeletal Muscle ◽

Muscle Cells ◽

Skeletal Muscle Cells

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Extracellular Matrix-Derived Hydrogels as Biomaterial for Different Skeletal Muscle Tissue Replacements

Materials ◽

10.3390/ma13112483 ◽

2020 ◽

Vol 13 (11) ◽

pp. 2483 ◽

Cited By ~ 1

Author(s):

Daniele Boso ◽

Edoardo Maghin ◽

Eugenia Carraro ◽

Mattia Giagante ◽

Piero Pavan ◽

...

Keyword(s):

Skeletal Muscle ◽

Extracellular Matrix ◽

Muscle Tissue ◽

Myogenic Differentiation ◽

Skeletal Muscle Tissue ◽

Tissue Microenvironment ◽

Topographical Cues

Recently, skeletal muscle represents a complex and challenging tissue to be generated in vitro for tissue engineering purposes. Several attempts have been pursued to develop hydrogels with different formulations resembling in vitro the characteristics of skeletal muscle tissue in vivo. This review article describes how different types of cell-laden hydrogels recapitulate the multiple interactions occurring between extracellular matrix (ECM) and muscle cells. A special attention is focused on the biochemical cues that affect myocytes morphology, adhesion, proliferation, and phenotype maintenance, underlining the importance of topographical cues exerted on the hydrogels to guide cellular orientation and facilitate myogenic differentiation and maturation. Moreover, we highlight the crucial role of 3D printing and bioreactors as useful platforms to finely control spatial deposition of cells into ECM based hydrogels and provide the skeletal muscle native-like tissue microenvironment, respectively.

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Validation of exercise-response genes in skeletal muscle cells of Thoroughbred racing horses

Animal Bioscience ◽

10.5713/ajas.18.0749 ◽

2021 ◽

Vol 34 (1) ◽

pp. 134-142

Author(s):

Doh Hoon Kim ◽

Hyo Gun Lee ◽

Nipin Sp ◽

Dong Young Kang ◽

Kyoung-Jin Jang ◽

...

Keyword(s):

Skeletal Muscle ◽

Muscle Tissue ◽

Gene Networks ◽

Muscle Cells ◽

Skeletal Muscle Tissue ◽

High Throughput Analysis ◽

Qrt Pcr ◽

Upregulated Genes ◽

Six Genes

Objective: To understand the athletic characteristics of Thoroughbreds, high-throughput analysis has been conducted using horse muscle tissue. However, an <i>in vitro</i> system has been lacking for studying and validating genes from in silico data. The aim of this study is to validate genes from differentially expressed genes (DEGs) of our previous RNA-sequencing data <i>in vitro</i>. Also, we investigated the effects of exercise-induced stress including heat, oxidative, hypoxic and cortisol stress on horse skeletal muscle derived cells with the top six upregulated genes of DEGs.Methods: Enriched pathway analysis was conducted using the Database for Annotation, Visualization, and Integrated Discovery (DAVID) tool with upregulated genes in horse skeletal muscle tissue after exercise. Among the candidates, the top six genes were analysed through geneMANIA to investigate gene networks. Muscle cells derived from neonatal horse skeletal tissue were maintained and subjected to exercise-related stressors. Transcriptional changes in the top six genes followed by stressors were investigated using quantitative reverse transcription-polymerase chain reaction (qRT-PCR).Results: The inflammation response pathway was the most commonly upregulated pathway after horse exercise. Under non-cytotoxic conditions of exercise-related stressors, the transcriptional response of the top six genes was different among types of stress. Oxidative stress yielded the most similar expression pattern to DEGs.Conclusion: Our results indicate that transcriptional change after horse exercise in skeletal muscle tissue strongly relates to stress response. The qRT-PCR results showed that stressors contribute differently to the transcriptional regulation. These results would be valuable information to understand horse exercise in the stress aspect.

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Recycled algae-based carbon materials as electroconductive 3D printed skeletal muscle tissue engineering scaffolds

Journal of Materials Science Materials in Medicine ◽

10.1007/s10856-021-06534-6 ◽

2021 ◽

Vol 32 (7) ◽

Author(s):

Selva Bilge ◽

Emre Ergene ◽

Ebru Talak ◽

Seyda Gokyer ◽

Yusuf Osman Donar ◽

...

Keyword(s):

Skeletal Muscle ◽

Tissue Engineering ◽

Electrical Stimulation ◽

Muscle Tissue ◽

Carbonaceous Material ◽

Hydrothermal Carbonization ◽

Skeletal Muscle Tissue ◽

Myotube Formation ◽

3D Printed

AbstractSkeletal muscle is an electrically and mechanically active tissue that contains highly oriented, densely packed myofibrils. The tissue has self-regeneration capacity upon injury, which is limited in the cases of volumetric muscle loss. Several regenerative therapies have been developed in order to enhance this capacity, as well as to structurally and mechanically support the defect site during regeneration. Among them, biomimetic approaches that recapitulate the native microenvironment of the tissue in terms of parallel-aligned structure and biophysical signals were shown to be effective. In this study, we have developed 3D printed aligned and electrically active scaffolds in which the electrical conductivity was provided by carbonaceous material (CM) derived from algae-based biomass. The synthesis of this conductive and functional CM consisted of eco-friendly synthesis procedure such as pre-carbonization and multi-walled carbon nanotube (MWCNT) catalysis. CM obtained from biomass via hydrothermal carbonization (CM-03) and its ash form (CM-03K) were doped within poly(ɛ-caprolactone) (PCL) matrix and 3D printed to form scaffolds with aligned fibers for structural biomimicry. Scaffolds were seeded with C2C12 mouse myoblasts and subjected to electrical stimulation during the in vitro culture. Enhanced myotube formation was observed in electroactive groups compared to their non-conductive counterparts and it was observed that myotube formation and myotube maturity were significantly increased for CM-03 group after electrical stimulation. The results have therefore showed that the CM obtained from macroalgae biomass is a promising novel source for the production of the electrically conductive scaffolds for skeletal muscle tissue engineering.

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