scholarly journals Electrophysiological analysis of healthy and dystrophic 3D bioengineered skeletal muscle tissues

2021 ◽  
Author(s):  
Christine T. Nguyen ◽  
Majid Ebrahimi ◽  
Penney M. Gilbert ◽  
Bryan Andrew Stewart
Keyword(s):  
Skeletal Muscle ◽  
Muscle Cell ◽  
Muscle Tissue ◽  
Three Dimensional ◽  
Dystrophin Gene ◽  
Muscle System ◽  
Electrophysiological Analysis ◽  
Cytoskeletal Network ◽  
Muscle Tissues

Recently, methods for creating three-dimensional (3D) human skeletal muscle tissues from myogenic cell lines have been reported. Bioengineered muscle tissues are contractile and respond to electrical and chemical stimulation. In this study we provide an electrophysiological analysis of healthy and dystrophic 3D bioengineered skeletal muscle tissues. We focus on Duchenne muscular dystrophy (DMD), a fatal muscle disorder involving the skeletal muscle system. The dystrophin gene, which when mutated causes DMD, encodes for the Dystrophin protein, which anchors the cytoskeletal network inside of a muscle cell to the extracellular matrix outside the cell. Here, we enlist a 3D in vitro model of DMD muscle tissue, to evaluate an understudied aspect of DMD, muscle cell electrical properties uncoupled from presynaptic neural inputs. Our data shows that electrophysiological aspects of DMD are replicated in the 3D bioengineered skeletal muscle tissue model. Furthermore, we test a block co-polymer, poloxamer 188, and demonstrate capacity for improving the membrane potential in DMD muscle. Therefore, this study serves as the baseline for a new in vitro method to examine potential therapies directed at muscular disorders.

scholarly journals Electrophysiological analysis of healthy and dystrophic 3D bioengineered skeletal muscle tissues

2020 ◽  
Author(s):  
Christine T Nguyen ◽  
Majid Ebrahmi ◽  
Penney M Gilbert ◽  
Bryan A Stewart
Keyword(s):  
Skeletal Muscle ◽  
Muscle Cell ◽  
Muscle Tissue ◽  
Three Dimensional ◽  
Muscle System ◽  
Electrophysiological Analysis ◽  
Cytoskeletal Network ◽  
Muscle Tissues ◽  
Dystrophin Protein

AbstractRecently, methods for creating three-dimensional (3D) human skeletal muscle tissues from myogenic cell lines have been reported. Bioengineered muscle tissues are contractile and respond to electrical and chemical stimulation. In this study we provide an electrophysiological analysis of healthy and dystrophic 3D bioengineered skeletal muscle tissues. We focus on Duchenne muscular dystrophy (DMD), a fatal muscle disorder involving the skeletal muscle system. The dystrophin gene, which when mutated causes DMD, encodes for the Dystrophin protein, which anchors the cytoskeletal network inside of a muscle cell to the extracellular matrix outside the cell. Here, we enlist a 3D in vitro model of DMD muscle tissue, to evaluate an understudied aspect of DMD, muscle cell electrical properties uncoupled from presynaptic neural inputs. Our data shows that electrophysiological aspects of DMD are replicated in the 3D bioengineered skeletal muscle tissue model. Furthermore, we test a block co-polymer, poloxamer 188, and demonstrate capacity for improving the membrane potential in DMD muscle.Therefore, this study serves as the baseline for a new in vitro method to examine potential therapies directed at muscular disorders.

scholarly journals Skeletal muscle-on-a-chip: an in vitro model to evaluate tissue formation and injury

Lab on a Chip ◽  
2017 ◽  
Vol 17 (20) ◽  
pp. 3447-3461 ◽  
Author(s):  
Gaurav Agrawal ◽  
Aereas Aung ◽  
Shyni Varghese
Keyword(s):  
Skeletal Muscle ◽  
Muscle Injury ◽  
In Vitro Model ◽  
Three Dimensional ◽  
Tissue Formation ◽  
Microfluidic Platform ◽  
Muscle Tissues ◽  
Vitro Model

We introduce a microfluidic platform in which we culture three-dimensional skeletal muscle tissues, while evaluating tissue formation and toxin-induced muscle injury.

scholarly journals A Cylindrical Molding Method for the Biofabrication of Plane-Shaped Skeletal Muscle Tissue

Micromachines ◽  
2021 ◽  
Vol 12 (11) ◽  
pp. 1411
Author(s):  
Minghao Nie ◽  
Ai Shima ◽  
Kenta Fukushima ◽  
Yuya Morimoto ◽  
Shoji Takeuchi
Keyword(s):  
Skeletal Muscle ◽  
Muscle Tissue ◽  
Myogenic Differentiation ◽  
Skeletal Muscle Tissue ◽  
Differentiation Markers ◽  
Animal Products ◽  
Molding Method ◽  
Muscle Tissues ◽  
Central Pillar

Muscle tissues can be fabricated in vitro by culturing myoblast-populated hydrogels. To counter the shrinkage of the myoblast-populated hydrogels during culture, a pair of anchors are generally utilized to fix the two ends of the hydrogel. Here, we propose an alternative method to counter the shrinkage of the hydrogel and fabricate plane-shaped skeletal muscle tissues. The method forms myoblast-populated hydrogel in a cylindrical cavity with a central pillar, which can prevent tissue shrinkage along the circumferential direction. By eliminating the usages of the anchor pairs, our proposed method can produce plane-shaped skeletal muscle tissues with uniform width and thickness. In experiments, we demonstrate the fabrication of plane-shaped (length: ca. 10 mm, width: 5~15 mm) skeletal muscle tissue with submillimeter thickness. The tissues have uniform shapes and are populated with differentiated muscle cells stained positive for myogenic differentiation markers (i.e., myosin heavy chains). In addition, we show the assembly of subcentimeter-order tissue blocks by stacking the plane-shaped skeletal muscle tissues. The proposed method can be further optimized and scaled up to produce cultured animal products such as cultured meat.

scholarly journals Trauma induced tissue survival in vitro with a muscle-biomaterial based osteogenic organoid system: a proof of concept study

2019 ◽  
Author(s):  
Tao He ◽  
Jörg Hausdorf ◽  
Yan Chevalier ◽  
Roland Manfred Klar
Keyword(s):  
Skeletal Muscle ◽  
Bone Tissue ◽  
Muscle Tissue ◽  
Three Dimensional ◽  
Good Alternative ◽  
Clinical Environment ◽  
In Vitro Systems ◽  
Osteogenic Gene Expression

Abstract Background The translation from animal research into the clinical environment remains problematic, as animal systems do not adequately replicate the human in vivo environment. Bioreactors have emerged as a good alternative that can reproduce part of the human in vivo processes at an in vitro level. However, in vitro bone formation platforms primarily utilizes stem cells only, with tissue based in vitro systems remaining poorly investigated. As such, the present pilot study explored the tissue behavior and cell survival capability within a new in vitro skeletal muscle tissue-based biomaterial organoid bioreactor system to maximize future bone tissue engineering prospects. Results Three dimensional printed β-tricalcium phosphate/hydroxyapatite devices were either wrapped in a sheet of rat muscle tissue or first implanted in a heterotopic muscle pouch that was then excised and cultured in vitro for up to 30 days. Devices wrapped in muscle tissue showed cell death by day 15. Contrarily, devices in muscle pouches showed angiogenic and limited osteogenic gene expression tendencies with consistent TGF-ß 1 , COL4A1 , VEGF-A , RUNX-2 , and BMP-2 upregulation, respectively. Histologically, muscle tissue degradation and fibrin release was seen being absorbed by devices acting possibly as a support for new tissue formation in the bioceramic scaffold that supports progenitor stem cell osteogenic differentiation.Conclusions These results therefore demonstrate that the skeletal muscle pouch-based biomaterial culturing system can support tissue survival over a prolonged culture period and represents a novel organoid tissue model that with further adjustments could generate bone tissue for direct clinical transplantations.

Lipid peroxidation of skeletal muscle: an in vitro study

1983 ◽  
Vol 3 (7) ◽  
pp. 609-619 ◽  
Author(s):  
M. J. Jackson ◽  
D. A. Jones ◽  
R. H. T. Edwards
Keyword(s):  
Skeletal Muscle ◽  
Lipid Peroxidation ◽  
Free Radical ◽  
Vitamin E ◽  
Hydroxyl Radicals ◽  
Muscle Tissue ◽  
In Vitro Study ◽  
Muscle Tissues ◽  
In Vitro Technique

The process of lipid peroxidation of skeletal muscle has been examined in vitro by monitoring the autoxidation of skeletal-muscle homogenates. Skeletal-muscle tissue has been shown to have considerable capacity for autoxidation and the process has been found to be initiated by a free-radical-mediated mechanism, critically dependent on the presence of free iron in the homogenate. The initiating radicals have not been firmly identified, but the results suggest that neither superoxide or hydroxyl radicals are involved. An in vitro technique for assessment of the antioxidant capacity of muscle tissue has also been developed which is capable of demonstrating differences between muscle tissues with differing vitamin E contents.

scholarly journals Three-Dimensional Bioprinting of Functional Skeletal Muscle Tissue Using GelatinMethacryloyl-Alginate Bioinks

Micromachines ◽  
2019 ◽  
Vol 10 (10) ◽  
pp. 679 ◽  
Author(s):  
Seyedmahmoud ◽  
Çelebi-Saltik ◽  
Barros ◽  
Nasiri ◽  
Banton ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Tissue Engineering ◽  
Muscle Tissue ◽  
Three Dimensional ◽  
Skeletal Muscle Tissue ◽  
3D Bioprinting ◽  
Uv Crosslinking ◽  
Tissue Constructs ◽  
Muscle Tissues ◽  
Alginate Concentration

Skeletal muscle tissue engineering aims to fabricate tissue constructs to replace or restore diseased or injured skeletal muscle tissues in the body. Several biomaterials and microscale technologies have been used in muscle tissue engineering. However, it is still challenging to mimic the function and structure of the native muscle tissues. Three-dimensional (3D) bioprinting is a powerful tool to mimic the hierarchical structure of native tissues. Here, 3D bioprinting was used to fabricate tissue constructs using gelatin methacryloyl (GelMA)-alginate bioinks. Mechanical and rheological properties of GelMA-alginate hydrogels were characterized. C2C12 myoblasts at the density 8 × 106 cells/mL were used as the cell model. The effects of alginate concentration (0, 6, and 8% (w/v)) and crosslinking mechanism (UV crosslinking or ionic crosslinking with UV crosslinking) on printability, cell viability, proliferation, and differentiation of bioinks were studied. The results showed that 10% (w/v) GelMA-8% (w/v) alginate crosslinked using UV light and 0.1 M CaCl2 provided the optimum niche to induce muscle tissue formation compared to other hydrogel compositions. Furthermore, metabolic activity of cells in GelMA bioinks was improved by addition of oxygen-generating particles to the bioinks. It is hoped that such bioprinted muscle tissues may find wide applications in drug screening and tissue regeneration.

scholarly journals Bioengineered Fascicle-Like Skeletal Muscle Tissue Constructs

2012 ◽  
Author(s):  
Devin Neal ◽  
Mahmut Selman Sakar ◽  
H. Harry Asada
Keyword(s):  
Skeletal Muscle ◽  
Cell Density ◽  
Muscle Cell ◽  
Muscle Tissue ◽  
Skeletal Muscle Tissue ◽  
Muscle Injuries ◽  
Tissue Constructs

Tissue engineered skeletal muscle constructs have and will continue to be valuable in treating, and testing various muscle injuries and diseases. However a significant drawback to numerous methods of producing 3D skeletal muscle constructs grown in vitro is that muscle cell density as a fraction of total volume or mass, is often significantly lower than muscle found in vivo. Therefore a method to increase muscle cell density within a construct is needed.

Micro-Topography Enhances Directional Myogenic Differentiation of Skeletal Precursor Cells

2007 ◽  
Vol 1052 ◽  
Author(s):  
Yi Zhao
Keyword(s):  
Skeletal Muscle ◽  
Myogenic Differentiation ◽  
Three Dimensional ◽  
Skeletal Myoblasts ◽  
Aspect Ratios ◽  
Cell Proliferation And Differentiation ◽  
Muscle Tissues ◽  
Chip Fabrication ◽  
On Chip

AbstractSkeletal muscle tissues were constructed using an in vitro model, by differentiating skeletal myoblasts using an array of linear microstructures with the medium aspect ratios. The adaptation of skeletal myoblasts has been characterized with immunoflurescence microscopy during cell proliferation and differentiation. In particular, the dependence of the alignment efficiency on the dimensions of the microstructures was studied. The morphology difference of the myotubes in the three-dimensional tissues was reported. This paper holds the promise of efficient on-chip fabrication of skeletal muscle tissues and has an important implication in direct muscle repair and muscular mechanics.

scholarly journals Trauma induced tissue survival in vitro with a muscle-biomaterial based osteogenic organoid system: a proof of concept study

2019 ◽  
Author(s):  
Tao He ◽  
Jörg Hausdorf ◽  
Yan Chevalier ◽  
Roland Manfred Klar
Keyword(s):  
Skeletal Muscle ◽  
Tissue Engineering ◽  
Bone Tissue Engineering ◽  
Bone Tissue ◽  
Muscle Tissue ◽  
Three Dimensional ◽  
Good Alternative ◽  
Clinical Environment

Abstract Background: The translation from animal research into the clinical environment remains problematic, as animal systems do not adequately replicate the human in vivo environment. Bioreactors have emerged as a good alternative that can reproduce part of the human in vivo processes at an in vitro level. Bone tissue-engineering bioreactors, however, still are cell based with tissue based in vitro systems remaining poorly investigated. As such, the present pilot study explored the tissue behavior and cell survival capability within a new in vitro skeletal muscle tissue-based biomaterial organoid bioreactor system to maximize future bone tissue engineering prospects. Results: Three dimensional printed β-tricalcium phosphate/hydroxyapatite devices were either wrapped in a sheet of rat muscle tissue or first implanted in a heterotopic muscle pouch that was then excised and cultured in vitro for up to 30 days. Devices wrapped in muscle tissue showed cell death by day 15. Contrarily, devices in muscle pouches showed angiogenic and limited osteogenic gene expression tendencies with consistent TGF-ß1, COL4A1, VEGF-A, RUNX-2, and BMP-2 upregulation, respectively. Histologically, muscle tissue degradation and fibrin release was seen being absorbed by devices acting possibly as a support for new tissue formation in the bioceramic scaffold that supports progenitor stem cell osteogenic differentiation.Conclusions: These results therefore demonstrate that the skeletal muscle pouch-based biomaterial culturing system can support tissue survival over a prolonged culture period and represents a novel organoid tissue model that with further adjustments could generate bone tissue for direct clinical transplantations.

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