Effects of the tetrahedral framework nucleic acids on the skeletal muscle regeneration in vitro and in vivo

In response injury, intrinsic repair mechanisms are activated in skeletal muscle to replace the damaged muscle fibers with new muscle fibers. The regeneration process starts with the proliferation of satellite cells to give rise to myoblasts, which subsequently differentiate terminally into myofibers. Here, we investigated the promotion effect of pigment epithelial-derived factor (PEDF) on muscle regeneration. We report that PEDF and a synthetic PEDF-derived short peptide (PSP; residues Ser93-Leu112) induce satellite cell proliferation in vitro and promote muscle regeneration in vivo. Extensively, soleus muscle necrosis was induced in rats by bupivacaine, and an injectable alginate gel was used to release the PSP in the injured muscle. PSP delivery was found to stimulate satellite cell proliferation in damaged muscle and enhance the growth of regenerating myofibers, with complete regeneration of normal muscle mass by 2 wk. In cell culture, PEDF/PSP stimulated C2C12 myoblast proliferation, together with a rise in cyclin D1 expression. PEDF induced the phosphorylation of ERK1/2, Akt, and STAT3 in C2C12 myoblasts. Blocking the activity of ERK, Akt, or STAT3 with pharmacological inhibitors attenuated the effects of PEDF/PSP on the induction of C2C12 cell proliferation and cyclin D1 expression. Moreover, 5-bromo-2′-deoxyuridine pulse-labeling demonstrated that PEDF/PSP stimulated primary rat satellite cell proliferation in myofibers in vitro. In summary, we report for the first time that PSP is capable of promoting the regeneration of skeletal muscle. The signaling mechanism involves the ERK, AKT, and STAT3 pathways. These results show the potential utility of this PEDF peptide for muscle regeneration.

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Conditional Deletion of Dicer in Adult Mice Impairs Skeletal Muscle Regeneration

International Journal of Molecular Sciences ◽

10.3390/ijms20225686 ◽

2019 ◽

Vol 20 (22) ◽

pp. 5686 ◽

Cited By ~ 2

Author(s):

Satoshi Oikawa ◽

Minjung Lee ◽

Takayuki Akimoto

Keyword(s):

Skeletal Muscle ◽

Muscle Regeneration ◽

Myogenic Differentiation ◽

Tibialis Anterior Muscle ◽

Critical Factor ◽

Skeletal Muscle Regeneration ◽

Small Noncoding Rnas ◽

Adult Mice

Skeletal muscle has a remarkable regenerative capacity, which is orchestrated by multiple processes, including the proliferation, fusion, and differentiation of the resident stem cells in muscle. MicroRNAs (miRNAs) are small noncoding RNAs that mediate the translational repression or degradation of mRNA to regulate diverse biological functions. Previous studies have suggested that several miRNAs play important roles in myoblast proliferation and differentiation in vitro. However, their potential roles in skeletal muscle regeneration in vivo have not been fully established. In this study, we generated a mouse in which the Dicer gene, which encodes an enzyme essential in miRNA processing, was knocked out in a tamoxifen-inducible way (iDicer KO mouse) and determined its regenerative potential after cardiotoxin-induced acute muscle injury. Dicer mRNA expression was significantly reduced in the tibialis anterior muscle of the iDicer KO mice, whereas the expression of muscle-enriched miRNAs was only slightly reduced in the Dicer-deficient muscles. After cardiotoxin injection, the iDicer KO mice showed impaired muscle regeneration. We also demonstrated that the number of PAX7+ cells, cell proliferation, and the myogenic differentiation capacity of the primary myoblasts did not differ between the wild-type and the iDicer KO mice. Taken together, these data demonstrate that Dicer is a critical factor for muscle regeneration in vivo.

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Dietary Flaxseed Mitigates Impaired Skeletal Muscle Regeneration: in Vivo, in Vitro and in Silico Studies

International Journal of Medical Sciences ◽

10.7150/ijms.13268 ◽

2016 ◽

Vol 13 (3) ◽

pp. 206-219 ◽

Cited By ~ 4

Author(s):

Felicia Carotenuto ◽

Alessandra Costa ◽

Maria Cristina Albertini ◽

Marco Bruno Luigi Rocchi ◽

Alexander Rudov ◽

...

Keyword(s):

Skeletal Muscle ◽

Muscle Regeneration ◽

In Silico ◽

Skeletal Muscle Regeneration ◽

In Silico Studies

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IL-4 and SDF-1 Increase Adipose Tissue-Derived Stromal Cell Ability to Improve Rat Skeletal Muscle Regeneration

International Journal of Molecular Sciences ◽

10.3390/ijms21093302 ◽

2020 ◽

Vol 21 (9) ◽

pp. 3302

Author(s):

Małgorzata Zimowska ◽

Karolina Archacka ◽

Edyta Brzoska ◽

Joanna Bem ◽

Areta M. Czerwinska ◽

...

Keyword(s):

Skeletal Muscle ◽

Adipose Tissue ◽

Muscle Regeneration ◽

Structure And Function ◽

Skeletal Muscle Regeneration ◽

Stem Cell Markers ◽

Myogenic Factors ◽

And Function

Skeletal muscle regeneration depends on the satellite cells, which, in response to injury, activate, proliferate, and reconstruct damaged tissue. However, under certain conditions, such as large injuries or myopathies, these cells might not sufficiently support repair. Thus, other cell populations, among them adipose tissue-derived stromal cells (ADSCs), are tested as a tool to improve regeneration. Importantly, the pro-regenerative action of such cells could be improved by various factors. In the current study, we tested whether IL-4 and SDF-1 could improve the ability of ADSCs to support the regeneration of rat skeletal muscles. We compared their effect at properly regenerating fast-twitch EDL and poorly regenerating slow-twitch soleus. To this end, ADSCs subjected to IL-4 and SDF-1 were analyzed in vitro and also in vivo after their transplantation into injured muscles. We tested their proliferation rate, migration, expression of stem cell markers and myogenic factors, their ability to fuse with myoblasts, as well as their impact on the mass, structure and function of regenerating muscles. As a result, we showed that cytokine-pretreated ADSCs had a beneficial effect in the regeneration process. Their presence resulted in improved muscle structure and function, as well as decreased fibrosis development and a modulated immune response.

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Plasmin activity is required for myogenesis in vitro and skeletal muscle regeneration in vivo

Blood ◽

10.1182/blood.v99.8.2835 ◽

2002 ◽

Vol 99 (8) ◽

pp. 2835-2844 ◽

Cited By ~ 62

Author(s):

Mònica Suelves ◽

Roser López-Alemany ◽

Frederic Lluı́s ◽

Gloria Aniorte ◽

Erika Serrano ◽

...

Keyword(s):

Skeletal Muscle ◽

Muscle Regeneration ◽

Myoblast Fusion ◽

Skeletal Muscle Regeneration ◽

Wild Type ◽

Plasmin Activity ◽

Type Plasminogen Activator ◽

Deficient Mice

Abstract Plasmin, the primary fibrinolytic enzyme, has a broad substrate spectrum and is implicated in biologic processes dependent upon proteolytic activity, such as tissue remodeling and cell migration. Active plasmin is generated from proteolytic cleavage of the zymogen plasminogen (Plg) by urokinase-type plasminogen activator (uPA) and tissue-type plasminogen activator (tPA). Here, we have investigated the role of plasmin in C2C12 myoblast fusion and differentiation in vitro, as well as in skeletal muscle regeneration in vivo, in wild-type and Plg-deficient mice. Wild-type mice completely repaired experimentally damaged skeletal muscle. In contrast, Plg−/− mice presented a severe regeneration defect with decreased recruitment of blood-derived monocytes and lymphocytes to the site of injury and persistent myotube degeneration. In addition, Plg-deficient mice accumulated fibrin in the degenerating muscle fibers; however, fibrinogen depletion of Plg-deficient mice resulted in a correction of the muscular regeneration defect. Because we found that uPA, but not tPA, was induced in skeletal muscle regeneration, and persistent fibrin deposition was also reproducible in uPA-deficient mice following injury, we propose that fibrinolysis by uPA-dependent plasmin activity plays a fundamental role in skeletal muscle regeneration. In summary, we identify plasmin as a critical component of the mammalian skeletal muscle regeneration process, possibly by preventing intramuscular fibrin accumulation and by contributing to the adequate inflammatory response after injury. Finally, we found that inhibition of plasmin activity with α2-antiplasmin resulted in decreased myoblast fusion and differentiation in vitro. Altogether, these studies demonstrate the requirement of plasmin during myogenesis in vitro and muscle regeneration in vivo.

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uPA, uPAR and muscle regeneration in vitro and in vivo

Fibrinolysis and Proteolysis ◽

10.1016/0268-9499(94)90702-1 ◽

1994 ◽

Vol 8 ◽

pp. 148

Author(s):

P. Muñoz-Canoves ◽

D. Talaricol ◽

J. Felez ◽

F. Blasil

Keyword(s):

Muscle Regeneration ◽

Regeneration In Vitro

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In Vivo Activation of STAT3 Signaling in Satellite Cells and Myofibers in Regenerating Rat Skeletal Muscles

Journal of Histochemistry & Cytochemistry ◽

10.1177/002215540205001202 ◽

2002 ◽

Vol 50 (12) ◽

pp. 1579-1589 ◽

Cited By ~ 65

Author(s):

Katsuya Kami ◽

Emiko Senba

Keyword(s):

Skeletal Muscle ◽

Satellite Cells ◽

Muscle Regeneration ◽

Intracellular Signaling ◽

Skeletal Muscles ◽

Early Stage ◽

Skeletal Muscle Regeneration ◽

Stat3 Signaling

Although growth factors and cytokines play critical roles in skeletal muscle regeneration, intracellular signaling molecules that are activated by these factors in regenerating muscles have been not elucidated. Several lines of evidence suggest that leukemia inhibitory factor (LIF) is an important cytokine for the proliferation and survival of myoblasts in vitro and acceleration of skeletal muscle regeneration. To elucidate the role of LIF signaling in regenerative responses of skeletal muscles, we examined the spatial and temporal activation patterns of an LIF-associated signaling molecule, the signal transducer and activator transcription 3 (STAT3) proteins in regenerating rat skeletal muscles induced by crush injury. At the early stage of regeneration, activated STAT3 proteins were first detected in the nuclei of activated satellite cells and then continued to be activated in proliferating myoblasts expressing both PCNA and MyoD proteins. When muscle regeneration progressed, STAT3 signaling was no longer activated in differentiated myoblasts and myotubes. In addition, activation of STAT3 was also detected in myonuclei within intact sarcolemmas of surviving myofibers that did not show signs of necrosis. These findings suggest that activation of STAT3 signaling is an important molecular event that induces the successful regeneration of injured skeletal muscles.

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Scaffolds containing chitosan, gelatin and graphene oxide for bone tissue regeneration in vitro and in vivo

International Journal of Biological Macromolecules ◽

10.1016/j.ijbiomac.2017.01.034 ◽

2017 ◽

Vol 104 ◽

pp. 1975-1985 ◽

Cited By ~ 78

Author(s):

S. Saravanan ◽

Anjali Chawla ◽

M. Vairamani ◽

T.P. Sastry ◽

K.S. Subramanian ◽

...

Keyword(s):

Graphene Oxide ◽

Bone Tissue ◽

Tissue Regeneration ◽

Bone Tissue Regeneration ◽

Regeneration In Vitro

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Matured Myofibers in Bioprinted Constructs with In Vivo Vascularization and Innervation

Gels ◽

10.3390/gels7040171 ◽

2021 ◽

Vol 7 (4) ◽

pp. 171

Author(s):

Catherine G. Y. Ngan ◽

Anita Quigley ◽

Richard J. Williams ◽

Cathal D. O’Connell ◽

Romane Blanchard ◽

...

Keyword(s):

Neuromuscular Diseases ◽

Structural Development ◽

Muscle Loss ◽

Test Drug ◽

Scaffold Material ◽

Volumetric Muscle Loss ◽

Primary Mouse ◽

First Time

For decades, the study of tissue-engineered skeletal muscle has been driven by a clinical need to treat neuromuscular diseases and volumetric muscle loss. The in vitro fabrication of muscle offers the opportunity to test drug-and cell-based therapies, to study disease processes, and to perhaps, one day, serve as a muscle graft for reconstructive surgery. This study developed a biofabrication technique to engineer muscle for research and clinical applications. A bioprinting protocol was established to deliver primary mouse myoblasts in a gelatin methacryloyl (GelMA) bioink, which was implanted in an in vivo chamber in a nude rat model. For the first time, this work demonstrated the phenomenon of myoblast migration through the bioprinted GelMA scaffold with cells spontaneously forming fibers on the surface of the material. This enabled advanced maturation and facilitated the connection between incoming vessels and nerve axons in vivo without the hindrance of a scaffold material. Immunohistochemistry revealed the hallmarks of tissue maturity with sarcomeric striations and peripherally placed nuclei in the organized bundles of muscle fibers. Such engineered muscle autografts could, with further structural development, eventually be used for surgical reconstructive purposes while the methodology presented here specifically has wide applications for in vitro and in vivo neuromuscular function and disease modelling.

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Persistent COUP-TFII Expression Underlies the Myopathy and Impaired Muscle Regeneration Observed in Resistance to Thyroid Hormone-Alpha

Journal of the Endocrine Society ◽

10.1210/jendso/bvab048.1658 ◽

2021 ◽

Vol 5 (Supplement_1) ◽

pp. A814-A814

Author(s):

Paola Aguiari ◽

Yan-Yun Liu ◽

Astgik Petrosyan ◽

Sheue-Yann Cheng ◽

Gregory A Brent ◽

...

Keyword(s):

Skeletal Muscle ◽

Thyroid Hormone ◽

Muscle Regeneration ◽

Myogenic Differentiation ◽

Skeletal Muscle Regeneration ◽

Hormone Signaling ◽

Muscle Loss ◽

Resistance To Thyroid Hormone ◽

Skeletal Muscle Loss

Abstract Myopathic changes, including muscular dystrophy and weakness, are commonly described in hypothyroid and hyperthyroid patients. Thyroid hormone signaling, via activation of thyroid nuclear receptor alpha (THRA), plays an essential role in maintaining muscle mass, function, and regeneration. A mouse model of resistance to thyroid hormone carrying a frameshift mutation in the THRA gene (THRA-PV) is associated with accelerated skeletal muscle loss with aging and impaired regeneration after injury(1,2). We previously demonstrated that the expression of nuclear orphan receptor chicken ovalbumin upstream promoter-factor II (COUP-TFII, or Nr2f2) persists during myogenic differentiation in THRA-PV myoblasts and skeletal muscle of aged THRA- PV mice. COUP-TFII is known to regulate myogenesis negatively and has a role in Duchenne-like Muscular Dystrophies(3). COUP-TFII physically and functionally interacts with THRA in primary myoblasts isolated from WT and THRA-PV mice, as demonstrated via co-immunoprecipitation and chromatin-immunoprecipitation. We observed that satellite cells from THRA-PV mice display impaired myoblast proliferation and in vitro myogenic differentiation compared to WT cells. However, the silencing of COUP-TFII expression using siRNA probes restores in vitro myogenic potential of THRA-PV myoblasts and shifts the mRNA expression profile closer to WT myoblasts, with a higher proliferation of myoblasts and a higher number of fully differentiated myotubes after 5 days of myogenic induction. Moreover, RNAseq analysis on myoblasts from THRA-PV mice after COUP-TFII knockdown shows that COUP-TFII silencing reverses the transcriptomic profile of THRA-PV myoblasts and results in reactivation of pathways involved in muscle function and extracellular matrix remodeling/deposition. These findings indicate that the persistent COUP-TFII expression in THRA-PV mice is responsible for the abnormal muscle phenotype. In conclusion, COUP-TFII and THRA cooperate during murine post-natal myogenesis, and COUP-TFII is critical for the accelerated skeletal muscle loss with aging and impaired muscle regeneration after injury in THRA-PV mice. These studies can help increase our knowledge of the mechanisms involved in thyroid hormone signaling during skeletal muscle regeneration, ultimately increasing the possibility of designing more specific treatments for patients with thyroid hormone-induced myopathies. References: 1. Milanesi, A., et al., Endocrinology 2016; 2. Kaneshige, M. et al., Proc Natl Acad Sci U S 2001; 3. Lee HJ, et al, Sci Rep. 2017.

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