Viability Of C2C12 Myoblasts Research Articles

Ginsenoside Rc prevents dexamethasone-induced muscle atrophy and enhances muscle strength and motor function

BackgroundA decline in muscle mass and function can impact the health, disease vulnerability, and mortality of older adults. Prolonged use of high doses of glucocorticoids, such as dexamethasone (DEX), can cause muscle wasting and reduced strength. Ginsenoside Rc (gRc) has been shown to protect muscles by activating the PGC-1α pathway and improving mitochondrial function. The effects of gRc on muscle atrophy and function in mice are not fully understood. Methods and resultsThe study discovered that gRc prevented the DEX-induced decrease in viability of C2C12 myoblasts and myotubes. Furthermore, gRc inhibited myotube degradation and the upregulation of muscle degradation proteins induced by DEX. Transcriptome analysis of myotubes showed that gRc enhances muscle generation processes while suppressing the TGF-β pathway and oxidative stress response. In mice, gRc effectively reversed the reductions in body weight, muscle mass, and muscle fibers caused by DEX. Furthermore, gRc significantly enhanced muscle strength and exercise capacity. Docking and transcriptome analyses indicated that gRc may act as a competitive inhibitor of DEX at the glucocorticoid receptor, potentially preventing muscle loss. ConclusionThe study suggests that gRc can prevent DEX-induced muscle wasting and weakness. Consequently, it may be a viable treatment option for sarcopenia and muscle-related disorders in various medical conditions.

Biomimetic sponges for regeneration of skeletal muscle following trauma.

Skeletal muscle is inept in regenerating after traumatic injuries due to significant loss of basal lamina and the resident satellite cells. To improve regeneration of skeletal muscle, we have developed biomimetic sponges composed of collagen, gelatin, and laminin (LM)-111 that were crosslinked with 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC). Collagen and LM-111 are crucial components of the muscle extracellular matrix and were chosen to impart bioactivity whereas gelatin and EDC were used to provide mechanical strength to the scaffold. Morphological and mechanical evaluation of the sponges showed porous structure, water-retention capacity and a compressive modulus of 590-808 kPa. The biomimetic sponges supported the infiltration and viability of C2 C12 myoblasts over 5 days of culture. The myoblasts produced higher levels of myokines such as VEGF, IL-6, and IGF-1 and showed higher expression of myogenic markers such as MyoD and myogenin on the biomimetic sponges. Biomimetic sponges implanted in a mouse model of volumetric muscle loss (VML) supported satellite, endothelial, and inflammatory cell infiltration but resulted in limited myofiber regeneration at 2 weeks post-injury. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 92-103, 2019.

Change in viability of C2C12 myoblasts under compression, shear and oxidative challenges

Skeletal and epidermal loadings can damage muscle cells and contribute to the development of deep tissue injury (DTI) – a severe kind of pressure ulcers affecting many people with disability. Important predisposing factors include the multiaxial stress and strain fields in the internal tissues, particularly the vulnerable muscles around bony prominences. A careful study of the mechanical damage thresholds for muscle cell death is critical not only to the understanding of the formation of DTI, but also to the design of various body support surfaces for prevention. In this paper, we measured the mechanical damage thresholds of C2C12 myoblasts under prescribed compressive strains (15% and 30%) and shear strains (from 0% to 100%), and studied how oxidative stress, as caused potentially by reperfusion or inflammation, may affect such damage thresholds. A flat plate was used to apply a uniform compressive strain and a radially increasing shear strain on disks of Gelatin–methacrylate (GelMA) hydrogel with myoblasts encapsulated within. The percentages of cell death were estimated with propidium iodide (PI) and calcein AM staining. Results suggested that cell death depended on both the level and duration of the applied strain. There seemed to be a non-linear coupling between compression and shear. Muscle cells often need to function biomechanically in challenging oxidative environments. To study how oxidative stress may affect the mechanical damage thresholds of myoblasts, cell viability under compressive and shear strains was also studied after the cells were pre-treated for different durations (1h and 20h) with different concentrations (0.1mM and 0.5mM) of hydrogen peroxide (H2O2). Oxidative stress can either compromise or enhance the cellular resistance to shear damage, depending on the level and duration of the oxidative exposure.

Effect of Low-Energy Gallium-Aluminum-Arsenide and Aluminium Gallium Indium Phosphide Laser Irradiation on the Viability of C2C12 Myoblasts in a Muscle Injury Model

To evaluate the effect of phototherapy on the viability of cultured C2C12 myoblasts under different nutritional conditions (muscle injury model) using low-energy gallium-aluminum-arsenide (GaAlAs) and aluminium-gallium-indium-phosphide (InGaAlP) lasers with different wavelengths and powers. The beneficial effects of phototherapy using low-energy lasers depend on irradiation parameters and type of laser used, but there are no data in the literature on C2C12 myoblasts proliferation after phototherapy with GaAlAs and InGaAlP lasers. A C2C12 cell line cultured in regular (10% fetal bovine serum, FBS) and nutrient-deficient (5% FBS) media were irradiated with low-energy GaAlAs (660 nm) and InGaAlP (780 nm) lasers with energy densities of 3.8, 6.3, and 10 J/cm2, and 3.8, 10, and 17.5 J/cm2, respectively. Cell proliferation was assessed indirectly 24 h after irradiation by measuring the mitochondrial activity and using the crystal violet assay. There were no significant differences in cell viability between laser-treated myoblasts and control cultures for all tested parameters after 24 h of cell culture, although cell cultures grown in regular nutrient medium supplemented with 10% FBS exhibited higher growth rates than cultures, irradiated or not, grown in nutrient-deficient medium. Laser phototherapy did not improve C2C12 viability under regular or nutrient-deficient (muscle injury model) conditions using the above parameters.