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The muscular-skeletal system is derived from the mesoderm, and in humans is called an endoskeleton. Cartilage is secreted by chondrocytes, which consist of a firm but elastic matrix called chondrin and is what composes the majority of the fetal skeleton. Axial Skeleton consists of the skull, the vertebrae column, and the rib cage, which act as a support structure for the core of the body.
Appendicular Skeleton consists of all of the limbs, the arms and the legs, and the pelvic and pectoral girdles.
Long Bones consist of two dilated ends called the epiphysis, which have a spongy bone core inside of a compact bone sheath. Red Marrow contains hematopoitic stem cells, which are used in the production of red blood cells and immune system cells.
Click here to learn about what kind of immune system cells hematopoitic stem cells differentiate into.
Joints are how the 206 bones of the adult skeleton interact and come in two varieties movable and immovable.
Articular Cartilage coats the articular surfaces of bones so impact is restricted to joint cartilage and not the bone.
Muscles work in conjunction with the nervous system in order to perform almost all of the essential tasks that keep humans alive such as digestion, circulation and voluntary muscle movement. Smooth Muscle is non-striated, with one centrally located nucleus per cell and is under control by the involuntary or autonomic nervous system. Cardiac Muscle consists of cardiac fibers, which are striated, with one to two nucleuses per cell and under control from the involuntary or autonomic nervous system. Myofibrils which are simply a bunch of sarcomeres put together, is surrounded by the sarcoplasmic reticulum, which is a modified endoplasmic reticulum containing Ca2+. T-tubules are an extension of the cell membrane that are connected perpendicularly to myofibrils, through which ions flow.
The sarcoplasmic reticulum (SR) is a specialized smooth endoplasmic reticulum that works to store calcium and then release it in response to an action potential. Red Muscle (Slow Twitch) is composed of more mitochondria, which aid in aerobic respiration and a greater number of myoglobin. The sarcomere is the basic unit of a muscle fiber and is made of either thick or thin filaments. Contraction occurs when the nervous system sends a signal from a motor neuron, which travels until it reaches the nerve terminal (synaptic bouton) where the release of a neurotransmitter such as acetylcholine into the synapse at the neuromuscular junction results in contraction when it connects with a specific receptor. An action potential generated at the neuromuscular junction is conducted along the sarcolemma and the t-system and transported into the fiber as it passes. The release of a large amount of calcium binds to troponin, which causes tropomyosin to shift out of the way, exposing myosin-binding sites to actin.
As the free globular heads of myosin bind are exposed to actin sites, cross bridges are formed and the myosin head bends in something called the power stroke towards the M line. ATPase activity in the myosin heads provides the energy for the power stroke, and results in the dissociation of actin from myosin. Once the sarcoplasmic reticulum stimulation is lost, the concentration of calcium ions decreases and the products of ATP hydrolysis from the power stroke free space allowing for actin and myosin dissociation. Muscle cells have an all or nothing response, which is related to the amount of energy emitted by a neuron. Simple Twitches are the response of a single fiber to a brief stimulus at or above the threshold levels.
Latent Period is the period between stimuli and contractions created from a spreading action potential.
Summation (Tetanus) occurs when fibers are exposed to frequent and prolonged stimulation and therefore cannot relax.
We have partnered with Amazon to provide you with the lowest prices on the highest quality textbooks and MCAT study resources. This information is intended for physicians and related personnel, who understand that medical information is often imperfect, and must be interpreted in the context of a patient's clinical data using reasonable medical judgment. We will be provided with an authorization token (please note: passwords are not shared with us) and will sync your accounts for you. Bioengineering skeletal muscle often requires customized equipment and intricate casting techniques.
An assortment of methods for the generation of bio-artificial skeletal muscle have been previously described (Table 1), with variations on aspects including chamber construction, matrix composition and ultimate tissue size generated. Even though reproducibility is improved, these casting methods and inserts demand specialized equipment, which commonly translates into an increase in cost.
Currently, various hydrogel components are routinely used, both individually and in combination, to successfully engineer muscle tissue containing striated and aligned myotubes. Although numerous methods for bioengineering skeletal muscle exist, currently no standardized culture vessel and protocol has been proposed. An adaptable chamber system was generated by using 18 mm sections of biological grade silicone tubing (outer diameter: 5 mm) which was cut in half (lengthwise) (Figure 1A). HSKM cells are larger than C2C12 myoblasts; this accounts for the lower number of human myoblasts in the hydrogel mix when compared to mouse myoblasts. Brightfield images were captured at various stages of muscle development using a Motic 3.0 MP camera and an Olympus stereo microscope (VMZ, Japan).
Muscle tissue engineering is no longer in its infancy, nor is it the prerogative of only a few laboratories.
Imaging of myoblasts functioning in a three-dimensional space is also more closely aligned to in vivo behavior.

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The work was supported by the South African National Research Foundation, South African Medical Research Council and University of KwaZulu-Natal.
It works by having smooth, continuous contracts and exhibits myogenic properties, which means it can function without nervous system input. It is under control from both the involuntary and the somatic nervous system and also consists of strong, forceful contractions. Outside the sarcolemma is the sarcoplasm, which is a modified cytoplasm that can propagate an action potential. T-tubules run deep into the cell so the action potential can propagate successfully through the muscle. This in turn pulls on the actin, which draws thin filaments to the H-zone center, which shortens the sarcomere. Myosin resets itself by binding another molecule of ATP, which is then free to bind with actin again.
Once disconnected, sarcomeres return to original width, and without the calcium ions, myosin-binding sites covered by tropomyosin will prevent contraction. After contraction a muscle becomes unresponsive for a short period of time, and is unresponsive to stimuli. During times of plenty, the body stores creatine phosphate by transferring a phosphate from an ATP molecule to a creative molecule, which is an easily reversible reaction and allows for immediate conversion into energy.
They are composed of scattered cells and an amorphous base (liquid, jelly, solid), which attaches epithelium to underlying tissues. Loose fibers have a lot of ground substance and extracellular matrix and is often found in fat and the support around organs. By purchasing your products through our website links to Amazon you help support the content development for future generations and it costs you nothing.
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One of the major hurdles when initially trying to establish in vitro tissue engineered muscle constructs is the lack of consistency across published methodology. 3D models have advantages over 2D cell cultures in mimicking in vivo conditions as they allow for the study of dimensionality, cellular architecture, cell polarity and function. Some models require post-modification with custom-made inserts that replace pins or Velcro adhesion points. In other more recently-developed models these anchorage points and the distance between them in custom-built models varied from 4 to 50 mm, according to the size of the wells and aspects required for the experiment.
In addition the system may not easily be adaptable to different well-types or tissue sizes.
Also, many are prohibitive due to the required customized equipment and intricate casting techniques.
For histochemical investigation of desmin expression and the actin cytoskeleton, muscle constructs were fixed for 2 h in the wells in paraformaldehyde (4% prepared in PBS).
After 12–15 days in culture, differentiated C2C12 myotubes showed clear formation of actin fibers (Figure 2D). Successful generation of mouse and human skeletal muscle constructs using the simple silicone chamber system.
This model is adaptable to fit into any existing culture dish or chamber used in most laboratories. Biological grade silicone is inexpensive, readily available and allows for ease of pin insertion and subsequent tissue manipulation. With this model we have described the expression of desmin, an intermediate filament that plays a key role in the integration of striated muscle morphology and function (Capetanaki et al., 2007).
The authors also thank the UKZN Microanalysis and Microscopy Unit (Pietermaritzburg) as well as Graeme Marwick (Denel Dynamics) for all their assistance.
Many myofibrils is called a myocyte (muscle cell) A muscle is simply the parallel arrangement of these myocytes.
For this reason, when humans die the myosin heads cannot break away from the myosin because no new ATP is generated, thus setting the body into rigor mortis. After death, because there is no ATP generation, the muscles set into rigor mortis, which is permanent contraction that occurs because without ATP, myosin cannot detach from actin. Connective tissue is essentially just cells along with a secreted extracellular matrix by fibroblasts in order to provide support.
Although this diversity allows for specialization according to specific research goals, lack of standardization hampers comparative efforts. Confluent myoblast monolayers cultured in a matrix-coated petri dish under differentiating conditions may also form scaffold-free 3D muscle tissue due to contractility of the differentiating fibers (Table 1C). The flexibility of the cantilever posts in the more advanced models is an improvement from the originally employed metal pins or large, fixed Velcro pads or metal mesh (Vandenburgh et al., 2008). The use of an array of posts in a wafer pattern rather than simply two adhesion points permits the formation of a sheet-like culture that allows for the investigation of muscle cell alignment. This is followed by replacement of the growth media (GM) with differentiation media (DM) to stimulate differentiation into muscle fibers. Below we describe an inexpensive, accessible hydrogel-based system that may be readily standardized, yet is easily customized to reflect desired matrix combinations and tissue size.

The tubes were fitted into each well of a 4- or 24-well-culture plate and secured within the wells using Sylgard 182 (Dow Corning Corporation, cat.
In addition, aligned myotubes expressed desmin (Figure 2E) and longitudinal sections showed evidence of organization into multinucleated myotubes (Figure 2F), which is an initial requirement for functionality.
Despite this simplicity, variations in the distance between pins, as well as cell number and matrix-cell volume can be readily achieved.
In addition, after initial use, the various components of the chamber system may also be reused following de-cellularization with ammonium hydroxide and cleaning with alcohol and sonication.
We also showed the transition of myoblasts into elongated, multi-nucleated myotubes, one of the relevant steps during myoblast differentiation. Application of a cell sheet-polymer film complex with temperature sensitivity for increased mechanical strength and cell alignment capability. Novel method for fabrication of skeletal muscle construct from the C2C12 myoblast cell line using serum-free medium AIM-V.
These muscles contain actin and myosin, which enable this type of muscle to have longer and more sustained contractions.
Tetanus occurs when more frequent than normal contractions combined with a lack of relaxation, create strong twitches, which if prolonged lead to fatigue.
The base or ground substance is essentially the glue that holds the fibers and matrix together. Differences in cell type, number and density, variability in matrix and scaffold usage as well as inconsistency in the distance between and type of adhesion posts complicates initial establishment of the technique with confidence.
If genetically modified to express recombinant protein, these constructs can be used for therapeutic protein delivery (Vandenburgh et al., 1996).
Surgical grade stainless steel pins are inserted through the silicone wall at predefined distances from each other (2). Such lack of uniformity was highlighted by a summary of the range of moulds already employed in muscle tissue engineering (Table 1). In addition, the use of appropriate pins as anchor points allows for future mechanical stimulation to investigate contractile forces and allows for the study of internal stresses during muscle differentiation in 3D cultures. This model, with standardized parameters, may be used as an optimized system for initial evaluation of factors involved in skeletal muscle generation from both primary cultured myoblasts and established cell lines. We describe an inexpensive, but readily adaptable silicone chamber system for the generation of skeletal muscle constructs that can readily be standardized and used to elucidate myoblast behavior in a three-dimensional space. While each model has specific advantages, key methodological aspects differ considerably between the various models which may potentially hamper efficient comparison. Silk sutures pinned into the Sylgard base act as handling points and mimic flexible tendons for the cylindrical myooid. Sylgard was allowed to cure for 24 h and the plates sterilized overnight under an ultraviolet light. The silicone tube is secured in place within the well with Sylgard 182 to form a chamber (3). It is also useful for investigations into genetic manipulation, drug therapy or co-culture of complimentary cell phenotypes.
Engineered tissue may further be processed for histological purposes or immunocytochemical investigation of transcription factors and expressed proteins.
Excitability and isometric contractile properties of mammalian skeletal muscle constructs engineered in virto. Matrigel, but not collagen I, maintains the differentiation capacity of muscle derived cells in vitro. Muscle generation, regeneration and adaptation can also be investigated in this model, which is more advanced than differentiated myotubes. A critical overview of the various models is required before describing our simple chamber system.
The laminin matrix employed merely forms a separating layer between the Sylgard coating and the cultured cells, while the final histology and molecular characteristics reflect that of skeletal muscle.
The hydrogel-cell suspension is pipetted into the silicone chamber around the pins (4) and the well is flooded with growth media once the gel construct has set (5). Constructs cultured for 15 days in DM were also fixed with glutaraldehyde (2%, 2 h at room temperature), dehydrated with a graded series of alcohols and embedded in Spurr's resin.
We propose that the method we describe may allow skeletal muscle research groups utilizing 2D cell culture models to move into 3D tissue models with relative ease. Further development of a tissue engineered muscle repair construct in vitro for enhanced functional recovery following implantation in vivo in a murine model of volumetric muscle loss injury. Effect of precise mechanical loading on fibroblast populated collagen lattices: morphological changes.
Finally, the matrix combinations as reported for the different models vary considerably (Table 1).
Thin sections were cut and DIC images were obtained with the 710 Zeiss confocal microscope. In the current study we describe an inexpensive, readily adaptable silicone chamber system for the generation of skeletal muscle constructs. This will be important to enable more rapid enhancement of our understanding of muscle synthesis, repair and adaptation in vitro, in a model more advanced than differentiated myotubes. The culture period to establish this model is, however, considerably longer than the previously-mentioned models.

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