Intramuscular Connective Tissue Research Articles

Extramuscular myofascial force transmission within the rat anterior tibial compartment: proximo-distal differences in muscle force.

Intramuscular connective tissues are continuous to extramuscular connective tissues. If force is transmitted there, differences should be present between force at proximal and distal attachments of muscles. Extensor digitorum longus (EDL), tibialis anterior (TA), and extensor hallucis longus muscles (EHL) were excited simultaneously and maximally. Only EDL length was changed, exclusively by moving the position of its proximal tendon. Distal force exerted by TA + EHL complex was not affected significantly. Proximal and distal EDL isometric force were not equal for most EDL lengths: Fprox - Fdist ranged from 0 to approximately +22.7% of Fprox at higher lengths and from 0 to approximately -24.5% at the lowest lengths. It is concluded that extramuscular connections transmit force from muscle. Significant proximo-distal differences of EDL force remained after repeated measurements, regardless of length order, although their length dependence was altered. Measurements of both proximal and distal EDL force were highly reproducible, if EDL did not attain higher lengths than target length. After being active at high lengths, proximal and distal length-force curves were altered at low lengths but not for the highest length range. Extensor digitorum longus length-active force hysteresis was present for proximal as well as distal EDL measurements with increasing and decreasing isometric length order. Further isolating EDL removed the proximo-distal difference for active EDL force. However a decreased difference for passive EDL force remained, which was ascribed to remaining extramuscular connective tissue linkages. It is concluded that extramuscular myofascial force transmission is an important feature of muscle that is not isolated from its surrounding tissues.

The influence of marinating with weak organic acids and salts on the intramuscular connective tissue and sensory properties of beef

Longissimus dorsi muscles obtained from middle-aged bovine carcasses were used in this project. Initially, 2%, 4%, and 6% NaCl, 50 mM, 100 mM, and 150 mM CaCl2, and 0.5%, 1.0%, and 1.5% lactic and citric acids solutions were prepared and then treated 1:1 with (w/v) meat pieces in polyethylene bags at 4 °C for 24 h for marinating. Some physical and organoleptical changes in the meat treated with the above-mentioned solutions were determined. According to control samples, the highest lightness (L*) value was found in meat marinated with CaCl2 while the lowest L* value was in meat marinated with lactic acid. No positive correlation was found between penetrometer values and tenderness except in meat treated with lactic acid. Panel tests showed that all procedures increased tenderness and juiciness to a certain extent. All concentrations of NaCl and CaCl2 had a positive effect on taste and aroma of meat while acid concentrations up to 1.0% were acceptable. Increased water binding capacity was found in all samples. Electrophoresis of connective tissue proteins obtained from the same muscle showed that NaCl and CaCl2 did not cause any change in the structure, but that both lactic and citric acid increased tissue breakdown.

The extracellular matrix of muscle--implications for manipulation of the craniofacial musculature.

Successful adaptation of craniofacial skeletal muscle is dependent upon the connective tissue component of the muscle. This is exemplified by procedures such as distraction histo/osteogenesis. The mechanisms underlying remodelling of intramuscular connective tissue are complex and multifactorial and involve extracellular matrix (ECM) molecules, receptors for the ECM (integrins) and enzymes that remodel the ECM (MMPs). This review discusses the current state of knowledge and clinical implications of connective tissue biology as applied to craniofacial skeletal muscle.

Influence of weak organic acids and salts on the denaturation characteristics of intramuscular connective tissue. A differential scanning calorimetry study

Intramuscular connective tissue obtained from Longissimus dorsi muscle of a 4-year-old beef carcass was treated with NaCl solutions of 2, 4, and 6% (w/v), and CaCl 2 solutions of 50, 100, and 150 mM, and citric and lactic acid solutions of 0.5, 1.0, and 1.5% for three different marinating periods (24, 46, and 72 h). Changes in denaturation characteristics were investigated using differential scanning calorimetry and it was found that the denaturation onset temperature ( T o) and the denaturation peak temperature ( T p) increased as the NaCl concentration increased but decreased as the CaCl 2 concentration increased, irrespective of marinating time. Lactic and citric acid decreased T o to about 39°C, from over 60°C breaking the structure of fibrils.

Tenderization of Heated Sliced Beef by Succinylated Glycerol Monostearate, a Novel Meat Tenderizer

ABSTRACT Succinylated glycerol monostearate (SGMS) was most effective in reducing shear‐force value when used as a surfactant to tenderize meat using sheep casings as a model of intramuscular connective tissue. Roasted beef slices that had been treated with 2% granular SGMS showed a marked decrease in toughness due to the penetration of SGMS along the perimysium during heating. Collagen fibrils in the perimysium were transformed into a sheet‐like structure that seemed very likely to be a complex of SGMS and thermally denatured collagen. These structural changes could account for the tenderness of the roasted beef slices.

Exercise With Lack of Motor Control: Lessons From Studies on Spinal Cord Patients

Study of cardiovascular and metabolic regulation can be performed using electrical stimulation on paralyzed muscle. Simultaneously, electrical induced training of spinal cord injured individuals can beneficially improve their aerobic capacity and thus lower risk for inactivity associated diseases. The normal correlation between oxygen uptake and cardiac output is maintained during electrical involuntary exercise, and venous blood return is most likely a candidate for this relation during exercise. With regards to metabolism, substrate mobilization of glucose and fatty acids is impaired in the absence of motor control and neural feedback from muscle, during electrical exercise in spinal cord injured muscle glycogenolysis dominates. Electrical stimulation ergometer training in spinal cord injured rapidly increases glycolytic and oxidative enzymes, and with a more delayed response results in a fiber-type transformation from dominating myosin heavy chain isoform IIx to IIa and ultimately towards type I. Intramuscular connective tissue, collagen type IV content did not change in response to training, whereas degradation rate rose, indicating an increased turnover rate in connective tissue with training.

Inferences on force transmission from muscle fiber architecture of the canine diaphragm.

Functional properties of the diaphragm are mediated by muscle structure. Modeling of force transmission necessitates a precise knowledge of muscle fiber architecture. Because the diaphragm experiences loads both along and transverse to the long axes of its muscle fibers in vivo, the mechanism of force transmission may be more complex than in other skeletal muscles that are loaded uniaxially along the muscle fibers. Using a combination of fiber microdissections and histological and morphological methods, we determined regional muscle fiber architecture and measured the shape of the cell membrane of single fibers isolated from diaphragm muscles from 11 mongrel dogs. We found that muscle fibers were either spanning fibers (SPF), running uninterrupted between central tendon (CT) and chest wall (CW), or were non-spanning fibers (NSF) that ended within the muscle fascicle. NSF accounted for the majority of fibers in the midcostal, dorsal costal, and lateral crural regions but were only 25-41% of fibers in the sternal region. In the midcostal and dorsal costal regions, only approximately 1% of the NSF terminated within the fascicle at both ends; the lateral crural region contained no such fibers. We measured fiber length, tapered length, fiber diameters along fiber length, and the taper angle for 271 fibers. The lateral crural region had the longest mean length of SPF, which is equivalent to the mean muscle length, followed by the costal and sternal regions. For the midcostal and crural regions, the percentage of tapered length of NSF was 45.9 +/- 5.3 and 40.6 +/- 7.5, respectively. The taper angle was approximately 0.15 degrees for both, and, therefore, the shear component of force was approximately 380 times greater than the tensile component. When the diaphragm is submaximally activated, as during normal breathing and maximal inspiratory efforts, muscle forces could be transmitted to the cell membrane and to the extracellular intramuscular connective tissue by shear linkage, presumably via structural transmembrane proteins.

Myofascial Force Transmission Causes Interaction between Adjacent Muscles and Connective Tissue: Effects of Blunt Dissection and Compartmental Fasciotomy on Length Force Characteristics of Rat Extensor Digitorum Longus Muscle

Muscles within the anterior tibial compartment (extensor digitorum longus: EDL, tibialis anterior: TA, and extensor hallucis longus muscles: EHL) and within the peroneal compartment were excited simultaneously and maximally. The ankle joint was fixed kept at 90°. For EDL length force characteristics were determined. This was performed first with the anterior tibial compartment intact (1), and subsequently after: (2) blunt dissection of the anterior and lateral interface of EDL and TA. (3) Full longitudinal lateral fasciotomy of the anterior tibial compartment. (4) Full removal of TA and EHL muscles. Length-force characteristics were changed significantly by these interventions. Blunt dissection caused a force decrease of approximately 10% at all lengths, i.e., without changing EDL optimum or active slack lengths. This indicates that intermuscular connective tissue mediates significant interactions between adjacent muscles. Indications of its relatively stiff mechanical properties were found both in the physiological part of the present study, as well as the anatomical survey of connective tissue. Full lateral compartmental fasciotomy increased optimum length and decreased active slack length, leading to an increase of length range (by ˜47%), while decreasing optimal force. As a consequence an increase in force for the lower length range was found. Such changes of length force characteristics are compatible with an increased distribution of fiber mean sarcomere length. On the basis of these results, it is concluded that extramuscular connective tissue has a sufficiently stiff connection to intramuscular connective tissue to be able to play a role in force transmission. Therefore, in addition to intramuscular myofascial force transmission, extramuscular force transmission has to be considered within intact compartments of limbs. A survey of connective tissue structures within the compartment indicated sheet-like neuro-vascular tracts to be major components of extramuscular connective tissue with connections to intramuscular connective tissue stroma. Removal of TA and EHL yielded yet another decrease of force (mean for optimal force ˜10%). No significant changes of optimum and active slack lengths could be shown in this case. It is concluded that myofascial force transmission should be taken into account when considering muscular function and its coordination, and in clinical decisions regarding fasciotomy and repetitive strain injury.

SHEAR FORCE ANALYSIS AS AN INDICATOR OF CHANGES IN MECHANICAL STRENGTH OF ISOLATED BEEF INTRAMUSCULAR CONNECTIVE TISSUE DURING AGING

Intramuscular connective tissue (IMCT) was isolated from bovine semitendinosus (ST) muscles aged 3, 5, 7, 10, 14, 21, 28, and 35 days postmortem. Shear force values were collected from raw ST muscle and IMCT preparations to determine changes in mechanical strength (resistance to shearing) of the connective tissue fibers during postmortem aging. Statistical analysis indicated significant differences in mean shear force values of isolated IMCT preparations between aging periods (P 0.05) was observed. Other investigators have incorporated the same procedure and reported decreasing shear force values with increased aging times for IMCT preparations and raw ST samples. Our results were not in agreement with these findings.

Immunochemical and Immunohistochemical Identification of a Minor Collagen in Raw Muscles of Decapod Mollusks

ABSTRACT: Approximately 10% of total muscle collagen in 6 decapod molluskan species was recovered as minor collagenous fractions by limited pepsin digestion and differential salt precipitation. The main alpha components (α1) in these fractions showed similar peptide maps of V-8 protease and lysyl endopeptidase digests to each other and had reactivity to the antiserum against the al component of the minor collagen (named Type SQ-II) from Todarodes pacificus muscle in immunoblot analysis. These results suggested that the minor molecular species of collagen corresponding to Type SQ-II of Todarodes pacificus was widely distributed in the mantle muscle of decapod mollusks. In addition, immunohistochemical experiments revealed that Type SQ-II collagen distributed mainly in intramuscular thin connective tissue (endomysium), suggesting the functional importance of this molecule to the development of meat texture of decapod mollusks.

Molecular species of collagen in the intramuscular connective tissues of the marine crab, Scylla serrata

Type V like collagens are widely distributed in marine invertebrates, particularly crustaceans and molluscs. We have been investigating the nature of collagens in the muscular tissues of crustaceans. The presence of type V like homotrimeric collagen in prawn muscle was noted before. We report here a comparative analysis of collagens purified from the pepsin digest of abdominal and pereiopod muscle tissues of the crab, Scylla serrata. The major collagen in either muscle precipitated at 1.2 M NaCl at acid pH, suggestive of a type V like property. The homotrimeric collagen was then purified to near homogeneity by precipitation with 20% ammonium sulphate. Solubility characteristics and biochemical studies indicated the leg muscle collagens to be highly crosslinked and stabilised by more bound carbohydrates, as compared to the abdominal muscle collagen. Analysis of amino acid composition revealed a close similarity to known type V collagens and the leg muscle collagen was characterised by more lysine hydroxylation and slightly reduced glycine content. The leg muscle collagen had a higher denaturation temperature and intrinsic viscosity than the abdominal muscle collagen. Our results confirm the similarity of major crustacean muscle collagens to vertebrate type V collagen. Further, the relative complexity of leg muscle collagen, unlike the abdominal muscle collagen, correlates to the specific functional requirements, where the former is involved in locomotion and preying and the latter in normal growth and development.

NMR and DSC studies during thermal denaturation of collagen

Epimysial and intramuscular connective tissues from calf and cow muscle were studied by NMR and DSC. Water proton NMR transverse relaxation times ( T 2) were measured at 10°C for both native and thermally-denatured at 90°C for 30–360 min. DSC measurements were used to determine the temperature and the variation enthalpy of sol→gel transition. According to the heating time, significant differences were observed between tissues. NMR discriminated the type of collagen whereas DSC distinguished the age of tissue. Differences were related to the degree of protein hydration, emphasising the complementary information from these two analytical tools.

Scanning Electron Microscopic Study of the Neuromuscular Junctions of the Cricothyroid and Thyroarytenoid Muscles in Rats

Neuromuscular junctions were observed in the cricothyroid (CT) and thyroarytenoid (TA) muscles of adult rats by scanning electron microscopy after removing the intramuscular connective tissue components using the HCl hydrolysis method. Morphologically, the junctions were classified into three types in the CT muscle and two types in the TA muscle, based on the structural characteristics of the subneural apparatuses, including junctional folds. In the CT muscle, type 1 junctions (32%) consisted of more than 15 cup-like depressions with slit-like junctional folds. Type 2 junctions (20%) were characterized by approximately 10 cup-like depressions with a small number of pit- or slit-like junctional folds. Type 3 junctions (48%) had irregular labyrinthine gutters with slit-like junctional folds. In the TA muscle, type 1 (82%) and 2 (18%) junctions had similar structures to type 1 and 2 junctions in the CT muscle, respectively. Histochemical studies using myosin adenosine triphosphatase staining showed that both CT and TA muscles predominantly consisted of type II muscle fibers (78% and 82%, respectively), and that the diameter of type II fibers was larger than that of type I fibers. These findings suggest that the type 2 junction belongs to type I muscle fibers, while both type 1 and type 3 junctions belong to type II fibers, and that the type 3 junction is a structural variation of the type 1 junction. The significance of the structural differences of the subneural apparatuses in the intrinsic laryngeal muscles is discussed briefly.

Morphological Characteristics of Neuromuscular Junctions of the Opossum (Didelphis albiventris) Extraocular Muscles: A Scanning-Electron-Microscopic Study

Three types of neuromuscular junctions were described in the extraocular muscles of the opossum. The present study demonstrates the three-dimensional characteristics of these neuromuscular junctions after HCl connective tissue digestion. Adult opossum of both sexes were used and the neuromuscular junctions of the extraocular muscles were examined after removal of the intramuscular connective tissue and basal layer. This material was examined with a scanning electron microscope. Two types of ‘en plaque’ neuromuscular junction were described: the continuous type revealed elongated and branched primary synaptic grooves separated from each other by sarcolemma protuberances with different sizes, and the discontinuous or punctiform type which presents very shallow and discontinuous grooves when compared with the former. The multiple neuromuscular junctions were observed as two or three junctions associated with the same muscular fiber. The multiple junctions were present in thin fibers (around 11 μm caliber); the en plaque junctions were associated with large diameter fibers (around 21 μm). This study confirms and reveals the detailed morphological characteristics of the three neuromuscular junction types previously described by transmission electron microscope in the extraocular muscles of opossum.

Changes in intramuscular connective tissue and collagen solubility of bovine m. semitendinosus during retorting

Vacuum-packed slices of bovine m. semitendinosus (ST) muscle were retorted to internal temperatures of 50, 60, 70, 80, 90, 100 and 121°C. Changes in the microstructure of the endomysium and perimysium were evaluated using a scanning electron microscope (SEM). Changes in solubility of intramuscular collagen were also estimated. In samples heated to 50°C changes in intramuscular connective tissue were not seen. At 60°C granulation of the perimysium and sarcolemma was seen, and on longitudinal sections, as well as granulation, fibrous structures of perimysium were observed. At 70°C increased granulation was observed. In the range 80–121°C progressive compression of the meat structure was seen in the transverse sections. The percentage of soluble collagen in the meat changed little up to an internal temperature of 60°C, but at 70°C it almost doubled, and in the range 80–121°C drastically decreased.

Effect of rate of distraction on loss of range of joint movement, muscle stiffness, and intramuscular connective tissue content during surgical limb‐lengthening: A study in the rabbit

Surgical lengthening of limbs often results in loss of range of joint movement and this has been shown to be associated with an increase in passive tension and an increase in collagen content of the muscles. In this study, we have investigated the length/tension properties and the connective tissue component of muscle distracted at three different rates in order to determine whether low rates of distraction would enable the connective tissue component, as well as the contractile component (number of serial sarcomeres), to adapt more completely to the increased functional length of the muscle and thus lead to improved range of joint movement. It was found that loss of range of movement varied with rate of distraction. At the low rate, there was no change in the passive tension or collagen content compared to muscles from sham-operated animals, and range of movement was significantly greater than at the other rates. At the medium rate, although the muscles showed good adaptation in terms of serial sarcomere number, passive tension and collagen content was increased and range of movement reduced, indicating that changes in the connective tissue component are important factors in loss of joint movement. In the case of muscle distracted at a high rate, failure of the muscle fibres to add on sufficient sarcomeres, combined with changes in the connective tissue, resulted in almost total loss of joint movement. Anat Rec 255:78–83, 1999. © 1999 Wiley-Liss, Inc.

Effect of rate of distraction on loss of range of joint movement, muscle stiffness, and intramuscular connective tissue content during surgical limb-lengthening: a study in the rabbit.

Surgical lengthening of limbs often results in loss of range of joint movement and this has been shown to be associated with an increase in passive tension and an increase in collagen content of the muscles. In this study, we have investigated the length/tension properties and the connective tissue component of muscle distracted at three different rates in order to determine whether low rates of distraction would enable the connective tissue component, as well as the contractile component (number of serial sarcomeres), to adapt more completely to the increased functional length of the muscle and thus lead to improved range of joint movement. It was found that loss of range of movement varied with rate of distraction. At the low rate, there was no change in the passive tension or collagen content compared to muscles from sham-operated animals, and range of movement was significantly greater than at the other rates. At the medium rate, although the muscles showed good adaptation in terms of serial sarcomere number, passive tension and collagen content was increased and range of movement reduced, indicating that changes in the connective tissue component are important factors in loss of joint movement. In the case of muscle distracted at a high rate, failure of the muscle fibres to add on sufficient sarcomeres, combined with changes in the connective tissue, resulted in almost total loss of joint movement.

Transmission of forces within mammalian skeletal muscles

Most models of in vivo musculoskeletal function fail to take into account the diversity of force trajectories defined by muscle fiber architecture. It has been shown for many muscles, across species, that muscle fibers commonly end within muscle fascicles without reaching a myotendinous junction, and that many of these fibers show a progressive decline in cross-sectional area along the length of the muscle. The significance of these anatomical observations is that the tapering would seem to preclude forces generated at the largest cross-sectional area of the fibers being transmitted to the sarcomeres toward the ends of the tapered fiber. If all of the forces are transmitted via the sarcomeres arranged in series, those few sarcomeres at the smaller ends of the fibers must tolerate the stress exerted by the more numerous sarcomeres arranged in parallel at the portions of the fiber with larger cross-sectional areas. A logical alternative would be for forces to be transmitted laterally along the length of a fiber to the cell membrane and the extracellular matrix. Such a structural arrangement would permit an alternative force transmission vector and minimize the necessity for a precise level of force to be generated along the entire length of a fiber. There are cytoarchitectural and biochemical data demonstrating the presence of a subcellular network which is appropriately located to transmit forces from the active intracellular contractile elements to the extracellular intramuscular connective tissues. However, to fully comprehend how forces are transmitted from individual cross bridges to the tendon, it will be necessary to understand the interactions of all of the components of the muscle–tendon complex from the molecular to the multicellular level. It is insufficient to know the physiology of the individual components in a restricted experimental paradigm and assume that these conditions account for the functional characteristics in vivo. Thus, the challenge is to understand how the sarcomeres and all of the associated structures transmit the forces of the whole muscle to its attachments.

Muscle as a collagen fiber reinforced composite: a review of force transmission in muscle and whole limb

Even though no direct physiologic evidence proving that myo-tendinous junctions at the end of myofibers are sites of force transmission is available, these locations are accepted to support this function, because its specialized morphology resembles that of load-bearing membranes in structure and location: Its design is fit for force transmission of force exerted by myofibers to tendinous fibrous material. Shearing of the interface between these structures is thought to be stronger than direct tensile transmission. On the basis of morphological studies of ‘in-series fibered muscle’ and biomechanical modeling it has been argued previously that force could also be transmitted laterally from the tapered ends of myofibers onto in series myofiber via the intramuscular connective tissue component. Shearing of the interfaces between myofibers is hypothesized to be the mechanisms of transmission. The interfaces are made up of basal membranes of both myofibers and their common endomysium. The issue of lateral force transmission from myofibers has not been addressed for whole muscle, in which myofibers are attached at both ends to tendinous aponeuroses, nor is any direct experimental evidence available about possible functional importance of this phenomenon in whole muscle. The primary objective of this presentation is to review available literature on myo-tendinous and myo-fascial force transmission, present evidence from experiments involving tenotomy, fasciatomy and aponeurotomy regarding its importance and consider implications for our thinking about muscle(s) and movement.

Effects of high pressure treatments on intramuscular connective tissue

The changes in the morphological structure and proteoglycan (PGs) extractability of the intramuscular connective tissues from bovine skeletal muscle exposed to high pressure (100∼400 MPa) were investigated and compared to those in muscle during ageing. SDS-PAGE profiles of the PGs were also studied. During ageing, structural changes in the endomysium and perimysium occured, and disruption of the honeycomb-like structure of endomysium was observed. In the pressurized muscle, deformation of the honeycomb-like structure of endomysium was accelerated with increase of pressure applied to the muscle, and disruption of endomysium was observed in several places in the muscle pressurized at 400 MPa. Changes in the extractability of PGs were not observed in the pressurized muscle, whereas the extractability of PGs decreased with ageing. Changes in the density of the main component in the PGs with molecular mass of more than 3000 kDa on the SDS-PAGE were not observed in the PGs from the pressurized muscle, whereas the density of this band gradually decreased during the ageing. The decrease in the amounts of PGs extracted from the aged muscle seems to be due to degradation of the PGs into small dialysable molecules. From the results, it seems that high pressure treatment has some different effects on intramuscular connective tissue to ageing. ©