Connective tissue structures of human skeletal muscle and their significance in the biomechanics of the body

By means of light optic and electron microscopy (SAM, TAM) histoconstruction of the connective tissue structures of the human skeletal muscles have been investigated and its analysis has been performed from biomechanical point of view. Fibrillar elements of the connective tissue are demonstrated to play an important role in structural adaptation of the skeletal muscle, as the organ, performing certain mechanical functions. The data obtained makes it possible to formulate the state, that the fibrillar network of the connective tissue is a polyfunctional system, that ensures integration of the structural elements of the muscle, transmission of mechanical strains, is the carcass of the organ and participates in formation of its buffer and amortizational mechanisms. The integration mechanisms of the main functional elements of the muscle belly, tendons and fascia to a great extent are of a unification character.     

Periostin regulates collagen fibrillogenesis and the biomechanical properties of connective tissues

Periostin is predominantly expressed in collagen-rich fibrous connective tissues that are subjected to constant mechanical stresses including: heart valves, tendons, perichondrium, cornea, and the periodontal ligament (PDL). Based on these data we hypothesize that periostin can regulate collagen I fibrillogenesis and thereby affect the biomechanical properties of connective tissues. Immunoprecipitation and immunogold transmission electron microscopy experiments demonstrate that periostin is capable of directly interacting with collagen I. To analyze the potential role of periostin in collagen I fibrillogenesis, gene targeted mice were generated. Transmission electron microscopy and morphometric analyses demonstrated reduced collagen fibril diameters in skin dermis of periostin knockout mice, an indication of aberrant collagen I fibrillogenesis. In addition, differential scanning calorimetry (DSC) demonstrated a lower collagen denaturing temperature in periostin knockout mice, reflecting a reduced level of collagen cross-linking. Functional biomechanical properties of periostin null skin specimens and atrioventricular (AV) valve explant experiments provided direct evidence of the role that periostin plays in regulating the viscoelastic properties of connective tissues. Collectively, these data demonstrate for the first time that periostin can regulate collagen I fibrillogenesis and thereby serves as an important mediator of the biomechanical properties of fibrous connective tissues.     

Effects of structure and strain measurement technique on the material properties of young human tendons and fascia

It is generally recognized that the organization of collagen bundles in soft tissues strongly influences their material properties. To study this, sixty failure tests were conducted on double-layered fascia lata, 'isolated' parallel-bundled tendons (gracilis and semitendinosus) and parallel-bundled bone-patellar tendon-bone units taken from about the knees of eighteen young human donors (mean age of 26 yr). Surprisingly, most material parameters for the two-layered fascia lata did not differ significantly from corresponding values for the isolated tendons and tendon-bone preparations, suggesting their longitudinal fibers predominated during loading. Differences were present however between the gracilis tendon and all other tissues for both modulus and maximum stress. The large variations in reported maximum and failure strains for tendons, fascia and other collagenous tissues prompted the other phase of the study. During 15 of the 60 failure tests, surface markers were simultaneously filmed to determine; differences between local surface strains and grip to grip values; the amount of tissue slippage and/or failure occurring in the grips; and the effect of strain measurement technique on tissue moduli and failure energy densities. Maximum local strains were found to be 25-30% of grip strains for all tissues tested. Some slippage and/or failure could be seen in all isolated tissues which were gripped directly although their maximum grip strains were similar to values for tendon-bone units. For all tissues, two to three fold differences were also found in moduli and failure energy densities between grip and midregion measurements.     

Autophagy guards tendon homeostasis

Tendons are vital collagen-dense specialized connective tissues transducing the force from skeletal muscle to the bone, thus enabling movement of the human body. Tendon cells adjust matrix turnover in response to physiological tissue loading and pathological overloading (tendinopathy). Nevertheless, the regulation of tendon matrix quality control is still poorly understood and the pathogenesis of tendinopathy is presently unsolved. Autophagy, the major mechanism of degradation and recycling of cellular components, plays a fundamental role in the homeostasis of several tissues. Here, we investigate the contribution of autophagy to human tendons’ physiology, and we provide in vivo evidence that it is an active process in human tendon tissue. We show that selective autophagy of the endoplasmic reticulum (ER-phagy), regulates the secretion of type I procollagen (PC1), the major component of tendon extracellular matrix. Pharmacological activation of autophagy by inhibition of mTOR pathway alters the ultrastructural morphology of three-dimensional tissue-engineered tendons, shifting collagen fibrils size distribution. Moreover, autophagy induction negatively affects the biomechanical properties of the tissue-engineered tendons, causing a reduction in mechanical strength under tensile force. Overall, our results provide the first evidence that autophagy regulates tendon homeostasis by controlling PC1 quality control, thus potentially playing a role in the development of injured tendons.Subject terms: Physiology, Cell biology     

Double-layered free temporal fascia flap as a two-layered tendon-gliding surface

We report the use of a two-layered free fascial flap consisting of temporoparietal and deep temporal fascia based on a single vascular pedicle, the superficial temporal artery and vein. The flap was used to reconstruct an extensive degloving injury of the dorsum of the hand, in which multiple intact extensor tendons lay fully exposed on all sides, with exposed bone beneath them. By sandwiching the tendons between the layers of vascularized fascia, gliding surfaces were provided, both superficial and deep to the exposed tendons. The single-stage reconstruction was completed with a split-thickness skin graft. The patient returned to heavy manual work within 12 weeks of injury. He obtained an excellent range of movement without the need for tenolysis.     

Fibrous framework of human skeletal muscles, fasciae and tendons

By means of scanning and transmissive electron microscopy, the construction of the fibrous framework of the human skeletal muscles, fasciae and tendons has been investigated and its morphofunctional analysis has been performed. The fibrous framework of the endomysium is presented as a complexly organized system of anastomosing fibers of the connective tissue, forming a net-like construction. The fibrous structures of the framework are united into a whole construction by connecting fibers and fibrils. Different types of structural interconnection of collagenous fibers with sarcolemma are revealed. The structure of the fibrous framework both in different muscles and within one muscle has certain peculiarities. The main constructive element of the fascial fibrous framework make large anastomosing collagenous fibers, their architectonics is stabilized by connective fibers and fibrils. The construction of the tendinous fibrous framework is characterized by a pronounced anisotropia of the largest collagenous fibers and a developed network of connective structures both on the surface and inside the collagenous fibers. Structural mechanisms, interconnecting muscles and tendons, are demonstrated. Presence of anastomoses between the fibrils in the composition of the collagenous fibers in the fascia and Achilles tendon are stated. Together with the peculiarities existing, the general principle of the structural organization of the fibrous framework of the muscle system is the net-like constructure dependent on presence of anastomoses and elements of the connective system between the fibrous structures. Depending on the organ's function, the construction of the network acquires certain specific morphological forms.     

Influence of occlusal stabilization splint on the asymmetric activity of masticatory muscles in patients with temporomandibular dysfunction

The aim of present study was to evaluate the symmetry of masticatory muscles' activity at various clenching levels in the intercuspal position in patients with functional disorders and in healthy subjects. The purpose was also to determine the effect of full-arch maxillary stabilization splint on the asymmetry of masticatory muscle activity in patients with temporomandibular dysfunction. In this study 6 TMD patients and 12 healthy subjects were investigated. Surface EMG recordings were obtained from left and right anterior temporal, left and right masseter and from the sub-mandibular group in the region of the anterior belly of the digastric muscle on the left and right side during clenching with the maximum 100% voluntary contraction (MVC) as well as during clenching at 50% and 25% of the maximum activity in the position of maximal intercuspation of teeth. In order to quantify asymmetrical masticatory muscle activity, the asymmetry index (AI) was calculated for each subject and for each muscle from the average anterior temporal, masseter and digastric potentials recorded during each test (100% MVC, 50% MVC and 25% MVC). In the group of patients EMG recordings were repeated during and after the splint therapy. The asymmetries of masticatory muscle activity was present in both groups, but in the group of TMD patients the asymmetry indices for anterior temporal muscle at 100% MVC (p = 0.049) and 50% MVC (p = 0.031) were significantly higher. Results have shown that the use of splint suppressed the asymmetry of all muscles, as during the splint therapy the asymmetry indices were lowered. After the therapy, the level of temporal muscle symmetry during submaximal clenching in the intercuspal position increased significantly (p = 0.046). This investigation points out that electromyography may be a valuable method of documenting that asymmetric activity of masticatory muscles improves after occlusal splint therapy in patients with TMD.     

Local strain measurement reveals a varied regional dependence of tensile tendon mechanics on glycosaminoglycan content

Proteoglycans (PG) and their associated glycosaminoglycan (GAG) side chains are known to play a key role in the bearing of compressive loads in cartilage and other skeletal connective tissues. In tendons and connective tissues that are primarily loaded in tension, the influence of proteoglycans on mechanical behavior is debated due to conflicting experimental evidence that alternately supports or controverts a functional role of proteoglycans in bearing tensile load. In this study we sought to better reconcile these conflicting data by investigating the possibility that GAG content is differentially related to tensile tendon mechanics depending upon the anatomical subregion one considers. To test this hypothesis, we quantified the mechanical consequences of proteoglycan disruption within specific tendon anatomical subregions using an optical–mechanical measurement approach.Achilles tendons from adult mice were treated with chondroitinase ABC to obtain two groups consisting of native tendons and GAG-depleted tendons. All the tendons were mechanically tested and imaged with high-resolution digital video in order to optically quantify tendon strains. Tendon surface strains were locally analyzed in three main subregions: the central midsubstance, and the proximal and distal midsubstance near the muscle and bone insertions, respectively. Upon GAG digestion, the tendon midsubstance softened appreciably near the bone insertion, while elastic modulus in the central and proximal thirds was unchanged. Thus the contribution of PGs to tensile tendon mechanics is not straightforward and points to a heterogeneous and complex structure–function relationship in tendon. This study further highlights the importance of performing local strain analysis with regard to tensile tendon mechanics.     

Surgical anatomy and blood supply of the fascial layers of the temporal region

In 15 fresh cadavers (30 sides), we studied the two layers of fascia in the temporal region, with particular regard to their blood supply and to their usefulness--together or separately--as microvascular free-tissue autografts. The superficial temporal fascia (temporoparietal fascia, epicranial aponeurosis) lies immediately deep to the hair follicles. It is part of the subcutaneous musculoaponeurotic system and is continuous in all directions with other structures belonging to that layer--including the galea above and the SMAS layer of the face below. The deep temporal fascia (temporalis fascia, investing fascia of temporalis) is separated from the superficial fascia by an avascular plane of loose areolar tissue. It completely invests the superficial aspect of the temporalis muscle down to (but not beyond) the zygomatic arch. It is firmly attached to periosteum all around the margin of the muscles. Below it is attached to the upper border of the zygomatic arch. We found the deep temporal fascia to be supplied solely by the middle temporal artery, a constant branch of the superficial temporal. The middle temporal artery arises 1 to 3 cm below the upper border of the zygomatic arch, runs always superficial to the arch, and enters the deep temporal fascia immediately above that layer's attachment to the zygomatic arch. If the middle temporal vessels are protected, the two layers of temporal fascia can be raised together as a fully vascularized tissue island. This island can be fashioned as a bilobed or a double-layered flap, depending on the manner of dissection. The potential surgical usefulness of these findings is discussed.     

Muscle-driven finite element simulation of human foot movements

This paper describes a finite element scheme for realistic muscle-driven simulation of human foot movements. The scheme is used to simulate human ankle plantar flexion. A three-dimensional anatomically detailed finite element model of human foot and lower leg is developed and the idea of generating natural foot movement based entirely on the contraction of the plantar flexor muscles is used. The bones, ligaments, articular cartilage, muscles, tendons, as well as the rest soft tissues of human foot and lower leg are included in the model. A realistic three-dimensional continuum constitutive model that describes the biomechanical behaviour of muscles and tendons is used. Both the active and passive properties of muscle tissue are accounted for. The materials for bones and ligaments are considered as homogeneous, isotropic and linearly elastic, whereas the articular cartilage and the rest soft tissues (mainly fat) are defined as hyperelastic materials. The model is used to estimate muscle tissue deformations as well as stresses and strains that develop in the lower leg muscles during plantar flexion of the ankle. Stresses and strains that develop in Achilles tendon during such a movement are also investigated.     

Muscle-driven finite element simulation of human foot movements

This paper describes a finite element scheme for realistic muscle-driven simulation of human foot movements. The scheme is used to simulate human ankle plantar flexion. A three-dimensional anatomically detailed finite element model of human foot and lower leg is developed and the idea of generating natural foot movement based entirely on the contraction of the plantar flexor muscles is used. The bones, ligaments, articular cartilage, muscles, tendons, as well as the rest soft tissues of human foot and lower leg are included in the model. A realistic three-dimensional continuum constitutive model that describes the biomechanical behaviour of muscles and tendons is used. Both the active and passive properties of muscle tissue are accounted for. The materials for bones and ligaments are considered as homogeneous, isotropic and linearly elastic, whereas the articular cartilage and the rest soft tissues (mainly fat) are defined as hyperelastic materials. The model is used to estimate muscle tissue deformations as well as stresses and strains that develop in the lower leg muscles during plantar flexion of the ankle. Stresses and strains that develop in Achilles tendon during such a movement are also investigated.     

The effect of sterilization on mechanical properties of soft tissue allografts

One major concern regarding soft tissue allograft use in surgical procedures is the risk of disease transmission. Current techniques of tissue sterilization, such as irradiation have been shown to adversely affect the mechanical properties of soft tissues. Grafts processed using Biocleanse processing (a proprietary technique developed by Regeneration Technologies to sterilize human tissues) will have better biomechanical characteristics than tissues that have been irradiated. Fifteen pairs of cadaveric Achilles tendon allografts were obtained and separated into three groups of 10 each. Three treatment groups were: Biocleanse, Irradiated, and Control (untreated). Each specimen was tested to determine the biomechanical properties of the tissue. Specimens were cyclically preloaded and then loaded to failure in tension. During testing, load, displacement, and optical strain data were captured. Following testing, the cross sectional area of the tendons was determined. Tendons in the control group were found to have a higher extrinsic stiffness (slope of the load–deformation curve, p = .005), have a higher ultimate stress (force/cross sectional area, p = .006) and higher ultimate failure load (p = .003) than irradiated grafts. Biocleanse grafts were also found to be stiffer than irradiated grafts (p = .014) yet were not found to be statistically different from either irradiated or non-irradiated grafts in terms of load to failure. Biocleanse processing seems to be a viable alternative to irradiation for Achilles tendon allografts sterilization in terms of their biomechanical properties.     

Muscle performance during maximal isometric and dynamic contractions is influenced by the stiffness of the tendinous structures.

Contractile force is transmitted to the skeleton through tendons and aponeuroses, and, although it is appreciated that the mechanocharacteristics of these tissues play an important role for movement performance with respect to energy storage, the association between tendon mechanical properties and the contractile muscle output during high-force movement tasks remains elusive. The purpose of the study was to investigate the relation between the mechanical properties of the connective tissue and muscle performance in maximal isometric and dynamic muscle actions. Sixteen trained men participated in the study. The mechanical properties of the vastus lateralis tendon-aponeurosis complex were assessed by ultrasonography. Maximal isometric knee extensor force and rate of torque development (RTD) were determined. Dynamic performance was assessed by maximal squat jumps and countermovement jumps on a force plate. From the vertical ground reaction force, maximal jump height, jump power, and force-/velocity-related determinants of jump performance were obtained. RTD was positively related to the stiffness of the tendinous structures (r = 0.55, P < 0.05), indicating that tendon mechanical properties may account for up to 30% of the variance in RTD. A correlation was observed between stiffness and maximal jump height in squat jumps and countermovement jumps (r = 0.64, P < 0.05 and r = 0.55, P < 0.05). Power, force, and velocity parameters obtained during the jumps were significantly correlated to tendon stiffness. These data indicate that muscle output in high-force isometric and dynamic muscle actions is positively related to the stiffness of the tendinous structures, possibly by means of a more effective force transmission from the contractile elements to the bone.     

Three-dimensional dynamical capabilities of the human masticatory muscles

Many habitual human jaw movements are non-symmetrical. Generally, it is observed that when the lower incisors move to one side the contralateral condyle moves forwards onto the articular eminence, whereas the ipsilateral condyle stays in the mandibular fossa, moving slightly to the ipsilateral side. These jaw movements are the result of contractions of active masticatory muscles and guided by the temporomandibular joints, their ligaments and passive elastic properties of the muscles. It is not known whether the movements are primarily dependent on passive guidance, active muscle control or both. Therefore, the objective of this study was to analyse the interplay between these factors during non-symmetrical jaw movements. A six-degrees-of-freedom dynamical biomechanical model of the human masticatory system was used. The movements were not restricted to a priori defined joint axes. Jaw movement simulations were performed by unilateral activity of the muscles. The ligaments or the passive elastic properties of the muscles could be removed during these simulations. Laterodeviations conform to naturally observed ones could be generated by unilateral muscle contractions. The movement of the lower incisors was hardly affected by the absence of passive elastic muscle properties or temporomandibular ligaments. The latter, however, influenced the movement of the condyles. The movements could be understood by analysing the combination of forces and torques with respect to the centre of gravity of the lower jaw. In addition, the loading of the condyles appeared to be an important determinant for the movement. This analysis emphasizes that the movements of the jaw are primarily dependent on the orientation of the contributing muscles with respect to this centre of gravity and not on the temporomandibular ligaments or passive elastic muscle properties.     

Skeletal muscle-smooth muscle interaction: An unusual myoelastic system

The serratus superficailis metapatagialis (SSM) of pigeons is a skeletal muscle with unusual properties. It lies between the ribs and the trailing edge of the wing, where it is attached to the skin by a system of smooth muscles having elastic tendons. Wing movements during flight induce marked changes in this muscle's length. The SSM inserts onto the deep fascia, and at its termination the skeletal muscle contains large numbers of microtubules. Many myofibrils attach to leptomeric organelles, which then attach to the terminal end of the skeletal muscle fiber. The deep fascia next connects to the dermis of the skin by bundles of smooth muscles that have elastic tendons at both ends. This system allows large movements of the muscle while preventing its fibers from overstretching. The movements and presumed forces acting at this muscle make the presence of sensory receptors such as muscle spindles unlikely. Spindles are absent in this muscle.     

Stress analysis of the standing foot following surgical plantar fascia release

Plantar fascia release is a surgical alternative for patients who suffer chronic heel pain due to plantar fasciitis and are unaffected by conservative treatment. A computational (finite element) model for analysis of the structural behavior of the human foot during standing was utilized to investigate the biomechanical effects of releasing the plantar fascia. The model integrates a system of five planar structures in the directions of the foot rays. It was built according to accurate geometric data of MRI, and includes linear and non-linear elements that represent bony, cartilaginous, ligamentous and fatty tissues. The model was successfully validated by comparing its resultant ground reactions with foot-ground pressure measurements and its predicted displacements with those observed in radiological tests. Simulation of plantar fascia release (partial or total) was accomplished by gradually removing parts of the fascia in the model. The results showed that total fascia release causes extensive arch deformation during standing, which is greater than normal deformation by more than 2.5mm. Tension stresses carried by the long plantar ligaments increased significantly, and may exceed the normal average stress by more than 200%. Since the contribution of the plantar fascia to the foot's load-bearing ability is of major importance, its release must be very carefully considered, and the present model may be used to help surgeons decide upon the desired degree of release.     

Stomach stress and strain depend on location, direction and the layered structure

The stomach is as other parts of the gastrointestinal tract functionally subjected to dimensional change. Hence, the biomechanical properties are of functional importance. Our group has previously demonstrated that the stress–strain properties of the rat and rabbit stomach wall were species-, location- and direction-dependent. We further wanted to study the anisotropic biomechanical properties of the stomach wall in pigs. Furthermore, we made an in-depth biomechanical test on the layered wall of the stomach in different regions. Two stomach strips were cut both in longitudinal direction (parallel with the greater curvature) and circumferential direction (perpendicular to the greater curvature) from the gastric fundus, corpus and antrum. One strip was used for the non-separated (intact) wall test and the other one was separated for the test on the mucosa–submucosa and muscle layers individually. The length, thickness and width of each strip were measured from digital images. The uni-axial stress and strain were computed from the force generation and the tissue strip deformation during stretching. The muscle layer was the thickest in the antrum whereas the mucosal–submucosal layer was the thickest in the corpus of the stomach (P<0.01). The strips from the corpus were stiffest among the three regions in both longitudinal and circumferential directions (P<0.001). The longitudinal strips was stiffer than the circumferential strips in all three regions (P<0.001) and the mucosa–submucosa strips was stiffer than the intact wall and the muscle layer in both directions for the fundus and the corpus (P<0.001). The constant a of the intact wall and mucosa–submucosa layer was in both directions linearly associated with the mucosa–submucosa thickness. In conclusion, the uni-axial stress–strain curves of pig stomach were location-, direction- and layer-dependent. The stiffer wall in the corpus is likely due to its thicker mucosa, i.e., the stiffness of the mucosa–submucosa layer seems can explain the intact wall stiffness. Since the structure and function of the pig stomach are similar to the human stomach, we believe that the data obtained from this study can be extended to humans. Detailed biomechanical mapping of the stomach will likely help us to understand physiological functions of the different parts of the human stomach, such as gastric accommodation and mechanosensation.     

Invited Review: Role of mechanophysiology in aging of ECM: effects of changes in mechanochemical transduction.

Mechanical forces play a role in the development and evolution of extracellular matrices (ECMs) found in connective tissue. Gravitational forces acting on mammalian tissues increase the net muscle forces required for movement of vertebrates. As body mass increases during development, musculoskeletal tissues and other ECMs are able to adapt their size to meet the increased mechanical requirements. However, the control mechanisms that allow for rapid growth in tissue size during development are altered during maturation and aging. The purpose of this mini-review is to examine the relationship between mechanical loading and cellular events that are associated with downregulation of mechanochemical transduction, which appears to contribute to aging of connective tissue. These changes result from decreases in growth factor and hormone levels, as well as decreased activation of the phosphorelay system that controls cell division, gene expression, and protein synthesis. Studies pertaining to the interactions among mechanical forces, growth factors, hormones, and their receptors will better define the relationship between mechanochemical transduction processes and cellular behavior in aging tissues.     

Soft-tissue masses of the temporal fossa

An unusual problem, a soft-tissue mass of the left temporal fossa, is described. Differential diagnoses and pertinent studies for identifying the mass are reviewed. Our patient had a temporalis fascia defect with herniation of the temporal muscle causing his mass and associated symptoms. Surgical exploration, necessary for diagnosis, afforded us the opportunity for definitive treatment.