Evidence from surface microscopy for recognition of fleshy and tendinous muscle insertion in extant vertebrate femora: implications for muscle reconstruction in fossils

Recognition of muscle attachment sites and their modification has been an important tool in anthropologic and paleontologic research, but has been compromised by limited ability to recognise sites of tendinous attachments. We investigated bone–tendon (three sites) and bone–muscle (six sites) interfaces in six pairs of femora across a broad taxonomic spectrum of higher amniote archosaurs (both recent and fossil) by epi-illumination microscopy. Direct fleshy and indirect tendinous muscle attachments were identified by dissection of fresh specimens and examination of fossils and the surface microscopic changes identified at those locations. Examination revealed bone modifications specific to each type of muscle insertion, allowing them to be identified and distinguished. Application of a surface microscopy technique not only permits more confident localisation of tendinous attachments, but for the first time allows recognition of sites of direct fleshy muscle attachments – in a reproducible manner across phylogenetic lines.     

Metamorphosis of cells and intercellular substances in skeletogenesis

We investigated the attachment of tendons to the skeleton (tubercula humeri, patella) in regard to the chondrogenesis and osteogenesis in these areas. Then we compared our findings with those in other sites of skeletogenesis. A metamorphosis, transformation, of structure and nature of cells and intercellular susbstances in correlation with another by feedback characterized the transition from perichondrium and periost or tendon to the skeleton. The initial form of cells are in the case of tendons without doubt the special form of fibrocytes, the tendocytes. We believe that this finding confirm the earlier expressed opinion fibrocytes of the periost with the same structure as tendocytes metamorphose to osteoblasts. The metamorphosis of cells and intercellular substances at tendon attachments occur like in other regions of skeletogenesis (periost, epiphysis) in relative clearly definable stages: tendon, Ansatzstruktur (attachment structure), Ansatzknorpel (attachment cartilage) as a part of apophysis or Einstrahlungsknochen (invading bone). In regard to histogenesis periost and attachment structure are homologous formations. The latter one may be a reticular structure of collagen fibers or a fibrocartilage of which the cells are surrounded from a polysaccharide capsule and a fiber capsule partly accompanied by needle like minerals. The osseous attachment is composed from parallel running fibrils. This structure forms a Grenzstreifen (border sector) with a varied number of needle like crystals along the fibrils. The amount of minerals are increased in the Grenzlinie (border line) to bone and subsequently the crystals disappear. The tendinous mineralisation is a temporary one. However, the fibrils get the Fibrillencharakter (nature of fibrills) of the fibrils of lamellar bone. Later on the bone minerals appear. It is doubtful that osteogenesis occur always in the same manner. However, the vague term ossification can only concern the bone specific metamorphosis of fibrils, never the widely distributed mineralisation. Therefore, skeletogenesis ist one of the metamorphosis stages from the embryonal (!) determinated skeletal mesenchyme to mature and finally to old and atrophying bone. This metamorphosis may be a process probably following a controlled program. A developmental stage releases the development of the next one. A feedback influences the organisation of the different parts, e.g. cells and intercellularsubstances. The increasing diversity of the system developed in the way of Selbstorganisation (self-reproduction). With the exhaustion of the program informations started the ageing.     

Mechanical properties of tendon and aponeurosis of human gastrocnemius muscle in vivo.

Load-strain characteristics of tendinous tissues (Achilles tendon and aponeurosis) were determined in vivo for human medial gastrocnemius (MG) muscle. Seven male subjects exerted isometric plantar flexion torque while the elongation of tendinous tissues of MG was determined from the tendinous movements by using ultrasonography. The maximal strain of the Achilles tendon and aponeurosis, estimated separately from the elongation data, was 5.1 +/- 1.1 and 5.9 +/- 1.6%, respectively. There was no significant difference in strain between the Achilles tendon and aponeurosis. In addition, no significant difference in strain was observed between the proximal and distal regions of the aponeurosis. The results indicate that tendinous tissues of the MG are homogeneously stretched along their lengths by muscle contraction, which has functional implications for the operation of the human MG muscle-tendon unit in vivo.     

Evaluation of a subject-specific finite-element model of the equine metacarpophalangeal joint under physiological load

The equine metacarpophalangeal (MCP) joint is frequently injured, especially by racehorses in training. Most injuries result from repetitive loading of the subchondral bone and articular cartilage rather than from acute events. The likelihood of injury is multi-factorial but the magnitude of mechanical loading and the number of loading cycles are believed to play an important role. Therefore, an important step in understanding injury is to determine the distribution of load across the articular surface during normal locomotion. A subject-specific finite-element model of the MCP joint was developed (including deformable cartilage, elastic ligaments, muscle forces and rigid representations of bone), evaluated against measurements obtained from cadaver experiments, and then loaded using data from gait experiments. The sensitivity of the model to force inputs, cartilage stiffness, and cartilage geometry was studied. The FE model predicted MCP joint torque and sesamoid bone flexion angles within 5% of experimental measurements. Muscle–tendon forces, joint loads and cartilage stresses all increased as locomotion speed increased from walking to trotting and finally cantering. Perturbations to muscle–tendon forces resulted in small changes in articular cartilage stresses, whereas variations in joint torque, cartilage geometry and stiffness produced much larger effects. Non-subject-specific cartilage geometry changed the magnitude and distribution of pressure and the von Mises stress markedly. The mean and peak cartilage stresses generally increased with an increase in cartilage stiffness. Areas of peak stress correlated qualitatively with sites of common injury, suggesting that further modelling work may elucidate the types of loading that precede joint injury and may assist in the development of techniques for injury mitigation.     

Comparisons of antibody reactivity and enzyme sensitivity between small proteoglycans from bovine tendon, bone, and cartilage

Preparations of small proteoglycans from bovine tendon, bone, and cartilage have been compared for sensitivity to various enzymes and reactivity with different polyclonal antibodies. Chondroitinase ABC digestion of all proteoglycans generated a core protein preparation that migrated similarly in sodium dodecyl sulfate-polyacrylamide electrophoresis as a doublet band with Mr approximately equal to 45,000. The small proteoglycans of cartilage were divided into two populations based upon electrophoretic migration of the intact molecules (Rosenberg, L. C., Choi, H. U., Tank, L-H., Johnson, T. L., Pal, S., Webber, C., Reiner, A., and Poole, A. R. (1985) J. Biol. Chem. 260, 6304-6313). The core preparations of tendon, bone, and the faster-migrating (PG II) proteoglycans of cartilage all interacted in Western blot/enzyme-linked immunosorbent assay analysis with polyclonal antibody raised against either the tendon or bone proteoglycans. The slower-migrating (PG I) proteoglycans of cartilage did not react with these antibodies. Digestion of the tendon small proteoglycan with Staphylococcus aureus V8 protease released glycosaminoglycan chains from the molecule and generated a 40-kDa protein fragment that was resistant to further rapid degradation by the enzyme. This large digestion fragment was also prominent following V8 protease digestion of the faster-migrating (PG II) population of small cartilage proteoglycans, but not the small proteoglycan of bone. The N-terminal amino acid sequence of the tendon (PG II) proteoglycan was determined. These observations provide additional evidence for heterogeneity among the chemically similar small proteoglycans from different tissues.     

Differential x-ray diagnosis of tendinosis in the region of the greater trochanter of the femur and coxarthrosis and tubercular trochanteritis

The paper is concerned with the findings of examination of 1132 patients with complaints of pains in the hip joint. The cause of the pain symptoms was found on clinical and x-ray examination. In 28.8% the pain syndrome in the hip joint and disorder of its locomotor function were determined by tendinosis in the greater trochanter. Three stages in the development of a process were singled on the basis of morphological investigations: (I) the substitution of a part of the tendon for a fibrillar cartilage (roentgenonegative); (II) calcification of a fibrillar cartilage; (III) the development of a new spongy bone at the site of a calcified cartilage. Stages II and III have a clear x-ray image and are easily diagnosed. These signs differentiate tendinosis from coxarthrosis and tuberculous trochanteritis.     

Structure and proteoglycan composition of specialized regions of the elastic tendon of the chicken wing

The elastic tendon of the avian wing has been described by others as a unique structure with elastic properties due to the predominance of elastic fibers in the midsubstance. Further analyses of the tendon have shown it to possess five anatomically distinct regions. Besides the major elastic region, a distally located fibrocartilage and three tendinous regions are present. The tendinous regions connect: (1) the muscle to the elastic region, (2) the elastic region to the fibrocartilage and (3) the latter to the insertion site. The elastic region possesses thick and abundant elastic fibers and very thin, interconnecting collagen fibers. The collagen fibers in the sesamoid fibrocartilage are thick and interwoven, defining spaces occupied by fibrochondrocytes embedded in a non-fibrillar and highly metachromatic matrix. Biochemical analyses have shown that the fibrocartilage has about tenfold the amount of glycosaminoglycans (GAGs) found in the other regions. The main GAG in this region was chondroitin sulfate (CS) (plus keratan sulfate as detected immunocytochemically), while the other regions showed variable amounts of CS, dermatan sulfate (DS) and heparan sulfate. Further analyses have shown that a large CS-bearing proteoglycan is found in the fibrocartilage. The elastic region possesses two main proteoglycans, a large CS-bearing proteoglycan (which reacted with an antibody against keratan sulfate after chondroitinase ABC treatment) and a predominant DS-bearing proteoglycan, which showed immunoreactivity when assayed with an anti-biglycan antibody. The results demonstrate that the elastic tendon is a complex structure with complex regional structural and compositional adaptations, suited to different biomechanical roles.     

A monoclonal antibody distinguishes growth cartilage from other types of cartilage: a new probe for osteogenic cartilage

Summary Monoclonal antibodies (mAbs) were raised by injection of a homogenate of cultured growth cartilage (GC) cells from young rabbit ribs. These mAbs were examined by immunohistochemical staining for their reactivity to paraffin sections of rabbit tissues. The results showed that an mAb reacted preferentially with late hypertrophic and calcified costal GC zones. The mAb also reacted with hypertrophic GC adjacent to bone that existed in sternum and femur, but not to other cartilages, including resting cartilage, articular cartilage, auricular cartilage, nasal cartilage, tracheal cartilage and meniscus cartilage, or with other tissues, including tendon, skin, muscles, lung, liver, heart, thymus, spleen, eye and gut. It reacted with a wider area of the GC zone when the sections were decalcified, although its reactivity with the extended area was much less intensive than that with late hypertrophic and calcified GC zones. On treatment of the sections with bacterial collagenase, neither the reactive area nor its intensity were changed, while when treated with trypsin the reactivity was lost.These results suggest the existence of a certain molecule which distinguishes GC (osteogenic cartilage) from other (non-osteogenic) cartilage. This mAb is a useful probe for distinguishing osteogenic cartilage from non-osteogenic cartilage, and for studying differentiation steps of cartilage cells in endochondral bone formation. The mAb can also be used as a probe for clinical and stored specimens because it reacts with decalcified and paraffin-embedded human specimens.     

Decellularized tissue and cell-derived extracellular matrices as scaffolds for orthopaedic tissue engineering

The reconstruction of musculoskeletal defects is a constant challenge for orthopaedic surgeons. Musculoskeletal injuries such as fractures, chondral lesions, infections and tumor debulking can often lead to large tissue voids requiring reconstruction with tissue grafts. Autografts are currently the gold standard in orthopaedic tissue reconstruction; however, there is a limit to the amount of tissue that can be harvested before compromising the donor site. Tissue engineering strategies using allogeneic or xenogeneic decellularized bone, cartilage, skeletal muscle, tendon and ligament have emerged as promising potential alternative treatment. The extracellular matrix provides a natural scaffold for cell attachment, proliferation and differentiation. Decellularization of in vitro cell-derived matrices can also enable the generation of autologous constructs from tissue specific cells or progenitor cells. Although decellularized bone tissue is widely used clinically in orthopaedic applications, the exciting potential of decellularized cartilage, skeletal muscle, tendon and ligament cell-derived matrices has only recently begun to be explored for ultimate translation to the orthopaedic clinic.     

Breast reconstruction with latissimus dorsi flap: improved aesthetic results after transection of its humeral insertion.

Does transecting the tendinous insertion of the latissimus dorsi on the humerus improve aesthetic results and avoid the displeasing bulge in the armpit that sometimes occurs when the latissimus dorsi is used for breast reconstruction? In a prospective study, 60 patients who were having breast cancer surgery and simultaneous breast reconstruction using the latissimus dorsi flap were randomized for cutting (n = 29) or leaving intact (n = 31) the tendinous muscle insertion on the humerus. The cosmetic outcome was evaluated by patients and surgeons 6 to 12 months postoperatively. Patients reported good cosmetic results in 29 of 31 cases with the humeral insertion left intact and in 26 of 29 cases when the tendon was cut (p = 0.59), as compared with 21 of 31 cases versus 25 of 29 cases (p = 0.091), according to the surgeon's evaluation. A lateral bulge was more frequently observed by the surgeons in the group with intact insertion (10 of 31 patients), as compared with the group with a transected humeral insertion (2 of 29 patients). Discomfort caused by this bulge was reported by 19 of 31 patients with intact insertion, but only 3 of 29 patients with the tendon cut (p < 0.0001). The additional transection of the tendon was not associated with any complications. The additional transection of the tendinous humeral insertion of the latissimus dorsi muscle improves aesthetic results and avoids a displeasing bulge in the axilla when the latissimus dorsi flap is used for breast reconstruction.     

Persistence of Meckel's cartilage in sub-adult Struthio camelus and Dromaius novaehollandiae

This study describes the persistence of an embryonal structure through to sub-adulthood in the ostrich and emu. Mandibles from sub-adult ostrich and emu were subjected to special staining, light microscopy and dissected to reveal and describe Meckel's cartilage. Meckel's cartilage, composed of hyaline cartilage, was present within the neurovascular canal of both species. The persistence through to sub-adulthood of Meckel's cartilage in the ostrich and emu is a feature not previously reported in any other avian species. The proximal end of Meckel's cartilage was ossified in the region of the articular bone and the distal end was ossified in some specimens. Although this structure may ossify at a much later stage in life, the function in young and sub-adult birds may be to dampen shockwaves along the intramandibular nerve that result from the action of pecking. In the ostrich, the M. pseudotemporalis superficialis tendon inserted onto the supra-angular bone and Meckel's cartilage. In the emu, a small portion of the tendon was attached to the supra-angular bone and the main part to Meckel's cartilage. The persistence of Meckel's cartilage in adult lepidosaurs, crocodilians and ratites represents an unusual shared trait between the extant members of the above groups.     

Collagen crosslinking and cartilage glycosaminoglycan composition in normal and scoliotic chickens

The amounts of lysine-derived crosslinks in collagens from tendon, cartilage, intervertebral disc, and bone and changes in the composition of sternal cartilage glycosaminoglycans were estimated in two lines of chickens, a control-isogenic line and a line that develops scoliosis. In the scoliotic line, scoliosis first appears at 3-4 weeks and progressively increases in severity and incidence so that 90% of the birds express the lesion by week 10. We have reported previously that cartilage, tendon, and bone collagens from scoliotic birds are more soluble than corresponding collagens from normal birds. Herein, collagen crosslinking and altered proteoglycan metabolism are examined as possible mechanisms for the differences in collagen solubility. At 1 week of age there were fewer reducible crosslinking amino acids (hydroxylysinonorleucine, dihydroxylysinonorleucine, and lysinonorleucine) in collagens from sternal cartilage and tendon in the scoliotic line than in the isogenic line. However, by week 3 and at weeks 5 or 7 values were similar in both groups. The amounts of hydroxypyridinium in vertebral bone and intervertebral disc collagen were also similar in both groups of birds. Consequently, differences in collagen crosslinking do not appear to be a persistent developmental defect underlying the expression of scoliosis in the model. However, differences were observed in cartilage proteoglycans and glycosaminoglycans from the scoliotic line that were not present in cartilage from the isogenic line. The average molecular weight of the uronide-containing glycosaminoglycans was 30% less in the scoliotic line than in the isogenic line, i.e., 12,000 compared to 18,000. The size distribution of cartilage proteoglycans from the scoliotic line also differed from that of proteoglycans from the isogenic line.(ABSTRACT TRUNCATED AT 250 WORDS)     

Hypoxia upregulates angiogenesis and synovial cell migration in rheumatoid arthritis

Introduction  

Rheumatoid arthritis (RA) is characterised by invasion of cartilage, bone and tendon by inflamed synovium. Previous studies in our laboratory have shown that hypoxia is a feature of RA synovitis. In the present study, we investigated the consequences of hypoxia on angiogenesis and synovial fibroblast migration in RA.     

Low-intensity tensile loading increases intratendinous glucose uptake in the Achilles tendon.

The metabolic activity of tendinous tissues has traditionally been considered to be of limited magnitude. However, recent studies have suggested that glucose uptake increases in the force-transmitting tissues as a response to contractile loading, which in turn indicates an elevated tissue metabolism. The purpose of the present study was to investigate whether such a mechanism could be observed for the human Achilles tendon following tensile loading. Six subjects participated in the study. Unilateral Achilles tendon loading was applied by 25-min intermittent voluntary plantar flexor contractions. A radioactive tracer ([18F]-2-fluoro-2-deoxy-D-glucose) was administered during muscle action, and glucose uptake was measured by use of PET. Regions of interest were defined on the PET images corresponding to the cross section of Achilles tendon at two longitudinally separated sites (insertion and free tendon). Glucose uptake index was determined within respective regions of interest for the active and resting leg. Tendon force during voluntary contractions was approximately 13% of maximal voluntary contraction force. Tendon loading induced an elevated glucose uptake index compared with that of the contralateral resting tendon in the region of tendon insertion (0.13 +/- 0.05 vs. 0.09 +/- 0.02; P < 0.05) and at the free tendon (0.12 +/- 0.01 vs. 0.08 +/- 0.02; P < 0.05). The present data suggest that tissue metabolism is elevated in the human Achilles tendon in response to low-intensity loading.     

On foundations of discrete element analysis of contact in diarthrodial joints

Information about the stress distribution on contact surfaces of adjacent bones is indispensable for analysis of arthritis, bone fracture and remodeling. Numerical solution of the contact problem based on the classical approaches of solid mechanics is sophisticated and time-consuming. However, the solution can be essentially simplified on the following physical grounds. The bone contact surfaces are covered with a layer of articular cartilage, which is a soft tissue as compared to the hard bone. The latter allows ignoring the bone compliance in analysis of the contact problem, i.e. rigid bones are considered to interact through a compliant cartilage. Moreover, cartilage shear stresses and strains can be ignored because of the negligible friction between contacting cartilage layers. Thus, the cartilage can be approximated by a set of unilateral compressive springs normal to the bone surface. The forces in the springs can be computed from the equilibrium equations iteratively accounting for the changing contact area. This is the essence of the discrete element analysis (DEA). Despite the success in applications of DEA to various bone contact problems, its classical formulation required experimental validation because the springs approximating the cartilage were assumed linear while the real articular cartilage exhibited non-linear mechanical response in reported tests. Recent experimental results of Ateshian and his co-workers allow for revisiting the classical DEA formulation and establishing the limits of its applicability. In the present work, it is shown that the linear spring model is remarkably valid within a wide range of large deformations of the cartilage. It is also shown how to extend the classical DEA to the case of strong nonlinearity if necessary.     

Adult mesenchymal stem cells for tissue engineering versus regenerative medicine

Adult mesenchymal stem cells (MSCs) can be isolated from bone marrow or marrow aspirates and because they are culture-dish adherent, they can be expanded in culture while maintaining their multipotency. The MSCs have been used in preclinical models for tissue engineering of bone, cartilage, muscle, marrow stroma, tendon, fat, and other connective tissues. These tissue-engineered materials show considerable promise for use in rebuilding damaged or diseased mesenchymal tissues. Unanticipated is the realization that the MSCs secrete a large spectrum of bioactive molecules. These molecules are immunosuppressive, especially for T-cells and, thus, allogeneic MSCs can be considered for therapeutic use. In this context, the secreted bioactive molecules provide a regenerative microenvironment for a variety of injured adult tissues to limit the area of damage and to mount a self-regulated regenerative response. This regenerative microenvironment is referred to as trophic activity and, therefore, MSCs appear to be valuable mediators for tissue repair and regeneration. The natural titers of MSCs that are drawn to sites of tissue injury can be augmented by allogeneic MSCs delivered via the bloodstream. Indeed, human clinical trials are now under way to use allogeneic MSCs for treatment of myocardial infarcts, graft-versus-host disease, Crohn's Disease, cartilage and meniscus repair, stroke, and spinal cord injury. This review summarizes the biological basis for the in vivo functioning of MSCs through development and aging.     

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.     

Morphological analysis of tendinous structure in the American alligator jaw muscles

In the American alligator, the jaw muscles show seven bundles of tendinous structure: cranial adductor tendon, mandibular adductor tendon, lamina anterior inferior, trap-shaped lamina lateralis, lamina intramandibularis, lamina posterior, and depressor mandibular tendon (originating from the musculus depressor mandibulae, m. pseudotemporalis, m. adductor mandibulae posterior, m. adductor mandibulae externus, m. intramandibularis, m. pterygoideus anterior, and m. pterygoideus posterior). These tendinous structures are composed of many collagen fibrils and elastic fibers; however, the distributions and sizes of the fibers in these tendinous components differ in comparison with those of other masticatory muscles. The differences of these properties reflect the kinetic forces or the stretch applied to each tendon by the muscle during jaw movements in spite of the simple tendon-muscle junctions. © 1993 Wiley-Liss, Inc.