Tendon conditioning: artefact or property?

Isolated tendons subjected to cyclic tensile loads higher than those experienced in the tendons' recent history exhibit 'conditioning', i.e. gradually increasing elongations upon loading and gradually increasing residual elongations after unloading in the first few loading-unloading cycles. The present study examines whether this behaviour is a measurement artefact or an actual time-dependent property. The gastrocnemius tendons of six men who refrained from rigorous physical activities prior to the experiment were loaded cyclically by 10 repeated isometric plantarflexion contractions at 80% of the moment generated during plantarflexion maximal voluntary contraction (MVC). In each contraction, the elongation of the gastrocnemius tendon at 80% of MVC and the residual tendon elongation after relaxation were obtained from the analysis of sonographs recorded during the test. The tendon elongation during activation and the residual tendon elongation after relaxation increased by ca. 5 mm from the first contraction to the tenth contraction, with no changes obtained after the fifth contraction. The behaviour of the tendon in the first five contractions indicates the presence of conditioning. It is therefore concluded that conditioning is a relevant property and not an artefact associated with in vitro testing. This has implications for joint kinematics and muscle excursion.     

Effect of supraspinatus tendon repair technique on the infraspinatus tendon

Supraspinatus tendon tears are common and often propagate into larger tears that include the infraspinatus tendon, resulting in loss of function and increased pain. Previously, we showed that the supraspinatus and infraspinatus tendons mechanically interact through a range of rotation angles, potentially shielding the torn supraspinatus tendon from further injury while subjecting the infraspinatus tendon to increased risk of injury. Surgical repair of torn supraspinatus tendons is common, yet the effect of the repair on the infraspinatus tendon is unknown. Since we have established a relationship between strain in the supraspinatus and infraspinatus tendons the success of a supraspinatus tendon repair depends on its effect on the loading environment in the infraspinatus tendon. More specifically, the effect of transosseous supraspinatus tendon repair in comparison to one that utilizes suture anchors, as is commonly done with arthroscopic repairs, on this interaction through these joint positions will be evaluated. We hypothesize that at all joint positions evaluated, both repairs will restore the interaction between the two tendons. For both repairs, (1) increasing supraspinatus tendon load will increase infraspinatus tendon strain and (2) altering the rotation angle from internal to external will increase strain in the infraspinatus tendon. Strains were measured in the infraspinatus tendon insertion through a range of joint rotation angles and supraspinatus tendon loads, for the intact, transosseous, and suture anchor repaired supraspinatus tendons. Images corresponding to specific supraspinatus tendon loads were isolated for the infraspinatus tendon insertion for analysis. The effect of supraspinatus tendon repair on infraspinatus tendon strain differed with joint position. Altering the joint rotation did not change strain in the infraspinatus tendon for any supraspinatus tendon condition. Finally, increasing supraspinatus tendon load resulted in an increase in average maximum and decrease in average minimum principal strain in the infraspinatus tendon. There is a significant difference in infraspinatus tendon strain between the intact and arthroscopically (but not transosseous) repaired supraspinatus tendons that increases with greater loads. Results suggest that at low loads neither supraspinatus tendon repair technique subjects the infraspinatus tendon to potentially detrimental loads; however, at high loads, transosseous repairs may be more advantageous over arthroscopic repairs for the health of the infraspinatus tendon. Results emphasize the importance of limiting loading of the repaired supraspinatus tendon and that at low loads, both repair techniques restore the interaction to the intact supraspinatus tendon case.     

A non-invasive method of tendon force measurement

The ability to measure the forces exerted in vivo on tendons and, consequently, the forces produced by muscles on tendons, offers a unique opportunity to investigate questions in disciplines as varied as physiology, biomechanics, orthopaedics and neuroscience. Until now, tendon loads could be assessed directly only by means of invasive sensors implanted within or attached to these collagenous structures. This study shows that the forces acting on tendons can be measured, in a non-invasive way, from the analysis of the propagation of an acoustic wave. Using the equine superficial digital flexor tendon as a model, it is demonstrated that the velocity of an ultrasonic wave propagating along the main axis of a tendon increases with the force applied to this tendon. Furthermore, we show that this velocity measurement can be performed even in the presence of skin overlying the tendon. To validate this measurement technique in vivo, the ultrasonic velocity plots obtained in the Achilles tendon at the walk were compared to the loads plots reported by other authors using invasive transducers.     

Hysteresis measurements in intact human tendon

Mechanical hysteresis in tendons has traditionally been quantified from tensile testing of isolated specimens. Limitations associated with tendon displacement measurement and clamping, and uncertainties as to whether in vitro material represents intact tendon function necessitate measuring hysteresis under in vivo conditions. In the present study such measurements were taken in the human tibialis anterior (TA) tendon. Having the foot fixed on a dynamometer footplate, the displacement of the TA tendon during stimulation and relaxation of the TA muscle was recorded by means of ultrasonography in six men. Combining moment data corresponding to 0, 20, 40, 60, 80 and 100% of maximum voltage moment and the respective tendon-displacement data, a hysteresis loop was obtained between the load–displacement curves during contraction and relaxation. Measurement of the hysteresis loop area yielded a value of 19%. This value agrees with results from in vitro tensile tests of low-stress tendons, suitable for tensile force transmission and joint displacement control. In fact, the human TA tendon has such functional characteristics. The methodology presented allows design of longitudinal and cross-sectional experimental protocols, and in vivo assessment of tendon function and propensity to overheat.     

Effect of hydrogen peroxide on human tendon allograft

Bacterial contamination of tendon allografts at the completion of processing has historically been about 2 %, with tendons that are found to be culture positive being discarded. Treatment of tendon allograft with hydrogen peroxide at the beginning of tissue processing may reduce bacterial contamination, however, the potential side effects of hydrogen peroxide treatment include hydrolysis of the collagen and this may alter the mechanical properties of the graft. Pairs of human tendons were used. One was washed in 3 % hydrogen peroxide for 5 min and the untreated tendon was used as a control. The ultimate tensile strength of the tendons was determined using a material testing machine. A freeze clamp technique was used to hold the tendons securely at the high loads required to cause tendon failure. There was no statistical difference in the ultimate tensile strength between the treated and untreated tendons. Mean strength ranged from Extensor Hallucis Longus at 588 Newtons to Tibialis Posterior at 2,366 Newtons. Hydrogen peroxide washing may reduce bacterial contamination of tendon allograft and does not affect the strength of the tendon.     

Mechanical properties of human patellar tendon at the hierarchical levels of tendon and fibril

Tendons are strong hierarchical structures, but how tensile forces are transmitted between different levels remains incompletely understood. Collagen fibrils are thought to be primary determinants of whole tendon properties, and therefore we hypothesized that the whole human patellar tendon and its distinct collagen fibrils would display similar mechanical properties. Human patellar tendons (n = 5) were mechanically tested in vivo by ultrasonography. Biopsies were obtained from each tendon, and individual collagen fibrils were dissected and tested mechanically by atomic force microscopy. The Young's modulus was 2.0 ± 0.5 GPa, and the toe region reached 3.3 ± 1.9% strain in whole patellar tendons. Based on dry cross-sectional area, the Young's modulus of isolated collagen fibrils was 2.8 ± 0.3 GPa, and the toe region reached 0.86 ± 0.08% strain. The measured fibril modulus was insufficient to account for the modulus of the tendon in vivo when fibril content in the tendon was accounted for. Thus, our original hypothesis was not supported, although the in vitro fibril modulus corresponded well with reported in vitro tendon values. This correspondence together with the fibril modulus not being greater than that of tendon supports that fibrillar rather than interfibrillar properties govern the subfailure tendon response, making the fibrillar level a meaningful target of intervention. The lower modulus found in vitro suggests a possible adverse effect of removing the tissue from its natural environment. In addition to the primary work comparing the two hierarchical levels, we also verified the existence of viscoelastic behavior in isolated human collagen fibrils.     

Analysis of cumulative strain in tendons and tendon sheaths

Twenty-five fresh frozen flexor digitorum profundus tendons stratified by sex were subjected to uniaxial step stress and cyclic loads in twelve intact human cadaver hands. By attaching specially designed clip strain gage transducers on tendons just proximal and distal to an undisrupted carpal tunnel, the interactions of the tendons, tendon sheath and retinacula were measured. The elastic and viscous response of the tendon composites to step stresses were found to fit fractional power functions of stress and time respectively. A significant and quantifiable decrease in strain from the proximal to the distal tendon segment was found to be a function of wrist deviation. The results indicate that an accumulation of strain does occur in tendinous tissues during physiologic loading.     

Dynamic viscoelastic behavior of lower extremity tendons during simulated running.

The aim of this project was to see whether the tendon would show creep during long-term dynamic loading (here referred to as dynamic creep). Pig tendons were loaded by a material-testing machine with a human Achilles tendon force profile (1.37 Hz, 3% strain, 1,600 cycles), which was obtained in an earlier in vivo experiment during running. All the pig tendons showed some dynamic creep during cyclic loading (between 0.23 +/- 0.15 and 0.42 +/- 0.21%, means +/- SD). The pig tendon data were used as an input of a model to predict dynamic creep in the human Achilles tendon during running of a marathon and to evaluate whether there might consequently be an influence on group Ia afferent-mediated length and velocity feedback from muscle spindles. The predicted dynamic creep in the Achilles tendon was considered to be too small to have a significant influence on the length and velocity feedback from soleus during running. In spite of the characteristic nonlinear viscoelastic behavior of tendons, our results demonstrate that these properties have a minor effect on the ability of tendons to act as predictable, stable, and elastic force transmitters during long-term cyclic loading.     

A dynamic simulator to evaluate distal radio-ulnar joint kinematics

In order to perform cadaveric biomechanical studies of the human forearm and distal radio-ulnar joint, a dynamic simulator has been constructed. The device is based upon a Plexiglas frame, to which the ulna is secured in a vertical orientation and the humerus in a horizontal orientation. The hand is secured in a sliding bar linkage to a stepper-motor that is used to rotate the forearm. The tendons to be loaded are connected to pneumatic actuators that provide agonist and antagonist muscle loading resulting in torque along the forearm axis. The muscle loading profiles and magnitudes are programmable as a function of the pronation-supination position and direction. A magnetic tracking system is used to collect three-dimensional kinematics data of up to four segments, in conjunction with the muscle tendon loads, forearm torque and other prescribed experimental measures. All functions are under PC control using custom software written with LabVIEW (National Instruments, Austin, TX). For the DRUJ testing, the validity of the tendon loading protocol to produce physiologic torque/rotation patterns was verified using in vivo data. The relationship of individual muscle forces to forearm torque was determined by a cadaveric study.     

Effects of rotator cuff tears on muscle moment arms: a computational study

Rotator cuff tears cause morphologic changes to cuff tendons and muscles, which can alter muscle architecture and moment arm. The effects of these alterations on shoulder mechanical performance in terms of muscle force and joint strength are not well understood. The purpose of this study was to develop a three-dimensional explicit finite element model for investigating morphological changes to rotator cuff tendons following cuff tear. The subsequent objectives were to validate the model by comparing model-predicted moment arms to empirical data, and to use the model to investigate the hypothesis that rotator cuff muscle moment arms are reduced when tendons are divided along the force-bearing direction of the tendon. The model was constructed by extracting tendon, cartilage, and bone geometry from the male Visible Human data set. Infraspinatus and teres minor muscle and tendon paths were identified relative to the humerus and scapula. Kinetic and kinematic boundary conditions in the model replicated experimental protocols, which rotated the humerus from 45 degrees internal to 45 degrees external rotation with constant loads on the tendons. External rotation moment arms were calculated for two conditions of the cuff tendons: intact normal and divided tendon. Predicted moment arms were within the 1-99% confidence intervals of experimental data for nearly all joint angles and tendon sub-regions. In agreement with the experimental findings, when compared to the intact condition, predicted moment arms were reduced for the divided tendon condition. The results of this study provide evidence that one potential mechanism for the reduction in strength observed with cuff tear is reduction of muscle moment arms. The model provides a platform for future studies addressing mechanisms responsible for reduced muscle force and joint strength including changes to muscle length-tension operating range due to altered muscle and tendon excursions, and the effects of cuff tear size and location on moment arms and muscle forces.     

Effects of supercritical fluid CO<Subscript>2</Subscript> and 25 kGy gamma irradiation on the initial mechanical properties and histological appearance of tendon allograft

Tendon allografts, when autograft options are limited or when obtaining an autograft is not aligned with the patients’ best interest, play an important role in tendon and ligament reconstruction. To minimize the risk of infectious disease transmission tissue banks perform screening tests and the allografts cleaned are sterilized. The current study examines and compares the initial mechanical properties and histological appearance of supercritical CO₂ (SCCO₂)-treated and gamma-irradiated porcine extensor tendons. Thirty intact porcine forelimb extensor tendons randomized equally into three groups: control group, gamma-irradiation group, and SCCO₂-treated group. Once treated, histological assessment and histomorphologic measurements were made on the histological sections obtained from each tendon while stiffness and ultimate failure loads were evaluated from tensile testing. Histological evaluation of gamma-irradiated tendons showed significant disruption to the hierarchical morphology of the fascicle bundles, which was not evident in SCCO₂-treated specimens. Histomorphologic measurements showed a significant increase for measured dead space (void) between tendon fibrils of the gamma-irradiated group comparing to both control and SCCO₂ treated groups (p?<?0.01). There was a significant reduction in the ultimate failure load for tendons treated by gamma-irradiation compared to the control group (p?<?0.05). No statistically significant difference was detected between control and SCCO₂-treated tendons in the ultimate failure load. Stiffness values were not significantly different between three-study groups. This study suggests that while gamma-irradiation has a deleterious effect on mechanical properties of tendon tissue, SCCO₂ does not alter the biomechanical properties and the histological structure of porcine extensor tendons.     

Bridging tendon defects using autologous tenocyte engineered tendon in a hen model

Tendon defects remain a major concern in plastic surgery because of the limited availability of tendon autografts. Whereas immune rejection prohibits the use of tendon allografts, most prosthetic replacements also fail to achieve a satisfactory long-term result of tendon repair. The tissue engineering technique, however, can generate different tissues using autologous cells and thus may provide an optimal approach to address this concern. The purpose of this study was to test the feasibility of engineering tendon tissues with autologous tenocytes to bridge a tendon defect in either a tendon sheath open model or a partial open model in the hen. In a total of 40 Leghorn hens, flexor tendons were harvested from the left feet and were digested with 0.25% type II collagenase. The isolated tenocytes were expanded in vitro and mixed with unwoven polyglycolic acid fibers to form a cell-scaffold construct in the shape of a tendon. The constructs were wrapped with intestinal submucosa and then cultured in Dulbecco's Modified Eagle Medium plus 10% fetal bovine serum for 1 week before in vivo transplantation. On the feet, a defect of 3 to 4 cm was created at the second flexor digitorum profundus tendon by resecting a tendon fragment. The defects were bridged either with a cell-scaffold construct in the experimental group ( n= 20) or with scaffold material alone in the control group ( n= 20). Specimens were harvested at 8, 12, and 14 weeks postrepair for gross and histologic examination and for biomechanical analysis. In the experimental group, a cordlike tissue bridging the tendon defect was formed at 8 weeks postrepair. At 14 weeks, the engineered tendons resembled the natural tendons grossly in both color and texture. Histologic examination at 8 weeks showed that the neo-tendon contained abundant tenocytes and collagen; most collagen bundles were randomly arranged. The undegraded polyglycolic acid fibers surrounded by inflammatory cells were also observed. At 12 weeks, tenocytes and collagen fibers became longitudinally aligned, with good interface healing to normal tendon. At 14 weeks, the engineered tendons displayed a typical tendon structure hardly distinguishable from that of normal tendons. Biomechanical analysis demonstrated increased breaking strength of the engineered tendons with time, which reached 83 percent of normal tendon strength at 14 weeks. In the control group, polyglycolic acid constructs were mostly degraded at 8 weeks and disappeared at 14 weeks. However, the breaking strength of the scaffold materials accounted for only 9 percent of normal tendon strength. The results of this study indicated that tendon tissue could be engineered in vivo to bridge a tendon defect. The engineered tendons resembled natural tendons not only in gross appearance and histologic structure but also in biomechanical properties.     

Quadriceps tendon allografts as an alternative to Achilles tendon allografts: a biomechanical comparison

Quadriceps tendon with a patellar bone block may be a viable alternative to Achilles tendon for anterior cruciate ligament reconstruction (ACL-R) if it is, at a minimum, a biomechanically equivalent graft. The objective of this study was to directly compare the biomechanical properties of quadriceps tendon and Achilles tendon allografts. Quadriceps and Achilles tendon pairs from nine research-consented donors were tested. All specimens were processed to reduce bioburden and terminally sterilized by gamma irradiation. Specimens were subjected to a three phase uniaxial tension test performed in a custom environmental chamber to maintain the specimens at a physiologic temperature (37 ± 2 °C) and misted with a 0.9 % NaCl solution. There were no statistical differences in seven of eight structural and mechanical between the two tendon types. Quadriceps tendons exhibited a significantly higher displacement at maximum load and significantly lower stiffness than Achilles tendons. The results of this study indicated a biomechanical equivalence of aseptically processed, terminally sterilized quadriceps tendon grafts with bone block to Achilles tendon grafts with bone block. The significantly higher displacement at maximum load, and lower stiffness observed for quadriceps tendons may be related to the failure mode. Achilles tendons had a higher bone avulsion rate than quadriceps tendons (86 % compared to 12 %, respectively). This was likely due to observed differences in bone block density between the two tendon types. This research supports the use of quadriceps tendon allografts in lieu of Achilles tendon allografts for ACL-R.     

In vivo human tendon mechanical properties: effect of resistance training in old age

Recent advances in ultrasound scanning have made it possible to obtain the mechanical properties of human tendons in vivo. Application of the in vivo method in elderly individuals showed that their patellar tendons stiffened in response to a 14-week resistance training program by approximately 65% both structurally and materially. The rate of muscle torque development increased by approximately 27%, indicating faster contractile force transmission to the skeleton. The present findings suggest that strength training in old age can, at least partly, reverse the deteriorating effect of ageing on tendon properties and function.     

Elastic properties of human Achilles tendon are correlated to muscle strength.

The purpose of this study was to investigate whether the mechanical properties of the Achilles tendon were correlated to muscle strength in the triceps surae in humans. Twenty-four men and twelve women exerted maximal voluntary isometric plantar flexion (MVIP) torque. The elongation (DeltaX) and strain of the Achilles tendon (epsilon), the proximal part of which is the composite of the gastrocnemius tendon and the soleus aponeurosis, at MVIP were determined from the displacement of the distal myotendinous junction of the medial gastrocnemius using ultrasonography. The Achilles tendon force at MVIP (F) was calculated from the MVIP torque and the Achilles tendon moment arm. There were no significant differences in either the F-DeltaX or F-epsilon relationships between men and women. DeltaX and epsilon were 9.8 +/- 2.6 mm and 5.3 +/- 1.6%, respectively, and were positively correlated to F (r = 0.39, P < 0.05; r = 0.39, P < 0.05), which meant that subjects with greater muscle strength could store more elastic energy in the tendon. The regression y-intercepts for the F-DeltaX (P < 0.01) and F-epsilon (P < 0.05) relationship were significantly positive. These results might indicate that the Achilles tendon was stiffer in subjects with greater muscle strength, which may play a role in reducing the probability of tendon strain injuries. It was suggested that the Achilles tendon of subjects with greater muscle strength did not impair the potential for storing elastic energy in tendons and may be able to deliver the greater force supplied from a stronger muscle more efficiently. Furthermore, the difference in the Achilles tendon mechanical properties between men and women seemed to be correlated to the difference in muscle strength rather than gender.     

Human patellar tendon stiffness is restored following graft harvest for anterior cruciate ligament surgery

Minimising post-operative donor site morbidity is an important consideration when selecting a graft for surgical reconstruction of the torn anterior cruciate ligament (ACL). One of the most common procedures, the bone-patellar tendon-bone (BPTB) graft involves removal of the central third from the tendon. However, it is unknown whether the mechanical properties of the donor site (patellar tendon) recover. The present study investigated the mechanical properties of the human patellar tendon in 12 males (mean±S.D. age: 37±14 years) who had undergone surgical reconstruction of the ACL using a BPTB graft between 1 and 10 years before the study (operated knee; OP). The uninjured contralateral knee served as a control (CTRL). Patellar tendon mechanical properties were assessed in vivo combining dynamometry with ultrasound imaging. Patellar tendon stiffness was calculated from the gradient of the tendon's force–elongation curve. Tendon stiffness was normalised to the tendon's dimensions to obtain the tendon's Young's modulus. Cross-sectional area (CSA) of OP patellar tendons was larger by 21% than CTRL tendons (P<0.01). Patellar tendon stiffness was not significantly different between OP and CTRL tendons, but the Young's modulus was lower by 24% in OP tendons (P<0.01). A compensatory enlargement of the patellar tendon CSA, presumably due to scar tissue formation, enabled a recovery of tendon stiffness in the OP tendons. The newly formed tendon tissue had inferior properties as indicated by the reduced tendon Young's modulus, but it increased to a level that enabled recovery of tendon stiffness.     

Correlations between mean echogenicity and material properties of normal and diseased equine superficial digital flexor tendons: an in vitro segmental approach

The objective of this study was to test the hypothesis that tendon echogenicity is associated with the material properties of the corresponding tendon site, especially in case of lesions, due to local changes in tendon matrix composition. Four normal and nine spontaneously injured equine superficial digital flexor tendons (SDFT) were isolated then ultrasonographically examined under tension, in a special device placed in a water bath. Ultrasonographic transversal images (7.5MHz linear transducer) of five segments along each tendon were digitized, and analyzed in order to measure the mean cross-sectional area (MCSA) and mean echogenicity (ME) of each segment. The tendons were then tested in traction until rupture in a testing machine. For each segment, stress and strain were determined throughout the test, and the elastic modulus (EM) was evaluated. The tendon lesions were also documented by histology. No correlation was found between ME and the material properties of normal tendon segments. At the rupture sites of the nine diseased tendons, ME was positively correlated with maximal stress and EM, whereas no correlation was demonstrated with maximal strain. Besides, a positive correlation was demonstrated between ME and both MCSA and EM, when the three metacarpal segments of the diseased tendons were considered. Although ME gives only rough information about tendon matrix structure, it does show, under these in vitro conditions, significant correlations with material properties of pathological tendon segments, which may improve the functional significance and therefore the prognostic value of the ultrasonographic examination of tendon lesions.     

Mechanical response of individual collagen fibrils in loaded tendon as measured by atomic force microscopy

A precise analysis of the mechanical response of collagen fibrils in tendon tissue is critical to understanding the ultrastructural mechanisms that underlie collagen fibril interactions (load transfer), and ultimately tendon structure–function. This study reports a novel experimental approach combining macroscopic mechanical loading of tendon with a morphometric ultrascale assessment of longitudinal and cross-sectional collagen fibril deformations. An atomic force microscope was used to characterize diameters and periodic banding (D-period) of individual type-I collagen fibrils within murine Achilles tendons that were loaded to 0%, 5%, or 10% macroscopic nominal strain, respectively. D-period banding of the collagen fibrils increased with increasing tendon strain (2.1% increase at 10% applied tendon strain, p < 0.05), while fibril diameter decreased (8% reduction, p < 0.05). No statistically significant differences between 0% and 5% applied strain were observed, indicating that the onset of fibril (D-period) straining lagged macroscopically applied tendon strains by at least 5%. This confirms previous reports of delayed onset of collagen fibril stretching and the role of collagen fibril kinematics in supporting physiological tendon loads. Fibril strains within the tissue were relatively tightly distributed in unloaded and highly strained tendons, but were more broadly distributed at 5% applied strain, indicating progressive recruitment of collagen fibrils. Using these techniques we also confirmed that collagen fibrils thin appreciably at higher levels of macroscopic tendon strain. Finally, in contrast to prevalent tendon structure–function concepts data revealed that loading of the collagen network is fairly homogenous, with no apparent predisposition for loading of collagen fibrils according to their diameter.     

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.     

In situ fibril stretch and sliding is location-dependent in mouse supraspinatus tendons

Tendons are able to transmit high loads efficiently due to their finely optimized hierarchical collagen structure. Two mechanisms by which tendons respond to load are collagen fibril sliding and deformation (stretch). Although many studies have demonstrated that regional variations in tendon structure, composition, and organization contribute to the full tendon׳s mechanical response, the location-dependent response to loading at the fibril level has not been investigated. In addition, the instantaneous response of fibrils to loading, which is clinically relevant for repetitive stretch or fatigue injuries, has also not been studied. Therefore, the purpose of this study was to quantify the instantaneous response of collagen fibrils throughout a mechanical loading protocol, both in the insertion site and in the midsubstance of the mouse supraspinatus tendon. Utilizing a novel atomic force microscopy-based imaging technique, tendons at various strain levels were directly visualized and analyzed for changes in fibril d-period with increasing tendon strain. At the insertion site, d-period significantly increased from 0% to 1% tendon strain, increased again from 3% to 5% strain, and decreased after 5% strain. At the midsubstance, d-period increased from 0% to 1% strain and then decreased after 7% strain. In addition, fibril d-period heterogeneity (fibril sliding) was present, primarily at 3% strain with a large majority occurring in the tendon midsubstance. This study builds upon previous work by adding information on the instantaneous and regional-dependent fibrillar response to mechanical loading and presents data proposing that collagen fibril sliding and stretch are directly related to tissue organization and function.