Geometric and mechanical properties of human cervical spine ligaments

This study characterized the geometry and mechanical properties of the cervical ligaments from C2-T1 levels. The lengths and cross-sectional areas of the anterior longitudinal ligament, posterior longitudinal ligament, joint capsules, ligamentum flavum, and interspinous ligament were determined from eight human cadavers using cryomicrotomy images. The geometry was defined based on spinal anatomy and its potential use in complex mathematical models. The biomechanical force-deflection, stiffness, energy, stress, and strain data were obtained from 25 cadavers using in situ axial tensile tests. Data were grouped into middle (C2-C5) and lower (C5-T1) cervical levels. Both the geometric length and area of cross section, and the biomechanical properties including the stiffness, stress, strain, energy, and Young's modulus, were presented for each of the five ligaments. In both groups, joint capsules and ligamentum flavum exhibited the highest cross-sectional area (p < 0.005), while the longitudinal ligaments had the highest length measurements. Although not reaching statistical significance, for all ligaments, cross-sectional areas were higher in the C5-T1 than in the C2-C5 group; and lengths were higher in the C2-C5 than in the C5-T1 group with the exception of the flavum (Table 1 in the main text). Force-deflection characteristics (plots) are provided for all ligaments in both groups. Failure strains were higher for the ligaments of the posterior (interspinous ligament, joint capsules, and ligamentum flavum) than the anterior complex (anterior and posterior longitudinal ligaments) in both groups. In contrast, the failure stress and Young's modulus were higher for the anterior and posterior longitudinal ligaments compared to the ligaments of the posterior complex in the two groups. However, similar tendencies in the structural responses (stiffness, energy) were not found in both groups. Researchers attempting to incorporate these data into stress-analysis models can choose the specific parameter(s) based on the complexity of the model used to study the biomechanical behavior of the human cervical spine.     

Measurements of cross-sectional area of collagen structures (knee ligaments) by means of an optical method

Measurement methods of cross-sectional area of collagen tissues and structures, so far chosen, are critically reviewed. A new optical method, founded on section outline plotting, by which it is easy to compute the cross-sectional area, is suggested and shown in particular. Experimental results, achieved by applying the method to the measurement of bovine knee ligament sections, are reported; these results are compared with those taken from samples coming from the same ligaments.     

Anatomy of the cruciate ligaments and their function in extension and flexion of the human knee joint

The areas of the femoral origin of the cruciate ligaments have approximately the shape of sectors of ellipses, the one for the anterior ligament on the lateral condyle posteroproximally and the one for the posterior ligament on the medial condyle distally. By means of a new technique of dissection, combined with the use of X-rays, the change in distance between the origin and insertion and so the change of tension of single bundles of the ligaments could be analyzed. Only a rather thin bundle in each cruciate ligament is in constant tension: "guiding bundles." The maximal diminution of distance between the origin and insertion for some bundles is 65%. In the anterior cruciate ligament the majority of fibres are taut in extreme extension: "limiting bundles." The same is true in the posterior cruciate ligament in extreme flexion. There are also some fibres, especially in the posterior cruciate ligament, that are taut only in an intermediate position. The geometric analysis of the function of different groups of fibers was performed by a modification of Menschik's concept of a four-bar link.     

A new method of measuring the cross-sectional area of connective tissue structures

There appears to be no generally accepted method of measuring in-situ the cross-sectional area of connective tissues, particularly small ones, before mechanical testing. An instrument has therefore been devised to measure the cross-sectional area of one such tissue, the rabbit medial collateral ligament, directly and nondestructively. However, the methodology is general and could be applied to other tissues with appropriate changes in detail. The concept employed in the instrument is to measure the thickness of the tissue as a function of position along the width of the tissue. The plot obtained of thickness versus width position is integrated to provide the cross-sectional area. This area is accurate to within 5 percent, depending mainly on alignment of the instrument and pre-load of the ligament. Results on the mid-substance of the rabbit medial collateral ligaments are repeatable and reproducible. Values of maximum width and thickness are less variable than those obtained with a vernier caliper. The measured area is considerably less than that estimated assuming rectangular cross-section and slightly less than that estimated on the assumption of elliptical cross-section.     

Ligament fibre recruitment at the human ankle joint complex in passive flexion

Knowledge of ligament fibre recruitment at the human ankle joint complex is a fundamental prerequisite for analysing mobility and stability. Previous experimental and modelling studies have shown that ankle motion must be guided by fibres within the calcaneofibular and tibiocalcaneal ligaments, which remain approximately isometric during passive flexion. The purpose of this study was to identify these fibres.

Three below-knee amputated specimens were analysed during passive flexion with combined radiostereometry for bone pose estimation and 3D digitisation for ligament attachment area identification. A procedure based on singular value decomposition enabled matching bone pose with digitised data and therefore reconstructing position in space of ligament attachment areas in each joint position. Eleven ordered fibres, connecting corresponding points on origin and insertion curves, were modelled for each of the following ligaments: posterior talofibular, calcaneofibular, anterior talofibular, posterior tibiotalar, tibiocalcaneal, and anterior tibiotalar.

The measured changes in length for the ligament fibres revealed patterns of tightening and slackening. The most anterior fibre of the calcaneofibular and the medio-anterior fibre of the tibiocalcaneal ligament exhibited the most isometric behaviour, as well as the most posterior fibre of the anterior talofibular ligament. Fibres within the calcaneofibular ligament remain parallel in the transverse plane, while those within the tibiocalcaneal ligament become almost parallel in joint neutral position. For both these ligaments, fibres maintain their relative inclination in the sagittal plane throughout the passive flexion range.

The observed significant change in both shape and orientation of the ankle ligaments suggest that this knowledge is fundamental for future mechanical analysis of their response to external forces.     

New aspects of the morphology and function of the human hip joint ligaments.

The capsular ligaments of the human hip joint were submitted to exact morphological analysis, and they proved to be multiple and numerous. We have described various ligamentous systems and their interconnections, and have suggested new terminologies and systematics. The ligaments were subjected to functional analysis by means of measuring strips to determine the positions in which the ligaments are taut. The ligament systems were all found to serve a restrictive function, and various parts of the apparatus restricted all possible movements in the hip joint. Extension is restricted by the medial iliofemoral complex, abduction by the pubofemoral ligament, and adduction by the posterior coxal ligaments and by the superior ischiofemoral ligament. Flexion is restricted by the inferior ischiofemoral ligaments, inward rotation by the superior ischiofemoral ligament, and outward rotation by the lateral iliofemoral complex. Only the ligament of the femoral head is unable to exert a restricting function, despite reaching a state of tension in extreme adduction.     

Specializations within the lumbosacral spinal cord of the pigeon

The prominent accessory lobes of Lachi in birds are considered to be marginal nuclei; similar nuclei have been implicated in mechanoreceptive functions in snakes and lampreys. Reptile studies emphasized the involvement of the denticulate ligament with this mechanoreceptive function. This investigation examines the fine structure of the accessory lobes of Lachi in pigeons and their interaction with ligaments for features which might support such a mechanoreceptive function. In the lumbosacral area of the spinal cord, the lateral longitudinal ligaments and the ventral longitudinal ligament are hypertrophied. The ventral transverse ligaments are present only within the lumbosacral segments of the spinal cord and they interconnect with the lateral and ventral longitudinal ligaments. The lateral longitudinal ligament makes intimate contact with the spinal cord, and many glial processes from the spinal cord mingle with and are firmly attached to collagenous fibers of the ligament. The lobes lie dorsal to the lateral longitudinal ligament in the exact area where it interconnects with the transverse ligament. The lobe's multipolar neurons have a number of synaptic contacts but no unusual specializations were noted. Most of each lobe is composed of interdigitating saccular structures filled to varying degrees with flocculent material. The sacs are extensions of the cytoplasm of neuroglial cells, which also give rise to membranes surrounding neuronal processes and the sacs themselves. A possible functional relationship of the lobes and the ligaments of the lumbosacral spinal segments within the vertebral column is described.     

Geometrical changes of knee ligaments and patellar tendon during passive flexion

Patterns of fibre elongation and orientation for the cruciate and collateral ligaments of the human knee joint and for the patellar tendon have not yet been established in three-dimensions. These patterns are essential for understanding thoroughly the contribution of these soft tissues to joint function and of value in surgical treatments for a more conscious assessment of the knee status. Measurements from 10 normal cadaver knees are here reported using an accurate surgical navigation system and consistent anatomical references, over a large flexion arc, and according to current recommended conventions. The contours of relevant sub-bundles were digitised over the corresponding origins and insertions on the bones. Representative fibres were calculated as the straight line segments joining the centroids of these attachment areas. The most isometric fibre was also taken as that whose attachment points were at the minimum change in length over the flexion arc. Changes in length and orientation of these fibres were reported versus the flexion angle. A good general repeatability of intra- and inter-specimens was found. Isometric fibres were found in the locations reported in the literature. During knee flexion, ligament sub-bundles slacken in the anterior cruciate ligament, and in the medial and lateral collateral ligaments, whereas they tighten in the posterior cruciate ligament. In each cruciate ligament the two compounding sub-bundles have different extents for the change in fibre length, and also bend differently from each other on both tibial planes. In the collateral ligaments and patellar tendon all fibres bend posteriorly. Patellar tendon underwent complex changes in length and orientation, on both the tibial sagittal and frontal planes. For the first time thorough and consistent patterns of geometrical changes are provided for the main knee ligaments and tendons after careful fibre mapping.     

The insertion sites of selected human carpal ligaments. A quantitative morphological study using three-dimensional computer reconstructions

The shape and size of insertion sites associated to the relevant carpal ligaments are given in terms of outlines and surface areas. The sites are reconstructed as areas on the contours of the carpal bones with the use of three-dimensional reconstruction of serially sectioned wrists. The computer-models of the wrist-joint show accurately the sofar uncharted three-dimensional orientations of the insertions. The documentation of the surface areas is a first approach to the quantitative analysis of the bone-ligament complex of the wrist-joint. This is needed in future studies about the role of the ligament behavior in wrist-joint motion.     

The development and validation of a charge-coupled device laser reflectance system to measure the complex cross-sectional shape and area of soft tissues

Determination of the stresses in soft tissues such as ligaments and tendons under uniaxial tension require accurate measurement of their cross-sectional area. Of the many methods available, there are concerns regarding contact methods which exert external loads and deform the cross-sectional shape of soft tissues. Hence, the area measurements are affected. On the other hand, non-contact methods have difficulties in dealing with complex shapes, especially with concavities. To address these problems, a new measurement system using a charge-coupled device (CCD) laser displacement sensor has been developed and tested. This system measures the complete surface profile of the object by rotating the laser 360° around the soft tissue. Then, the cross-sectional shape is reconstructed and the cross-sectional area determined via Simpson's rule. The system's accuracy was first verified with objects of various cross-sectional shapes and areas (cylinder: 23.1, 76.5, 510.3 mm²; cuboid: 34.3, 163.8, 316.7 mm², and cylinder with concavities: 121.4 mm²). The CCD laser reflectance system's accuracy was within 2.0% for these objects. To test biological application, the goat Achilles tendon and the anteromedial bundle of the porcine anterior cruciate ligament specimens were measured and compared to values obtained using another accepted technique, the laser micrometer system. The areas obtained using the CCD laser reflectance system were 4.4% and 9.7% lower than those obtained with the laser micrometer system respectively. These differences could be mainly attributed to concavities. Thus, the CCD laser reflectance system is an improved method for measuring the cross-sectional shape and area of soft tissues since it can detect and account for concavities without physically contacting the specimen.     

Analysis of simulated single ligament transection on the mechanical behaviour of a lumbar functional spinal unit.

The influence of the different lumbar spinal ligaments on intersegmental rotation is not fully understood. In order to explore this effect, a finite element model of the functional spinal unit L3/L4 was loaded with pure moments in the three main anatomic planes. The two extremes--minimum and maximum--ligament stiffness values reported in the literature were applied. After virtual transection of each of the spinal ligaments in turn, the intersegmental rotation and forces in the remaining ligaments were calculated. On flexion, the highest force was found for the posterior longitudinal ligament; on extension and lateral bending for the anterior longitudinal ligament; and on axial rotation for the facet capsular ligament. The strongest influence on intersegmental rotation is exerted by the interspinous ligament on flexion, by the anterior longitudinal ligament on extension and lateral bending, and by the facet capsular ligaments on axial rotation. Ligament stiffness has a strong influence on intersegmental rotation and forces in the ligaments, so that finite element models of spinal segments must be validated by experimental data. This study should help to elucidate the role of the various ligaments.     

Ligament structure, physiology and function

Ligaments are specialized connective tissues with very interesting biomechanical properties. They have the ability to adapt to the complex functions that each are required to perform. While ligaments were once thought to be inert, they are in fact responsive to many local and systemic factors that influence their function within the organism. Injury to a ligament results in a drastic change in its structure and physiology and creates a situation where ligament function is restored by the formation of scar tissue that is biologically and biomechanically inferior to the tissue it replaces. This article will briefly review the basic structure, physiology and function of normal versus healing knee ligaments, referring specifically to what is known about two of the most extensively studied and clinically relevant knee ligaments, the anterior cruciate (ACL) and medial collateral (MCL) ligaments of the knee. Those readers wishing for more comprehensive sources of information on ligament biology and biomechanics are referred to many excellent reviews on these topics.     

Characteristics of the Three Ligaments of Human Spring Ligament Complex from a Viewpoint of Elements

To elucidate characteristics of the three ligaments constituting the spring ligament complex from a viewpoint of elements, the authors investigated age-related changes of elements, relationships among their elements, relationships among ligaments in the elements, and gender differences in the three ligaments of the spring ligament complex, the superomedial calcaneonavicular (SMCN), inferoplantar longitudinal calcaneonavicular (ICN), and third or medioplantar oblique calcaneonavicular (TCN) ligaments. After ordinary dissection at Nara Medical University was finished, the SMCN, ICN, and TCN ligaments of the spring ligament complex were removed from the subjects. The subjects consisted of 10 men and 12 women, ranging in age from 62 to 99 years (average age = 80.5 ± 9.7 years). After incineration with nitric acid and perchloric acid, the element contents were determined by inductively coupled plasma-atomic emission spectrometry. It was found that although the Ca and P content hardly changed in the SMCN ligament with aging, the Ca and P content in the ICN ligament increased to about three and five times higher in the 80s in comparison with the 60s, respectively, whereas in the TCN ligament, it increased about 40% and 90% higher in the 80s compared with the 60s, respectively. Regarding the relationships among elements, significant direct correlations were found among the contents of Ca, P, and Mg in all the three ligaments of the spring ligament complex. This finding was in agreement with the previous finding obtained with the three ligaments of the anterior cruciate ligament, posterior longitudinal ligament, and ligamentum capitis femoris. Whether there were significant correlations among the three ligaments of the spring ligament complex with regard to the Ca, P, S, Mg, Zn, and Fe contents was examined using Pearson’s correlation. It was found that there were significant direct correlations between the SMCN and TCN ligaments in all the Ca, P, Mg, and Zn contents and also between the SMCN and ICN ligaments in both the Mg and Fe contents but not between the TCN and ICN ligaments in the six element contents. Regarding the gender difference in elements, a significant gender difference was found only in the Mg content of the SMCN ligament, being significantly higher in men than in women.     

Recruitment of knee joint ligaments

On the basis of earlier reported data on the in vitro kinematics of passive knee-joint motions of four knee specimens, the length changes of ligament fiber bundles were determined by using the points of insertion on the tibia and femur. The kinematic data and the insertions of the ligaments were obtained by using Roentgenstereophotogrammetry. Different fiber bundles of the anterior and posterior cruciate ligaments and the medial and lateral collateral ligaments were identified. On the basis of an assumption for the maximal strain of each ligament fiber bundle during the experiments, the minimal recruitment length and the probability of recruitment were defined and determined. The motions covered the range from extension to 95 degrees flexion and the loading conditions included internal or external moments of 3 Nm and anterior or posterior forces of 30 N. The ligament length and recruitment patterns were found to be consistent for some ligament bundles and less consistent for other ligament bundles. The most posterior bundle of each ligament was recruited in extension and the lower flexion angles, whereas the anterior bundle was recruited for the higher flexion angles. External rotation generally recruited the collateral ligaments, while internal rotation recruited the cruciate ligaments. However, the anterior bundle of the posterior cruciate ligament was recruited with external rotation at the higher flexion angles. At the lower flexion angles, the anterior cruciate and the lateral collateral ligaments were recruited with an anterior force. The recruitment of the posterior cruciate ligament with a posterior force showed that neither its most anterior nor its most posterior bundle was recruited at the lower flexion angles. Hence, the posterior restraint must have been provided by the intermediate fiber bundles, which were not considered in the experiment. At the higher flexion angles, the anterior bundles of the anterior cruciate ligament and the posterior cruciate ligament were found to be recruited with anterior and posterior forces, respectively. The minimal recruitment length and the recruitment probability of ligament fiber bundles are useful parameters for the evaluation of ligament length changes in those experiments where no other method can be used to determine the zero strain lengths, ligament strains and tensions.     

Variable tensility of the oral arm plate ligaments of the brittlestar Ophiura ophiura (Echinodermata: Ophiuroidea)

The oral arm plates of the brittlestar Ophiura ophiura L. are connected to lateral arm plates at distal and proximal ligamentous junctions. The distal junction is mobile and is disrupted during arm autotomy; the proximal junction is more rigid and does not participate in autotomy. Aspects of the morphology and mechanical properties of the distal and proximal oral arm plate ligaments have been investigated in order to determine if their tensility is under physiological control. By means of creep tests it was found that elevation of the external potassium (K⁺) ion concentration causes a decrease in the viscosity of the distal ligament which is either transient or continues until rupture intervenes. In forced vibration tests the distal ligament often shows a biphasic stiffening then softening response to excess K ^- ions. Anaesthetics block the softening phase but enhance the stiffening component of this response. This ligament is also softened by repetitive electrical stimuli but stiffened by excess calcium ions and by acetylcholine. The proximal ligament appears to have the capacity for only transient changes in mechanical properties. Both ligaments are penetrated by the processes of juxtaligamental cells whose perikarya are arranged in clusters innervated by hyponeural nerves. These cells are thought to modulate the interfibrillar cohesion of the ligaments. It is concluded that the distal and proximal ligaments are mutable collagenous structures which in their stiffened condition help to maintain arm posture without the need for continuous muscular activity, and that at autotomy the distal ligament undergoes a profound loss of tensile strength which facilitates arm detachment.     

Biomechanics of changes in ACL and PCL material properties or prestrains in flexion under muscle force-implications in ligament reconstruction

The effects of changes in cruciate ligament material and prestrain on knee joint biomechanics following ligament reconstruction surgery by a tendon are not adequately known. A 3D nonlinear finite element model of the entire knee joint was used to investigate the joint response at different flexion angles under a quadriceps force while varying ACL and PCL initial strains or material properties. The ACL and PCL forces as well as tibiofemoral contact forces/areas substantially increased with greater ACL or PCL initial strains or stiffness. The patellofemoral contact force slightly increased whereas the tibial extensor moment slightly decreased with tenser or stiffer ACL. Reverse trends were predicted with slacker ACL. Results confirm the hypotheses that changes in the prestrain of one cruciate ligament substantially influence the force in the other cruciate ligament and the entire joint and that the use of the patellar tendon (PT) as a replacement for cruciate ligaments markedly alters the joint biomechanics with trends similar to those predicted when increasing prestrains. Forces in both ACL and PCL ligaments increased as one of them became tenser or stiffer and diminished as it became slacker. These results have important consequences in joint biomechanics following ligament injuries or replacement and tend to recommend the use of grafts with smaller prestrains (i.e. slacker than intact) when using the PT as the replacement material with stiffness greater than that of replaced ligament itself.     

Effect of changes in cruciate ligaments pretensions on knee joint laxity and ligament forces

The knee joint cruciate ligaments are reconstructed with the rationale to avoid joint instability, recurrent injury, damage to soft tissues and osteoarthritis. Wide range of procedures with different stiffness, pretension, orientation and insertion locations have been proposed with the primary goal to restore the joint laxity. Apart from the general lack of success in preservation of force in the reconstructed ligament itself, the concern, not yet addressed, arises as to the effect of such perturbation on the other intact cruciate ligament. The interaction between cruciate ligament forces is hypothesized in this work. Using a 3-D nonlinear finite element model of the tibiofemoral joint, we examined this hypothesis by quantifying the extent of coupling between cruciate ligaments while varying the prestrain in each ligament under flexion with and without anterior-posterior (A-P) loads. A remarkable coupling was predicted between cruciate ligament forces in flexion thus confirming the hypothesis; forces in both cruciate ligaments increased as initial strain or pretension in one of them increased whereas they both diminished as one of them became slack. Moreover, changes in laxity and in ligament forces as a cruciate ligament prestrained or pretensioned varied with flexion angle and external loads. These findings have important consequences in joint functional biomechanics following a ligament injury or replacement surgery and in selection of laxity matched or ligament force matched pretensioning protocols.     

Biomechanics of changes in ACL and PCL material properties or prestrains in flexion under muscle force-implications in ligament reconstruction

The effects of changes in cruciate ligament material and prestrain on knee joint biomechanics following ligament reconstruction surgery by a tendon are not adequately known. A 3D nonlinear finite element model of the entire knee joint was used to investigate the joint response at different flexion angles under a quadriceps force while varying ACL and PCL initial strains or material properties. The ACL and PCL forces as well as tibiofemoral contact forces/areas substantially increased with greater ACL or PCL initial strains or stiffness. The patellofemoral contact force slightly increased whereas the tibial extensor moment slightly decreased with tenser or stiffer ACL. Reverse trends were predicted with slacker ACL. Results confirm the hypotheses that changes in the prestrain of one cruciate ligament substantially influence the force in the other cruciate ligament and the entire joint and that the use of the patellar tendon (PT) as a replacement for cruciate ligaments markedly alters the joint biomechanics with trends similar to those predicted when increasing prestrains. Forces in both ACL and PCL ligaments increased as one of them became tenser or stiffer and diminished as it became slacker. These results have important consequences in joint biomechanics following ligament injuries or replacement and tend to recommend the use of grafts with smaller prestrains (i.e. slacker than intact) when using the PT as the replacement material with stiffness greater than that of replaced ligament itself.     

A role for calcitonin gene-related peptide in rabbit knee joint ligament healing

Knee joint ligament healing has been shown to be improved when the torn ligament ends remain in contact, however, the rationale for these effects is unknown. The sensory neuropeptide calcitonin gene related peptide (CGRP) has potent trophic and vasodilatatory properties and as such is thought to be advantageous in wound repair. In ascertaining a role for CGRP in rabbit medial collateral ligament healing, the present study examined changes in CGRP-like immunoreactivity (CGRP-LI) and CGRP-mediated vasomotor responses in gap injured (non-contact), Z-plasty apposed (contact), and sham operated control medial collateral ligaments. At 6 weeks post-trauma, CGRP-LI decreased in the healing zone of gap injured and Z-plasty apposed medial collateral ligaments compared with controls, and non-contact ligament nerve fibres exhibited an abnormal morphology. Topical administration of CGRP (10(-13) to 10(-9) mol) caused a dose-dependent increase in ligament perfusion in each experimental group of knees. The CGRP-mediated vasodilatation associated with gap injured ligaments was not significantly different from controls (P = 0.06), whereas apposed medial collateral ligaments showed an augmented response to the peptide (P < 0.0005). These findings indicate that the beneficial effects of ligament interposition post-trauma may be related to an enhanced responsiveness to CGRP in conjunction with a more typical re-innervation profile. Conversely, the aberrant characteristics of CGRP-LI nerves occurring in gap injured tissue is suggestive of impaired CGRP release which may explain the poor functional recovery associated with these ligaments.     

Intrinsic innervation of the rat knee joint articular capsule and ligaments

In spite of the practical importance of having a detailed knowledge of knee joint innervation to understand the pathophysiologic aspects, little information is now available concerning the density and pattern of the nerve fibres which are distributed to it. The present study has been designed to investigate the density and distribution of nerve fibres and receptor corpuscles in the knee joint articular capsule, cruciate and collateral ligaments in the rat, using the acetylcholinesterase (AChE) histochemical in toto staining technique. The investigation was performed on male Wistar rats of 3 months of age, some of which had been treated with capsaicin to deplete their afferent 'C' fibres of their content of neuropeptides. AChE-positive nerve fibres and different types of receptor corpuscle endings were found within articular capsule and ligaments. The highest density of AChE-positive nerve fibres was noticeable in the fibular collateral ligament followed by the tibial collateral ligament, the posterior cruciate ligament, the anterior cruciate ligament and the articular capsule. In the articular capsule the number of type I endings was higher than in the ligaments. The opposite is true for the other type of receptor corpuscles found as well as for nerve endings. Capsaicin treatment significantly reduced the density of AChE-positive nerve fibres in knee joint ligaments but did not affect nerve fibres in the articular capsule. Moreover, it caused the disappearance of some kind of receptor corpuscles within the collateral and cruciate ligaments. The above data collectively suggest that the AChE in toto staining technique may represent a good method for investigating joint innervation and that a significant percentage of nerve fibres supplying knee joint ligaments is represented by C fibre afferents.