Knee ligament mechanical properties are not influenced by estrogen or its receptors

Women are at greater risk of tearing their knee anterior cruciate ligament (ACL) than men participating in similar athletic activities. There is currently no conclusive explanation for this disparity; however, as ACL injuries in women have been linked with estrogen fluctuations during the menstrual cycle, one hypothesis is that estrogen has a direct detrimental effect on knee ligament mechanical properties. This study investigated the influence of estrogen and its receptors (ER alpha and ER beta) on knee ligament mechanical properties. This was achieved by testing the viscoelastic and tensile mechanical properties of knee medial collateral ligaments (MCL) and ACLs from: 1) male Sprague-Dawley rats treated with either estrogen (17alpha-ethynylestradiol; 0.03 mg/kg) or an ER alpha-specific agonist (propyl pyrazole triol; 2 mg/kg), and 2) female mice with a null mutation of the gene encoding for ER beta. Estrogen treatment had no significant effects on the viscoelastic or tensile mechanical properties of the rat MCL or ACL. Similarly, pharmacological stimulation of ER alpha using a selective agonist in rats and genetic modulation of ER beta by null mutation of its gene in mice did not influence MCL or ACL properties. These data indicate that estrogen does not have a major direct effect on ligament mechanical properties. Energies for the prevention of the disproportionately high rate of knee ligament injuries in women may be better spent focusing on more established and modifiable risk factors, such as abnormalities in neuromuscular control about the knee.     

Effects of postmortem storage by freezing on ligament tensile behavior

The purpose of this study is to examine the effect of prolonged postmortem freezing storage (between 1 1/2 and 3 months at -20 degrees C) on the structural properties of the medial collateral ligament (MCL)-bone complex as well as the mechanical properties of the MCL substance from the rabbit knee. Tensile testing of the femur-MCL-tibia specimen was performed and no statistically significant changes were noted between the fresh and stored samples in terms of the cyclic stress relaxation, the load-deformation characteristics, as well as the load, deformation and energy absorbing capability at failure. The area of hysteresis of the stored samples was significantly reduced in the first few cycles, however. The mechanical properties of the MCL substance, as represented by the stress-strain curves, tensile strength and ultimate strain also did not change following storage. We conclude, therefore, proper and careful storage by freezing would have little or no effect on the biomechanical properties of the ligaments.     

Water content alters viscoelastic behaviour of the normal adolescent rabbit medial collateral ligament.

Testing environment is an important factor in the outcome of mechanical tests on connective tissue. The purpose of this investigation was to determine the effect of ligament water content on ligament mechanical behaviour by altering the test environment. Water content of medial collateral ligament (MCLs) from 19 three-month-old New Zealand White rabbits was varied in subsets of ligaments pairs by means of immersion in 2, 10 or 25% sucrose or 0.9% phosphate-buffered saline (PBS) solutions for 1 h. One knee joint was cycled 50 times in the designated solution (experimental), while the contralateral knee (uncycled control) was simultaneously soaked in the same tank. Following cycling, the water contents of both test and control ligaments were determined. Water contents of 22 normal MCLs were determined immediately post-sacrifice and served as 'normal water content' controls. Normalized peak cyclic load changes were used as a measure of the viscoelastic behaviour of each MCL. Results demonstrated that only ligaments soaked (but not cycled) in a 10% sucrose solution had water contents (60.5 +/- 2.5%) which were statistically similar to the 22 fresh normal MCLs (63.9 +/- 6.0%). Ligaments soaked in PBS (74.0 +/- 1.3%) or 2% sucrose (69.2 +/- 2.3%) had significantly higher water contents compared to fresh normal MCLs. Ligaments with higher water contents (e.g. soaked in PBS or 2% sucrose) demonstrated greater cyclic load relaxation compared to ligaments with lower contents (e.g. soaked in 25 or 10% sucrose). Different fluid test environments can significantly alter ligament water content and, in turn, significantly affect ligament viscoelastic behaviour.     

Comparison of elastic,viscoelastic and failure tensile material properties of knee ligaments and patellar tendon

The knee ligaments and patellar tendon function in concert with each other and other joint tissues, and are adapted to their specific physiological function via geometry and material properties. However, it is not well known how the viscoelastic and quasi-static material properties compare between the ligaments. The purpose of this study was to characterize and compare these material properties between the knee ligaments and patellar tendon.Dumbbell-shaped tensile test samples were cut from bovine knee ligaments (ACL, LCL, MCL, PCL) and patellar tendon (PT) and subjected to tensile testing (n = 10 per ligament type). A sinusoidal loading test was performed at 8% strain with 0.5% strain amplitude using 0.1, 0.5 and 1 Hz frequencies. Subsequently, an ultimate tensile test was performed to investigate the stress-strain characteristics.At 0.1 Hz, the phase difference between stress and strain was higher in LCL compared with ACL, PCL and PT (p < 0.05), and at 0.5 Hz that was higher in LCL compared with all other ligaments and PT (p < 0.05). PT had the longest toe-region strain (p < 0.05 compared with PCL and MCL) and MCL had the highest linear and strain-dependent modulus, and toughness (p < 0.05 compared with ACL, LCL and PT).The results indicate that LCL is more viscous than other ligaments at low-frequency loads. MCL was the stiffest and toughest, and its modulus increased most steeply at the toe-region, possibly implying a greater amount of collagen. This study improves the knowledge about elastic, viscoelastic and failure properties of the knee ligaments and PT.     

Successful fertilization of Varicorhinus macrolepis eggs with sperm subjected to two freeze-thaw cycles

Although sperm from several fish species have been successfully cryopreserved, few studies have been done in small and/or endangered species. The aim of the present work was to develop a method of freezing and refreezing Varicorhinus macrolepis semen in 1.8 mL cryovials. The effect of extenders and cryoprotectants on the motility of post-thaw sperm was examined. The motility of frozen-thawed sperm in extender D-15 was higher than that in MPRS and fish Ringer solution (P<0.05). Dimethyl sulfoxide (DMSO) and glycerol provided greater protection to sperm than methanol during freezing and thawing; the most effective concentration of DMSO and glycerol was 10%. The fertilization rate of frozen-thawed sperm was not significantly different from that of fresh sperm. Furthermore, mean (+/-S.D.) hatching rate did not differ significantly between frozen-thawed (82.7+/-12.4%) and fresh sperm (90.7+/-4.5%). Although frozen-thawed sperm that was immediately refrozen had 0% post-thaw motility, frozen semen that was refrozen after dilution with D-15 (containing DMSO at a ratio of 1:2) had post-thaw motility of 38.3+/-2.9%. Motility was lower for refrozen than for frozen sperm (P<0.05). Furthermore, fertilization and hatching rates of refrozen sperm were 42.9+/-6.7 and 34.1+/-10.5%, respectively, which were lower than that of fresh sperm (P<0.05).     

Strain-rate sensitivity of the rabbit MCL diminishes at traumatic loading rates

This study was performed to determine whether the viscoelastic behavior of ligaments persists at high rates of loading, such as those associated with sports-related trauma or motor vehicle accidents. Medial collateral ligaments (MCLs) from 22 skeletally mature New Zealand White rabbits were tensile tested quasi-statically and via two impact conditions at displacement rates of 0.17 mm/s (n=22), 640+/-160 mm/s (n=10) and 2500+/-270 mm/s (n=12) (corresponding to strain rates of approximately 1.0%/s, 3660%/s and 14,000%/s, respectively). Despite dramatic increases in displacement rate, only a modest strain-rate effect was observed when the specimens tested quasi-statically were compared to those tested via impact (24% and 37% increases in stiffness and failure load, respectively). There were no differences in the structural (e.g. 145+/-30 and 136+/-29 N/mm stiffness values, respectively) or failure properties (e.g. 434+/-91 and 443+/-154 N failure load values, respectively) of the two impact-tested groups. Our findings suggest that the rabbit MCL is not viscoelastic at loading rates approximating those associated with high-energy trauma.     

Structural properties of the medial collateral ligament complex of the human knee

The anatomy of the medial collateral ligament (MCL) complex consists of three identifiable passive restraining structures: the longitudinal fibres of the superficial medial collateral ligament (sMCL), the deep medial collateral ligament (dMCL), and the posteromedial capsule (PMC). The purpose of this study was to measure and compare the structural properties of these three individual structures. Eight human cadaveric knees (age 72-89 years, mean = 77 years, S.D. 5.3) were harvested and bone-ligament-bone tensile testing specimens prepared. After preconditioning, the specimens were extended to failure at 1000 mm/min in an Instron tensile testing machine. Ligament bundles failed either mid-substance or at their bony attachments. The ligament bundles had maximum loads of 534 N (sMCL), 194 N (dMCL), 425 N (PMC) and failed at 10.2, 7.1, and 12.0 mm mean extension, respectively. The maximum load and linear stiffness of the sMCL were significantly higher than those of the dMCL but not the PMC. The maximum load of the PMC was significantly higher than that of the dMCL; the linear stiffness of the PMC was higher than that of the dMCL but this did not reach statistical significance. The dMCL failed at a significantly lower extension than the other structures. The sMCL bundles that failed at their bony attachment were remounted using a freezing clamp fixture and again extended to failure, resulting in mid-substance failure at 884 N (74% higher). This study has shown that the PMC of the knee has comparable structural properties to the long superficial MCL and the short, deep MCL. In summary, the structural properties of the different component structures of the medial ligament complex indicate possible functional significance.     

Herbal remedies improve the strength of repairing ligament in a rat model.

Herbal remedies have been reported to be effective in controlling inflammation for acute soft tissue injuries. There exist, however, no reports of their effects on collagen production and remodeling; thus mechanical strength studies of the tissues have not been reported. This study tested the effects of a herbal remedy on the strength of healing medial collateral ligaments (MCL) in rats. Sixteen rats receiving surgical transection to their right MCLs and eight receiving sham operation were tested. Eight of the MCL-injured animals were treated with an adhesive herbal plaster application to their right knees, while the other eight in the MCL injured group and the sham group were treated with plain adhesive plaster to their right knees. The MCLs were harvested and tested at either 3 or 6 weeks post-operation. The ultimate tensile strength (UTS) and stiffness normalized to the uninjured side of each animal of the herb and sham groups were significantly larger than those of the control at both 3 and 6 weeks (p = 0.001). No significant difference was found in stiffness between the herb and sham groups (p > 0.05). We concluded that the herbal remedy improves the UTS and stiffness of repairing MCLs at 3 and 6 weeks after injury.     

The symmetry of the medial collateral and anterior cruciate ligament properties: a biochemical study in the rat hind limb

The medial collateral (MCL) and the anterior cruciate ligament (ACL) of the rat's knee are frequently used in biomedical research and occasionally in ligament healing studies. The contralateral normal ligament serves as a control. In this study the presence of symmetry in the biomechanical properties of the MCL and the ACL was investigated. Bilateral femur-MCL-tibia and femur-ACL-tibia preparations were obtained from the hind limbs of sixty rats and were subjected to tensile testing to failure under the same loading conditions. Tensile load to failure, stiffness and energy absorption capacity were measured and the mode of failure was recorded. All biomechanical parameters were not significantly different between the two knees of the same animal, although significant individual variation was evident. The most common mechanism of failure was mid-substance tear. Symmetry seems to exist in the biomechanical properties of the MCL and the ACL in the rat knee. When ligament healing is evaluated, increased group size is necessary and the use of a normal control group may be advisable. The contralateral normal knee ligament may serve as a control when the properties of an injured ligament are evaluated and when the parameters of tensile testing failure under similar load conditions are applied.     

Ligamentous versus physeal failure in murine medial collateral ligament biomechanical testing

This study examines the age at which a femoral physeal failure ceased to occur in a mouse model of medial collateral ligament (MCL) testing. Biomechanical testing of the MCL with load to failure can result in physeal failure rather than MCL failure in skeletally immature animals. Failure mode depended significantly on age (p<0.05). Sixty percent of the knees tested at 4 months failed at the physis rather than at the ligament, whereas, only ten percent of the knees tested at 5 and 6 months failed at the physis. The mean ultimate force to failure for the specimens in which the failure occurred at the ligament was 8.1 N with a higher values for the right side versus the left (p<0.05). For the specimens in which the failure occurred at the physis, the mean ultimate force to failure was 11.2 N. We now consider that 5 month old mice are functionally skeletally mature and old enough to be tested biomechanically with few failures at the physis.     

Maximal peak torque as a predictor of angle-specific torques of hamstring and quadriceps muscles in man.

This study assessed the relationship between the isokinetic peak torque (PT) (speed of movement 1.05 and 3.14 rads-1) and the angle-specific torques (ASTs) at 0.26 and 1.31 rad of knee flexion in multiple contractions of the quadriceps and hamstrings in 70 individuals with a chronic anterior cruciate ligament (ACL) insufficiency and 78 individuals with a chronic medial collateral ligament (MCL) insufficiency in one knee. At every test speed, the Pearson product moment correlation coefficients (r) between the PT and ASTs were highly significant (P less than 0.001) in the uninjured knees (r = 0.61-0.93) as well as in the knees with ACL (r = 0.61-0.87) and MCL (r = 0.74-0.91) insufficiency. In addition, in both groups the majority of the correlation coefficients exceeded 0.80, which is generally regarded as the threshold for the relationship to be considered clinically significant. Furthermore, using regression analysis, both extremities showed completely non-systematic distribution of the residuals. It is concluded that in healthy knees or knees with ACL or MCL insufficiency, the predictability of ASTs from PT was good, and, therefore, that AST analyses may offer little additional information about thigh muscle function to that obtained from a simpler and more commonly used measurement, the PT analysis.     

A biomechanical assessment to evaluate breed differences in normal porcine medial collateral ligaments

Little information is available on the role of genetic factors and heredity in normal ligament behaviour and their ability to heal. Assessing these factors is challenging because of the lack of suitable animal models. Therefore, the purpose of this study was to develop a porcine model in order to evaluate and compare the biomechanical differences of normal medial collateral ligaments (MCLs) between Yorkshire (YK) and red Duroc (RD) breeds. It was hypothesized that biomechanical differences would not exist between normal YK and RD MCLs. Comparisons between porcine and human MCL were also made. A biomechanical testing apparatus and protocol specific to pig MCL were developed. Ligaments were subjected to cyclic and static creep tests and then elongated to failure. Pig MCL morphology, geometry, and low- and high-load mechanical behaviour were assessed. The custom-designed apparatus and protocol were sufficiently sensitive to detect mechanical property differences between breeds as well as inter-leg differences. The results reveal that porcine MCL is comparable in both shape and size to human MCL and exhibits similar structural and material failure properties, thus making it a feasible model. Comparisons between RD and YK breeds revealed that age-matched RD pigs weigh more, have larger MCL cross-sectional area, and have lower MCL failure stress than YK pigs. The effect of weight may have influenced MCL geometrical and biomechanical properties, and consequently, the differences observed may be due to breed type and/or animal weight. In conclusion, the pig serves as a suitable large animal model for genetic-related connective tissue studies.     

Medical collateral ligament healing subsequent to different treatment regimens

The response of transected canine medical collateral ligaments (MCL) to clinical treatment regimens was investigated. These regimens included no surgical repair with no immobilization and surgical repair with various periods of immobilization. The biomechanical, biochemical, and histological properties of the healing MCL were examined 6 and 12 wk postoperatively. At 6 wk, all healing MCLs had increased cellularity with decreased levels of total collagen and increased amounts of reducible Schiff base cross-links and type III collagen. Biomechanically, the varus-valgus (V-V) knee laxity was significantly increased, and no group achieved normal structural or mechanical properties. At 12 wk the histological appearance of the MCL became more normal but still had increased cellularity. Biochemically, the total collagen levels in experimental MCLs were not statistically different from the controls, but these MCLs still had high amounts of type III collagen and an even higher number of reducible cross-links. From knees in which the MCL was not treated, the V-V knee laxity and the ultimate loads of the femur-MCL-tibia complex achieved normal values. However, the stress-strain properties for these MCLs and those treated with repair and immobilization did not completely recover.     

The influence of ligament modelling strategies on the predictive capability of finite element models of the human knee joint

In finite element (FE) models knee ligaments can represented either by a group of one-dimensional springs, or by three-dimensional continuum elements based on segmentations. Continuum models closer approximate the anatomy, and facilitate ligament wrapping, while spring models are computationally less expensive. The mechanical properties of ligaments can be based on literature, or adjusted specifically for the subject. In the current study we investigated the effect of ligament modelling strategy on the predictive capability of FE models of the human knee joint. The effect of literature-based versus specimen-specific optimized material parameters was evaluated. Experiments were performed on three human cadaver knees, which were modelled in FE models with ligaments represented either using springs, or using continuum representations. In spring representation collateral ligaments were each modelled with three and cruciate ligaments with two single-element bundles. Stiffness parameters and pre-strains were optimized based on laxity tests for both approaches. Validation experiments were conducted to evaluate the outcomes of the FE models.Models (both spring and continuum) with subject-specific properties improved the predicted kinematics and contact outcome parameters. Models incorporating literature-based parameters, and particularly the spring models (with the representations implemented in this study), led to relatively high errors in kinematics and contact pressures. Using a continuum modelling approach resulted in more accurate contact outcome variables than the spring representation with two (cruciate ligaments) and three (collateral ligaments) single-element-bundle representations. However, when the prediction of joint kinematics is of main interest, spring ligament models provide a faster option with acceptable outcome.     

A comparative evaluation of the mechanical properties of the rabbit medial collateral and anterior cruciate ligaments.

The biomechanical properties of the medial collateral and anterior cruciate ligaments from 30 New Zealand White rabbits were measured. Because of its complex geometry, the ACL was divided into two portions (medial and lateral) to provide uniform loading. This allowed an examination of the intra-ligamentous properties. A laser micrometer system was used to measure the cross-sectional area for tensile stress and a video dimension analyzer was used to measure the strain. The mechanical properties (stress-strain curves) of the MCL and ACL were different, with the modulus (determined between 4 and 7% strain) in the MCL (1120 +/- 153 MPa) more than twice that of either portion of the ACL (516 +/- 64 and 516 +/- 69 MPa for the medial and lateral portions, respectively). This higher modulus correlated with the more uniform and dense appearance of the collagen fibrils examined with scanning electron microscopy (SEM).     

Effect of ACL deficiency on MCL strains and joint kinematics

The knee joint is partially stabilized by the interaction of multiple ligament structures. This study tested the interdependent functions of the anterior cruciate ligament (ACL) and the medial collateral ligament (MCL) by evaluating the effects of ACL deficiency on local MCL strain while simultaneously measuring joint kinematics under specific loading scenarios. A structural testing machine applied anterior translation and valgus rotation (limits 100 N and 10 N m, respectively) to the tibia of ten human cadaveric knees with the ACL intact or severed. A three-dimensional motion analysis system measured joint kinematics and MCL tissue strain in 18 regions of the superficial MCL. ACL deficiency significantly increased MCL strains by 1.8% (p<0.05) during anterior translation, bringing ligament fibers to strain levels characteristic of microtrauma. In contrast, ACL transection had no effect on MCL strains during valgus rotation (increase of only 0.1%). Therefore, isolated valgus rotation in the ACL-deficient knee was nondetrimental to the MCL. The ACL was also found to promote internal tibial rotation during anterior translation, which in turn decreased strains near the femoral insertion of the MCL. These data advance the basic structure-function understanding of the MCL, and may benefit the treatment of ACL injuries by improving the knowledge of ACL function and clarifying motions that are potentially harmful to secondary stabilizers.     

Comparison of material properties in fascicle-bone units from human patellar tendon and knee ligaments

The fascicle material properties in bone-fascicle-bone units were determined for the anterior and posterior cruciate ligaments (ACL, PCL), the lateral collateral ligament (LCL) and the patellar tendon (PT) from three young human donor knees. Groups of fascicles from each tissue were isolated with intact bone ends and failed at a high strain rate in a saline bath at 37 degrees C. In each knee tested the load related material properties (linear modulus, maximum stress and energy density to maximum stress) for the patellar tendon were significantly larger than corresponding values for the cruciate and collateral ligaments. Bundles from different ligaments in the same knee were similar to each other in their mechanical behavior. In addition, no significant differences were present in the maximum strains recorded for any of the four tissue types examined. The results presented have implications in studies of ligament injury. They are also important in the design and use of synthetic and biological ligament replacements and in tissue and whole knee modeling.     

An improved OpenSim gait model with multiple degrees of freedom knee joint and knee ligaments

Musculoskeletal models are widely used to investigate joint kinematics and predict muscle force during gait. However, the knee is usually simplified as a one degree of freedom joint and knee ligaments are neglected. The aim of this study was to develop an OpenSim gait model with enhanced knee structures. The knee joint in this study included three rotations and three translations. The three knee rotations and mediolateral translation were independent, with proximodistal and anteroposterior translations occurring as a function of knee flexion/extension. Ten elastic elements described the geometrical and mechanical properties of the anterior and posterior cruciate ligaments (ACL and PCL), and the medial and lateral collateral ligaments (MCL and LCL). The three independent knee rotations were evaluated using OpenSim to observe ligament function. The results showed that the anterior and posterior bundles of ACL and PCL (aACL, pACL and aPCL, pPCL) intersected during knee flexion. The aACL and pACL mainly provided force during knee flexion and adduction, respectively. The aPCL was slack throughout the range of three knee rotations; however, the pPCL was utilised for knee abduction and internal rotation. The LCL was employed for knee adduction and rotation, but was slack beyond 20° of knee flexion. The MCL bundles were mainly used during knee adduction and external rotation. All these results suggest that the functions of knee ligaments in this model approximated the behaviour of the physical knee and the enhanced knee structures can improve the ability to investigate knee joint biomechanics during various gait activities.     

ACL deficiency impairs the vasoconstrictive efficacy of neuropeptide Y and phenylephrine in articular tissues: a laser speckle perfusion imaging study.

Sympathetic-derived neuropeptide Y (NPY) helps regulate inflammatory responses in injury and disease, is a vasoconstrictor, and stimulates angiogenesis. Rupture of the anterior cruciate ligament (ACL) is a common clinical presentation that results in tissue inflammation, hyperemia, and angiogenesis in the intact medial collateral ligament (MCL). This study is the first to examine the vasoregulatory role of NPY in ACL-deficient knee joints by using the newly developed technique of laser speckle perfusion imaging (LSPI). MCL blood flow was measured in two groups of adult rabbits: unoperated control (n = 6), and 6-wk ACL transected (n = 5). Under anesthesia, the MCL was surgically exposed and tissue blood flow was imaged at high resolution using LSPI. NPY was applied to the MCL vasculature in topical boluses of 100 mul (dose range 10(-14) to 10(-9) mol), and the alpha-adrenoceptor agonist phenylephrine was applied in doses of 10(-14), 10(-10), and 10(-7) mol. In control rabbits, topical administration of NPY or phenylephrine produced dose-dependent vasopressor responses (maximal effect at 10(-9) mol NPY and 10(-7) mol phenylephrine). In ACL-transected knees, there was little or no vasoconstrictive response to NPY at any dose. The response to phenylephrine was significantly reduced compared with control ligaments. Possible causes of the reduced vasoconstrictive response to NPY in the MCL after 6 wk of ACL deficiency include development of tolerance to the peptide due to a prolonged increase in sympathetic nerve activity or change in the distribution or functionality of the NPY Y(1) receptors. Chronic ACL deficiency leads to profound and protracted hyperemia in associated articular tissues. Abrogation of a vasoconstrictor response to both NPY and phenylephrine in the MCL indicates that ACL deficiency induces major changes in the vascular physiological homeostasis.     

Endothelial dysfunction and decreased vascular responsiveness in the anterior cruciate ligament-deficient model of osteoarthritis.

Chronic inflammation associated with osteoarthritis (OA) may alter normal vascular responses and contribute to joint degradation. Vascular responses to vasoactive mediators were evaluated in the medial collateral ligament (MCL) of the anterior cruciate ligament (ACL)-deficient knee. Chronic joint instability and progressive OA were induced in rabbit knees by surgical transection of the ACL. Under halothane anesthesia, laser speckle perfusion imaging (LSPI) was used to measure MCL blood flow in unoperated control (n = 12) and 6-wk ACL-transected knees (n = 12). ACh, bradykinin, histamine, substance P (SP), and prostaglandin E(2) (PGE(2)) were applied to the MCL vasculature in topical boluses of 100 microl (dose range 10(-14) to 10(-8) mol). In normal joints, ACh, bradykinin, histamine, and PGE(2) evoked a dilatory response. Substance P caused a biphasic response that was dilatory from 10(-14) to 10(-11) mol and constricting at higher doses. In ACL-deficient knees, ACh, bradykinin, histamine, and SP decreased perfusion, whereas PGE(2) had a biphasic response that decreased perfusion at 10(-14) to 10(-11) mol and was dilatory at higher concentrations. Sodium nitroprusside increased perfusion in resting and phenylephrine-precontracted vessels with no significant differences between ACL-transected and control knees. Femoral artery occlusion and release increased perfusion by 74.3 +/- 11.1% in control knees but only by 25.8 +/- 4.4% in ACL-deficient knees. The altered responsiveness of the MCL vasculature to these inflammatory mediators may indicate endothelial dysfunction in the MCL, which may contribute to the progression and severity of OA and to the adaptation of the joint in an altered mechanical environment.