Structure and Function Are Not the Same: The Case for Restoring Mechanoreceptor Continuity Following Anterior Cruciate Ligament Injury.

Anterior cruciate ligament (ACL) injury, particularly in increasingly young and active adolescents, continues to pose a clinical challenge with re-injury rates reported as high as 30%. Evidence also suggests that current standard-of-care ACL reconstruction (ACLR) does not mitigate post-traumatic osteoarthritis (PTOA) risk. Bridge- enhanced ACL restoration (BEAR) is a recently developed and tested ACL surgery that promotes primary healing of the native ACL with excellent early results. BEAR has shown to reduce signs of early PTOA compared to ACLR in an animal model. Here, we describe a theoretical framework related to re-innervation that can clarify why the outcomes of ACLR and BEAR surgeries differ. We also discuss how ongoing and new challenges in determining return-to-sport readiness following the competing surgeries may differ, and how emerging imaging tools and measures of neuromuscular function may aid in clinical decision-making to decrease the likelihood of re-injury and PTOA risk.

Assessment of ultrashort echo time (UTE) T2* mapping at 3T for the whole knee: repeatability, the effects of fat suppression, and knee position.

Background: Knee tissues such as tendon, ligament and meniscus have short T2* relaxation times and tend to show little to no signal in conventional magnetic resonance acquisitions. An ultrashort echo time (UTE) technique offers a unique tool to probe fast-decaying signals in these tissues. Clinically relevant factors should be evaluated to quantify the sensitivity needed to distinguish diseased from control tissues. Therefore, the objectives of this study were to (I) quantify the repeatability of UTE-T2* relaxation time values, and (II) evaluate the effects of fat suppression and (III) knee positioning on UTE-T2* relaxation time quantification. Methods: A dual-echo, three-dimensional center-out radially sampling UTE and conventional gradient echo sequences were utilized to image gadolinium phantoms, one ex-vivo specimen, and five in-vivo subjects on a clinical 3T scanner. Scan-rescan images from the phantom and in-vivo experiments were used to evaluate the repeatability of T2* relaxation time values. Fat suppressed and non-suppressed images were acquired for phantoms and the ex-vivo specimen to evaluate the effect of fat suppression on T2* relaxation time quantifications. The effect of knee positioning was evaluated by imaging in-vivo subjects in extended and flexed positions within the knee coil and comparing T2* relaxation times quantified from tissues in each position. Results: Phantom and in-vivo measurements demonstrated repeatable T2* mapping, where the percent difference between T2* relaxation time quantified from scan-rescan images was less than 8% for the phantom and knee tissues. The coefficient of variation across fat suppressed and non-suppressed images was less than 5% for the phantoms and ex-vivo knee tissues, showing that fat suppression had a minimal effect on T2* relaxation time quantification. Knee position introduced variability to T2* quantification of the anterior cruciate ligament, posterior cruciate ligament, and patellar tendon, with percent differences exceeding 20%, but the meniscus showed a percent difference less than 10%. Conclusions: The 3D radial UTE sequence presented in this study could potentially be used to detect clinically relevant changes in mean T2* relaxation time, however, reproducibility of these values is impacted by knee position consistency between scans.

A novel large animal model of posttraumatic osteoarthritis induced by inflammation with mechanical stability.

OBJECTIVES: Animal models are needed to reliably separate the effects of mechanical joint instability and inflammation on posttraumatic osteoarthritis (PTOA) pathogenesis. We hypothesized that our modified intra-articular drilling (mIAD) procedure induces cartilage damage and synovial changes through increased inflammation without causing changes in gait. METHODS: Twenty-four Yucatan minipigs were randomized into the mIAD (n=12) or sham control group (n=12). mIAD animals had two osseous tunnels drilled into each of the tibia and femur adjacent to the anterior cruciate ligament (ACL) attachment sites on the left hind knee. Surgical and contralateral limbs were harvested 15 weeks post-surgery. Cartilage degeneration was evaluated macroscopically and histologically. Synovial changes were evaluated histologically. Interleukin-1 beta (IL-1ß), nuclear factor kappa B (NF-κB), and tumor necrosis factor alpha (TNF-α) mRNA expression levels in the synovial membrane were measured using quantitative real-time polymerase chain reaction. IL-1ß and NF-κB levels in chondrocytes were assessed using immunohistochemistry. Load asymmetry during gait was recorded by a pressure-sensing walkway system before and after surgery. RESULTS: The mIAD surgical knees demonstrated greater gross and histological cartilage damage than contralateral (P<.01) and sham knees (P<.05). Synovitis was present only in the mIAD surgical knee. Synovial inflammatory marker (IL-1ß, NF-κB, and TNF-α) expression was three times higher in the mIAD surgical knee than the contralateral (P<.05). Chondrocyte IL-1ß and NF-κB levels were highest in the mIAD surgical knee. In general, there were no significant changes in gait. CONCLUSIONS: The mIAD model induced PTOA through inflammation without affecting gait mechanics. This large animal model has significant applications for evaluating the role of inflammation in PTOA and for developing therapies aimed at reducing inflammation following joint injury.

Long-Term Bilateral Neuromuscular Function and Knee Osteoarthritis after Anterior Cruciate Ligament Reconstruction.

Neuromuscular function is thought to contribute to posttraumatic osteoarthritis (PTOA) risk in anterior cruciate ligament (ACL)-reconstructed (ACLR) patients, but sensitive and easy-to-use tools are needed to discern whether complex muscle activation strategies are beneficial or maladaptive. Using an electromyography (EMG) signal analysis technique coupled with a machine learning approach, we sought to: (1) identify whether ACLR muscle activity patterns differed from those of healthy controls, and (2) explore which combination of patient outcome measures (thigh muscle girth, knee laxity, hop distance, and activity level) predicted the extent of osteoarthritic changes via magnetic resonance imaging (MRI) in ACLR patients. Eleven ACLR patients 10-15 years post-surgery and 12 healthy controls performed a hop activity while lower limb muscle EMG was recorded bilaterally. Osteoarthritis was evaluated based on MRI. ACLR muscle activity patterns were bilaterally symmetrical and differed from those of healthy controls, suggesting the presence of a global adaptation strategy. Smaller ipsilateral thigh muscle girth was the strongest predictor of inferior MRI scores. The ability of our EMG analysis approach to detect meaningful neuromuscular differences that could ultimately be related to thigh muscle girth provides the foundation to further investigate a direct link between muscle activation patterns and PTOA risk.

Predicting anterior cruciate ligament failure load with T2* relaxometry and machine learning as a prospective imaging biomarker for revision surgery.

Non-invasive methods to document healing anterior cruciate ligament (ACL) structural properties could potentially identify patients at risk for revision surgery. The objective was to evaluate machine learning models to predict ACL failure load from magnetic resonance images (MRI) and to determine if those predictions were related to revision surgery incidence. It was hypothesized that the optimal model would demonstrate a lower mean absolute error (MAE) than the benchmark linear regression model, and that patients with a lower estimated failure load would have higher revision incidence 2 years post-surgery. Support vector machine, random forest, AdaBoost, XGBoost, and linear regression models were trained using MRI T2* relaxometry and ACL tensile testing data from minipigs (n = 65). The lowest MAE model was used to estimate ACL failure load for surgical patients at 9 months post-surgery (n = 46) and dichotomized into low and high score groups via Youden's J statistic to compare revision incidence. Significance was set at alpha = 0.05. The random forest model decreased the failure load MAE by 55% (Wilcoxon signed-rank test: p = 0.01) versus the benchmark. The low score group had a higher revision incidence (21% vs. 5%; Chi-square test: p = 0.09). ACL structural property estimates via MRI may provide a biomarker for clinical decision making.

Wavelet analysis reveals differential lower limb muscle activity patterns long after anterior cruciate ligament reconstruction.

The purpose of this study was to test whether differences in muscle activity patterns between anterior cruciate ligament-reconstructed patients (ACLR) and healthy controls could be detected 10 to 15 years post-surgery using a machine learning classification approach. Eleven ACLR subjects and 12 healthy controls were recruited from an ongoing prospective randomized clinical trial. Surface electromyography (EMG) signals were recorded from gastrocnemius medialis and lateralis, tibialis anterior, vastus medialis, rectus femoris, biceps femoris, and semitendinosus muscles. Muscle activity was analyzed using wavelet analysis and examined within four sub-phases of the hop test, as well as an average of the task as a whole. K-nearest neighbor machine learning combined with a leave-one-out validation was used to classify the muscle activity patterns as either ACLR or Control. When muscle activity was averaged across the whole hop task, activity patterns for all muscles except the tibialis anterior were identified as being different between the study cohorts. ACLR patients demonstrated continuous muscle activities that spanned take-off, airborne, and landing hop phases versus healthy controls who displayed timed and regulated islets of muscle activities specific to each hop phase. The most striking features were 25-50% greater relative quadriceps intensity and approximately 66% diminished biceps femoris intensity in ACLR patients. The current findings are in contrast to previous work using conventional co-contraction and muscle activation onset EMG measures of the same dataset, underscoring the sensitivity and potential of the wavelet approach coupled with machine learning to reveal meaningful adaptation strategies in this at-risk population.

A transfer learning approach for automatic segmentation of the surgically treated anterior cruciate ligament.

Quantitative magnetic resonance imaging enables quantitative assessment of the healing anterior cruciate ligament or graft post-surgery, but its use is constrained by the need for time consuming manual image segmentation. The goal of this study was to validate a deep learning model for automatic segmentation of repaired and reconstructed anterior cruciate ligaments. We hypothesized that (1) a deep learning model would segment repaired ligaments and grafts with comparable anatomical similarity to intact ligaments, and (2) automatically derived quantitative features (i.e., signal intensity and volume) would not be significantly different from those obtained by manual segmentation. Constructive Interference in Steady State sequences were acquired of ACL repairs (n = 238) and grafts (n = 120). A previously validated model for intact ACLs was retrained on both surgical groups using transfer learning. Anatomical performance was measured with Dice coefficient, sensitivity, and precision. Quantitative features were compared to ground truth manual segmentation. Automatic segmentation of both surgical groups resulted in decreased anatomical performance compared to intact ACL automatic segmentation (repairs/grafts: Dice coefficient = .80/.78, precision = .79/.78, sensitivity = .82/.80), but neither decrease was statistically significant (Kruskal-Wallis: Dice coefficient p = .02, precision p = .09, sensitivity p = .17; Dunn post-hoc test for Dice coefficient: repairs/grafts p = .054/.051). There were no significant differences in quantitative features between the ground truth and automatic segmentation of repairs/grafts (0.82/2.7% signal intensity difference, p = .57/.26; 1.7/2.7% volume difference, p = .68/.72). The anatomical similarity performance and statistical similarities of quantitative features supports the use of this automated segmentation model in quantitative magnetic resonance imaging pipelines, which will accelerate research and provide a step towards clinical applicability.

Cartilage Damage Is Related to ACL Stiffness in a Porcine Model of ACL Repair.

Inferior anterior cruciate ligament (ACL) structural properties may inadequately restrain tibiofemoral joint motion following surgery, contributing to the increased risk of post-traumatic osteoarthritis. Using both a direct measure of ACL linear stiffness and an in vivo magnetic resonance imaging (MRI) T2 *-based prediction model, we hypothesized that cartilage damage and ACL stiffness would increase over time, and that an inverse relationship between cartilage damage and ACL stiffness would emerge at a later stage of healing. After either 6, 12, or 24 weeks (w) of healing after ACL repair, ACL linear stiffness was determined from the force-displacement relationship during tensile testing ex vivo and predicted in vivo from the MRI T2 *-based multiple linear regression model in 24 Yucatan minipigs. Tibiofemoral cartilage was graded postmortem. There was no relationship between cartilage damage and ACL stiffness at 6 w (R2 = 0.04; p = 0.65), 12 w (R2 = 0.02; p = 0.77), or when the data from all animals were pooled (R2 = 0.02; p = 0.47). A significant inverse relationship between cartilage damage and ACL stiffness based on both ex vivo measurement (R2 = 0.90; p < 0.001) and in vivo MRI prediction (R2 = 0.78; p = 0.004) of ACL stiffness emerged at 24 w. This result suggests that 90% of the variability in gross cartilage changes is associated with the repaired ACL linear stiffness at 6 months of healing. Clinical Significance: Techniques that provide a higher stiffness to the repaired ACL may be required to mitigate the post-traumatic osteoarthritis commonly seen after ACL injury, and MRI T2 * can be used as a noninvasive estimation of ligament stiffness. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:2249-2257, 2019.

Osteoarthritis develops in the operated joint of an ovine model following ACL reconstruction with immediate anatomic reattachment of the native ACL.

We tested the hypothesis that immediate reattachment of the native anterior cruciate ligament (ACL) can prevent kinematic changes and the development of osteoarthritis (OA). Five sheep underwent anatomic unilateral ACL reconstruction (ACL-R). Animals from a previous study served as sham (n = 7) or non-operated (n = 17) controls. At 4 points of walking gait, 6 degrees of freedom stifle joint kinematics of ACL-R animals were compared with sham controls at 4 and 20 weeks post-surgery. Gross cartilage, bone, and meniscal changes were graded at euthanasia; paired and differential scores were compared. Inter-animal differences were noted in all groups. Of 48 points of gait comparison between ACL-R and sham operated groups, 42 points showed no difference (p > 0.05). Of the six significant differences (p < 0.05), internal rotation in ACL-R animals accounted for three. At 20 weeks, differential scores showed that sham operated joints were morphologically indistinguishable from non-operated controls (p ≥ 0.129) while ACL-R joints had significantly higher combined cartilage and osteophyte scores than those controls (p ≤ 0.003). This method of ACL reconstruction in sheep did not restore normal walking gait kinematics completely and allowed some OA to develop in operated joints. OA may result from relatively subtle mechanical abnormalities, apparently more so in some individuals than others.

Complete ACL/MCL deficiency induces variable degrees of instability in sheep with specific kinematic abnormalities correlating with degrees of early osteoarthritis.

People are not equally disabled by combined anterior cruciate ligament (ACL)/medial collateral ligament (MCL) injuries, nor do they all develop osteoarthritis (OA). Although biological/biomechanical causes are not clear, some association presumably exists between joint instability and OA development. We hypothesized that degree of OA development following standardized complete ACL/MCL injuries will vary directly with the degree of biomechanical abnormality between individuals. Three groups of sheep were used to test the hypothesis: 17 normal, 9 ACL/MCL transected, and 7 sham animals. Normal joints were assessed morphologically while sham and experimental animals had gait assessment pre- and at 4 and 20 weeks post-surgery, with cartilage and bone changes being mapped and graded at sacrifice at 20 weeks. Sham joints were morphologically normal and had only one minor kinematic change at 20 weeks. Although variable, ACL/MCL deficient animals showed significant kinematic abnormalities in 4/6 degrees of freedom (DOFs), as well as cartilage/bone damage by 20 weeks (p < 0.05). Linear regression analysis revealed that changes in medial-lateral (ML) translation were related to the current level of joint degradation as represented by total gross OA score (p = 0.0044, R(2) = 0.71) in the ACL/MCL transected group. Even identical ACL/MCL injuries result in inter-animal variations in instability and OA, however significant kinematic abnormalities in ML translation do relate to early OA in sheep.

Meniscectomy causes significant in vivo kinematic changes and mechanically induced focal chondral lesions in a sheep model.

Significant meniscal loss with progression to osteoarthritis is common in humans. In vitro work suggests that meniscectomy causes increased joint contact stress, but what other alterations in dynamic joint actions actually occur remains unknown. In a sheep model, we tested the hypothesis that complete lateral meniscectomy increases joint abduction, shifting the in vivo locations of tibiofemoral contact to regions that qualitatively correspond to locations of chondral damage. Nine sheep underwent unilateral arthrotomy (n = 4) or arthrotomy plus complete lateral meniscectomy (n = 5). Kinematics were collected prior to surgery and serially up to 20 weeks post-surgery. Gross cartilage damage was mapped in each joint, graded using a published scoring scheme used in goats, and compared to the locations of minimum tibiofemoral distance. Over the 20 weeks, meniscectomy caused increased stifle abduction and medial tibial translation, shifting the points of minimum tibiofemoral distance 7.5 ± 2.1 mm laterally and 3.3 ± 1.1 mm anteriorly (mean ± SEM), which corresponded to the locations of focal chondral damage. Locations of new tibiofemoral contact in the meniscectomized compartment qualitatively correspond to subject-specific locations of early chondral damage in an ovine model.