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Background: Posttraumatic osteoarthritis (PTOA) is a common sequela of anterior cruciate ligament (ACL) injury, even when surgical treatment is selected. The effect of patient sex on cartilage health after ACL injury and surgical treatment has been less studied. Purpose/Hypothesis: The study objective was to compare the macroscopic cartilage damage that develops after ACL surgery in male and female Yucatan minipigs. It was hypothesized that after ACL surgery, the macroscopic cartilage damage of the tibiofemoral joints from female animals would be greater than that from male animals. Additionally, it was hypothesized that the effect of sex on the macroscopic cartilage damage would depend on surgical treatment. Study Design: Controlled laboratory study Methods: Twelve-month follow-up data were obtained for 55 adolescent Yucatan minipigs (22 female/33 male) that were randomized to 1 of 3 experimental groups: no treatment (ACL transection [ACLT]), ACL reconstruction, and bridge-enhanced ACL restoration. The Osteoarthritis Research Society International guidelines were used to determine a standardized macroscopic cartilage damage score on 5 surfaces of the knee joint. Results: Females had significantly worse mean total macroscopic cartilage damage scores on the surgical side (adjusted P value [ P adj] = .04) and significantly better scores on the contralateral side ( P adj = .01) when compared with males. The trochlear damage scores were also significantly worse in females for surgical limbs ( P adj = .009) and significantly better for the contralateral limbs ( P adj < .001) when compared with males. Although there were no significant differences in total macroscopic cartilage damage scores between sexes within treatment groups on the surgical limbs (ACLT, P adj = 0.45; ACL reconstruction, P adj = .56; bridge-enhanced ACL restoration, P adj = .23), the mean trochlear scores on the surgical limb of females were significantly worse than those of the males in the ACLT group ( P adj = .003). Conclusion: Mean total macroscopic cartilage damage scores of Yucatan minipigs were significantly worse in females than males, regardless of treatment. These differences were predominantly found in the trochlear scores across all treatment groups. Clinical Relevance: These data suggest that patient sex could be more influential in the progression of PTOA than surgical treatment after ACL injury. Identifying factors responsible for this discrepancy may prove valuable to identify targets to slow PTOA progression in male and female ACL-injured populations.

Background: Anterior cruciate ligament (ACL) revision surgery is challenging for both patients and surgeons. Understanding the risk factors for failure after bridge-enhanced ACL restoration (BEAR) may help with patient selection for ACL restoration versus ACL reconstruction. Purpose: To identify the preoperative risk factors for ACL revision surgery within the first 2 years after BEAR. Study Design: Case-control study; Level of evidence, 3. Methods: Data from the prospective BEAR I, II, and III trials were used to determine the preoperative risk factors for ACL revision surgery. All patients with a complete ACL tear (aged 13-47 years, depending on the trial), who met all other inclusion/exclusion criteria and underwent a primary BEAR procedure within 30 to 50 days from the injury (dependent on the trial), were included. Demographic data (age, sex, body mass index), baseline patient-reported outcomes (International Knee Documentation Committee [IKDC] subjective score, Marx activity score), preoperative imaging results (ACL stump length, notch size, tibial slope), and intraoperative findings (knee hyperextension, meniscal status) were evaluated to determine their contribution to the risk of ipsilateral ACL revision surgery. Results: A total of 123 patients, with a median age of 17.6 years (interquartile range, 16-23 years), including 67 (54%) female patients, met study criteria. Overall, 18 (15%) patients required ACL revision surgery in the first 2 years after the BEAR procedure. On bivariate analyses, younger age ( P = .011), having a contact injury at the time of the initial tear ( P = .048), and increased medial tibial slope (MTS; P = .029) were associated with a higher risk of ipsilateral revision surgery. Multivariable logistic regression analyses identified 2 independent predictors of revision: patient age and MTS. The odds of ipsilateral revision surgery were decreased by 32% for each 1-year increase in age (odds ratio, 0.684 [95% CI, 0.517-0.905]; P = .008) and increased by 28% for each 1° increase in MTS (odds ratio, 1.280 [95% CI, 1.024-1.601]; P = .030). Sex, baseline IKDC or Marx score, knee hyperextension, and meniscal status were not significant predictors of revision. Conclusion: Younger age and higher MTS were predictors of ipsilateral ACL revision surgery after the BEAR procedure. Younger patients with higher tibial slopes should be aware of the increased risk for revision surgery when deciding to undergo ACL restoration.

Anterior cruciate ligament (ACL) injuries occur at a high frequency in the United States with approximately 400,000 ACL reconstructions being performed each year. While ACL reconstruction is our current gold standard of treatment, it does not restore joint motion, or prevent the premature development of posttraumatic osteoarthritis (PTOA) in many patients. Thus, new treatments for an ACL injury, which are less invasive and minimize patient morbidity, including cartilage damage, are highly desirable. We have used a tissue-engineered approach to stimulate ligament healing, to improve upon current treatment options. In this review, we describe and discuss our work moving a tissue engineering strategy from the concept to bench, preclinical, clinical trials and ultimately FDA 510(k) de Novo approval, providing clinicians and patients with a viable alternative to ACL reconstruction.

Smaller anterior cruciate ligament (ACL) size in females has been hypothesized to be a key contributor to a higher incidence of ACL tears in that population, as a lower cross-sectional area (CSA) directly corresponds to a larger stress on the ligament for a given load. Prior studies have used a mid-length CSA measurement to quantify ACL size. In this study, we used magnetic resonance imaging to quantify the CSA along the entire length of the intact ACL. We hypothesized that changes in the ACL CSA along its length would have different patterns in males and females. We also hypothesized that changes in ACL CSA along its length would be associated with body size or knee size with different associations in females and males. MR images of contralateral ACL-intact knees of 108 patients (62 females, 13-35 years) undergoing ACL surgery were used to measure the CSA along the ACL length, using a custom program. For both females and males, the largest CSA was located at 37%-39% of ACL length from the tibial insertion. Compared to females, males had a significantly larger CSA only within the distal 41% of the ACL (p 0.001). ACL CSA was associated with patient height and weight in males (r 0.3; p 0.05), whereas it was associated with intercondylar notch width in females (r 0.3; p 0.05). These findings highlight the importance of standardizing the location of measurement of ACL CSA.

Background:Preclinical studies suggest that for complete midsubstance anterior cruciate ligament (ACL) injuries, a suture repair of the ACL augmented with a protein implant placed in the gap between the torn ends (bridge-enhanced ACL repair [BEAR]) may be a viable alternative to ACL reconstruction (ACLR).Hypothesis:We hypothesized that patients treated with BEAR would have a noninferior patient-reported outcomes (International Knee Documentation Committee [IKDC] Subjective Score; prespecified noninferiority margin, –11.5 points) and instrumented anteroposterior (AP) knee laxity (prespecified noninferiority margin, +2-mm side-to-side difference) and superior muscle strength at 2 years after surgery when compared with patients who underwent ACLR with autograft.Study Design:Randomized controlled trial; Level of evidence, 1.Methods:One hundred patients (median age, 17 years; median preoperative Marx activity score, 16) with complete midsubstance ACL injuries were enrolled and underwent surgery within 45 days of injury. Patients were randomly assigned to receive either BEAR (n = 65) or autograft ACLR (n = 35 [33 with quadrupled semitendinosus-gracilis and 2 with bone–patellar tendon–bone]). Outcomes—including the IKDC Subjective Score, the side-to-side difference in instrumented AP knee laxity, and muscle strength—were assessed at 2 years by an independent examiner blinded to the procedure. Patients were unblinded after their 2-year visit.Results:In total, 96% of the patients returned for 2-year follow-up. Noninferiority criteria were met for both the IKDC Subjective Score (BEAR, 88.9 points; ACLR, 84.8 points; mean difference, 4.1 points [95% CI, –1.5 to 9.7]) and the side-to-side difference in AP knee laxity (BEAR, 1.61 mm; ACLR, 1.77 mm; mean difference, –0.15 mm [95% CI, –1.48 to 1.17]). The BEAR group had a significantly higher mean hamstring muscle strength index than the ACLR group at 2 years (98.2% vs 63.2%; P < .001). In addition, 14% of the BEAR group and 6% of the ACLR group had a reinjury that required a second ipsilateral ACL surgical procedure ( P = .32). Furthermore, the 8 patients who converted from BEAR to ACLR in the study period and returned for the 2-year postoperative visit had similar primary outcomes to patients who had a single ipsilateral ACL procedure.Conclusion:BEAR resulted in noninferior patient-reported outcomes and AP knee laxity and superior hamstring muscle strength when compared with autograft ACLR at 2-year follow-up in a young and active cohort. These promising results suggest that longer-term studies of this technique are justified.Registration:NCT02664545 (ClinicalTrials.gov identifier)

To determine the transcriptomic changes seen in early- to mid-stage posttraumatic osteoarthritis (PTOA) development, 72 Yucatan minipigs underwent transection of the anterior cruciate ligament. Subjects were randomized to no further intervention, ligament reconstruction, or ligament repair, followed by articular cartilage harvesting and RNA-sequencing at three different postoperative timepoints (1, 4, and 52 weeks). Six additional subjects received no ligament transection and provided cartilage tissue to serve as controls. Differential gene expression analysis between post-transection cartilage and healthy cartilage revealed an initial increase in transcriptomic differences at 1 and 4 weeks followed by a stark reduction in transcriptomic differences at 52 weeks. This analysis also showed how different treatments genetically modulate the course of PTOA following ligament disruption. Specific genes (e.g., MMP1, POSTN, IGF1, PTGFR, HK1) were identified as being upregulated in the cartilage of injured subjects across all timepoints regardless of treatment. At the 52-week timepoint, 4 genes (e.g., A4GALT, EFS, NPTXR, ABCA3) that—as far as we know—have yet to be associated with PTOA were identified as being concordantly differentially expressed across all treatment groups when compared to controls. Functional pathway analysis of injured subject cartilage compared to control cartilage revealed overarching patterns of cellular proliferation at 1 week, angiogenesis, ECM interaction, focal adhesion, and cellular migration at 4 weeks, and calcium signaling, immune system activation, GABA signaling, and HIF-1 signaling at 52 weeks.

Quantitative magnetic resonance imaging has been used to evaluate the structural integrity of knee joint structures. However, variations in acquisition parameters between scanners pose significant challenges. Understanding the effect of small differences in acquisition parameters for quantitative sequences is vital to the validity of cross-institutional studies, and for the harmonization of large, heterogeneous datasets to train machine learning models. The study objective was to assess the reproducibility of T

Quantitative magnetic resonance imaging (qMRI) has been used to determine the failure properties of ACL grafts and native ACL repairs and/or restorations. How these properties relate to future clinical, functional, and patient-reported outcomes remain unknown. The study objective was to investigate the relationship between non-contemporaneous qMRI measures and traditional outcome measures following Bridge-Enhanced ACL Restoration (BEAR). It was hypothesized that qMRI parameters at 6 months would be associated with clinical, functional, and/or patient-reported outcomes at 6 months, 24 months, and changes from 6 to 24 months post-surgery.Data of BEAR patients (n = 65) from a randomized control trial of BEAR versus ACL reconstruction (BEAR II Trial; NCT02664545) were utilized retrospectively for the present analysis. Images were acquired using the Constructive Interference in Steady State (CISS) sequence at 6 months post-surgery. Single-leg hop test ratios, arthrometric knee laxity values, and International Knee Documentation Committee (IKDC) subjective scores were determined at 6 and 24 months post-surgery. The associations between traditional outcomes and MRI measures of normalized signal intensity, mean cross-sectional area (CSA), volume, and estimated failure load of the healing ACL were evaluated based on bivariate correlations and multivariable regression analyses, which considered the potential effects of age, sex, and body mass index.CSA (r = 0.44, p = 0.01), volume (r = 0.44, p = 0.01), and estimated failure load (r = 0.48, p = 0.01) at 6 months were predictive of the change in single-leg hop ratio from 6 to 24 months in bivariate analysis. CSA (βThe qMRI-based measures of CSA, volume, and estimated failure load were predictive of a positive functional outcome trajectory from 6 to 24 months post-surgery. These variables measured using qMRI at 6 months post-surgery could serve as prospective markers of the functional outcome trajectory from 6 to 24 months post-surgery, aiding in rehabilitation programming and return-to-sport decisions to improve surgical outcomes and reduce the risk of reinjury.Level II.

The primary source of synovial fluid inflammatory mediators is currently unknown and may include different tissues comprising the joint, including the synovium and articular cartilage. Prior work in a porcine model has demonstrated that anterior cruciate ligament (ACL) surgery leads to significant changes in early gene expression in the synovium and articular cartilage, which are the same whether concomitant ligament restoration is performed or not. In this study, 36 Yucatan minipigs underwent ACL surgery, and a custom multiplex assay was used to measure synovial fluid protein levels of MMP-1, MMP-2, MMP-3, MMP-7, MMP-9, MMP-12, MMP-13, IL-1α, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-18, GM-CSF, and TNFα in 18 animals at 1 and 4 weeks after surgery. Linear regressions were used to evaluate the relationships between synovial fluid protein levels and the previously reported gene expression levels in the articular cartilage and synovium from the same animal cohort. Synovial fluid levels of MMP-13 and IL-6 were significantly correlated with synovial gene expression (P=.003 and P

Background: Magnetic resonance–based measurements of signal intensity have been used to track healing of surgically treated anterior cruciate ligaments (ACLs). However, it is unknown how the signal intensity values in different regions of the ligament or graft change during healing. Hypotheses: (1) Normalized signal intensity of the healing graft or repaired ACL is heterogeneous; (2) temporal changes in normalized signal intensity values differ among the tibial, middle, and femoral regions; and (3) there are no differences in regional normalized signal intensity values 2 years postoperatively among grafts, repaired ACLs, and contralateral native ACLs. Study Design: Cohort study; Level of evidence, 2. Methods: Magnetic resonance imaging scans were analyzed from patients in a trial comparing ACL reconstruction (n = 35) with bridge-enhanced ACL repair (n = 65). The ACLs were segmented from images acquired at 6, 12, and 24 months postoperatively and were partitioned into 3 sections along the longitudinal axis (femoral, middle, and tibial). Linear mixed modeling was used to compare location-specific differences in normalized ligament signal intensity among time points (6, 12, and 24 months) and groups (ACL reconstruction, repair, and contralateral native ACL). Results: For grafts, the middle region had a higher mean normalized signal intensity when compared with the femoral region at all time points ( P < .01) but compared with the tibial region only at 6 months ( P < .01). For repaired ACLs, the middle region had a higher mean normalized signal intensity versus the femoral region at all time points ( P < .01) but versus the tibial region only at 6 and 12 months ( P < .04). From 6 to 24 months, the grafts showed the greatest reduction in normalized signal intensity in the femoral and middle regions (vs tibial regions; P < .01), while there were no regional differences in repaired ACLs. At 2 years after surgery, repaired ACLs had a lower normalized signal intensity in the tibial region as compared with reconstructed grafts and contralateral native ACLs ( P < .01). Conclusion: The results suggest that graft remodeling is location specific. Repaired ACLs were more homogeneous, with lower or comparable normalized signal intensity values at 2 years as compared with the contralateral native ACL and reconstructed grafts.

INTRODUCTION Adolescent athletes' and their guardians' preferences for sports medicine surgeon characteristics are unknown. METHODS Unique, anonymous surveys regarding preferences in characteristics of sports medicine surgeons were given to both patients (aged 10 to 18 years) and their guardians before being seen by a sports medicine surgeon. RESULTS Patients and their guardians reported shared decision making as the most important surgeon characteristic, followed by understanding patients' sports and goals. A higher percentage of male patients than female patients had a surgeon sex preference (P = 0.005); however, for both the groups, this ranked lower than other surgeon qualities. Nearly all respondents reported that the adolescent patient should be involved in medical decision making; yet, physician selection was determined by the guardian 65% of the time. CONCLUSIONS Both adolescent patients and their guardians reported shared decision making and understanding patients' sports and goals as surgeon qualities that were more important to them than surgeon sex. LEVEL OF EVIDENCE Level IV, cross-sectional study.

Background: Primary repair of the anterior cruciate ligament (ACL) augmented with a tissue engineered scaffold to facilitate ligament healing is a technique under development for patients with ACL injuries. The size (the amount of tissue) and signal intensity (the quality of tissue) of the healing ligament as visualized on magnetic resonance imaging (MRI) have been shown to be related to its strength in large animal models. Hypothesis: Both modifiable and nonmodifiable risk factors could influence the size and signal intensity of the repaired ligament in patients at 6 months after surgery. Study Design: Case series; Level of evidence, 4. Methods: 62 patients (mean age, 19.4 years; range, 14-35 years) underwent MRI of the knee 6 months after ACL repair augmented with an extracellular matrix scaffold. The signal intensity (normalized to cortical bone) and average cross-sectional area of the healing ligament were measured from the MRI stack obtained by use of a gradient echo sequence. Associations between these 2 measures and patient characteristics, which included demographic, clinical, and anatomic features, were determined by use of multivariable regression analysis. Results: A larger cross-sectional area of the repaired ligament at 6 months was associated with male sex, older age, and the performance of a larger notchplasty ( P < .05 for all associations). A lower signal intensity at 6 months, indicating greater similarity to normal ligament, was associated with a smaller tibial slope and greater side-to-side difference in quadriceps strength 3 months after surgery. Other factors, including preoperative body mass index, mechanism of injury, tibial stump length, and Marx activity score, were not significantly associated with either MRI parameter at 6 months. Conclusion: Modifiable factors, including surgical notchplasty and slower recovery of quadriceps strength at 3 months, were associated with a larger cross-sectional area and improved signal intensity of the healing ACL after bridge-enhanced ACL repair in this preliminary study. Further studies to determine the optimal size of the notchplasty and the most effective postoperative rehabilitation strategy after ACL repair augmented by a scaffold are justified. Registration: NCT02664545 (ClinicalTrials.gov identifier).

Background: The cross-sectional area (CSA) of the anterior cruciate ligament (ACL) and reconstructed graft has direct implications on its strength and knee function. Little is known regarding how the CSA changes along the ligament length and how those changes vary between treated and native ligaments over time. Hypothesis: It was hypothesized that (1) the CSA of reconstructed ACLs and restored ACLs via bridge-enhanced ACL restoration (BEAR) is heterogeneous along the length. (2) Differences in CSA between treated and native ACLs decrease over time. (3) CSA of the surgically treated ACLs is correlated significantly with body size (ie, height, weight, body mass index) and knee size (ie, bicondylar and notch width). Study Design: Cohort study; Level of evidence, 2. Methods: Magnetic resonance imaging scans of treated and contralateral knees of 98 patients (n = 33 ACL reconstruction, 65 BEAR) at 6, 12, and 24 months post-operation were used to measure the ligament CSA at 1% increments along the ACL length (tibial insertion, 0%; femoral insertion, 100%). Statistical parametric mapping was used to evaluate the differences in CSA between 6 and 24 months. Correlations between body and knee size and treated ligament CSA along its length were also assessed. Results: Hamstring autografts had larger CSAs than native ACLs at all time points ( P < .001), with region of difference decreasing from proximal 95% of length (6 months) to proximal 77% of length (24 months). Restored ACLs had larger CSAs than native ACLs at 6 and 12 months, with larger than native CSA only along a small midsubstance region at 24 months ( P < .001). Graft CSA was correlated significantly with weight (6 and 12 months), bicondylar width (all time points), and notch width (24 months). Restored ACL CSA was significantly correlated with bicondylar width (6 months) and notch width (6 and 12 months). Conclusion: Surgically treated ACLs remodel continuously within the first 2 years after surgery, leading to ligaments/grafts with heterogeneous CSAs along the length, similar to the native ACL. While reconstructed ACLs remained significantly larger, the restored ACL had a CSA profile comparable with that of the contralateral native ACL. In addition to size and morphology differences, there were fundamental differences in factors contributing to CSA profile between the ACL reconstruction and BEAR procedures. Registration: NCT 02664545 ( ClinicalTrials.gov identifier).

Previous clinical studies have shown that psychological factors have significant effects on an athlete's readiness to return to sport after anterior cruciate ligament (ACL) reconstruction (ACLR).We hypothesized that patients who underwent bridge-enhanced ACL restoration (BEAR) would have higher levels of psychological readiness to return to sport compared with patients who underwent ACLR.Randomized controlled trial; Level of evidence, 1.A total of 100 patients (median age, 17 years; median preoperative Marx activity score, 16) with complete midsubstance ACL injuries were randomized to either the BEAR procedure (n = 65) or autograft ACLR (n = 35 [33 hamstring and 2 bone--patellar tendon-bone]) and underwent surgery within 45 days of injury. Objective, functional, and patient-reported outcomes, including the ACL--Return to Sport after Injury (ACL-RSI) scale, were assessed at 6, 12, and 24 months postoperatively.Patients who underwent the BEAR procedure had significantly higher ACL-RSI scores at 6 months compared with those who underwent ACLR (71.1 vs 58.2;Patients who underwent the BEAR procedure had higher ACL-RSI scores at 6 months postoperatively. Better ACL-RSI scores at 6 months were related most strongly to higher IKDC scores at 6 months and were also associated with lower pain levels, better muscle recovery, and less knee laxity at 6 months.NCT02664545 (ClinicalTrials.gov identifier).

Background: Little is known about sex-based differences in anterior cruciate ligament (ACL) tissue quality in vivo or the association of ACL size (ie, volume) and tissue quality (ie, normalized signal intensity on magnetic resonance imaging [MRI]) with knee anatomy. Hypothesis: We hypothesized that (1) women have smaller ACLs and greater ACL normalized signal intensity compared with men, and (2) ACL size and normalized signal intensity are associated with age, activity levels, body mass index (BMI), bicondylar width, intercondylar notch width, and posterior slope of the lateral tibial plateau. Study Design: Cross-sectional study; Level of evidence, 3. Methods: Knee MRI scans of 108 unique ACL-intact knees (19.7 ± 5.5 years, 62 women) were used to quantify the ACL signal intensity (normalized to cortical bone), ligament volume, mean cross-sectional area, and length. Independent t tests were used to compare the MRI-based ACL parameters between sexes. Univariate and multivariate linear regression analyses were used to investigate the associations between normalized signal intensity and size with age, activity levels, BMI, bicondylar width, notch width, and posterior slope of the lateral tibial plateau. Results: Compared with men, women had significantly smaller mean ACL volume (men vs women: 2028 ± 472 vs 1591 ± 405 mm3), cross-sectional area (49.4 ± 9.6 vs 41.5 ± 8.6 mm2), and length (40.8 ± 2.8 vs 38.1 ± 3.1 mm) ( P < .001 for all), even after adjusting for BMI and bicondylar width. There was no difference in MRI signal intensity between men and women (1.15 ± 0.24 vs 1.12 ± 0.24, respectively; P = .555). BMI, bicondylar width, and intercondylar notch width were independently associated with a larger ACL ( R 2 > 0.16, P < .001). Younger age and steeper lateral tibial slope were independently associated with shorter ACL length ( R 2 > 0.03, P < .04). The combination of BMI and bicondylar width was predictive of ACL volume and mean cross-sectional area ( R 2 < 0.3). The combination of BMI, bicondylar width, and lateral tibial slope was predictive of ACL length ( R 2 = 0.39). Neither quantified patient characteristics nor anatomic variables were associated with signal intensity. Conclusion: Men had larger ACLs compared with women even after adjusting for BMI and knee size (bicondylar width). No sex difference was observed in signal intensity, suggesting no difference in tissue quality. The association of the intercondylar notch width and lateral tibial slope with ACL size suggests that the influence of these anatomic features on ACL injury risk may be partially explained by their effect on ACL size. Registration: NCT02292004 and NCT02664545 ( ClinicalTrials.gov identifier).

Anterior cruciate ligament injuries result in posttraumatic osteoarthritis in the medial compartment of the knee, even after surgical treatment. How the chondrocyte distribution within the articular cartilage changes early in this process is currently unknown. The study objective was to investigate the chondrocyte distribution within the medial femoral condyle after an anterior cruciate ligament transection in a preclinical model. Forty-two adolescent Yucatan minipigs were allocated to receive unilateral anterior cruciate ligament surgery (n = 36) or no surgery (n = 6). Central coronal sections of the medial femoral condyle were obtained at 1- and 4 weeks after surgery, and the chondrocyte distribution was measured via whole slide imaging and a cell counting batch processing tool utilized in ImageJ. Ki-67 immunohistochemistry was performed to identify proliferating cells. Empty lacunae, karyolysis, karyorrhexis, and pyknosis were used to identify areas of irreversible cell injury. The mean area of irreversible cell injury was 0% in the intact controls, 13.4% (95% confidence interval: 6.4, 20.3) at 1-week post-injury and 19.3% (9.7, 28.9) at 4 weeks post-injury (p < .015). These areas occurred closest to the femoral intra-articular notch. The remaining areas containing viable chondrocytes had Ki-67-positive cells (p < .02) and increased cell density in the middle (p < .03) and deep zones (p = .001). For the entire section, the total chondrocyte number did not change significantly post-operatively; however, the density of cells in the peripheral regions of the medial femoral condyle increased significantly at 1- and 4 weeks post-injury relative to the intact control groups (p = .032 and .004, respectively). These data demonstrate a peripheral shift in the viable chondrocyte population of the medial femoral condyle after anterior cruciate ligament injury and further suggest that chondrocytes with the capacity to proliferate are not confined to one particular cartilage layer.

The objective was to determine if an intra-articular injection of an extracellular matrix (ECM) powder and blood composite (ECM-B) after anterior cruciate ligament (ACL) injury would have a mitigating effect on posttraumatic osteoarthritis and if that effect would be different with terminal sterilization of the ECM powder before use. Eighty Lewis rats underwent ACL transection and were divided into four groups: (1) intra-articular injection with phosphate-buffered saline (PBS; n = 20), (2) intra-articular injection of ECM-B using aseptically processed ECM (ASEPTIC; n = 20), (3) intra-articular injection of the ECM-busing ECM powder sterilized with 15 kGy electron beam irradiation (EBEAM; n = 20), and (4) intra-articular injection of the ECM-B using ECM powder sterilized with ethylene oxide (EO; n = 20). Twenty additional animals received capsulotomy only (SHAM). The animals were followed for 6 weeks and evaluations of gait, radiographs, and joint cartilage histology were performed. At 6 weeks, when compared to the SHAM group, the group treated with PBS had significantly worse gait and histologic changes, while the ASEPTIC group was not different from SHAM for either of these outcomes. When compared to the SHAM group, the EO group had similar gait outcomes, but greater histologic damage, and the EBEAM group had significantly worse gait and histological outcomes. The ECM-B composite produced using aseptically processed ECM powder mitigated the gait and histologic changes associated with osteoarthritis after ACL transection in the rat; however, care must be taken when selecting a terminal sterilization method as this may affect the effectiveness of treatment.

Background: The extent of posttraumatic osteoarthritis (PTOA) in the porcine anterior cruciate ligament (ACL) transection model is dependent on the surgical treatment selected. In a previous study, animals treated with bridge-enhanced ACL repair using a tissue-engineered implant developed less PTOA than those treated with ACL reconstruction (ACLR). Alterations in gait, including asymmetric weightbearing and shorter stance times, have been noted in clinical studies of subjects with osteoarthritis. Hypothesis: Animals receiving a surgical treatment that results in less PTOA (ie, bridge-enhanced ACL repair) would exhibit fewer longitudinal postoperative gait asymmetries over a 1-year period when compared with treatments that result in greater PTOA (ie, ACLR and ACL transection). Study Design: Controlled laboratory study. Methods: Thirty-six Yucatan minipigs underwent ACL transection and were randomized to receive (1) no further treatment, (2) ACLR, or (3) bridge-enhanced ACL repair. Gait analyses were performed preoperatively, and at 4, 12, 26, and 52 weeks postoperatively. Macroscopic cartilage assessments were performed at 52 weeks. Results: Knees treated with bridge-enhanced ACL repair had less macroscopic damage in the medial tibial plateau than those treated with ACLR or ACL transection (adjusted P = .03 for both comparisons). The knees treated with bridge-enhanced ACL repair had greater asymmetry in hindlimb maximum force and impulse loading at 52 weeks than the knees treated with ACL transection (adjusted P < .05 for both comparisons). Although not significant, there was a trend that knees treated with bridge-enhanced ACL repair had greater asymmetry in hindlimb maximum force and impulse loading (adjusted P < .10 for both comparisons) compared with ACLR. Conclusion: Contrary to our hypothesis, the surgical treatment resulting in less macroscopic cartilage damage (ie, bridge-enhanced ACL repair) exhibited greater asymmetry in load-related gait parameters than the other surgical groups. This finding suggests that increased offloading of the surgical knee may be associated with a slower rate of PTOA development. Clinical Relevance: Less cartilage damage at 52 weeks was found in the surgical group that continued to protect the limb from full body weight during gait. This finding suggests that protection of the knee from maximum stresses may be important in minimizing the development of PTOA in the ACL-injured knee within 1 year.

The roles that cytokines and matrix metalloproteinases play in the onset and progression of posttraumatic osteoarthritis (PTOA) remain a topic of debate. The study objective was to evaluate the concentrations of these inflammatory mediators during the development of mild to moderate PTOA in the porcine anterior cruciate ligament (ACL) surgical model. We hypothesized that there would be more animals with detectable mediators in the pigs that develop moderate PTOA (those receiving ACL reconstruction or untreated ACL transection) compared to those that develop mild PTOA (those receiving scaffold-enhanced ACL repair). 36 Yucatan minipigs underwent ACL transection and were randomized to: 1) no further treatment, 2) ACL reconstruction, or 3) scaffold-enhanced ACL repair. Synovial fluid samples were obtained pre-operatively, and at 1, 4, 12, 26 and 52 weeks post-operatively. The concentrations of inflammatory mediator in the synovial fluid samples were evaluated via multiplex assay. Macroscopic cartilage assessments were performed following euthanasia at 52 weeks. As found in prior studies, the repair group had significantly less cartilage damage than either the ACL transected or ACL reconstruction groups (P

The objective of this study was to develop an automated segmentation method for the anterior cruciate ligament that is capable of facilitating quantitative assessments of the ligament in clinical and research settings. A modified U-Net fully convolutional network model was trained, validated, and tested on 246 Constructive Interference in Steady State magnetic resonance images of intact anterior cruciate ligaments. Overall model performance was assessed on the image set relative to an experienced (>5 years) "ground truth" segmenter in two domains: anatomical similarity and the accuracy of quantitative measurements (i.e., signal intensity and volume) obtained from the automated segmentation. To establish model reliability relative to manual segmentation, a subset of the imaging data was resegmented by the ground truth segmenter and two additional segmenters (A, 6 months and B, 2 years of experience), with their performance evaluated relative to the ground truth. The final model scored well on anatomical performance metrics (Dice coefficient = 0.84, precision = 0.82, and sensitivity = 0.85). The median signal intensities and volumes of the automated segmentations were not significantly different from ground truth (0.3% difference, p = .9; 2.3% difference, p = .08, respectively). When the model results were compared with the independent segmenters, the model predictions demonstrated greater median Dice coefficient (A = 0.73, p = .001; B = 0.77, p = NS) and sensitivity (A = 0.68, p = .001; B = 0.72, p = .003). The model performed equivalently well to retest segmentation by the ground truth segmenter on all measures. The quantitative measures extracted from the automated segmentation model did not differ from those of manual segmentation, enabling their use in quantitative magnetic resonance imaging pipelines to evaluate the anterior cruciate ligament.

Background: A bridge-enhanced anterior cruciate ligament (ACL) repair (BEAR) procedure places an extracellular matrix implant, combined with autologous whole blood, in the gap between the torn ends of the ligament at the time of suture repair to stimulate healing. Prior studies have suggested that white blood cell (WBC) and platelet concentrations significantly affect the healing of other musculoskeletal tissues. Purpose/Hypothesis: The purpose of this study was to determine whether concentrations of various blood cell types placed into a bridging extracellular matrix implant at the time of ACL repair would have a significant effect on the healing ligament cross-sectional area or tissue organization (as measured by signal intensity). We hypothesized that patients with higher physiologic platelet and lower WBC counts would have improved healing of the ACL on magnetic resonance imaging (MRI) (higher cross-sectional area and/or lower signal intensity) 6 months after surgery. Study Design: Cohort study; Level of evidence, 2. Methods: A total of 61 patients underwent MRI at 6 months after bridge-enhanced ACL repair as part of the BEAR II trial. The normalized signal intensity and average cross-sectional area of the healing ligament were measured from a magnetic resonance stack obtained using a gradient echo sequence. The results were stratified by sex, and univariate and multivariate regression analyses determined significant correlations between blood cell concentrations on these 2 magnetic resonance parameters. Results: In unadjusted analyses, older age and male sex were associated with greater healing ligament cross-sectional area ( P < .04) but not signal intensity ( P > .15). Adjusted multivariable analyses indicated that in female patients, a higher monocyte concentration correlated with a higher ACL cross-sectional area (β = 1.01; P = .049). All other factors measured, including the physiologic concentration of platelets, neutrophils, lymphocytes, basophils, and immunoglobulin against bovine gelatin, were not significantly associated with either magnetic resonance parameter in either sex ( P > .05 for all). Conclusion: Although older age, male sex, and monocyte concentration in female patients were associated with greater healing ligament cross-sectional area, signal intensity of the healing ligament was independent of these factors. Physiologic platelet concentration did not have any significant effect on cross-sectional area or signal intensity of the healing ACL at 6 months after bridge-enhanced ACL repair in this cohort. Given these findings, factors other than the physiologic platelet concentration and total WBC concentration may be more important in the rate and amount of ACL healing after bridge-enhanced ACL repair.

Objectives: Meniscal injuries are extremely common. Several anatomical features of the knee, including the tibial plateau morphology, have been shown to influence knee biomechanics and the risk of ligamentous injuries. Little is known, however, how these morphological features influence the risk of isolated meniscal injuries in the ACL intact knee. In the current study, we used MRI and 3D image analysis to investigate anatomical variables of knees in patients with and without isolated meniscus tears. We hypothesized that there are differences in slopes and concavity of the tibial plateau between patients with isolated meniscus tears and matched normal controls. Methods: 65 subjects with first-instance isolated medial or lateral meniscal injuries requiring surgical fixation (Age: 15 +/- 2 years, BMI: 23.2 +/- 3.7 Kg/m2; 43% females) were matched to 65 subjects with normal knees and no prior injuries (Age: 15 +/- 2 years, BMI: 23.1 +/- 3.8 Kg/m2; 43% females) based on age, BMI and sex. Sagittal Proton Density SPACE MR images (preoperative for injured group) were used to measure the posterior slope of the medial (MTS) and lateral (LTS) tibial plateau, coronal slope of the tibial plateau (CTS), and the maximum depth of the medial tibia plateau (MTD, as a measure of medial tibial plateau concavity), following established techniques-Figure 1. Two-sample t-test was used to compare the quantified anatomical features between the cases and matched controls. Results: There were no differences in age (p = 0.999), sex distribution (p = 0.999) and BMI (p = 0.963) between the two cohorts. Compared to matched controls, patients with isolated meniscal tears had lower LTS (3.6 +/- 3.2 vs 5.6 +/- 3.0 degrees; p < 0.001), lower MTS (3.4 +/- 2.9 vs 4.9 +/- 2.5 degrees; p = 0.001) and deeper MTD (2.4 +/- 0.8 vs 1.5 +/- 0.8 mm; p < 0.001). There was no difference in CTS between the groups (3.6 +/- 1.6 vs 3.9 +/- 2.2 degrees; p = 0.300). Conclusion: This study suggests that subjects with isolated meniscus tear have a lower posterior tibial slope and a deeper MTD (more concave medial tibial plateau) than matched population who do not have a meniscus tear. This is contrary to what is known for ACL tears, where a higher posterior tibial slope and a shallower MTD have been associated with an increased risk of ACL tear. During load-bearing activities, in particular high impact movements such as jumping, increased tibial slope and decreased MTD have shown to result in greater anterior shear forces across the knee. In contrast, the less sloped and more concave plateau will experience lower shear forces but more compression, which could be detrimental to the meniscus. This study provides preliminary evidence suggesting a link between tibial plateau morphology and risk of isolated meniscal injuries. Further mechanistic studies are required to better understand the interaction between knee morphology, meniscal loading and subsequent risk of injury. [Figure: see text]

Objectives: Preclinical studies and an early clinical cohort study have suggested that for complete midsubstance ACL injuries, the technique of suture repair of the anterior cruciate ligament (ACL) augmented with a protein implant placed in the gap between the torn ends (bridge-enhanced ACL repair) may be a viable alternative to ACL reconstruction. We hypothesized that patients treated with bridge-enhanced ACL repair would have non-inferior patient reported outcomes and anteroposterior (AP) laxity, and superior hamstring strength at 2 years after surgery when compared with subjects who had ACL reconstruction with autograft. Study Design: Prospective Randomized Control Trial (NCT02664545). Methods: One hundred patients, ages 13 to 35, with complete mid-substance ACL injuries, were enrolled and underwent surgery within 45 days of injury. Sixty-five patients were randomized to receive a bridge-enhanced ACL repair (BEAR group) and 35 were randomized to receive autograft ACL reconstruction (ACLR group). Outcomes, including the International Knee Documentation Committee (IKDC) Subjective score, instrumented AP laxity and muscle strength were assessed at 2 years post-operatively. Both patients and assessors were blinded as to surgical assignment until after the 2-year assessment. Results: Ninety-six percent of the subjects returned for follow-up at 2 years. The median age of the subjects in both groups at enrollment was 17 years and the median pre-operative Marx score in both groups was 16 points. Non-inferiority criteria were met for both the IKDC subjective score and AP laxity. The BEAR group had significantly better recovery of hamstring strength than the ACLR group (pConclusion: Bridge-enhanced ACL repair using a specific protein-based implant led to non-inferior outcomes to that of ACL reconstruction for patient reported and instrumented knee laxity measures at 2 years after surgery in a young and active cohort of patients with acute mid-substance ACL tears.

Background: While a sex effect on outcomes following anterior cruciate ligament (ACL) reconstruction surgery has been previously documented, less is known following bridge-enhanced ACL repair (BEAR). We hypothesized that female sex would have significantly worse early functional outcomes and higher retear rates following primary repair of the ACL enhanced with a tissue-engineered scaffold. Methods: Sixty-five patients (28 males and 37 females), age 14–35 with a complete ACL tear underwent primary repair of the ACL enhanced with a tissue-engineered scaffold (bridge-enhanced ACL repair) within 45 days of injury. International Knee Documentation Committee (IKDC) and Knee Injury and Osteoarthritis Outcome (KOOS) scores, as well as instrumented anteroposterior (AP) laxity through KT-1000 testing and functional outcome measures were obtained at time points up to 2 years postoperatively and compared between males and females using mixed model repeated measures analyses and chi square tests. Results: There was no significant sex difference on the postoperative IKDC Subjective Score at 3, 6, 12, or 24 months or any of the five KOOS scores at 12 and 24 months. Instrumented AP laxity testing demonstrated mean (standard deviation) side-to-side differences that were similar in the two sexes at 2 years; 1.7 (2.7) mm and 1.5 (3.7) mm in females and males, respectively, p = 0.72. At 6 months postoperatively, males had a larger deficit in hamstring strength on the operated leg (14.0% vs. 1.7%; p = 0.03) and a larger deficit in quadriceps strength on the operated leg (11.3% vs. 2.0%; p = 0.004); however, no sex difference was noted at 12 or 24 months. Females demonstrated superior single leg hop testing at 6 and 12 months ([91.3% vs. 78.1%, p = 0.001], [96.9% vs. 87.0%, p = 0.01] respectively). There were no significant sex differences on ipsilateral (males; 14.3% vs. females; 13.9%, p = 1.00) or contralateral (males; 3.6% vs. females; 2.8%, p = 1.00) ACL reinjury rates. Conclusions: Female subjects had better hamstring and quadriceps strength indices at 6 months than males as well as better hop test results at the 6 and 12-month time period. Despite this, there was no significant sex difference on patient-reported outcomes and objective AP laxity testing at time points up to 2 years postoperatively. IMPACT STATEMENT: This is the first study comparing sex specific outcomes following the bridge-enhanced ACL repair technique (BEAR). The results of this study suggest that females have earlier recovery of both muscle strength and functional outcomes compared to their male counterparts. This is an important finding when considering future modifications to postoperative care and rehabilitation in females and males following this tissue-engineered BEAR technique.

Background: Multiple anatomic features of the femoral condyles and tibial plateau have been shown to influence knee biomechanics and risk of anterior cruciate ligament (ACL) injury. However, it remains unclear how these anatomic factors affect the midterm outcomes of ACL reconstruction. Hypothesis: Decreased femoral notch width, increased posterior and coronal slopes, and decreased concavity of the tibial plateau are associated with inferior clinical, patient-reported, and osteoarthritis-related outcomes 7 years after ACL reconstruction. Study Design: Cohort study; Level of evidence, 3. Methods: Prospectively collected data from 44 patients who did not have a subsequent graft or contralateral ACL failure within 7 years after unilateral ACL reconstruction were reviewed (mean ± SD age, 23.7 ± 9.2 years; 64% women). Notch width (after notchplasty), posterior slopes of the medial and lateral tibial plateau, maximum depth of the medial tibial plateau, and coronal tibial slope were measured from magnetic resonance images. Anatomic predictors of side-to-side differences in anterior-posterior knee laxity, Knee injury and Osteoarthritis Outcome Score (KOOS), medial joint space width, and side-to-side differences in Osteoarthritis Research Society International (OARSI) x-ray score, measured at 7 years, were identified with linear regression (bivariate) and stepwise regression (multivariate). Results: Increased posterior slope of the lateral tibial plateau was associated with increased side-to-side difference in knee laxity (bivariate model only), increased side-to-side difference in the OARSI score, and decreased KOOS subscores ( R2 > .10, P < .05). Increased posterior slope of the medial tibial plateau was associated with a higher side-to-side difference in the OARSI x-ray score (bivariate model only) and lower KOOS subscores ( R2 > .11, P < .03). Increased coronal tibial slope was associated with lower KOOS subscores ( R2 > .11, P < .03). Decreased medial tibial depth was associated with increased knee laxity as well as decreased KOOS subscores (ie, quality of life and symptoms; R2 > .12, P < .03). Postoperative notch width was not a significant predictor for any surgical outcome. None of the anatomic features were predictive of medial joint space width narrowing. Conclusion: Results partially support the hypothesis and highlight the importance of knee anatomy on several outcomes of ACL reconstruction among patients without subsequent graft or contralateral ACL injures. Increasing slopes in the coronal and sagittal planes with decreasing concavity of the medial tibial plateau lead to less favorable outcomes 7 years after surgery.

To determine the transcriptional profile of synovium during the molecular phase of post-traumatic osteoarthritis, anterior cruciate ligament transections (ACL) were performed in 36 Yucatan minipigs. Equal numbers were randomly assigned to no further treatment, ACL reconstruction or repair. Perimeniscal synovium for histopathology and RNA-sequencing was harvested at 1 and 4 weeks post-operatively and from six healthy control animals. Microscopic synovitis scores significantly worsened at 1 (p

Background: Bridge-enhanced anterior cruciate ligament repair (BEAR) has noninferior patient-reported outcomes when compared with autograft anterior cruciate ligament reconstruction (ACLR) at 2 years. However, the comparison of BEAR and autograft ACLR at earlier time points—including important outcomes such as resolution of knee pain and symptoms, recovery of strength, and return to sport—has not yet been reported. Hypothesis: It was hypothesized that the BEAR group would have higher outcomes on the International Knee Documentation Committee and Knee injury and Osteoarthritis Outcome Score, as well as improved muscle strength, in the early postoperative period. Study Design: Randomized controlled trial; Level of evidence, 1. Methods: A total of 100 patients aged 13 to 35 years with complete midsubstance anterior cruciate ligament injuries were randomized to receive a suture repair augmented with an extracellular matrix implant (n = 65) or an autograft ACLR (n = 35). Outcomes were assessed at time points up to 2 years postoperatively. Mixed-model repeated-measures analyses were used to compare BEAR and ACLR outcomes. Patients were unblinded after their 2-year visit. Results: Repeated-measures testing revealed a significant effect of group on the International Knee Documentation Committee Subjective Score ( P = .015), most pronounced at 6 months after surgery (BEAR = 86 points vs ACLR = 78 points; P = .001). There was a significant effect of group on the Knee injury and Osteoarthritis Outcome Score-Symptoms subscale scores ( P = .010), largely attributed to the higher BEAR scores at the 1-year postoperative time point (88 vs 82; P = .009). The effect of group on hamstring strength was significant in the repeated-measures analysis ( P < .001), as well as at all postoperative time points ( P < .001 for all comparisons). At 1 year after surgery, approximately 88% of the patients in the BEAR group and 76% of the ACLR group had been cleared for return to sport ( P = .261). Conclusion: Patients undergoing the BEAR procedure had earlier resolution of symptoms and increased satisfaction about their knee function, as well as improved resolution of hamstring muscle strength throughout the 2-year follow-up period. Registration: NCT02664545 (ClinicalTrials.gov identifier)

Anterior cruciate ligament (ACL) injuries are one of the most well-known orthopaedic injuries and are treated with one of the most common orthopaedic procedures performed in the United States. This surgical procedure, ACL reconstruction, is successful at restoring the gross stability of the knee. However, the outcomes of ACL reconstruction can be limited by short and long-term complications, including muscle weakness, graft rupture, and premature osteoarthritis. Thus, new methods of treating this injury are being explored. This review details the pathway of how a tissue engineering strategy can be used to improve the healing of the ACL in preclinical studies and then translated to patients in an FDA-approved clinical study. This review paper will outline the clinical importance of ACL injuries, history of primary repair, the pathology behind failure of the ACL to heal, pre-clinical studies, the FDA approval process for a high risk medical device, and the preliminary results from a first-in-human study. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:2606-2612, 2017.

Anterior cruciate ligament (ACL) volume and T2∗ relaxation times from magnetic resonance (MR) images have been previously shown to predict the structural properties of healing ligaments. We investigated whether MR imaging scan resolution and condition (in vivo, in situ, or ex vivo) affected ACL volume and T2∗ relaxation times in intact ligaments. ACLs of 14 pigs were imaged using a 3 T scanner and a six-channel flexcoil using at least two of four possible scan conditions: (1) in vivo moderate resolution (n = 14); (2) in vivo high resolution (n = 7); (3) in situ high resolution acquired within 60 minutes of euthanasia (n = 6); and (4) ex vivo high resolution following hind limb disarticulation and one freeze-thaw cycle (n = 7). T2∗ relaxation times were mapped to the ACL voxels. The total ACL volume was then divided into four sub-volumes (Vol1–4) based on predetermined increasing ranges of T2∗ times. ACL T2∗ statistics (first quartile, median, and standard deviation (SD)) were computed. Scan resolution had no effect on the total ACL volume, but Vol1 and first quartile T2∗ times decreased with high resolution and in situ/ex vivo scan conditions. The most dramatic differences in T2∗ summary statistics were between in vivo moderate and ex vivo high resolution scan conditions that included a freeze-thaw cycle: ACL T2∗ SD increased by over 50% in 9 animals, and more than 90% in 4 animals. Our results indicated that T2∗-based prediction models to quantify in vivo structural properties of healing ligaments should be based on high resolution in vivo MR scan conditions.

Background:There is a high incidence of a secondary anterior cruciate ligament (ACL) injury in unbraced adolescent athletes. Little is known about the effect of functional bracing with regard to the risk of secondary ACL injuries among adolescents.Hypothesis:Our primary hypothesis was that adolescents would have a high rate of secondary ACL injury even with brace use. A secondary hypothesis was that the reinjury rate with brace use would be lower than that of a historical control group of unbraced patients.Study Design:Case series; Level of evidence, 4.Methods:A group of 219 patients (age Results:The overall follow-up rate was 65% (142/219) at a mean 5.6 years after surgery. In the braced cohort, the overall graft retear rate was 10%, with the highest retear rates observed in male patients 17 years of age and younger (18%). The overall contralateral ACL injury and combined (graft or contralateral ACL) secondary injury rates were 13% and 23%, respectively, with female patients younger than 18 years having the highest contralateral and combined injury rates (17% and 26%, respectively). Younger age (odds ratio [OR], 0.70; P = .021), family history of ACL injury (OR, 2.81; P = .015), and higher lateral tibial slope (OR, 1.25; P = .016) were associated with increased risk of secondary knee injury in the braced cohort. Compared with the unbraced cohort, the braced cohort had a lower overall graft retear rate ( P = .028), a lower graft retear rate in patients younger than 18 years ( P = .012), lower early graft retear rate (within the first year after surgery) ( P = .011), and lower early graft retear rate in subjects younger than 18 years ( P = .003).Conclusion:Postoperative use of functional bracing can result in reduced risk of graft retear and no change in contralateral injury rates. Clinicians may want to consider the use of postoperative functional bracing in adolescent patients.

Anterior cruciate ligament (ACL) transection surgery in the minipig induces post-traumatic osteoarthritis (PTOA) in a pattern similar to that seen in human patients after ACL injury. Prior studies have reported the presence of cartilage matrix-degrading proteases, such as Matrix metalloproteinase-1 (MMP-1) and A disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS-4), in the synovial fluid of injured or arthritic joints; however, the tissue origin of these proteases is unknown. The objective of this study was to identify transcriptional processes activated in the synovium after surgical induction of PTOA with ACL transection, and to determine if processes associated with proteolysis were enriched in the synovium after ACL transection. Unilateral ACL transection was performed in adolescent Yucatan minipigs and synovium samples were collected at 1, 5, 9, and 14 days post-injury. Transcriptome-wide gene expression levels were determined using bulk RNA-Sequencing in the surgical animals and control animals with healthy knees. The greatest number of transcripts with significant changes was observed 1 day after injury. These changes were primarily associated with cellular proliferation, consistent with measurements of increased cellularity of the synovium at the two-week time point. At five to 14 days, the expression of transcripts relating to proteolysis and cartilage development was significantly enriched. While protease inhibitor-encoding transcripts (TIMP2, TIMP3) represented the largest fraction of protease-associated transcripts in the uninjured synovium, protease-encoding transcripts (including MMP1, MMP2, ADAMTS4) predominated after surgery. Cartilage development-associated transcripts that are typically not expressed by synovial cells, such as ACAN and COMP, were enriched in the synovium following ACL-transection. The upregulation in both catabolic processes (proteolysis) and anabolic processes (cartilage development) suggests that the synovium plays a complex, balancing role in the early response to PTOA induction.

Background: The quality of a repaired anterior cruciate ligament (ACL) or reconstructed graft is typically quantified in clinical studies by evaluating knee, lower extremity, or patient performance. However, magnetic resonance imaging of the healing ACL or graft may provide a more direct measure of tissue quality (ie, signal intensity) and quantity (ie, cross-sectional area). Hypotheses: (1) Average cross-sectional area or signal intensity of a healing ACL after bridge-enhanced ACL repair (BEAR) or a hamstring autograft (ACL reconstruction) will change postoperatively from 3 to 24 months. (2) The average cross-sectional area and signal intensity of the healing ligament or graft will correlate with anatomic features of the knee associated with ACL injury. Study Design: Cohort study; Level of evidence, 2. Methods: Patients with a complete midsubstance ACL tear who were treated with either BEAR (n = 10) or ACL reconstruction (n = 10) underwent magnetic resonance imaging at 3, 6, 12, and 24 months after surgery. Images were analyzed to determine the average cross-sectional area and signal intensity of the ACL or graft at each time point. ACL orientation, stump length, and bony anatomy were also assessed. Results: Mean cross-sectional area of the grafts was 48% to 98% larger than the contralateral intact ACLs at all time points ( P < .01). The BEAR ACLs were 23% to 28% greater in cross-sectional area than the contralateral intact ACLs at 3 and 6 months ( P < .02) but similar at 12 and 24 months. The BEAR ACLs were similar in sagittal orientation to the contralateral ACLs, while the grafts were 6.5° more vertical ( P = .005). For the BEAR ACLs, a bigger notch correlated with a bigger cross-sectional area, while a shorter ACL femoral stump, steeper lateral tibial slope, and shallower medial tibial depth were associated with higher signal intensity ( R2 > .40, P < .05). Performance of notchplasty resulted in an increased ACL cross-sectional area after the BEAR procedure ( P = .007). No anatomic features were correlated with ACL graft size or signal intensity. Conclusion: Hamstring autografts were larger in cross-sectional area and more vertically oriented than the native ACLs at 24 months after surgery. BEAR ACLs had a cross-sectional area, signal intensity, and sagittal orientation similar to the contralateral ACLs at 24 months. The early signal intensity and cross-sectional area of the repaired ACL may be affected by specific anatomic features, including lateral tibial slope and notch width—observations that deserve further study in a larger cohort of patients. Registration: NCT02292004 (ClinicalTrials.gov identifier)

OBJECTIVE To test whether intraarticular corticosteroid injection mitigates injury-induced synovitis and collagen degradation after anterior cruciate ligament transection (ACLT) and to characterize the synovial response using a functional genomics approach in a preclinical model of posttraumatic osteoarthritis. METHODS Yorkshire pigs underwent unilateral ACLT without subsequent corticosteroid injection (the ACLT group; n = 6) or ACLT with immediate injection of 20 mg triamcinolone acetonide (the steroid group; n = 6). A control group of pigs (the intact group; n = 6) did not undergo surgery. Total synovial membrane cellularity and synovial fluid concentration of C1,2C neoepitope-bearing collagen fragments 14 days after injury were primary end points and were compared between the ACLT, steroid, and intact groups. Cells were differentiated by histologic phenotype and counted, while RNA sequencing was used to quantify transcriptome-wide gene expression and monocyte, macrophage, and lymphocyte markers. RESULTS In the intact group, total cellularity was 13% (95% confidence interval [95% CI] 9-16) and the C1,2C concentration was 0.24 μg/ml (95% CI 0.08-0.39). In the ACLT group, significant increases were observed in total cellularity (to 21% [95% CI 16-27]) and C1,2C concentration (to 0.49 μg/ml [95% CI 0.39-0.59]). Compared to values in the ACLT group, total cellularity in the steroid group was nonsignificantly decreased to 17% (95% CI 15-18) (P = 0.26) and C1,2C concentration in the steroid group was significantly decreased to 0.29 μg/ml (95% CI 0.23-0.35) (P = 0.04). A total of 255 protein-coding transcripts were differentially expressed between the ACLT group and the intact group. These genes mainly enriched pathways related to cellular immune response, proteolysis, and angiogenesis. Mononuclear leukocytes were the dominant cell type in cell-dense areas. MARCO, SOCS3, CCR1, IL4R, and MMP2 expression was significantly associated with C1,2C levels. CONCLUSION Early intraarticular immunosuppression mitigated injury-induced increases in collagen fragments, an outcome better predicted by specific marker expression than by histologic measures of synovitis.

Since the 1970’s, ACL reconstruction has been the gold standard of treatment for ACL injuries in patients wanting to regain functional stability, and has been the focus of a comprehensive body of literature that has worked to improve this technique. While primary repair of the ACL had been found previously to fail in a high percentage of patients, some patients did well with suture repair of the ligament. This observation has led to interest in evaluating ways to improve techniques for repair of the ligament, rather than reconstruction, in the past decade. These more recent efforts have focused on augmentation with prosthetic or organic materials and biologic enhancement strategies in order to promote natural healing of the torn ligament. We present here the history and an update on primary repair of the ACL.

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

The objective of this study was to determine if an injection of a novel extracellular matrix scaffold and blood composite (EMBC) after anterior cruciate ligament (ACL) injury would have a mitigating effect on post-traumatic osteoarthritis (PTOA) development in rat knees. Lewis rats underwent unilateral ACL transection and were divided into three groups as follows: (1) no further treatment (ACLT; n = 10); (2) an intra-articular injection of EMBC on day 0 (INJ0; n = 11); and (3) an intra-articular injection of EMBC on day 14 (INJ14; n = 11). Ten additional animals received capsulotomy only (n = 10, SHAM group). The OARSI histology scoring of the tibial cartilage and micro-CT of the tibial epiphysis were performed after 35 days. The ratio of intact/treated hind limb forces during gait was determined using a variable resistor walkway. The OARSI cartilage degradation sum score and total degeneration width were significantly greater in the ACLT group when compared to the INJ0 (p = 0.031, and p = 0.005) and INJ14 (p = 0.022 and p = 0.04) group. Weight bearing on the operated limb only decreased significantly in the ACLT group (p = 0.048). In the rat ACL transection model, early or delayed injection of EMBC ameliorated the significant decrease in weight bearing and cartilage degradation seen in knees subjected to ACL transection without injection. The results indicate that the injection of EMBC may slow the process of PTOA following ACL injury and may provide a promising treatment for PTOA. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:995-1003, 2016.

The addition of platelet-rich plasma (PRP) to rotator cuff repair has not translated into improved outcomes after surgery. However, recent work stimulating ligament healing has demonstrated improved outcomes when PRP or whole blood is combined with an extracellular matrix carrier. The objective of this study was to evaluate the effect of three components of blood (plasma, platelets, and macrophages) on the in vitro activity of ovine rotator cuff cells cultured in an extracellular matrix environment. Tenocytes were obtained from six ovine infraspinatus tendons and cultured over 14 days in an extracellular matrix scaffold with the following additives: (1) plasma (PPP), (2) plasma and platelets (PAP), (3) plasma and macrophages (PPPM), (4) plasma, platelets and macrophages (PAPM), (5) phosphate buffered saline (PBS), and (6) PBS with macrophages (PBSM). Assays measuring cellular metabolism (AlamarBlue), proliferation (Quantitative DNA assay), synthesis of collagen and cytokines (SIRCOL, TNF-α and IL-10 ELISA, and MMP assay), and collagen gene expression (qPCR) were performed over the duration of the experiment, as well as histology at the conclusion. Plasma was found to stimulate cell attachment and spreading on the scaffold, as well as cellular proliferation. Platelets also stimulated cell proliferation, cellular metabolism, transition of cells to a myofibroblast phenotype, and contraction of the scaffolds. The addition of macrophages did not have any significant effect on the sheep rotator cuff cells in vitro. In vivo studies are needed to determine whether these changes in cellular function will translate into improved tendon healing.

Background: The risk of anterior cruciate ligament (ACL) injury is 2 to 10 times greater in women than men. While the effect of sex on injury risk is well established, its effects on surgical outcomes remain controversial. Purpose/Hypothesis: To investigate whether the biomechanical outcomes of ACL reconstruction are affected by sex using an established porcine model that displays similar sex-specific differences in knee anatomy and ligament structural properties to humans. The hypothesis was that there will be sex differences in ACL reconstruction outcomes with regard to the graft structural properties, knee laxity, and cartilage damage. Study Design: Controlled laboratory study. Methods: A total of 41 adolescent Yucatan minipigs (23 male, 18 female) underwent unilateral ACL transection and ACL reconstruction using sex-matched bone–patellar tendon–bone allografts (with or without additional bioenhancement). Graft biomechanical and histological properties, knee laxity, and cartilage damage were assessed after 15 weeks. A 2-factor analysis of variance was used to investigate the effect of sex on all the measured outcomes after adjusting for the treatment effect. Results: After 15 weeks of healing, female pigs had a significantly lower mean normalized graft yield load (by 18.5% ± 7.7%; P = .023) and linear stiffness (by 11.9% ± 5.6%; P = .043) compared with male pigs. Female pigs had significantly greater side-to-side differences in anteroposterior knee laxity at 30° (by 1.4 ± 0.6 mm; P = .028) and 90° (by 1.8 ± 0.8 mm; P = .032). Female pigs had a lower graft vascular density (by 0.8 ± 0.3 [analog scoring]; P = .021) with similar cellular and collagen-based histologic scores in both sexes ( P > .6). Female pigs also had a significantly larger area of cartilage damage (by 43.3 ± 14.8 mm2; P = .014) after conventional ACL reconstruction compared with their male counterparts. Conclusion: Female pigs had significantly worse outcomes (ie, graft structural properties, knee laxity, and cartilage damage) compared with male pigs in this translational model after 15 weeks of healing. Clinical Relevance: These data suggest that further optimization of ACL injury treatments may be needed to accommodate each sex instead of using a “one fits all” approach to improve surgical outcomes, decrease incidence of reinjury, and decrease posttraumatic osteoarthritis risk after ACL reconstruction.

Suture repair of the anterior cruciate ligament (ACL) has been widely abandoned in favor of ACL reconstruction, largely because of the high rates of failure and unreliability of the outcomes after suture repair. However, there have been recent basic science studies that suggest that combining a suture repair with a biological adjunct may improve the results of suture repair of the ACL, with several studies in large animal models showing equivalent strength of an ACL treated with bio-enhanced repaired to that of an ACL graft at 3, 6, and 12 months after surgery. In addition, the groups treated with bio-enhanced repair had significantly less osteoarthritis when compared with the animals undergoing ACL reconstruction. These findings have led to a renewed interest in bio-enhanced primary repair as a way to make repair of the ACL a viable option for a select group of patients in the future.

Magnetic resonance imaging (MRI) variables, such as T2* and volume, can predict the healing ligament structural properties. How these MR variables relate to semi-quantitative histology of the healing ACL is yet unknown. We hypothesized that T2* and volume would predict the histological scoring of a healing ACL. Yucatan minipigs underwent ACL transection and received bridge-enhanced ACL repair or no treatment. The surgical legs were harvested after 52 weeks and imaged using a high resolution 2-echo sequence. For each ligament, the volume and median T2* values were determined. The ACL specimens were then histologically analyzed using the advanced Ligament Maturity Index (LMI). The T2* of the healing ligaments significantly predicted the total LMI score as well as the cell, collagen and vessel sub-scores; R2 = 0.78, 0.67, 0.65, and 0.60, respectively (p ≤ 0.001). The ligament volume also predicted the total LMI score, cell, and collagen sub-scores; R2 = 0.39, 0.33, 0.37, and 0.60, respectively (p ≤ 0.001). A lower ligament T2* or a higher volume was associated with higher histological scores of the healing ligaments. This study provides a critical step in the development of a non-invasive method to evaluate ligament healing on a microscopic scale. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 33:1180–1187, 2015.

The number of publications on treatment of anterior cruciate ligament (ACL) injuries in the skeletally immature population has increased through the past decade [1-6]. However, opinions on whether pediatric ACL injuries should primarily be surgically reconstructed or conservatively treated are still divided within the pediatric orthopedic community [7, 8]. Evidence from high-level studies and randomized controlled trials is lacking [9], which leaves the field open for various treatment algorithms due to the lack of a solid scientific knowledge base. Risk factors for ACL injuries in skeletally immature patients are unknown, although it seems that boys may be more prone to rupturing their ACL before skeletal maturity, while girls have an increased risk through and after puberty [10, 11]. Many authors argue that the incidence of pediatric ACL injuries is rising. However, no epidemiological studies are available to support this statement. Increased awareness and advances in diagnostic methods, in addition to higher participation rates and earlier specialization in sports, may have led to an increase in the incidence of pediatric ACL tears.

Animal models have long been considered an important modality for studying ACL injuries. However, to our knowledge, the value of these preclinical models to study sex-related phenomena associated with ACL injury and recovery has not been evaluated.We asked whether (1) prominent anatomic and (2) biomechanical factors differ between female and male porcine knees, particularly those known to increase the risk of ACL injury.Eighteen intact minipig knees (nine males, nine females) underwent MRI to determine the femoral bicondylar width, intercondylar notch size (width, area and index), medial and lateral tibial slope, ACL size (length, cross-sectional area, and volume), and medial compartment tibiofemoral cartilage thickness. AP knee laxity at 30°, 60°, and 90° flexion and ACL tensile structural properties were measured using custom-designed loading fixtures in a universal tensile testing apparatus. Comparisons between males and females were performed for all anatomic and biomechanical measures. The findings then were compared with published data from human knees.Female pigs had smaller bicondylar widths (2.9 mm, ratio=0.93, effect size=-1.5) and intercondylar notches (width: 2.0 mm, ratio=0.79, effect size=-2.8; area: 30.8 mm2, ratio=0.76, effect size=-2.1; index: 0.4, ratio=0.84, effect size=-2.0), steeper lateral tibial slope (4.3°, ratio=1.13, effect size=1.1), smaller ACL (length: 2.7 mm, ratio=0.91, effect size=-1.1; area: 6.8 mm2, ratio=0.74, effect size=-1.5; volume: 266.2 mm3, ratio=0.68, effect size=-1.5), thinner medial femoral cartilage (0.4 mm, ratio=0.8, effect size=-1.1), lower ACL yield load (275 N, ratio=0.81, effect size=-1.1), and greater AP knee laxity at 30° (0.7 mm, ratio=1.32, effect size=1.1) and 90° (0.5 mm, ratio=1.24, effect size=1.1) flexion compared with their male counterparts. These differences were significant for all parameters (p≤0.04). Observed sex-related differences were similar to those reported for the human knee.Significant differences exist between knees of male and female pigs with respect to prominent anatomic and biomechanical factors. Our findings strongly agreed with published data regarding human knees.The findings highlight the use of the porcine large animal model to study the role of sex on ACL injuries and surgical outcome. This validated preclinical model may facilitate the development of novel, sex-specific interventions to prevent and treat ACL injuries for male and female patients.

Injuries to the anterior cruciate ligament (ACL) are currently treated with replacement of the torn ligament with a graft of tendon harvested from elsewhere in the knee. This procedure, called "ACL reconstruction," is excellent for restoring gross stability to the knee; however, there are relatively high graft failure rates in adolescent patients (Barber et al. in Arthroscopy 30(4):483-491, (2014); Engelman et al. in Am J Sports Med, (2014); Webster et al. in Am J Sports Med 42(3):641-647, (2014)), and the ACL reconstruction procedure does not prevent the premature osteoarthritis seen in patients after an ACL injury (Ajuied et al. in Am J Sports Med, (2013); Song et al. in J Sports Med 41(10):2340-2346, (2013); Tourville et al. Am J Sports Med 41(4):769-778, (2013)) .Thus, new solutions are needed for ACL injuries. Researchers have been investigating the use of scaffolds, growth factors and cells to supplement a suture repair of the ACL (bridge-enhanced repair; also called bio-enhanced repair in prior publications). In this paper, we will review the varied approaches which have been investigated for stimulating ACL healing and repair in preclinical models and how one of these technologies was able to move from promising preclinical results to FDA acceptance of an investigational device exemption application for a first-in-human study.

To determine the transcriptional profile of synovium during the molecular phase of post-traumatic osteoarthritis, anterior cruciate ligament transections (ACL) were performed in 36 Yucatan minipigs. Equal numbers were randomly assigned to no further treatment, ACL reconstruction or repair. Perimeniscal synovium for histopathology and RNA-sequencing was harvested at 1 and 4 weeks post-operatively and from six healthy control animals. Microscopic synovitis scores significantly worsened at 1 (p 0.001) and 4 weeks (p = 0.003) post-surgery relative to controls, and were driven by intimal hyperplasia and increased stromal cellularity without inflammatory infiltrates. Synovitis scores were similar between no treatment, reconstruction, and repair groups (p ≥ 0.668). Relative to no treatment at 1 week, 88 and 367 genes were differentially expressed in the reconstruction and repair groups, respectively (227 and 277 at 4 weeks). Relative to controls and with the treatment groups pooled, 1,683 transcripts were concordantly differentially expressed throughout the post-surgery time-course. Affected pathways included, proteolysis_connective tissue degradation (including upregulations of protease-encoding MMP1, MMP13, and ADAMTS4), and development_cartilage development (including upregulations of ACAN, SOX9, and RUNX2), among others. Using linear regression, significant associations of post-surgery synovial expression levels of 20 genes with the articular cartilage glycosaminoglycan loss were identified. These genes were predominantly related to embryonic skeletal system development and included RUNX2. In conclusion, this study confirmed an increased synovial expression of genes that may serve as targets to prevent cartilage degradation, including MMP1, MMP13, and ADAMTS4, in knees with microscopic synovitis and cartilage proteoglycan loss. Attractive novel targets include regulators of embryonic developmental processes in synovium. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res.