Several studies have highlighted the relationship between anterior cruciate (ACL) injury and knee geometry particularly tibial slope (TS). However, clinical data are inconsistent, whether the lateral or medial or slopes have a different influence on ACL injury. Our goal was to assess whether the medial, lateral slopes are associated with ACL injury and whether meniscus geometry is associated with ACL injury. In addition, we sought to determine if lateral meniscal height could serve as a simple surrogate measurement for ACL injury risk.

A case-controlled study compared 68 patients with an ACL injury and 68 matched nested controls. Radiological analysis of MRI measured the anterior-posterior distance of the medial and lateral plateaus, the tibial slope of both plateaus and meniscus geometry. Groups were compared using a Mann-Whitney test and α < 0 .05.

The lateral tibial plateau slope was significantly higher in the ACL injured group (6.92 degrees ±5.8) versus the control group 2.68 ±5.26 (p 0.0001). In addition, the lateral meniscal slope was significantly steeper with (ACL injuries: −1 ±4.7 versus −4.73 ±4.4 (p 0.0001) in the control group. The ACL Injured group had a significantly lower lateral meniscal height 0.76 cm ±0.09, compared to the control group that has 0.88 cm ±0.12 (p 0.0001). The Lateral meniscal height had a sensitivity of 76.47% and specificity 75% for predicting ACL injury using a cut off of

Patients with ACL-injury had significantly higher lateral tibial plateau slope. Lateral meniscus height was found to be an easy measurement to make on MRI with a high specificity for predicting ACL injury. Lateral tibial slope and meniscal Geometry can be used to identify patients with high risk of an ACL injury, that might benefit from further surgery to optimize rotational stability in high-risk patients.

Osteoarthritis (OA) is a chronic degenerative joint disorder that affects millions of people. There are currently no therapies that reverse or repair cartilage degradation in OA patients. Link N (DHLSDNYTLDHDRAIH) is a naturally occurring peptide that has been shown to increase both collagen and proteoglycan synthesis in chondrocytes and intervertebral disc cells [1,2]. Recent evidence indicates that Link N activates Smad1/5 signaling in cultured rabbit IVD cells presumably by interacting with the bone morphogenetic protein (BMP) type II receptor [3], however, whether a similar mechanism exists in chondrocytes remains unknown. In this study we determined whether Link N can stimulate matrix production and reverse degradation of human OA cartilage under inflammatory conditions.

OA cartilage was obtained from donors undergoing total knee arthroplasty with informed consent. OA cartilage/bone explants and OA chondrocytes were prepared from each donor. Cells were prepared in alginate beads (2×106 cells/mL) for gene expression analysis using qPCR. Cells and cartilage explants were exposed to IL-1β (10ng/ml), human Link N (hLN) (1μg/ml) or co-incubated with IL-1β+hLN for 7 and 21 days, respectively. Media was supplemented every three days. Cartilage/bone explants were measured for total glycosaminoglycan (GAG) content (retained and released) using the dimethylmethylene blue (DMMB) assay. Western blotting was performed to determine aggrecan and collagen expression in cartilage tissue. To determine NFκB activation, Western blotting was performed for detection of P-p65 in chondrocytes cultured in 2D following 10 min exposure of IL-1β in the presence of 10, 100, or 1000 ng/mL hLN.

Link N significantly decreased in a dose-dependent manner IL-1β-induced NFκB activation in chondrocytes. Gene expression profiling of matrix proteins indicated that there was a trend towards increased aggrecan and decreased collagen type I expression following hLN and IL-1β co-incubation. HLN significantly decreased the IL-1β-induced expression of catabolic enzymes MMP3 and MMP13, and the neuronal growth factor NGF (p < 0 .0001, n=3). In OA cartilage/bone explants, hLN reversed the loss of proteoglycan in cartilage tissue and significantly increased its synthesis whilst in the presence of IL-1β.

Link N stimulated proteoglycan synthesis and decreased MMP expression in OA chondrocytes under inflammatory conditions. One mechanism for Link N in preserving matrix protein synthesis may, in part, be due to its ability in rapidly suppressing IL-1β-induced activation of NF-κB. Further work is needed to determine whether Link N directly inhibits the IL-1β receptor or interferes with NFκB activation through an independent pathway(s).

Anterior cruciate ligament (ACL) injuries are frequent among athletes and a leading cause of time away from competition. Stability of the knee involves the ACL for limiting anterior tibial translation and the ALL (anterolateral ligament) to restrain internal rotation of the tibia. Present indications for treatment with a combined ACL-ALL reconstruction remain unclear and mostly subjective. We mathematically modeled the tibial plateau geometry to try and identify patients at risk of ACL injury, and develop an objective trigger point for the decision to proceed with additional surgery to optimize rotational stability in these higher risk patients. We hypothesized that an increased convexity and steepness of the posterior aspect of the lateral plateau would subject knees to higher rotational torques leading to potentially a higher risk of ACL injury.

The study design was a case-control study involving ACL reconstruction cases (n=68) and matched controls (n=68) between 2008–2015 at our institution. We used a two-dimensional approach, evaluating sagittal MRI images of the knee to model the posterior convexity of the lateral tibial plateau. Points were selected along the articular surface, and a least-squares regression was used to curve-fit a power function (y = a xn). In the equation, larger coefficient a and n represented steeper slopes. The cases and controls were compared using a Mann-Whitney-U test, and the statistical significance was set at α < 0.05. A subgroup analysis for females and males was also performed for the curve-fit coefficients.

We observed a significant difference in the tibial surface geometry between our ACL reconstruction cases and matched controls (Figure 1). The modeled power equation for our ACL cases had larger coefficients compared to controls for all groups. For all pooled subjects, coefficient a (ACL recon cases = 0.90 vs controls = 0.68, p < 0.0001) and coefficient n (ACL recon cases = 0.34 vs controls = 0.30, p = 0.07) (Table 1). For the statistically significant coefficient a, we found it had a sensitivity of 78.9% and specificity of 77.5% for the statistically significant coefficient a, we found it had a sensitivity of 78.9% and specificity of 77.5% for predicting injury, using a cut off coefficient of a = 0.78. The odds ratio was 12.6 [5.5 – 29].

The posterolateral cartilaginous slope of the tibial plateau was mathematically modeled in patients with ACL injury. Patients with ACL injury demonstrated abnormally steep and fast slopes compared to controls that may play predispose to ACL injury by increasing anterior translation forces and internal rotation torques sustained by their knee joint. A steeper slope may also explain high-grade pivot shifts on physical exam that are thought to be a relative indication for adding an associated ALL reconstruction. Our findings are promising for adding more objectivity to surgical decision-making, especially with identifying high-risk patients that may be candidates for combined ACL-ALL reconstructions.

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Testing potential therapeutics in the regeneration of the disc requires the use of model systems. Although several animal models have been developed to test intervertebral disc (IVD) regeneration, application becomes costly when used as a screening method. The bovine IVD organ culture system offers an inexpensive alternative, however, in the current paradigm, the bony vertebrae is removed to allow for nutrient diffusion to disc cells. This provides limitations on the conditions and strategies one can employ in investigating IVD regeneration and mechanisms in degenerative disc disease (i.e. complex loading). Although one method has been attempted to extend the survival of bovine vertebrae containing IVDs (vIVD) cell viability declined after two weeks in culture. Our goal was to develop and validate a long-term organ culture model with vertebral bone, which could be used subsequently for studying biological repair of disc degeneration and biomechanics.

Preparation of vIVDs: Bovine IVDs from the tails of 22–28-month-old steers were prepared for organ culture by parallel cuts through the adjacent vertebral bodies at 1cm from the endplates using an IsoMet®1000 Buehler precision sectioning saw. vIVDs were split into two groups: IVDs treated with PrimeGrowth Media kit (developed by Intervertech and licensed to Wisent Bioproducts) and IVDs with DMEM. The PrimeGrowth group was incubated for 1h in PrimeGrowth Isolation Medium (Cat# 319–511-EL) and the DMEM group for 1h in DMEM. After isolation, IVDs were washed in PrimeGrowth Neutralisation Medium (Cat# 319–512-CL) while the other IVDs were washed in DMEM. The discs isolated with PrimeGrowth and DMEM were cultured for up to 5 months in sterile vented 60 ml Leakbuster™ Specimen Containers in PrimeGrowth Culture Medium (Cat# 319–510-CL) and DMEM with no mechanical load applied. Live/Dead Assay: vIVDs cultured for 1 or 5 months were dissected and cell viability was assessed in different regions by confocal microscopy using Live/Dead® (Invitrogen) fluorescence assay. Glucose Diffusion: After one month of culture, vIVDs were incubated for 72h in diffusion medium containing PBS (1x), CaCl2 (1mM), MgCl2 (0.5mM), KCl2 (5mM), 0.1% BSA and 150µM 2-NDBG, a D-glucose fluorescent analogue. Discs were dissected and IVD tissues were incubated in guanidinium chloride extraction buffer. Extracts were measured for fluorescence.

After 5 months in culture, vIVDs prepared with PrimeGrowth kit demonstrated approximately 95% cell viability in all regions of the disc. However, dramatic reductions (∼90%) in vIVD viability were measured in DMEM group after 1 month. vIVD viability was related to the amount of 2-NDBG incorporated into the disc tissue.

We have developed a novel method for isolating IVDs with vertebral bone capable of long-term viability. This method may not only help in the discovery of novel therapeutics in disc regeneration, but could also advance our understanding on complex loading paradigms in disc degeneration.