Christopher G. Salib

Intra-articular Injection of a Substance P Inhibitor Affects Gene Expression in a Joint Contracture Model†

Substance P (SP), a neurotransmitter released after injury, has been linked to deregulated tissue repair and fibrosis in musculoskeletal tissues and other organs. Although SP inhibition is an effective treatment for nausea, it has not been previously considered as an anti‐fibrotic therapy. Although there are extensive medical records of individuals who have used SP antagonists, our analysis of human registry data revealed that patients receiving these antagonists and arthroplasty are exceedingly rare, thus precluding a clinical evaluation of their potential effects in the context of arthrofibrosis. Therefore, we pursued in vivo studies to assess the effect of SP inhibition early after injury on pro‐fibrotic gene expression and contractures in an animal model of post‐traumatic joint stiffening. Skeletally mature rabbits (n = 24) underwent surgically induced severe joint contracture, while injected with either fosaprepitant (a selective SP antagonist) or saline (control) early after surgery (3, 6, 12, and 24 h). Biomechanical testing revealed that differences in mean contracture angles between the groups were not statistically significant (P = 0.27), suggesting that the drug neither mitigates nor exacerbates joint contracture. However, microarray gene expression analysis revealed that mRNA levels for proteins related to cell signaling, pro‐angiogenic, pro‐inflammatory, and collagen matrix production were significantly different between control and fosaprepitant treated rabbits (P < 0.05). Hence, our study demonstrates that inhibition of SP alters expression of pro‐fibrotic genes in vivo. This finding will motivate future studies to optimize interventions that target SP to reduce the formation of post‐traumatic joint contractures.

Molecular pathology of total knee arthroplasty instability defined by RNA-seq

Total knee arthroplasty (TKA) is a durable and reliable procedure to alleviate pain and improve joint function. However, failures related to flexion instability sometimes occur. The goal of this study was to define biological differences between tissues from patients with and without flexion instability of the knee after TKA. Human knee joint capsule tissues were collected at the time of primary or revision TKAs and analyzed by RT-qPCR and RNA-seq, revealing novel patterns of differential gene expression between the two groups. Interestingly, genes related to collagen production and extracellular matrix (ECM) degradation were higher in samples from patients with flexion instability. Partitioned clustering analyses further emphasized differential gene expression patterns between sample types that may help guide clinical interpretations of this complication. Future efforts to disentangle the effects of physical and biological (e.g., transcriptomic modifications) risk factors will aid in further characterizing and avoiding flexion instability after TKA.

Joint contracture is reduced by intra-articular implantation of rosiglitazone-loaded hydrogels in a rabbit model of arthrofibrosis: ROSIGLITAZONE-LOADED HYDROGELS IN A RABBIT MODEL OF ARTHROFIBROSIS

Trauma, surgery, and other inflammatory conditions can lead to debilitating joint contractures. Adjunct pharmacologic modalities may permit clinical prevention and treatment of recalcitrant joint contractures. We investigated the therapeutic potential of rosiglitazone by intra‐articular delivery via oligo[poly(ethylene glycol)fumarate] (OPF) hydrogels in an established rabbit model of arthrofibrosis. OPF hydrogels loaded with rosiglitazone were characterized for drug elution properties upon soaking in minimum essential media (MEM) with 10% fetal bovine serum and measurements of drug concentrations via High Performance Liquid Chromatography (HPLC). Drug‐loaded scaffolds were surgically implanted into 24 skeletally mature female New Zealand White rabbits that were divided into equal groups receiving OPF hydrogels loaded with rosiglitazone (1.67 mg), or vehicle control (10 µl DMSO). After 8 weeks of joint immobilization, rabbits were allowed unrestricted cage activity for 16 weeks. Contracture angles of rabbit limbs treated with rosiglitazone showed statistically significant improvements in flexion compared to control animals (mean angles, respectively, 64.4° vs. 53.3°, p < 0.03). At time of sacrifice (week 24), animals in the rosiglitazone group continued to exhibit less joint contracture than controls (119.0° vs. 99.5°, p = 0.014). The intra‐articular delivery of rosiglitazone using implanted OPF hydrogels decreases flexion contractures in a rabbit model of arthrofibrosis without causing adverse effects (e.g., gross inflammation or arthritis). Statement of Clinical Significance: Post‐traumatic joint contractures are common and debilitating, with limited available treatment options. Pharmacologic interventions can potentially prevent and treat such contractures. This study is translational in that a commercially approved medication has been repurposed through a novel delivery device.

Validation of a dynamic joint contracture measuring device in a live rabbit model of arthrofibrosis: NOVEL JOINT CONTRACTURE MEASURING DEVICE

The current method of measuring arthrofibrosis in live rabbits is critically limited. Specifically, this method involves radioactive fluoroscopy, error‐prone goniometric measurements, and static joint angle outcomes that fail to approximate the compliance of tissues surrounding the joint. This study aims to validate a novel method of capturing the compliance of contracted tissues surrounding the joint without the use of fluoroscopy or animal sacrifice. Surgically induced contractures of one‐hundred and eight rabbits were measured using the current standard of contracture measurement (a pulley system) as well as a newly designed dynamic load cell (DLC) device. The DLC device was highly reliable when compared to the pulley system (r = 0.907, p < 0.001). Finally, the DLC device produced joint stiffness hysteresis curves capable of approximating the compliance of stiff joint tissues, ultimately calculating a mean joint stiffness of 1.57 ± 1.31 N · m · rad−1 (range, 0.33–6.37 N · m · rad−1). In conclusion, the DLC device represents a valid method for measuring joint contractures. Further, the DLC device notably improves current techniques by introducing the capacity to approximate the compliance of contracted tissues in living rabbits.

An Evidence-Based Clinical Prediction Algorithm for the Musculoskeletal Infection Society Minor Criteria

BACKGROUND The diagnosis of a periprosthetic joint infection (PJI) remains a clinical challenge, as there is no uniformly accepted gold standard. In 2011, the Musculoskeletal Infection Society (MSIS) convened a work group to create a standardized definition for a PJI that could be universally adopted. Based on the MSIS criteria, the diagnosis of a PJI can be made with 1 of the 2 major criteria, or 3 of the 5 minor criteria. The purpose of this study was to determine the likelihood of having a PJI based on the number of positive minor criteria and thereby develop a prediction algorithm for differentiating between a chronic PJI and a non-PJI based on the number of positive MSIS minor criteria. METHODS We retrospectively reviewed 297 patients who presented to a tertiary care center between 2004 and 2014 with a failed total joint arthroplasty and subsequently underwent a PJI workup to exclude chronic PJI. Patients were divided into 2 groups: (1) PJI group and (2) non-PJI group. Patients who had a positive PJI workup and subsequently underwent a 2-stage revision for infection were included in the PJI group. Patients who had a negative clinical and diagnostic workup were included in the non-PJI group. One hundred eighty-two patients met the criteria for inclusion in the study, 91 in each group. Univariate and multiple logistic regression analyses were used to evaluate 21 independent variables in each of the 2 groups. A prediction algorithm for differentiating between a chronic PJI and a non-PJI based on independent multivariate variables was created. RESULTS Patients who had a PJI differed significantly (P < .05) from those who did not have a PJI with regard to 10 independent variables, which included all the MSIS minor criteria we evaluated. Five independent multivariate variables were identified to differentiate between the 2 groups: positive cultures, elevated synovial white blood cell count, elevated synovial polymorphonuclear neutrophil percentage, elevated erythrocyte sedimentation rate, and elevated C-reactive protein. The predictive probability of a PJI for all 32 combinations of these 5 variables was: 3.6% for 1 positive variable, 19.3% for 2, 58.7% for 3, 83.8% for 4, and 97.8% for 5. The chi-squared test for trend and the area under the receiver-operating characteristic curve (0.977) suggest that the model is highly predictive, with an excellent diagnostic performance in identifying a PJI. CONCLUSIONS Diagnosing a PJI remains a clinical challenge as there is no gold standard for diagnosis. The development of the MSIS criteria, which is based on a consensus of over 400 of the worlds experts in musculoskeletal infection, was a major step forward in defining the diagnosis of a PJI. However, to our knowledge, the likelihood of having a PJI based on the number of positive minor criteria has yet to be validated or quantified. Of the 20 independent variables that were evaluated, 10 were found to be significantly associated with a PJI, including all the MSIS minor criteria evaluated. In addition, a diagnostic prediction algorithm was constructed to determine the likelihood of a PJI based on 5 binary independent multivariate variables. The relationship was also examined with a receiver-operating characteristic curve analysis. The area under the curve was 0.98, indicating excellent diagnostic performance for the MSIS minor criteria in identifying a PJI. LEVEL OF EVIDENCE III.

Molecular pathology of adverse local tissue reaction caused by metal-on-metal implants defined by RNA-seq.

Total hip arthroplasty (THA) alleviates hip pain and improves joint function. Current implant design permits long-term survivorship of THAs, but certain metal-on-metal (MoM) articulations can portend catastrophic failure due to adverse local tissue reactions (ALTR). Here, we identified biological and molecular differences between periacetabular synovial tissues of patients with MoM THA failure undergoing revision THA compared to patients undergoing primary THA for routine osteoarthritis (OA). Analysis of tissue biopsies by RNA-sequencing (RNA-seq) revealed that MoM patient samples exhibit significantly increased expression of immune response genes but decreased expression of genes related to extracellular matrix (ECM) remodeling. Thus, interplay between local tissue inflammation and ECM degradation may account for the pathology and compromised clinical outcomes in select patients with MoM implants. We conclude that adverse responses of host tissues to implant materials result in transcriptomic modifications in patients with MoM implants that permit consideration of strategies that could mitigate ECM damage.

A dedicated anticoagulation clinic does not improve postoperative management of warfarin after total joint arthroplasty

Background Periprosthetic joint infections (PJIs) are devastating complications. Excessive anticoagulation with warfarin is an independent risk factor for PJIs. The use of a dedicated anticoagulation clinic to improve warfarin management has not been proven. Methods Between 2006 and 2014, we identified 92 patients who were placed on postoperative warfarin, and later developed PJI. These patients were compared to 313 patients who underwent total joint arthroplasty placed on warfarin without developing PJI. Patients were included if they had no history of a venous thromboembolic event, were warfarin naive, and enrolled in the anticoagulation clinic. A univariate analysis compared independent variables, and statistical analysis was performed using Students t-test and Pearson chi-square test for continuous and categorical variables. Results Thirty-six PJI patients and 297 control patients met the inclusion criteria. The venous thromboembolism rate was 2.1%. At discharge, 82% of all patients were subtherapeutic. Patients were within their target international normalized ratio (INR) range 26.7% of the time. The mean INR in the initial postoperative period for the PJI group was 1.46 and 1.29 in the control group (P < .001). In the acute postoperative period, 13.3% of the knee PJI group were therapeutic or supratherapeutic compared with 3.5% in the knee control group (P = .002). Conclusions Despite utilization of a dedicated anticoagulation clinic, patients were only within their target INR range 27% of the time. Total knee arthroplasty patients who developed a PJI were more likely to be therapeutic or supratherapeutic in the initial postoperative period. Consequently, the risks associated with warfarin as a venous thromboembolism prophylaxis may outweigh the potential benefits.