GenPup-M: A novel validated owner-reported clinical metrology instrument for detecting early mobility changes in dogs.

OBJECTIVE: To use a previously validated veterinary clinical examination sheet, Liverpool Osteoarthritis in Dogs (LOAD) questionnaire, combined with kinetic and kinematic gait analysis in dogs with/without mobility problems to demonstrate the capacity of a novel clinical metrology instrument ("GenPup-M") to detect canine mobility impairments. DESIGN: Quantitative study. ANIMALS: 62 dogs (31 with mobility impairments and 31 without mobility impairments). PROCEDURE: The dogs' clinical history was obtained from owners and all dogs underwent a validated orthopaedic clinical examination. Mobility impairments were diagnosed in the mobility impaired group based on clinical history and orthopaedic examination. Owners were asked to complete GenPup-M along with a previously validated mobility questionnaire (Liverpool Osteoarthritis in Dogs (LOAD)) to identify construct validity. As a test of criterion validity, the correlation between instrument scores and the overall clinical examination scores, along with force-platform obtained peak vertical forces (PVF) were calculated. GenPup-M underwent internal consistency and factor analysis. Spatiotemporal parameters were calculated for dogs with/without mobility impairments to define the gait differences between these two groups. RESULTS: Principal Component Analysis identified GenPup-M had two components with Eigenvalues >1 ("stiffness/ease of movement" and "willingness to be active/exercise"). Cronbach's α was used to test internal consistency of GenPup-M and was found to be "good" (0.87). There was a strong, positive correlation between GenPup-M and LOAD responses (r2 = 0.69, p<0.001) highlighting construct validity. Criterion validity was also shown when comparing GenPup-M to clinical examination scores (r2 = 0.74, p<0.001) and PVF (r2 = 0.43, p<0.001). Quantitative canine gait analysis showed that there were statistically significant differences between peak vertical forces (PVF) of mobility impaired and non-mobility impaired dogs (p<0.05). Analyses of PVF showed that non-mobility impaired dogs more evenly distributed their weight across all thoracic and pelvic limbs when compared to mobility impaired dogs. There were also consistent findings that mobility impaired dogs moved slower than non-mobility impaired dogs. CONCLUSION AND CLINICAL RELEVANCE: GenPup-M is a clinical metrology instrument (CMI) that can be completed by dog owners to detect all mobility impairments, including those that are early in onset, indicating the versatility of GenPup-M to assess dogs with and without mobility impairments. Results of the study found that GenPup-M positively correlated with all three objective measures of canine mobility and consequently showed criterion and construct validity. Owner-reported CMIs such as GenPup-M allow non-invasive scoring systems which veterinary surgeons and owners can use to allow communication and longitudinal assessment of a dog's mobility. It is anticipated that GenPup-M will be used by owners at yearly vaccinations/health checks, allowing identification of any subtle mobility changes, and enabling early intervention.

Recall Bias in Client-Reported Outcomes in Canine Orthopaedic Patients Using Clinical Metrology Instruments.

OBJECTIVE: The aim of this study was to determine the accuracy of client recollection of their dogs' preconsultation status using clinical metrology instruments such as the Liverpool Osteoarthritis in Dogs (LOAD) and Canine Brief Pain Inventory (CBPI) questionnaires in dogs presenting to a referral orthopaedic clinic. STUDY DESIGN: This is a longitudinal prospective cohort study of client-owned dogs presenting for investigations of lameness (n = 217). LOAD and CBPI questionnaires were completed by the owners at the first consultation (T0). Owners were contacted at 2 (T1), 6 (T2), and 12 (T3) months and asked to recall their dogs' T0 status by completing another LOAD and CBPI questionnaire. The agreement between the T0 and recalled LOAD and CBPI scores was determined using the two-way mixed effects intraclass correlation coefficient (ICC). The Wilcoxon signed-rank test was used to determine the difference between scores. RESULTS: For the LOAD scores, there was moderate agreement between T0 and T1 (ICC: 0.64) and T0 and T2 (ICC: 0.53) scores and poor agreement between T0 and T3 (ICC: 0.496). For the CBPI Pain Severity Scores, there was poor agreement between T0 and all three subsequent time points (ICC < 0.5). For the CBPI Pain Interference Scores, there was moderate agreement between T0 and T1 (ICC: 0.57) and T2 (ICC: 0.56) scores and poor agreement between T0 and T3 (ICC: 0.43). CONCLUSION: The LOAD and CBPI questionnaires are subject to recall bias. Studies reporting retrospectively acquired CMI data should be interpreted with caution.

Ultrasonographic monitoring of feline epaxial muscle height as part of an annual wellness examination to assess for the development of sarcopenia.

OBJECTIVES: The aim of this study was to determine if epaxial muscle height (EMH) could be reliably incorporated into annual routine wellness screenings, and also determine its relationship to age, body condition score (BCS), subjective muscle assessment (SMA), breed and sex in mature cats. METHODS: EMH was determined independently by three observers from ultrasonographic examinations - collected by an additional trained individual - of cats enrolled at the Feline Healthy Ageing Clinic, University of Liverpool, UK. Age, body weight, BCS and SMA data were also collected. RESULTS: A total of 92 cats were included, 35 of which had repeat ultrasonographic examinations 12 months apart. Enrolled cats were a median age of 8 years and 9 months at the time of the first measurement. Variation in the quality of ultrasonographic images collected did not affect muscle depth measurements (P = 0.974). Further, there was good intra- and inter-observer repeatability for all observations (intraclass correlation range 0.97-0.99). There was a moderate positive association between EMH and body weight (r = 0.49, P <0.001) but no association with age (r = -0.05, P = 0.680). There were also positive associations in EMH among cats with different BCSs (P = 0.001) and SMAs (thoracic spine, P = 0.021; lumbar spine, P = 0.014), but breed (P = 0.429) and sex (P = 0.187) had no effect. Finally, there was no change in EMH measurements in the paired samples (P = 0.145) or correlation between percentage weight and EMH change over 12 months. CONCLUSIONS AND RELEVANCE: The accuracy of EMH measurement using ultrasonographic imaging is good, irrespective of observer experience and provided that the ultrasonographer has some training. This suggests that ultrasonographic measurement of EMH could have a major practical impact as a non-invasive determination of muscle mass in pet cat populations. Further research is required to assess longitudinal changes in muscle mass over time in senior pet cats.

Outcomes and complications reported from a multiuser canine hip replacement registry over a 10-year period.

OBJECTIVE: To report outcomes and complications associated with total hip replacements (THR) using a multiuser canine hip registry (CHR) and owner-administered questionnaire. STUDY DESIGN: Prospective longitudinal clinical study. ANIMALS: Dogs (n = 1852). METHODS: Total hip replacement cases submitted to a CHR were reviewed. An online questionnaire including an adapted "Liverpool Osteoarthritis in Dogs" (LOAD) score was e-mailed to owners. Data were analyzed to determine associations between clinical variables and the agreement by veterinary surgeons and owners for complications. RESULTS: A group of 1329 (72%) dogs had unilateral THRs and another group of 523 (28%) dogs had bilateral THRs, resulting in 2375 THRs. Indications included hip dysplasia and osteoarthritis (n = 2028/2375, 85%). Implants were manufactured by Kyon (n = 1087, 46%), BioMedtrix CFX (n = 514, 22%), BioMedtrix hybrid (n = 264, 11%), BioMedtrix BFX (n = 221, 9%), and Helica (n = 107, 4.5%). Median veterinary surgeon and owner follow up were 1328 and 900 days respectively. Postoperative LOAD scores (21 ± 9) reported by 461 owners improved compared to preoperative scores (11 ± 9) (P < .001). Veterinary surgeons reported complications in 201/2375 (8.5%) THRs and owners in 107/461 (23%) THRs, with moderate agreement (weighted kappa = 0.44). No associations were identified between complications and weight, age, sex, or breed. BioMedtrix BFX and Helica implants were associated with increased complications (P = .031) when used for revisions of femoral head and neck excisions. CONCLUSION: Excellent outcomes, including improved canine mobility, were reported after THRs. Complications were underreported by veterinary surgeons compared to owners in this first multiuser CHR. CLINICAL SIGNIFICANCE: Canine THRs are safe, effective procedures but THR implants should be carefully selected when revising femoral head and neck excisions.

Small RNA signatures of the anterior cruciate ligament from patients with knee joint osteoarthritis.

Introduction: The anterior cruciate ligament (ACL) is susceptible to degeneration, resulting in joint pain, reduced mobility, and osteoarthritis development. There is currently a paucity of knowledge on how anterior cruciate ligament degeneration and disease leads to osteoarthritis. Small non-coding RNAs (sncRNAs), such as microRNAs and small nucleolar RNA (snoRNA), have diverse roles, including regulation of gene expression. Methods: We profiled the sncRNAs of diseased osteoarthritic ACLs to provide novel insights into osteoarthritis development. Small RNA sequencing from the ACLs of non- or end-stage human osteoarthritic knee joints was performed. Significantly differentially expressed sncRNAs were defined, and bioinformatics analysis was undertaken. Results and Discussion: A total of 184 sncRNAs were differentially expressed: 68 small nucleolar RNAs, 26 small nuclear RNAs (snRNAs), and 90 microRNAs. We identified both novel and recognized (miR-206, -365, and -29b and -29c) osteoarthritis-related microRNAs and other sncRNAs (including SNORD72, SNORD113, and SNORD114). Significant pathway enrichment of differentially expressed miRNAs includes differentiation of the muscle, inflammation, proliferation of chondrocytes, and fibrosis. Putative mRNAs of the microRNA target genes were associated with the canonical pathways "hepatic fibrosis signaling" and "osteoarthritis." The establishing sncRNA signatures of ACL disease during osteoarthritis could serve as novel biomarkers and potential therapeutic targets in ACL degeneration and osteoarthritis development.

Proteoglycans play a role in the viscoelastic behaviour of the canine cranial cruciate ligament.

Proteoglycans (PGs) are minor extracellular matrix proteins, and their contributions to the mechanobiology of complex ligaments such as the cranial cruciate ligament (CCL) have not been determined to date. The CCLs are highly susceptible to injuries, and their extracellular matrix comprises higher PGs content than the other major knee ligaments. Hence these characteristics make CCLs an ideal specimen to use as a model in this study. This study addressed the hypothesis that PGs play a vital role in CCL mechanobiology by determining the biomechanical behaviour at low strain rates before and after altering PGs content. For the first time, this study qualitatively investigated the contribution of PGs to key viscoelastic characteristics, including strain rate dependency, hysteresis, creep and stress relaxation, in canine CCLs. Femur-CCL-tibia specimens (n = 6 pairs) were harvested from canine knee joints and categorised into a control group, where PGs were not depleted, and a treated group, where PGs were depleted. Specimens were preconditioned and cyclically loaded to 9.9 N at 0.1, 1 and 10%/min strain rates, followed by creep and stress relaxation tests. Low tensile loads were applied to focus on the toe-region of the stress-strain curves where the non-collagenous extracellular matrix components take significant effect. Biochemical assays were performed on the CCLs to determine PGs and water content. The PG content was ∼19% less in the treated group than in the control group. The qualitative study showed that the stress-strain curves in the treated group were strain rate dependent, similar to the control group. The CCLs in the treated group showed stiffer characteristics than the control group. Hysteresis, creep characteristics (creep strain, creep rate and creep compliance), and stress relaxation values were reduced in the treated group compared to the control group. This study suggests that altering PGs content changes the microstructural organisation of the CCLs, including water molecule contents which can lead to changes in CCL viscoelasticity. The change in mechanical properties of the CCLs may predispose to injury and lead to knee joint osteoarthritis. Future studies should focus on quantitatively identifying the effect of PG on the mechanics of intact knee ligaments across broader demography.

Ligament mechanics of ageing and osteoarthritic human knees.

Knee joint ligaments provide stability to the joint by preventing excessive movement. There has been no systematic effort to study the effect of OA and ageing on the mechanical properties of the four major human knee ligaments. This study aims to collate data on the material properties of the anterior (ACL) and posterior (PCL) cruciate ligaments, medial (MCL) and lateral (LCL) collateral ligaments. Bone-ligament-bone specimens from twelve cadaveric human knee joints were extracted for this study. The cadaveric knee joints were previously collected to study ageing and OA on bone and cartilage material properties; therefore, combining our previous bone and cartilage data with the new ligament data from this study will facilitate subject-specific whole-joint modelling studies. The bone-ligament-bone specimens were tested under tensile loading to failure, determining material parameters including yield and ultimate (failure) stress and strain, secant modulus, tangent modulus, and stiffness. There were significant negative correlations between age and ACL yield stress (p = 0.03), ACL failure stress (p = 0.02), PCL secant (p = 0.02) and tangent (p = 0.02) modulus, and LCL stiffness (p = 0.046). Significant negative correlations were also found between OA grades and ACL yield stress (p = 0.02) and strain (p = 0.03), and LCL failure stress (p = 0.048). However, changes in age or OA grade did not show a statistically significant correlation with the MCL tensile parameters. Due to the small sample size, the combined effect of age and the presence of OA could not be statistically derived. This research is the first to report tensile properties of the four major human knee ligaments from a diverse demographic. When combined with our previous findings on bone and cartilage for the same twelve knee cadavers, the current ligament study supports the conceptualisation of OA as a whole-joint disease that impairs the integrity of many peri-articular tissues within the knee. The subject-specific data pool consisting of the material properties of the four major knee ligaments, subchondral and trabecular bones and articular cartilage will advance knee joint finite element models.

Age-related changes in microRNAs expression in cruciate ligaments of wild-stock house mice.

Cruciate ligaments (CL) of the knee joint are injured following trauma or aging. MicroRNAs (miRs) are potential therapeutic targets in musculoskeletal disorders, but there is little known about the role of miRs and their expression ligaments during aging. This study aimed to (1) identify if mice with normal physical activity, wild-stock house mice are an appropriate model to study age-related changes in the knee joint and (2) investigate the expression of miRs in aging murine cruciate ligaments. Knee joints were collected from 6 and 24 months old C57BL/6 and wild-stock house mice (Mus musculus domesticus) for ligament and cartilage (OARSI) histological analysis. Expression of miR targets in CLs was determined in 6-, 12-, 24-, and 30-month-old wild-stock house mice, followed by the analysis of predicted mRNA target genes and Ingenuity Pathway Analysis. Higher CL and knee OARSI histological scores were found in 24-month-old wild-stock house mice compared with 6- and 24-month-old C57BL/6 and 6-month-old wild-stock house mice (p < 0.05). miR-29a and miR-34a were upregulated in 30-month-old wild-stock house mice in comparison with 6-, 12-, and 24-month-old wild-stock house mice (p < 0.05). Ingenuity Pathway Analysis on miR-29a and 34a targets was associated with inflammation through interleukins, TGFß and Notch genes, and p53 signaling. Collagen type I alpha 1 chain (COL1A1) correlated negatively with both miR-29a (r = -0.35) and miR-34a (r = -0.33). The findings of this study support wild-stock house mice as an appropriate aging model for the murine knee joint. This study also indicated that miR-29a and miR-34a may be potential regulators of COL1A1 gene expression in murine CLs.

CT attenuation of the medial coronoid process is reduced in dogs with medial coronoid disease but independent of arthroscopic disease severity.

OBJECTIVE: To compare the attenuation of the medial coronoid process (MCP) in dogs with and without arthroscopically confirmed evidence of medial coronoid disease (MCD). ANIMALS: The database at our institution was searched for cases with thoracic limb lameness, diagnosed with MCD by arthroscopic examination that had CT as part of their investigation and compared with a control group of elbow joints from cadavers euthanized for reasons unrelated to MCD. A total of 84 elbow joints were included that met these criteria. PROCEDURES: Following CT, a standardized measurement of the MCP was obtained from apex to base and the mean attenuation, SD, and total area were recorded. A comparative measurement was obtained from the proximal radial cortex at the level of the nutrient foramen. Elbow joint arthroscopy was carried out using standard portals, and the modified Outerbridge score was (MOS) used to score elbow joint cartilage. Descriptive and inferential statistics were carried out using MLwiN and R. RESULTS: Attenuation of the MCP was reduced in dogs with MCD compared with those with no MCD (P < .002). No significant differences were observed in the attenuation between categories of severity (MOS). There was good inter- and intraobserver agreement between measurements (intraclass correlation coefficient = 0.89 and 0.95, respectively). CLINICAL RELEVANCE: MCP attenuation is reduced in dogs with MCD compared with dogs with no evidence of MCD. This finding may be a useful tool for early detection of MCD, but there is no relationship with arthroscopic lesion severity.

Long-term follow up of 44 cats undergoing total hip replacement: Cases from a feline hip registry (2010-2020).

OBJECTIVE: To report indications, complications, and long-term outcomes following feline total hip replacement (THR) using a client-based clinical metrology questionnaire, the Feline Musculoskeletal Pain Index (FMPI), and owner satisfaction. STUDY DESIGN: Multi-institutional retrospective cohort study. ANIMALS: Cats (n = 44) that underwent THR (n = 56). METHODS: Feline THRs submitted to a registry over a 10-year period were reviewed. The FMPI and owner satisfaction surveys were used to assess outcome. RESULTS: Forty-four cats met the inclusion criteria. Median age was 2 years (range: 0.9-11), and median bodyweight 5 kg (range: 3.3-7.6). British Shorthair and Domestic Shorthair were the most frequent breeds. Most cats were neutered males (33/44) and slipped capital femoral epiphysis (SCFE) was the most common surgical indication (34/56). All implants were cemented micro and nano hip implants. Overall complications (11/56) included 9 major complications. The median duration of follow up was 752 days (range: 102-3089). No association was found between clinical variables and complications. The FMPI score improved from 0.111 (range: 0-1.222) to 2.111 (range: 0.888-3.666) postoperatively (P < .001). Owner satisfaction was reported as "very good" in 30/33 cases (90.9%). CONCLUSION: A validated client metrology questionnaire showed clinical improvement in cats following THR. The most common indication for THR in cats was SCFE occurring in young male neutered cats. Complication rates were comparable to previous reports. CLINICAL SIGNIFICANCE: Total hip replacement appears to be a successful surgical treatment for feline hip disease with very good owner satisfaction and acceptable complication rates.

The anterior cruciate ligament in murine post-traumatic osteoarthritis: markers and mechanics.

BACKGROUND: Knee joint injuries, common in athletes, have a high risk of developing post-traumatic osteoarthritis (PTOA). Ligaments, matrix-rich connective tissues, play important mechanical functions stabilising the knee joint, and yet their role post-trauma is not understood. Recent studies have shown that ligament extracellular matrix structure is compromised in the early stages of spontaneous osteoarthritis (OA) and PTOA, but it remains unclear how ligament matrix pathology affects ligament mechanical function. In this study, we aim to investigate both structural and mechanical changes in the anterior cruciate ligament (ACL) in a mouse model of knee trauma. METHODS: Knee joints were analysed following non-invasive mechanical loading in male C57BL/6 J mice (10-week-old). Knee joints were analysed for joint space mineralisation to evaluate OA progression, and the ACLs were assessed with histology and mechanical testing. RESULTS: Joints with PTOA had a 33-46% increase in joint space mineralisation, indicating OA progression. Post-trauma ACLs exhibited extracellular matrix modifications, including COL2 and proteoglycan deposition. Additional changes included cells expressing chondrogenic markers (SOX9 and RUNX2) expanding from the ACL tibial enthesis to the mid-substance. Viscoelastic and mechanical changes in the ACLs from post-trauma knee joints included a 20-21% decrease in tangent modulus at 2 MPa of stress, a decrease in strain rate sensitivity at higher strain rates and an increase in relaxation during stress-relaxation, but no changes to hysteresis and ultimate load to failure were observed. CONCLUSIONS: These results demonstrate that ACL pathology and viscoelastic function are compromised in the post-trauma knee joint and reveal an important role of viscoelastic mechanical properties for ligament and potentially knee joint health.

Aging in Cats: Owner Observations and Clinical Finding in 206 Mature Cats at Enrolment to the Cat Prospective Aging and Welfare Study.

Two hundred and six cats, aged between 7 and 10 years, from the North-west of the UK, were enrolled in a cat aging and welfare study to determine the frequency of age-related conditions and associations with husbandry, owner observations of physical appearance, activity and behavior. This is the largest study to date of mature cats that includes data from an owner questionnaire and clinical examinations. At enrolment, owners frequently reported physical changes (53%), behavioral changes (47%), changes to eating patterns (41%), and activity changes (40%) in their mature cats. On physical examination, 45% cats were in overweight condition and 12% were obese. A heart murmur was detected in 29% cats, whilst indirect systolic blood pressure (SBP) was >160 mmHg in 5% cats. Dental disease was present in 54% cats and was associated with a matted hair coat (P = 0.01), increased sleeping (P = 0.02), absence of gray hairs (P = 0.03), and increased irritability to other pets (P = 0.04). Abnormalities were evident in 58% of cats that allowed an orthopedic examination (OE) to be performed. These cats were older than cats with a normal OE (P = 0.01), and abnormal OE findings were associated with a matted coat (P = 0.03) and increased grooming (P = 0.04). Aazotaemia was present in 10% cats, and this was associated with cats being observed to "sniff their food and then walk away" (P = 0.04). Hyperthyroidism was diagnosed in 3% cats, who were older (P = 0.02), had a leaner BCS (P = 0.02) and lesser blood creatinine concentrations (P = 0.01). Hyperthyroid cats were also more likely to have increased liver enzyme activity and increased SBP (P = <0.001) compared with non-hyperthyroid cats. Of the 176 cats where all clinical assessments were conducted, only 12% had no evidence of any disease. Clinical abnormalities are commonly identified when thorough, clinical assessments are performed in mature pet cats visiting primary care practice.

Analysis of feline humeral fracture morphology and a comparison of fracture repair stabilisation methods: 101 cases (2009-2020).

OBJECTIVES: The aims of this study were to describe the type, presentation and prognostic factors of feline humeral fractures over a 10-year period and to compare three stabilisation systems for feline humeral diaphyseal fractures. METHODS: In total, 101 cats with humeral fractures presenting to seven UK referral centres between 2009 and 2020 were reviewed. Data collected included signalment, weight at the time of surgery, fracture aetiology, preoperative presentation, fixation method, surgical details, perioperative management and follow-up examinations. Of these cases, 57 cats with humeral diaphyseal fractures stabilised using three different fixation methods were compared, with outcome parameters including the time to radiographic healing, time to function and complication rate. RESULTS: The majority of the fractures were diaphyseal (71%), with only 10% condylar. Of the known causes of fracture, road traffic accidents (RTAs) were the most common. Neutered males were over-represented in having a fracture caused by an RTA (P = 0.001) and diaphyseal fractures were significantly more likely to result from an RTA (P = 0.01). Body weight had a positive correlation (r = 0.398) with time to radiographic healing and time to acceptable function (r = 0.315), and was significant (P = 0.014 and P = 0.037, respectively). Of the 57 humeral diaphyseal fractures; 16 (28%) were stabilised using a plate-rod construct, 31 (54%) using external skeletal fixation and 10 (18%) using bone plating and screws only. Open diaphyseal fractures were associated with more minor complications (P = 0.048). There was a significant difference between fixation groups in terms of overall complication rate between groups (P = 0.012). There was no significant difference between fixation groups in time to radiographic union (P = 0.145) or time to acceptable function (P = 0.306). CONCLUSIONS AND RELEVANCE: All three fixation systems were successful in healing a wide variety of humeral diaphyseal fractures. There was a significantly higher overall complication rate with external skeletal fixators compared with bone plating; however, the clinical impact of these is likely low.

The effect of exercise on the protein profile of rat knee joint intra- and extra-articular ligaments.

Injuries to the intra-articular anterior cruciate ligament (ACL) and the extra-articular medial collateral ligament (MCL) result in significant knee joint instability, pain, and immobility. Moderate endurance-type exercise can increase ligament strength but little is known on the effect of short-term regular bouts of high-intensity exercise on the extracellular matrix (ECM) structure of knee ligaments. Therefore, this study aimed to identify the effect of short-term regular bouts high exercise on the proteome of the rat ACL and MCL using mass spectrometry. Sprague-Dawley male rats (n = 6) were split into control and exercise groups, and subjected to high-intensity training for four 4 weeks followed by proteomic analyses of the ACL and MCL. Knee joint health status was assessed using OARSI and a validated histological scoring system. Histopathological analyses demonstrated no significant changes in either in cruciate, collateral ligaments, or cartilage between the control and exercised knee joints. However, significant proteins were found to be more abundant in the exercised ACL compared to ACL control group but not between the exercised MCL and control MCL groups. The significant abundant proteins in ACL exercise groups were mostly cytoskeletal, ribosomal and enzymes with several abundant matrisomal proteins such as collagen proteins and proteoglycans being found in this group. In conclusion, our results indicate that short-term regular bouts of high-intensity exercise have an impact on the intra-articular ACL but not extra-articular MCL ECM protein expression.

Intrafascicular chondroid-like bodies in the ageing equine superficial digital flexor tendon comprise glycosaminoglycans and type II collagen.

The superficial digital flexor tendon (SDFT) is considered functionally equivalent to the human Achilles tendon. Circular chondroid depositions scattered amongst the fascicles of the equine SDFT are rarely reported. The purpose of this study was the detailed characterization of intrafascicular chondroid-like bodies (ICBs) in the equine SDFT, and the assessment of the effect of ageing on the presence and distribution of these structures. Ultrahigh field magnetic resonance imaging (9.4T) series of SDFT samples of young (1-9 years) and aged (17-25 years) horses were obtained, and three-dimensional reconstruction of ICBs was performed. Morphological evaluation of the ICBs included histology, immunohistochemistry and transmission electron microscopy. The number, size, and position of ICBs was determined and compared between age groups. There was a significant difference (p = .008) in the ICB count between young and old horses with ICBs present in varying number (13-467; median = 47, mean = 132.6), size and distribution in the SDFT of aged horses only. There were significantly more ICBs in the tendon periphery when compared with the tendon core region (p = .010). Histological characterization identified distinctive cells associated with increased glycosaminoglycan and type II collagen extracellular matrix content. Ageing and repetitive strain frequently cause tendon micro-damage before the development of clinical tendinopathy. Documentation of the presence and distribution of ICBs is a first step towards improving our understanding of the impact of these structures on the viscoelastic properties, and ultimately their effect on the risk of age-related tendinopathy in energy-storing tendons.

Multiomics analysis of the mdx/mTR mouse model of Duchenne muscular dystrophy.

PURPOSE/AIM: Duchenne muscular dystrophy (DMD) is a progressive neuromuscular disease characterized by extensive muscle weakness. Patients with DMD lack a functional dystrophin protein, which transmits force and organizes the cytoskeleton of skeletal muscle. Multiomic studies have been proposed as a way to obtain novel insight about disease processes from preclinical models, and we used this approach to study pathological changes in dystrophic muscles. MATERIALS AND METHODS: We evaluated hindlimb muscles of male mdx/mTR mice, which lack a functional dystrophin protein and have deficits in satellite cell abundance and proliferative capacity. Wild type (WT) C57BL/6 J mice served as controls. Muscle fiber contractility was measured, along with changes in the transcriptome using RNA sequencing, and in the proteome, metabolome, and lipidome using mass spectrometry. RESULTS: While mdx/mTR mice displayed gross pathological changes and continued cycles of degeneration and regeneration, we found no differences in permeabilized fiber contractility between strains. However, there were numerous changes in the transcriptome and proteome related to protein balance, contractile elements, extracellular matrix, and metabolism. There was only a 53% agreement in fold-change data between the proteome and transcriptome. Numerous changes in markers of skeletal muscle metabolism were observed, with dystrophic muscles exhibiting elevated glycolytic metabolites such as 6-phosphoglycerate, fructose-6-phosphate and glucose-6-phosphate, fructose bisphosphate, phosphorylated hexoses, and phosphoenolpyruvate. CONCLUSIONS: These findings highlight the utility of multiomics in studying muscle disease, and provide additional insight into the pathological changes in dystrophic muscles that might help to indirectly guide evidence-based nutritional or exercise prescription in DMD patients.

Feline musculoskeletal ageing: How are we diagnosing and treating musculoskeletal impairment?

PRACTICAL RELEVANCE: An understanding of the process of musculoskeletal ageing - which all senior and geriatric cats will experience - is vital to maintaining the health and welfare of our ageing cat population. CLINICAL CHALLENGES: Assessment of the feline musculoskeletal system is not always straightforward. Diagnosis of impairment relies on input from owners and veterinarians in terms of visual observation, and clinical and orthopaedic examination, in addition to diagnostic imaging. AUDIENCE: This review is written for the primary care veterinary team. AIMS: The goals are to raise awareness and improve clinical diagnosis of musculoskeletal impairment as a result of ageing. The article also reviews therapeutic options and considers the evidence available for the prevention/deceleration of musculoskeletal ageing and impairment. EVIDENCE BASE: There is good evidence of a high prevalence of osteoarthritis (OA) and degenerative joint disease (DJD) in older cats. There is also good evidence to indicate that functional impairment and chronic pain are sequelae of musculoskeletal disease. However, there is a paucity of information for what is best practice for the management and treatment of musculoskeletal impairment in a clinical situation. There is also a lack of evidence on how prevention of central stimulation of the nervous system caused by musculoskeletal impairment and, in turn the development of chronic pain, can be avoided.

Investigation of Variables Associated with Surgical Site Infection following the Management of Canine Cranial Cruciate Ligament Rupture with a Lateral Fabellotibial Suture.

OBJECTIVE: This study investigated variables associated with surgical site infection (SSI) in dogs with cranial cruciate ligament rupture managed with stifle joint examination and lateral fabellotibial suture stabilization. STUDY DESIGN: A retrospective study of dogs that had stifle arthroscopy, stifle arthrotomy, or a combination of both, followed by lateral fabellotibial suture stabilization for cranial cruciate ligament rupture. All cases had a minimum follow-up of 90 days. Lameness grades were recorded preoperatively, and at 6-week and final follow-up. RESULTS: One hundred fifty procedures in 130 dogs met the inclusion criteria. Overall, SSI rate was 17.3% and removal of the lateral fabellotibial suture was performed in 53% of SSI. Multivariable analysis showed significant association between SSI and bodyweight (p = 0.013), and induction using propofol (p = 0.029). Multilevel ordinal logistic regression analysis showed a greater proportion of dogs had a higher lameness grade at 6-week (p = 0.021) and final follow-up (p = 0.002) assessments in the infected compared with non-infected dogs. CONCLUSION: Our study demonstrated a higher SSI incidence than previously reported in dogs undergoing a lateral fabellotibial suture for cranial cruciate ligament rupture. Bodyweight and induction with propofol were identified as significant risk factors for postoperative SSI. Owners could be advised of an increased SSI risk in larger dogs and consideration should be given to selection of induction agent. Dogs that develop an SSI have a worse lameness grade at 6-week and final follow-up.

A full-field 3D digital image correlation and modelling technique to characterise anterior cruciate ligament mechanics ex vivo.

It is limiting to use conventional methods when characterising material properties of complex biological tissues with inhomogeneous and anisotropic structure, such as the anterior cruciate ligament (ACL) in the knee joint. This study aims to develop and utilise a three-dimensional digital image correlation method (3D DIC) for the purpose of determining material properties of femur-ACL-tibia complex across the surface without any contact between the tissue and the loading equipment. A full-field (360° view) 3D DIC test setup consisting of six digital single-lens reflex cameras was developed and ACL specimens from skeletally mature dog knee joints were tested. The six cameras were arranged into three pairs and the cameras within each pair were positioned with 25° in between to obtain the desired stereovision output. The test setup was calibrated twice: first to obtain the intrinsic and extrinsic parameters within camera pairs, and second to align the 3D surfaces from each camera pair in order to generate the full view of the ACLs. Using the undeformed 3D surfaces of the ligaments, ACL-specific finite element models were generated. Longitudinal deformation of ligaments under tensile loads obtained from the 3D DIC, and this was analysed to serve as input for the inverse finite element analysis. As a result, hyperelastic coefficients from the first-order Ogden model that characterise ACL behaviour were determined with a marginal error of <1.5%. This test setup and methodology provides a means to accurately determine inhomogeneous and anisotropic material properties of ACL. The methodology described in this study could be adopted to investigate other biological and cultured tissues with complex structure. STATEMENT OF SIGNIFICANCE: Determining the material properties of soft tissues with complex anatomical structure, such as the anterior cruciate ligament (ACL), is important to better understand their contribution to musculoskeletal biomechanics. Current conventional methods for characterising material properties of the ACL are often limited to a contact measurement approach, however an improved understanding of the mechanics of this complex tissue is vital in terms of preventing injury and developing novel therapies. This article reports the development and utilisation of non-contact optical methodology involving full-field three-dimensional digital image correlation and finite element analysis to accurately investigate material properties of the ACL, in a controlled environment. This technique reduces inaccuracies due to specimen clamping and more importantly considers the inhomogeneous nature of the examined tissue.

Meniscal and ligament modifications in spontaneous and post-traumatic mouse models of osteoarthritis.

BACKGROUND: Osteoarthritis (OA) is a whole joint disease that affects all joint tissues, with changes in the articular cartilage (AC), subchondral bone and synovium. Pathologies in menisci and ligaments, however, are rarely analysed, although both are known to play vital roles in the mechanical stability of the joint. The aim of our study was to describe the pathological changes in menisci and ligament during disease development in murine spontaneous and post-traumatic surgically induced OA and to quantify tissue mineralisation in the joint space using micro-computed tomography (µCT) imaging during OA progression. METHODS: Knees of Str/ort mice (spontaneous OA model; 26-40 weeks) and C57CBA F1 mice following destabilisation of medial meniscus (DMM) surgery (post-traumatic OA model; 8 weeks after DMM), were used to assess histological meniscal and ligament pathologies. Joint space mineralised tissue volume was quantified by µCT. RESULTS: Meniscal pathological changes in Str/ort mouse knees were associated with articular cartilage lesion severity. These meniscal changes included ossification, hyperplasia, cell hypertrophy, collagen type II deposition and Sox9 expression in the fibrous region near the attachment to the knee joint capsule. Anterior cruciate ligaments exhibited extracellular matrix changes and chondrogenesis particularly at the tibial attachment site, and ossification was seen in collateral ligaments. Similar changes were confirmed in the post-traumatic DMM model. µCT analysis showed increased joint space mineralised tissue volume with OA progression in both the post-traumatic and spontaneous OA models. CONCLUSIONS: Modifications in meniscal and ligament mineralisation and chondrogenesis are seen with overt AC degeneration in murine OA. Although the aetiology and the consequences of such changes remain unknown, they will influence stability and load transmission of the joint and may therefore contribute to OA progression. In addition, these changes may have important roles in movement restriction and pain, which represent major human clinical symptoms of OA. Description of such soft tissue changes, in addition to AC degradation, should be an important aspect of future studies in mouse models in order to furnish a more complete understanding of OA pathogenesis.