Peter D. Clegg

Methicillin-resistant staphylococci in companion animals

We determined the molecular characteristics of methicillin-resistant staphylococci from animals and staff at a small animal and equine hospital. Methicillin-resistant Staphylococcus aureus (MRSA) identical to human EMRSA-15 was found in dogs and hospital staff. In contrast, 5 distinct MRSA strains were isolated from horses but not from hospital staff.

Aspartic Acid Racemization and Collagen Degradation Markers Reveal an Accumulation of Damage in Tendon Collagen That Is Enhanced with Aging

Little is known about the rate at which protein turnover occurs in living tendon and whether the rate differs between tendons with different physiological roles. In this study, we have quantified the racemization of aspartic acid to calculate the age of the collagenous and non-collagenous components of the high strain injury-prone superficial digital flexor tendon (SDFT) and low strain rarely injured common digital extensor tendon (CDET) in a group of horses with a wide age range. In addition, the turnover of collagen was assessed indirectly by measuring the levels of collagen degradation markers (collagenase-generated neoepitope and cross-linked telopeptide of type I collagen). The fractional increase in d-Asp was similar (p = 0.7) in the SDFT (5.87 × 10−4/year) and CDET (5.82 × 10−4/year) tissue, and d/l-Asp ratios showed a good correlation with pentosidine levels. We calculated a mean (±S.E.) collagen half-life of 197.53 (±18.23) years for the SDFT, which increased significantly with horse age (p = 0.03) and was significantly (p < 0.001) higher than that for the CDET (34.03 (±3.39) years). Using similar calculations, the half-life of non-collagenous protein was 2.18 (±0.41) years in the SDFT and was significantly (p = 0.04) lower than the value of 3.51 (±0.51) years for the CDET. Collagen degradation markers were higher in the CDET and suggested an accumulation of partially degraded collagen within the matrix with aging in the SDFT. We propose that increased susceptibility to injury in older individuals results from an inability to remove partially degraded collagen from the matrix leading to reduced mechanical competence.

Gene expression in human chondrocytes in late osteoarthritis is changed in both fibrillated and intact cartilage without evidence of generalised chondrocyte hypertrophy

Objectives: To investigate changes in gene expression in fibrillated and intact human osteoarthritis (OA) cartilage for evidence of an altered chondrocyte phenotype and hypertrophy. Methods: Paired osteochondral samples were taken from a high-load site and a low-load site from 25 OA joints and were compared with eight similar paired samples from age-matched controls. Gene expression of key matrix and regulatory genes was analysed by quantitative real-time reverse transcription-polymerase chain reaction on total RNA extracted from the cartilage. Results: There was a major change in chondrocyte gene expression in OA cartilage. SOX9 (38-fold) and aggrecan (4-fold) gene expression were both lower in OA (p<0.001), and collagen I (17-fold) and II (2.5-fold) gene expression were each increased in a subset of OA samples. The major changes in gene expression were similar at the fibrillated high-loaded site and the intact low-loaded site. There was no evidence of a generalised change in OA to proliferative or hypertrophic phenotype as seen in the growth plate, as genes associated with either stage of differentiation were unchanged (PTHrPR), or significantly downregulated (collagen X (14-fold, p<0.002), VEGF (23-fold, p<0.02), BCL-2 (5.6-fold, p<0.001), matrilin-1 (6.5-fold, p<0.001)). In contrast MMP-13 was significantly upregulated in the OA cartilage samples (5.3-fold, p<0.003). Conclusions: The expression of key chondrocyte genes, including aggrecan and SOX9, was decreased in OA cartilage and the changes were similar in both fibrillated high-loaded and intact low-loaded cartilage on the same joint. However, there was no significant upregulation of type X collagen, and other genes associated with chondrocyte further differentiation and hypertrophy.

Proposed Definitions and Criteria for Reporting Time Frame, Outcome, and Complications For Clinical Orthopedic Studies in Veterinary Medicine

Outcome, and Complications For Clinical Orthopedic Studies in Veterinary Medicine James L. Cook, DVM, PhD, Diplomate ACVS, Richard Evans, PhD, Michael G. Conzemius, DVM, PhD, Diplomate ACVS, B. Duncan X. Lascelles, BVSc, PhD, Diplomate ECVS, Diplomate ACVS, C. Wayne McIlwraith, BVSc, PhD, Diplomate ACVS, Antonio Pozzi, DMV, MS, Diplomate ACVS, Peter Clegg, MA, VetMB, PhD, Diplomate ECVS, MRCVS, John Innes, BVSc, PhD, DSAS (Orth), MRCVS, Kurt Schulz, DVM, Diplomate ACVS, John Houlton, MA, VetMB, DVR, DSAO, MRCVS, Diplomate ECVS, Lisa Fortier, DVM, PhD, Diplomate ACVS, Alan R. Cross, DVM, Diplomate ACVS, Kei Hayashi, DVM, PhD, Diplomate ACVS, Amy Kapatkin, DVM, MS, Diplomate ACVS, Dorothy Cimino Brown, DVM, MSCE, Diplomate ACVS, and Allison Stewart, DVM, MS, Diplomate ACVS Comparative Orthopaedic Laboratory, University of Missouri, Columbia, MO, College of Veterinary Medicine, University of Illinois, Urbana, IL, College of Veterinary Medicine, University of Minnesota, St. Paul, MN, Comparative Pain Research Laboratory, North Carolina State University,

Specialization of tendon mechanical properties results from interfascicular differences

Tendons transfer force from muscle to bone. Specific tendons, including the equine superficial digital flexor tendon (SDFT), also store and return energy. For efficient function, energy-storing tendons need to be more extensible than positional tendons such as the common digital extensor tendon (CDET), and when tested in vitro have a lower modulus and failure stress, but a higher failure strain. It is not known how differences in matrix organization contribute to distinct mechanical properties in functionally different tendons. We investigated the properties of whole tendons, tendon fascicles and the fascicular interface in the high-strain energy-storing SDFT and low-strain positional CDET. Fascicles failed at lower stresses and strains than tendons. The SDFT was more extensible than the CDET, but SDFT fascicles failed at lower strains than CDET fascicles, resulting in large differences between tendon and fascicle failure strain in the SDFT. At physiological loads, the stiffness at the fascicular interface was lower in the SDFT samples, enabling a greater fascicle sliding that could account for differences in tendon and fascicle failure strain. Sliding between fascicles prior to fascicle extension in the SDFT may allow the large extensions required in energy-storing tendons while protecting fascicles from damage.

The role of endogenous and exogenous enzymes in chronic wounds: a focus on the implications of aberrant levels of both host and bacterial proteases in wound healing.

Cutaneous wound healing is orchestrated by a number of physiological pathways that ultimately lead to reformation of skin integrity and the production of functional scar tissue. The remodeling of a wound is significantly affected by matrix metalloproteinases (MMPs), which act to control the degradation of the extracellular matrix (ECM). Regulation of MMPs is imperative for wound healing as excessive levels of MMPs can lead to disproportionate destruction of the wound ECM compared to ECM deposition. In addition to human MMPs, bacterial proteases have been found to be influential in tissue breakdown and, as such, have a role to play in the healing of infected wounds. For example, the zinc‐metalloproteinase, elastase, produced by Pseudomonas aeruginosa, induces degradation of fibroblast proteins and proteoglycans in chronic wounds and has also been shown to degrade host immune cell mediators. Microbial extracellular enzymes have also been shown to degrade human wound fluid and inhibit fibroblast cell growth. It is now being acknowledged that host and bacterial MMPs may act synergistically to cause tissue breakdown within the wound bed. Several studies have suggested that bacterial‐derived secreted proteases may act to up‐regulate the levels of MMPs produced by the host cells. Together, these findings indicate that bacterial phenotype in terms of protease producing potential of bacteria should be taken into consideration during diagnostic and clinical intervention of infected wound management. Furthermore, both host MMPs and those derived from infecting bacteria need to be targeted in order to increase the healing capacity of the injured tissue. The aim of this review is to investigate the evidence suggestive of a relationship between unregulated levels of both host and bacterial proteases and delayed wound healing.

A review of tendon injury: why is the equine superficial digital flexor tendon most at risk?

Tendon injury is one of the most common causes of wastage in the performance horse; the majority of tendon injuries occur to the superficial digital flexor tendon (SDFT) whereas few occur to the common digital extensor tendon. This review outlines the epidemiology and aetiology of equine tendon injury, reviews the different functions of the tendons in the equine forelimb and suggests possible reasons for the high rate of failure of the SDFT. An understanding of the mechanisms leading to matrix degeneration and subsequent tendon gross failure is the key to developing appropriate treatment and preventative measures.

Race- and course-level risk factors for fatal distal limb fracture in racing Thoroughbreds

REASONS FOR PERFORMING STUDY Considerable variation in the rates of equine fatality at different racecourses draws attention to probable risk factors at the level of course or race that might be partly responsible. Distal limb fractures are the most common cause of equine fatality on UK racecourses and identification of risk factors for such injuries and subsequent implementation of intervention strategies could significantly reduce the total number of racecourse fatalities. OBJECTIVES To identify race- and course-level risk factors for fatal distal limb fracture in Thoroughbreds on UK racecourses. METHODS A case-control study design was used. Case races were defined as those in which one or more horses sustained fatal fracture of the distal limb. Controls were selected in 2 different ways. Firstly, 3 races in which no fracture occurred were selected from all races of the same type held within 5 days of the case race (Analysis 1). Secondly, 3 control races were selected for each case race from all races of the same type held in the same year (Analysis 2). One hundred and nine cases were included in the study. Information about the race and the racecourses was collected from Computer Raceform. Conditional logistic regression was used to identify the relationship between a number of independent variables and the likelihood of fracture in a race. RESULTS Longer races with a larger number of runners were more likely to contain a fracture. Firmer going and fewer days since the last race on the same course were associated with an increased risk of fracture. The going at the course at the previous race meeting was also associated with the likelihood of fracture. CONCLUSIONS Modifications to the going on the day of a race and greater emphasis on ground maintenance between race meetings may have an impact on the risk of fatal distal limb fracture during racing. POTENTIAL RELEVANCE Modification of risk factors such as the going and number of days since the last race meeting could reduce the number of equine fatalities on UK racecourses. The condition of the racecourse may be an important risk factor and future research should focus on the identification of course maintenance techniques that produce the safest possible racing surfaces.

Cell phenotypic variation in normal and damaged tendons

Injuries to tendons are common in both human athletes as well as in animals, such as the horse, which are used for competitive purposes. Furthermore, such injuries are also increasing in prevalence in the ageing, sedentary population. Tendon diseases often respond poorly to treatment and require lengthy periods of rehabilitation. The tendon has a unique extracellular matrix, which has developed to withstand the mechanical demands of such tensile‐load bearing structures. Following injury, any repair process is inadequate and results in tissue that is distinct from original tendon tissue. There is growing evidence for the key role of the tendon cell (tenocyte) in both the normal physiological homeostasis and regulation of the tendon matrix and the pathological derangements that occur in disease. In particular, the tenocyte is considered to have a major role in effecting the subclinical matrix degeneration that is thought to occur prior to clinical disease, as well as in the severe degradative events that occur in the tendon at the onset of clinical disease. Furthermore, the tenocyte is likely to have a central role in the production of the biologically inadequate fibrocartilaginous repair tissue that develops subsequent to tendinopathy. Understanding the biology of the tenocyte is central to the development of appropriate interventions and drug therapies that will either prevent the onset of disease, or lead to more rapid and appropriate repair of injured tendon. Central to this is a full understanding of the proteolytic response in the tendon in disease by such enzymes as metalloproteinases, as well as the control of the inappropriate fibrocartilaginous differentiation. Finally, it is important that we understand the role of both intrinsic and extrinsic cellular elements in the repair process in the tendon subsequent to injury.

Mesenchymal stem cell therapy in equine musculoskeletal disease: scientific fact or clinical fiction?

The goal in the therapeutic use of mesenchymal stem cells (MSCs) in musculoskeletal disease is to harness the regenerative nature of these cells focussing on their potential to grow new tissues and organs to replace damaged or diseased tissue. Laboratory isolation of MSCs is now well established and has recently been demonstrated for equine MSCs. Stem cell science has attracted considerable interest in both the scientific and clinical communities because of its potential to regenerate tissues. Research into the use of MSCs in tissue regeneration in general reflects human medical needs, however, the nature, prevalence and prognosis of superficial digital flexor tendonitis has put equine veterinary science at the forefront of tendon regeneration research. Much has been investigated and learnt but it must be appreciated that in spite of this, the field is still relatively young and both communities must prepare themselves for considerable time and effort to develop the technology into a highly efficient treatments. The promise of functional tissue engineering to replace old parts with new fully justifies the interest. At present, however, it is important to balance the understanding of our current limitations with a desire to progress the technology.