Osteoarthritis, adipokines and the translational research potential in small animal patients.

Osteoartritis (OA) is a debilitating disease affecting both humans and animals. In the early stages, OA is characterized by damage to the extracellular matrix (ECM) and apoptosis and depletion of chondrocytes. OA progression is characterized by hyaline cartilage loss, chondrophyte and osteophyte formation, thickening of the joint capsule and function loss in the later stages. As the regenerative potential of cartilage is very limited and osteoarthritic changes are irreversible, prevention of OA, modulation of existing osteoarthritic joint inflammation, reducing joint pain and supporting joint function are the only options. Progression of OA and pain may necessitate surgical intervention with joint replacement or arthrodesis as end-stage procedures. In human medicine, the role of adipokines in the development and progression of OA has received increasing interest. At present, the known adipokines include leptin, adiponectin, visfatin, resistin, progranulin, chemerin, lipocalin-2, vaspin, omentin-1 and nesfatin. Adipokines have been demonstrated to play a pivotal role in joint homeostasis by modulating anabolic and catabolic balance, autophagy, apoptosis and inflammatory responses. In small animals, in terms of dogs and cats, naturally occurring OA has been clearly demonstrated as a clinical problem. Similar to humans, the etiology of OA is multifactorial and has not been fully elucidated. Humans, dogs and cats share many joint related degenerative diseases leading to OA. In this review, joint homeostasis, OA, adipokines and the most common joint diseases in small animals leading to naturally occurring OA and their relation with adipokines are discussed. The purpose of this review is highlighting the translational potential of OA and adipokines research in small animal patients.

Assessment of Outcome of Arthroscopic Subtotal Coronoidectomy in Treating Medial Coronoid Disease and Effect of Concurrent Autologous Conditioned Plasma in Dogs Using Force Plate Analysis.

OBJECTIVE: The aim of this study was to assess the outcome of arthroscopic subtotal coronoidectomy in treating medial coronoid disease and the effect of concurrent autologous conditioned plasma (ACP) in dogs using force plate analysis. STUDY DESIGN: This study included 16 dogs with unilateral medial coronoid disease based on computed tomographic imaging. Treatment consisted of an arthroscopic subtotal coronoidectomy. As concurrent therapy, eight dogs received ACP at the end of the arthroscopy and eight dogs a placebo. Force plate analysis, including vertical forces, braking and propulsive forces, corresponding impulses and symmetry indices, was performed prior to arthroscopy and at 4, 12 and 26 weeks postoperatively. RESULTS: After an initial reduction of all force plate analysis parameters, vertical and propulsive parameters gradually improved to the level of healthy dogs at 26 weeks postoperatively. In contrast, braking parameters did not reach normal values at 26 weeks, demonstrating ongoing loss of elbow joint function. The ACP and placebo group did not show any significant difference in force plate analysis parameters during the 26 weeks study period. CONCLUSION: Arthroscopic subtotal coronoidectomy did not result in complete normalization of ground reaction forces, impulses and symmetry indices. Concurrent treatment with ACP, immediately after arthroscopy, did not improve outcome. Our results show that outcome of arthroscopic subtotal coronoidectomy in treating medial coronoid disease is less favourable than previously reported.

Apoptosis Regulation in Osteoarthritis and the Influence of Lipid Interactions.

Osteoarthritis (OA) is one of the most common chronic diseases in human and animal joints. The joints undergo several morphological and histological changes during the development of radiographically visible osteoarthritis. The most discussed changes include synovial inflammation, the massive destruction of articular cartilage and ongoing joint destruction accompanied by massive joint pain in the later stadium. Either the increased apoptosis of chondrocytes or the insufficient apoptosis of inflammatory macrophages and synovial fibroblasts are likely to underly this process. In this review, we discuss the current state of research on the pathogenesis of OA with special regard to the involvement of apoptosis.

Comparison of Single versus Double Lateral Plating in Treatment of Feline Ilial Fractures Using Veterinary Cuttable Plates.

OBJECTIVE: The aim of this study was to compare the frequency of implant failure and the extent of pelvic canal narrowing associated with the fixation of ilial fractures in cats with a single veterinary cuttable plate (SLP) or double veterinary cuttable plates (DLP) applied to the lateral surface of the ilium. STUDY DESIGN: Radiographic evaluation of feline ilial fractures plated laterally using SLP or DLP. Pelvic canal narrowing directly postoperatively and at 6 weeks follow-up was objectively measured using the sacral index (SI). Radiographs were evaluated for implant failure and fracture healing. RESULTS: Seventy-seven cats satisfied the inclusion criteria. Twenty-nine fractures were treated with a SLP and 48 with DLP. Implant failure occurred significantly more (p = 0.001) in the SLP group (14/29) compared with the DLP group (6/48). Follow-up SI was significantly different between the two groups (p = 0.048, SLP median: 1.0 range: 0.83-2.4, DLP median: 0.98; range: 0.76-1.45). Median change in SI was -0.04 (range: -1.4 to 0.05) in the SLP group and 0.0 (range: -0.23 to 0.23) in the DLP group. This difference was significantly different (p = 0.031). CONCLUSION: DLP leads to significantly less implant failure and significantly less pelvic canal narrowing compared with SLP. This difference in pelvic canal narrowing was small and the clinical relevance remains unclear.

Proximodistal and caudocranial position of the insertion of the patellar ligament on the tibial tuberosity and patellar ligament length of normal stifles and stifles with grade II medial patellar luxation in small-breed dogs.

OBJECTIVE: To assess the role of the proximodistal and caudocranial relative position of the patellar ligament insertion on the tibia and patellar ligament length-to-patellar length ratio (PLL:PL) in small-breed dogs with and without grade II medial patellar luxation (MPL). STUDY DESIGN: Retrospective study. SAMPLE POPULATION: Dogs weighing ≤15 kg, including 43 stifles with MPL and 34 control stifles. METHODS: The proximodistal and caudocranial relative position of the patellar ligament insertion was determined as a ratio using the vertical (VTT) and horizontal distance (HTT) between the tibial tuberosity insertion and the tibial plateau divided by the tibial plateau length (TPL). In addition, PLL:PL and tibial plateau angle (TPA) were determined. RESULTS: The VTT:TPL ratio was lower in affected stifles (95% CI: 0.86-0.94) than in the control group (0.93-1.01; p = .01). No other difference was identified between affected and normal stifles. CONCLUSION: The only difference identified in this study consisted of a more proximal position of the patellar ligament insertion in the stifles of small-breed dogs with grade II MPL. CLINICAL SIGNIFICANCE: The more proximal position of the patellar ligament insertion will result in a more proximal position of the patella in the trochlear groove and may contribute to the development of MPL. Potentially, this will also affect the risk of recurrence of MPL after surgical treatment.

Treatment of deep corneal ulcers with porcine small intestinal submusosa using a modified surgical technique in dogs.

Treatment of deep corneal ulcers using small intestinal submucosa (SIS) without corneal sutures in combination with a third eyelid flap seems a promising technical modification. Advantages are a reduction in surgery time and no additional damage of healthy corneal tissue.

Orthopedic surgery increases atherosclerotic lesions and necrotic core area in ApoE-/- mice.

BACKGROUND AND AIMS: Observational studies show a peak incidence of cardiovascular events after major surgery. For example, the risk of myocardial infarction increases 25-fold early after hip replacement. The acuteness of this increased risk suggests abrupt enhancement in plaque vulnerability, which may be related to intra-plaque inflammation, thinner fibrous cap and/or necrotic core expansion. We hypothesized that acute systemic inflammation following major orthopedic surgery induces such changes. METHODS: ApoE-/- mice were fed a western diet for 10 weeks. Thereafter, half the mice underwent mid-shaft femur osteotomy followed by realignment with an intramedullary K-wire, to mimic major orthopedic surgery. Mice were sacrificed 5 or 15 days post-surgery (n = 22) or post-saline injection (n = 13). Serum amyloid A (SAA) was measured as a marker of systemic inflammation. Paraffin embedded slides of the aortic root were stained to measure total plaque area and to quantify fibrosis, calcification, necrotic core, and inflammatory cells. RESULTS: Surgery mice showed a pronounced elevation of serum amyloid A (SAA) and developed increased plaque and necrotic core area already at 5 days, which reached significance at 15 days (p = 0.019; p = 0.004 for plaque and necrotic core, respectively). Macrophage and lymphocyte density significantly decreased in the surgery group compared to the control group at 15 days (p = 0.037; p = 0.024, respectively). The density of neutrophils and mast cells remained unchanged. CONCLUSIONS: Major orthopedic surgery in ApoE-/- mice triggers a systemic inflammatory response. Atherosclerotic plaque area is enlarged after surgery mainly due to an increase of the necrotic core. The role of intra-plaque inflammation in this response to surgical injury remains to be fully elucidated.

Radiographic, computed tomographic, and arthroscopic findings in labrador retrievers with medial coronoid disease.

OBJECTIVE: To describe the radiographic, computed tomographic (CT), and arthroscopic findings in different age groups of Labrador Retrievers diagnosed with medial coronoid disease (MCD), and to compare the ulnar subtrochlear sclerosis (STS) observed on radiographs with the ratio between the mean attenuation of the ulnar subtrochlear bone and the mean attenuation of the cortical bone measured on CT. STUDY DESIGN: Prospective clinical study. ANIMALS: Dogs (n = 31; 31 elbow joints) and 6 healthy Labrador Retrievers (6 elbow joints). METHODS: Radiographic, CT, and intraoperative arthroscopic images (2008-2012) were evaluated. Statistical analysis was performed for the descriptive study to evaluate the difference in findings between age groups and to investigate the correlation between radiographic and CT evaluated ulnar STS. RESULTS: Ulnar STS (87.6%) was the most common radiographic findings in dogs ≤12 months and blurring of the cranial edge of the medial coronoid process (MCP; 66.7%) was the most common radiographic findings in dogs >12 months. MCP fragmentation was the most common CT finding in both age groups (93.8% [≤12 months]; 66.7% [>12 months]). A displaced fragment (68.8%) was the most common arthroscopic finding in dogs ≤12 months whereas osteochondromalacia (53.3%) was the most common finding in dogs >12 months. Sensitivity of radiography in detecting MCD was 93.8% (≤12 months) and 73.3% (>12 months) and for CT was 93.8% (≤12 months) and 66.7% (>12 months). Radiographic evaluated ulnar STS was strongly correlated with CT evaluated ulnar STS. CONCLUSION: Wide ranges of radiographic, CT, and arthroscopic findings in Labrador Retrievers diagnosed with medial coronoid disease were identified.

Decellularized cartilage-derived matrix as substrate for endochondral bone regeneration.

Following an endochondral approach to bone regeneration, multipotent stromal cells (MSCs) can be cultured on a scaffold to create a cartilaginous callus that is subsequently remodeled into bone. An attractive scaffold material for cartilage regeneration that has recently regained attention is decellularized cartilage-derived matrix (CDM). Since this material has shown potential for cartilage regeneration, we hypothesized that CDM could be a potent material for endochondral bone regeneration. In addition, since decellularized matrices are known to harbor bioactive cues for tissue formation, we evaluated the need for seeded MSCs in CDM scaffolds. In this study, ectopic bone formation in rats was evaluated for CDM scaffolds seeded with human MSCs and compared with unseeded controls. The MSC-seeded samples were preconditioned in chondrogenic medium for 37 days. After 8 weeks of subcutaneous implantation, the extent of mineralization was significantly higher in the MSC-seeded constructs versus unseeded controls. The mineralized areas corresponded to bone formation with bone marrow cavities. In addition, rat-specific bone formation was confirmed by collagen type I immunohistochemistry. Finally, fluorochrome incorporation at 3 and 6 weeks revealed that the bone formation had an inwardly directed progression. Taken together, our results show that decellularized CDM is a promising biomaterial for endochondral bone regeneration when combined with MSCs at ectopic locations. Modification of current decellularization protocols may lead to enhanced functionality of CDM scaffolds, potentially offering the prospect of generation of cell-free off-the-shelf bone regenerative substitutes.

Programmatic assessment of competency-based workplace learning: when theory meets practice.

BACKGROUND: In competency-based medical education emphasis has shifted towards outcomes, capabilities, and learner-centeredness. Together with a focus on sustained evidence of professional competence this calls for new methods of teaching and assessment. Recently, medical educators advocated the use of a holistic, programmatic approach towards assessment. Besides maximum facilitation of learning it should improve the validity and reliability of measurements and documentation of competence development. We explored how, in a competency-based curriculum, current theories on programmatic assessment interacted with educational practice. METHODS: In a development study including evaluation, we investigated the implementation of a theory-based programme of assessment. Between April 2011 and May 2012 quantitative evaluation data were collected and used to guide group interviews that explored the experiences of students and clinical supervisors with the assessment programme. We coded the transcripts and emerging topics were organised into a list of lessons learned. RESULTS: The programme mainly focuses on the integration of learning and assessment by motivating and supporting students to seek and accumulate feedback. The assessment instruments were aligned to cover predefined competencies to enable aggregation of information in a structured and meaningful way. Assessments that were designed as formative learning experiences were increasingly perceived as summative by students. Peer feedback was experienced as a valuable method for formative feedback. Social interaction and external guidance seemed to be of crucial importance to scaffold self-directed learning. Aggregating data from individual assessments into a holistic portfolio judgement required expertise and extensive training and supervision of judges. CONCLUSIONS: A programme of assessment with low-stakes assessments providing simultaneously formative feedback and input for summative decisions proved not easy to implement. Careful preparation and guidance of the implementation process was crucial. Assessment for learning requires meaningful feedback with each assessment. Special attention should be paid to the quality of feedback at individual assessment moments. Comprehensive attention for faculty development and training for students is essential for the successful implementation of an assessment programme.

Effects of observer on the diagnostic accuracy of low-field MRI for detecting canine meniscal tears.

Low-field MRI (lfMRI) has become increasingly accepted as a method for diagnosing canine meniscal tears in clinical practice. However, observer effects on diagnostic accuracy have not been previously reported. In this study, 50 consecutive stifle joints with clinical and radiologic evidence of cranial cruciate ligament insufficiency were investigated by lfMRI and arthroscopy. Fifteen observers who had varying levels of experience and who were unaware of arthroscopic findings independently reviewed lfMRI studies and recorded whether lateral and medial meniscal tears were present. Diagnostic accuracy (sensitivity, specificity, positive (PPV) and negative predictive value (NPV)) was determined for each observer and median values were calculated for all observers, using arthroscopy as the reference standard. Interrater agreement was determined based on intraclass correlation coefficient (ICC) analysis. Observer level of experience was compared with diagnostic sensitivity and specificity using correlation analysis. Based on pooled data for all observers, median sensitivity, specificity, PPV, and NPV for lfMRI diagnosis of lateral meniscal tears were 0.00, 0.94, 0.05, and 0.94, respectively. Median sensitivity, specificity, PPV, and NPV for medial meniscal tears were 0.74, 0.89, 0.83, and 0.79, respectively. Interrater agreement for all menisci was fair (0.51). Menisci were less consistently scored as having no tears (ICC = 0.13) than those scored as having tears (ICC = 0.50). No significant correlations between observer experience and diagnostic sensitivity/specificity were identified. Findings indicated that the accuracy of lfMRI for diagnosing canine meniscal tears was poor to fair and observer-dependent. Future studies are needed to develop standardized and widely accepted lfMRI criteria for diagnosing meniscal tears.

A differential effect of bone morphogenetic protein-2 and vascular endothelial growth factor release timing on osteogenesis at ectopic and orthotopic sites in a large-animal model.

In bone tissue engineering, growth factors are widely used. Bone morphogenetic proteins (BMPs) and vascular endothelial growth factor (VEGF) are the most well-known regulators of osteogenesis and angiogenesis. We investigated whether the timing of dual release of VEGF and BMP-2 influences the amount of bone formation in a large-animal model. Poly(lactic-co-glycolic acid) (PLGA) microparticles (MPs) were loaded with BMP-2 or VEGF to create sustained-release profiles, and rapidly degrading gelatin was loaded with either growth factor for fast-release profiles. To study in vivo osteogenicity, the two delivery vehicles were combined with biphasic calcium phosphate (BCP) scaffolds and implanted in 10 Beagle dogs for 9 weeks, at both ectopic (paraspinal muscles) and orthotopic sites (critical-size ulnar defect). The 9 ectopic groups contained combined or single BMP/VEGF dosage, in sustained- or fast-release profiles. In the ulnae of 8 dogs, fast VEGF and sustained BMP-2 were applied to one leg, and the other received the opposite release profiles. The two remaining dogs received bilateral control scaffolds. Bone growth dynamics was analyzed by fluorochrome injection at weeks 3, 5, and 7. Postoperative and posteuthanization X-rays of the ulnar implants were taken. After 9 weeks of implantation, bone quantity and bone growth dynamics were studied by histology, histomorphometry, and fluorescence microscopy. The release of the growth factors resulted in both enhanced orthotopic and ectopic bone formation. Bone formation started before 3 weeks and continued beyond 7 weeks. The ectopic BMP-2 fast groups showed significantly more bone compared to sustained release, independent of the VEGF profile. The ulna implants revealed no significant differences in the amount of bone formed. This study shows that timing of BMP-2 release largely determines speed and amount of ectopic bone formation independent of VEGF release. Furthermore, at the orthotopic site, no significant effect on bone formation was found from a timed release of growth factors, implicating that timed-release effects are location dependent.

Comparison of the effects of firocoxib, carprofen and vedaprofen in a sodium urate crystal induced synovitis model of arthritis in dogs.

A randomized, placebo-controlled, four-period cross-over laboratory study involving eight dogs was conducted to confirm the effective analgesic dose of firocoxib, a selective COX-2 inhibitor, in a synovitis model of arthritis. Firocoxib was compared to vedaprofen and carprofen, and the effect, defined as a change in weight bearing measured via peak ground reaction, was evaluated at treatment dose levels. A lameness score on a five point scale was also assigned to the affected limb. Peak vertical ground reaction force was considered to be the most relevant measurement in this study. The firocoxib treatment group performed significantly better than placebo at the 3 h post-treatment time point and significantly better than placebo and carprofen at the 7 h post-treatment time point. Improvement in lameness score was also significantly better in the dogs treated with firocoxib than placebo and carprofen at both the 3 and 7 h post-treatment time points.

Expression of osteotropic growth factors and growth hormone receptor in a canine distraction osteogenesis model.

Osteotropic growth factors play an important role in bone metabolism. Nevertheless, knowledge about their expression in relation to distraction osteogenesis remains limited. The aim of the present study was to determine the expression of growth hormone (GH), growth hormone receptor (GHR), insulin-like growth factor I (IGF-I), insulin-like growth factor II (IGF-II), and bone morphogenetic protein 2 (BMP-2) in distraction-induced bone regeneration. Expression of these factors was assessed during the consolidation phase, comparing distraction osteogenesis with osteotomy-induced bone formation. Real-time PCR was performed as a semiquantitative measurement of mRNA, and the relative expression levels of these factors were determined. In addition, plasma GH profiles and plasma concentrations of IGF-I, IGF-II, and insulin-like growth factor-binding protein 4 and -6 (IGFBP-4 and -6) were measured to assess their potential systemic role during bone formation. Expression of GHR, IGF-I, and BMP-2 had significantly increased in comparison with the expression of these factors in mature bone. Expression of GHR was significantly higher in distraction-induced bone regenerate than in osteotomy-induced bone. No significant differences were found for the expression of IGF-I and BMP-2 between distraction and osteotomy. Plasma concentrations of GH, IGF-I, IGF-II, IGFBP-4, and IGFBP-6 did not demonstrate any significant differences between treatment groups and controls. Upregulation of GHR expression in distraction osteogenesis may enhance sensitivity to endogenous systemic GH and thus promote consolidation of the regenerated bone. Changes in the systemic osteotropic growth factors GH, IGF-I, IGF-II, IGFBP-4, and IGFBP-6 do not seem to be of importance during distraction osteogenesis.

The efficacy of the bone markers osteocalcin and the carboxyterminal cross-linked telopeptide of type-I collagen in evaluating osteogenesis in a canine crural lengthening model.

The aim of the present study was to determine the efficacy of the bone markers osteocalcin (OC) and carboxyterminal cross-linked telopeptide of type-I collagen (ICTP) in evaluating new bone formation in the dog, using commercially available immunoassay kits. Dogs were randomly divided into three groups and a circular external skeletal fixation system (CESF) was mounted on the tibia. In the first group a distraction osteogenesis procedure of the crus was performed. The second group received an osteotomy without crural lengthening, whereas the third group served as a sham-operated control. Bone formation was assessed using densitometric image analysis of crural radiographs. Despite significant differences in the amount of newly formed bone, this finding was not reflected in the plasma levels of OC and ICTP. In conclusion, OC and ICTP were not efficacious as markers of bone formation and resorption during osteogenesis in this canine model.

Prognostic factors in treating antebrachial growth deformities with a lengthening procedure using a circular external skeletal fixation system in dogs.

OBJECTIVE: To evaluate treatment of antebrachial growth deformities (AGD) with a lengthening procedure using a circular external skeletal fixation (CESF) system and to determine prognostic factors. STUDY DESIGN: Prospective clinical study. ANIMALS: Thirty-four dogs with unilateral AGD. METHODS: Length deficits, angular and rotational deformities, elbow incongruity (EI), osteoarthritis (OA) of the elbow and carpal joint, function, and cosmesis were determined before and after a CESF lengthening procedure. RESULTS: On admission, EI (21 dogs; 62%), OA of the elbow joint (17 dogs; 50%), carpal OA (12 dogs; 35%), and concomitant elbow and carpal OA (5 dogs; 7%) were common findings. Treatment significantly improved function (normal, 20 dogs; 60%) and cosmesis (normal, 22 dogs; 65%). Angular and rotational deformities were almost completely corrected with small remaining length deficits. Elbow and carpal OA increased significantly during the follow-up period. Significant correlations were demonstrated between initial elbow OA and final function (R=0.42, P=.02), initial function and final function (R=0.41, P=.02), and initial ulnar and radial deficit and final cosmesis (R=0.58, P=.0001 and R=0.45, P=.008). CONCLUSIONS: Treatment of AGD by a CESF lengthening procedure was successful despite small remaining length deficits. Initial elbow OA, function, and ulnar and radial length deficits are prognostic factors in the treatment of AGD. CLINICAL RELEVANCE: Initial elbow OA and initial function are prognostic factors in predicting the functional outcome of treatment of AGD with a CESF lengthening procedure in the dog.