Dorsal-to-palmar branch neuroanastomosis in horses undergoing palmar digital neurectomy does not reduce neuroma formation or improve outcome.

OBJECTIVE: Chronic foot pain, a common cause of forelimb lameness, can be treated by palmar digital neurectomy (PDN). Complications include neuroma formation and lameness recurrence. In humans, neuroanastomoses are performed to prevent neuroma formation. The aim of the study was to evaluate the outcome of horses undergoing dorsal-to-palmar branch neuroanastomosis following PDN. STUDY DESIGN: Retrospective case series. ANIMALS: Eighty-five horses with PDN and dorsal-to-palmar branch neuroanastomosis. METHODS: Medical records for horses undergoing this procedure at two hospitals between 2015 and 2020 were reviewed. Palmar and dorsal nerve branches of the PDN were transected and end-to-end neuroanastomosis was performed by apposition of the perineurium. Follow-up was obtained from medical records and telephone interviews. Success was defined as resolution of lameness for at least one year. RESULTS: Lameness resolved following surgery in 81/85 (95%) horses with 57/84 (68%) sound at one year. Postoperative complications occurred in 19/85 (22%) cases. The main limitations of the study were an incomplete data set, inaccurate owner recall, and variations in procedure. CONCLUSION: Compared to previous studies, this technique resulted in similar numbers of horses sound immediately after surgery, a comparable rate of postoperative neuroma formation but a higher recurrence of lameness rate at 1 year postoperatively. CLINICAL SIGNIFICANCE: End-to-end neuroanastomosis of the dorsal and palmar branches of the PDN does not reduce the rate of neuroma formation in horses. Long-term outcome was less favorable compared to previously reported PDN techniques.

Serum amyloid A and fibrinogen as markers for early detection of surgical site infection associated with internal fixation in the horse.

The objective of this study was to determine the diagnostic ability of serum amyloid A (SAA) and fibrinogen for early detection of surgical site infection (SSI) after equine internal fixation. Horses undergoing internal fixation for fracture, arthrodesis, or osteotomy with internal fixation for limb deformity were included in the study. SAA and fibrinogen were measured on blood samples preoperatively and on days 1, 3, 5, 7, 10, and 14 postoperatively. Statistical analysis included use of Spearman's rank correlation, logistic regression, and calculating the area under the receiver operating characteristic (ROC) curve. SAA and fibrinogen measurements were both associated with SSI, with SAA being considered an excellent marker (area under the ROC curve 0.8) and fibrinogen being considered acceptable (<0.8). As the amount of time postoperatively increased, SAA elevations indicated a higher likelihood of SSI (area under the ROC curve 0.8 compared with fibrinogen 0.7). SAA and fibrinogen were predictive markers of SSI and SAA is of greater diagnostic utility when compared with fibrinogen. Persistent elevations of SAA postoperatively are associated with the development of SSI. Serial monitoring of SAA can be used to help predict the development of SSI in horses undergoing internal fixation. This may lead to earlier suspicion, and therefore recognition and treatment of SSI.

Computed tomographic assessment of fracture characteristics and subchondral bone injury in Thoroughbred racehorses with lateral condylar fractures and their relationship to outcome.

OBJECTIVE: To investigate the effects of fracture characteristics and concurrent subchondral bone pathology identified with computed tomography (CT) on the racing performance of Thoroughbred racehorses with fractures of the MC3/MT3 lateral condyle. STUDY DESIGN: Retrospective cohort study. SAMPLE POPULATION: Thoroughbred racehorses (n = 50) with a fracture of the MC3/MT3 lateral condyle, which had preoperative CT and internal fixation performed. METHODS: Medical records were reviewed for age, sex, limb, and surgical treatment. Computed tomography scans were evaluated to determine fracture characteristics including length, whether the fracture was incomplete or complete, and displacement. The presence of subchondral bone injury (SBI), sesamoid bone fracture, articular comminution, and fragmentation in the joint was noted. Racing data was obtained from an online database. Univariable and multivariable analyses determined associations between independent variables and outcomes. RESULTS: Thirty-three (66%) horses raced after surgery. Horses with sesamoid bone fractures (P = .021), MC3/MT3 comminution (P = .016) and intra-articular fragmentation (P = .015) were less likely to race postoperatively. Concurrent SBI did not affect outcome. In the final multivariable model, sex (P = .015) and whether a fracture was incomplete or complete (P = .007) were the most significant predictors of racing postoperatively with females and horses with complete fractures being less likely to race. CONCLUSION: The prognosis for racing after a lateral condylar fracture is favorable but is decreased in horses with complete fractures and certain concurrent joint pathology. CLINICAL SIGNIFICANCE: Horses presenting with lateral condylar fractures commonly have concurrent joint pathology. Computed tomography can aid in preoperative evaluation and prognostication by enabling more complete fracture assessment.

The clinical and histopathologic effects of potentiated chlorhexidine in the upper respiratory tract of horses.

OBJECTIVE: To describe the bactericidal and fungicidal properties of a 0.0005% chlorhexidine (CHD) solution potentiated with EDTA-Tris buffers (CHD-EDTA-Tris) and evaluate the safety of 0.0005% CHD-EDTA-Tris in the upper respiratory tract (URT) of normal horses. STUDY DESIGN: Clinical, prospective study. ANIMALS: Eight healthy, skeletally mature horses. METHODS: In vitro-serial dilutions of CHD-EDTA-Tris and EDTA-Tris alone were evaluated for bactericidal and fungicidal activity against Aspergillus fumigatus, Escherichia coli, Staphylococcus aureus, Streptococcus equi subspecies ssp. equi, Streptococcus equi ssp. zooepidemicus, and Pseudomonas aeruginosa. In vivo-eight healthy horses were topically treated twice with 30 ml of 0.0005% CHD-EDTA-Tris. Mucosal samples from each location were evaluated for the presence of inflammation or pathologic lesions. RESULTS: Solutions containing CHD were superior in fungal and bacterial killing to those without. In vitro-a 0.005% CHD-EDTA-Tris was 100% effective against all bacterial and fungal species evaluated, while a 0.0005% CHD-EDTA-Tris was less efficacious against A. fumigatus and S. equi ssp. equi. In vivo-a 0.0005% CHD-EDTA-Tris did not cause any clinical, gross, or histologic abnormalities when topically applied to the equine URT. CONCLUSIONS: A 0.0005% CHD-EDTA-Tris was highly effective for killing of common bacterial and fungal isolates in the equine upper respiratory tract. Short-term topical treatment of the equine URT with dilute CHD did not cause gross or histological inflammation in the tissue. CLINICAL SIGNIFICANCE: A 0.0005% CHD solution with EDTA-Tris should be considered for treatment of clinically relevant inflammatory or infectious conditions or in the URT of the horse.

A retrospective multicenter study on the use of locking compression plates for scapulohumeral arthrodesis in small equids.

OBJECTIVE: To report on technical aspects and outcomes after scapulohumeral arthrodesis (SHA) with a locking compression plate (LCP) in small equids. STUDY DESIGN: Retrospective multicenter case series. ANIMALS: Client-owned Shetland Ponies, Miniature Shetlands, and American Miniature Horses (n = 15). METHODS: Inclusion criteria were completed SHA with an LCP to treat scapulohumeral osteoarthritis/subluxation and availability of postoperative radiographs. Contributing surgeons completed a questionnaire to collect data. All radiographs were reviewed for this study. Follow-up information was obtained via re-admission to the hospital or telephone interview of the referring veterinarian or owner. Outcome was subjectively scored as excellent, good, moderate, or poor based on lameness and function. RESULTS: Sixteen SHAs were performed in 15 equids (body weight 65-145 kg) by nine different surgeons. A narrow 4.5/5.0 mm LCP (7-16 holes) was used in 14 of the 16 SHAs. Follow-up was collected 2.5 years after surgery (median; range: 9 weeks to 10 years). Major complications related to the SHAs were noted in 4 of the 13 horses alive long term, consisting of implant failure (n = 1), surgical site infection (n = 2), and scapular fracture (n = 1). The outcome was graded as excellent in 4, good in 5, and moderate in 4 horses. Two ponies were euthanized because scapulohumeral osteoarthritis and subluxation developed in the contralateral limb. CONCLUSION: Although major complications occurred in about one-third of ponies, SHA with LCP led to long-term survival with good function in most ponies. CLINICAL SIGNIFICANCE: SHA with an LCP offers a high chance for good long-term outcome in small equids although contralateral disease may affect outcome.

Comparative Analysis of Tenogenic Gene Expression in Tenocyte-Derived Induced Pluripotent Stem Cells and Bone Marrow-Derived Mesenchymal Stem Cells in Response to Biochemical and Biomechanical Stimuli.

The tendon is highly prone to injury, overuse, or age-related degeneration in both humans and horses. Natural healing of injured tendon is poor, and cell-based therapeutic treatment is still a significant clinical challenge. In this study, we extensively investigated the expression of tenogenic genes in equine bone marrow mesenchymal stem cells (BMSCs) and tenocyte-derived induced pluripotent stem cells (teno-iPSCs) stimulated by growth factors (TGF-ß3 and BMP12) combined with ectopic expression of tenogenic transcription factor MKX or cyclic uniaxial mechanical stretch. Western blotting revealed that TGF-ß3 and BMP12 increased the expression of transcription factors SCX and MKX in both cells, but the tenocyte marker tenomodulin (TNMD) was detected only in BMSCs and upregulated by either inducer. On the other hand, quantitative real-time PCR showed that TGF-ß3 increased the expression of EGR1, COL1A2, FMOD, and TNC in BMSCs and SCX, COL1A2, DCN, FMOD, and TNC in teno-iPSCs. BMP12 treatment elevated SCX, MKX, DCN, FMOD, and TNC in teno-iPSCs. Overexpression of MKX increased SCX, DCN, FMOD, and TNC in BMSCs and EGR1, COL1A2, DCN, FMOD, and TNC in teno-iPSCs; TGF-ß3 further enhanced TNC in BMSCs. Moreover, mechanical stretch increased SCX, EGR1, DCN, ELN, and TNC in BMSCs and SCX, MKX, EGR1, COL1A2, DCN, FMOD, and TNC in teno-iPSCs; TGF-ß3 tended to further elevate SCX, ELN, and TNC in BMSCs and SCX, MKX, COL1A2, DCN, and TNC in teno-iPSCs, while BMP12 further uptrended the expression of SCX and DCN in BMSCs and DCN in teno-iPSCs. Additionally, the aforementioned tenogenic inducers also affected the expression of signaling regulators SMAD7, ETV4, and SIRT1 in BMSCs and teno-iPSCs. Taken together, our data demonstrate that, in respect to the tenocyte-lineage-specific gene expression, BMSCs and teno-iPSCs respond differently to the tenogenic stimuli, which may affect the outcome of their application in tendon repair or regeneration.

Validation of standing cone beam computed tomography for diagnosing subchondral fetlock pathology in the Thoroughbred racehorse.

BACKGROUND: Subchondral bone pathology is common in Thoroughbred racehorses and believed to precede more serious injury. Early identification of pathology is critical to allow for intervention. OBJECTIVES: To determine interobserver variability of fetlock subchondral bone lesions using cone beam and fan beam computed tomography (CBCT, FBCT) and to validate a robotics-controlled CBCT to identify fetlock subchondral bone pathology in the Thoroughbred racehorse. STUDY DESIGN: Prospective cohort study. METHODS: FBCT and CBCT images were acquired of 25 metacarpo-/metatarsophalangeal joints of Thoroughbred racehorses. Images were analysed for subchondral bone lesions commonly identified in Thoroughbred fetlocks by an imaging specialist and surgery specialist. Interobserver and intermodality equivalence were determined with a Pearson correlation analysis and Bland-Altman equivalence test. RESULTS: Interobserver FBCT correlation was significant (P < .05) for 19 of 25 variables (Pearson R mean 0.77). Concordance was significant for all 25 variables (Bland Altman average difference 0.28 ± 0.21 mm). Interobserver CBCT correlation was significant for 21 of 25 variables (Pearson R mean 0.73). Concordance was significant for all variables (Bland Altman average difference 0.07 ± 1.90 mm). Intermodality (FBCT vs CBCT) correlation and concordance was significant for all variables as interpreted by the radiologist (Pearson R mean of 0.72, Bland Altman average difference 0.21 ± 0.47 mm). Intermodality correlation was significant for 19 of 25 variables as interpreted by the surgeon (Pearson R mean of 0.72). Concordance was significant for all variables (Bland Altman average difference 0.49 ± 0.52 mm). MAIN LIMITATIONS: Neither FBCT nor CBCT images were compared with other imaging modalities/histopathology; limited number of cases included; inconsistent agreement of small lesions in specific categories. CONCLUSIONS: Standing CBCT is a valid diagnostic modality to identify subchondral bone lesions in Thoroughbred fetlocks. This technology may provide valuable information regarding the development and progression of fetlock pathology and yield insight into predisposing factors leading to more severe pathology.

Standing Arthroscopic Treatment of Temporomandibular Joint Sepsis in a Horse.

A 15-year-old Thoroughbred gelding was presented for investigation of fever, right temporomandibular region swelling, and progressive pain when opening the mouth. Right temporomandibular joint (TMJ) sepsis was diagnosed based on synovial fluid analysis, sonographic imaging, and standing robotic cone-beam computed tomography. Concurrent otitis media and temporohyoid osteoarthropathy (THO) were also noted. The horse was treated with arthroscopic debridement and lavage during standing sedation followed by local and systemic antimicrobial therapy. There were no complications associated with the surgical procedure and the gelding's clinical signs resolved. Arthroscopy of the TMJ can be accomplished in the standing horse and should be considered when arthroscopic exploration or debridement of this joint is indicated. This is also the first case report of concurrent otitis media, TMJ sepsis, and THO; due to their close anatomic relationship, it is possible that septic otitis media could lead to pathology in the TMJ and temporohyoid joint, as described in humans.

Racing performance after surgical repair of medial condylar fracture of the third metacarpal/metatarsal bone in thoroughbred racehorses.

OBJECTIVE: To report the performance of thoroughbred racehorses after surgical repair of a medial condylar fracture of the third metacarpal/metatarsal bone. STUDY DESIGN: Retrospective cohort study. SAMPLE POPULATION: Forty-three horses surgically treated for medial condylar fractures, 30 with previous racing experience, 13 without previous racing experience (nonexperienced). METHODS: Medical records (2009-2017) were reviewed for signalment, radiographic fracture characteristics, repair technique, and postoperative morbidity and mortality. Each experienced horse was matched with two horses randomly selected from its most recent race to serve as healthy controls. Racing performance parameters (race rating, competitive level, speed rating, performance index) and career racing statistics were compared with multiple regression models between injured experienced horses and controls. The career racing statistics for nonexperienced horses were evaluated. Reasons for nonreturn to racing were obtained. RESULTS: The median duration of follow-up was 6 years (minimum 2, maximum 10). Twenty-one of 43 (49%) horses raced again as well as 18 of 30 (60%) experienced horses and three of 13 (23%) nonexperienced horses. Experienced horses were five times more likely than nonexperienced horses to return to racing (95% confidence interval = 0.07-0.58, P = .003). Higher preoperative racing performance parameters were associated with return to racing. Racing performance parameters were lower after the date of fracture in injured horses compared with controls. CONCLUSION: Experienced horses were more likely to return to racing after medial condylar fracture repair, although their performance was generally lower than that of comparably uninjured horses. CLINICAL SIGNIFICANCE: Owners should be aware that horses with medial condylar fractures likely will race at a lower level than their uninjured peers.

Regulation of the tenogenic gene expression in equine tenocyte-derived induced pluripotent stem cells by mechanical loading and Mohawk.

Cell-based therapeutic strategies afford major potential advantages in the repair of injured tendons. Generation of induced pluripotent stem cells (iPSCs) expands cell sources for "regenerative" therapy. However, its application in tendon repair is still limited and the effects remain unclear. In this study, equine tenocyte-derived iPSCs (teno-iPSCs) were generated by expressing four Yamanaka factors. Compared to parental tenocytes and bone marrow derived mesenchymal stem cells (BMSCs), the transcriptional activities of lineage-specific genes, including Mkx, Col1A2, Col14, DCN, ELN, FMOD, and TNC, were highly repressed in the resulting teno-iPSCs. Exposure to cyclic uniaxial mechanical loading increased the expression of Scx, Egr1, Col1A2, DCN, and TNC in teno-iPSCs and the expression of Scx, Egr1, DCN, and TNC in BMSCs. Reintroduction of tenogenic transcription factor Mohawk (Mkx) upregulated the expression of DCN in teno-iPSCs and the expression of Scx, Col14, and FMOD in BMSCs. Mechanical loading combined with ectopic expression of equine Mkx further enhanced the expression of Egr1, Col1A2, DCN, and TNC in teno-iPSCs and the expression of Scx, Egr1, and TNC in BMSCs. These data suggest that the repressed lineage-specific genes in the teno-iPSCs can be re-activated by mechanical loading and ectopic expression of Mkx. Our findings offer new insights into the application of iPSCs for basic and clinic research in tendon repair.

Arthrodesis of the metacarpophalangeal and metatarsophalangeal joints to treat osteoarthritis in 17 horses.

OBJECTIVE: To describe arthrodesis of the metacarpophalangeal (MCP) or metatarsophalangeal (MTP) joint with a locking compression plate (LCP) and a metallic tension band and report the outcome of horses treated for osteoarthritis (OA) with this technique. STUDY DESIGN: Retrospective case series. ANIMALS: Seventeen horses with OA of the MCP or MTP joint treated with arthrodesis. METHODS: Medical records (2004-2017) of horses treated for OA with arthrodesis of the MCP or MTP joint with an LCP and metallic tension band were reviewed. Preoperative variables included age, sex, breed, affected limb, and lameness grade. Surgical variables included implants used, surgery time, postoperative angle of joint, intraoperative complications, and anesthetic recovery method. Outcome was obtained from medical records and phone interviews. RESULTS: Lameness was scored as 4 out of 5 (range, 2-4) in 13 of 17 horses. All horses survived to discharge and were alive >6 months postoperatively, without any report of long-term complications. All horses were allowed unrestricted turnout, and 1 horse was lightly ridden. No postoperative infections or implant failures were reported. The only complications consisted of cast sores (n = 4). CONCLUSION: Arthrodesis of the MCP/MTP joint was associated with acceptable morbidity and good long-term outcomes in these 17 horses with OA refractory to medical management. CLINICAL SIGNIFICANCE: The morbidity and prognosis after fetlock arthrodesis for OA seem more favorable than reported in horses treated with traumatic disruption of the suspensory apparatus.

Surgical site infection associated with equine orthopedic internal fixation: 155 cases (2008-2016).

OBJECTIVE: To determine the prevalence of surgical site infection (SSI) after internal fixation and to identify risk factors for SSI and nonsurvival. STUDY DESIGN: Retrospective study. ANIMALS: One hundred fifty-five horses with long bone fractures or arthrodesis treated by internal fixation at 1 hospital between 2008-2016. METHODS: Signalment, diagnosis, surgical repair, surgeon, surgical time, antimicrobial use, SSI onset, bacterial identification, and adjunct treatments were recorded. Perioperative variables were analyzed to identify risk factors associated with outcomes. RESULTS: Surgical-site infection was reported in 22 of 155 (14.2%) horses, which is lower than what has been previously reported (P = .003). Horses with fetlock arthrodesis or ulnar fracture were more likely to develop SSI. Local prophylactic antimicrobial therapy was associated with an increased risk of SSI. Horses with SSI were 12 times (P < .0001) less likely to survive to discharge than horses without SSI. Horses with a fetlock or carpal arthrodesis or those with radial/humeral/femoral fractures were less likely to survive. No association was identified between open fractures, open reduction and internal fixation, or surgical times and SSI. CONCLUSION: The prevalence of SSI in this population was lower than what has been previously reported. Horses with fetlock or carpal arthrodesis or radial/humeral/femoral fractures were at increased risk for SSI and/or nonsurvival to discharge. A protective role of local antimicrobial therapy for SSI could not be established. CLINICAL RELEVANCE: The impact of SSI on outcomes of cases of equine internal fixation remains substantial. Identification of cases at higher risk of SSI should influence surgical technique, postoperative management, and early intervention when SSI is suspected. Additional investigation is warranted regarding local antimicrobial therapy.

Return to racing after surgical management of third carpal bone slab fractures in thoroughbred and standardbred racehorses.

OBJECTIVE: To determine the prognosis for racing of horses surgically treated for slab fractures of the third carpal bone (C3). STUDY DESIGN: Retrospective case study. ANIMALS: Horses (n = 125) surgically treated for C3 slab fractures. METHODS: Medical records of horses surgically treated for dorsal or sagittal C3 fractures were reviewed for age, sex, breed, limb, fracture type, degree of cartilage damage, and surgical treatment. Radiographs were evaluated to determine fracture depth, width, and displacement. Osteophytes, C3 lysis, and fragmentation were scored. Racing performance was obtained from online databases. Univariable and multivariable analyses were used to determine associations between independent variables and outcomes. RESULTS: Fifty-four (43%) horses raced postoperatively. Among thoroughbreds, 35% (30/86) with dorsal fractures and 63% (17/27) with sagittal fractures raced postoperatively. Among standardbreds, 77% (10/13) with dorsal fractures and 0% (0/2) with sagittal fractures raced postoperatively. Fracture displacement, C3 lysis, and cartilage damage affected the likelihood of racing postoperatively. Placement of 3.5-mm screws vs 4.5-mm screws and the placement of fewer screws were associated with improved likelihood of racing. CONCLUSION: The prognosis for postoperative racing of thoroughbreds with dorsal C3 fractures was less favorable than that previously reported. Concurrent joint pathology, such as cartilage damage at time of surgery, affected the ability of the horse to race postoperatively. CLINICAL SIGNIFICANCE: Although internal fixation of C3 slab fractures is required to restore joint congruity, return to racing should be expected in only 42% of thoroughbreds and 67% of standardbreds.

Pancarpal and partial carpal arthrodesis with 3 locking compression plates in 6 horses.

OBJECTIVE: To report the outcome of horses after pancarpal or partial carpal arthrodesis with 3 locking compression plates (LCP). STUDY DESIGN: Case series. ANIMALS: Six horses ranging in age from 8 months to 16 years and weighing 227-580 kg with severe carpal pathology including acute fractures, chronic osteoarthritis, and chronic angular limb deformity. METHODS: Pancarpal or partial carpal arthrodesis was performed with 3 LCP. Autologous cancellous bone grafts were used in 5 of 6 cases to facilitate joint arthrodesis. RESULTS: External coaptation was maintained for 4 to 6 weeks after surgery. Radiographic follow-up was available in all 6 cases, all of which reached arthrodesis and pasture soundness by 4-5 months postoperatively. One case required implant removal at 6 months because of implant exposure through the skin but returned to pasture soundness after removal. CONCLUSION: Carpal instability due to acute fractures or chronic disease was successfully stabilized with 3 short LCP, leading to pasture soundness in all 6 horses. CLINICAL SIGNIFICANCE: The use of 3 short LCP should be considered as a strategy to facilitate pancarpal or partial carpal arthrodesis by providing superior stability without placement of implants in the diaphysis of the radius and third metacarpus.

Use of locking compression plates in ulnar fractures of 18 horses.

OBJECTIVES: To describe the outcome, clinical findings, and complications associated with the use of the locking compression plate (LCP) for various types of ulnar fractures in horses. STUDY DESIGN: Retrospective case series. SAMPLE POPULATION: Client owned horses (n = 18). METHODS: Medical records, radiographs, and follow-up for horses having an ulnar fracture repaired using at least 1 LCP were reviewed. Fifteen of 18 horses had fractures of the ulna only, and 3 horses had fractures of the ulna and proximal radius. RESULTS: All 18 horses were discharged from the hospital. Complications occurred in 5 horses; incisional infection (n = 4, 22%), implant-associated infection (n = 2, 11%), and colic (n = 1, 6%). Follow-up was available for all horses at a range of 13-120 months and 15 horses (83%) were sound for their intended purpose and 3 horses (17%) were euthanatized. One horse was euthanatized for complications associated with original injury and surgery. CONCLUSIONS: The LCP is a viable method of internal fixation for various types of ulnar fractures, with most horses in this series returning to soundness.

Wnt10b and Dkk-1 gene therapy differentially influenced trabecular bone architecture, soft tissue integrity, and osteophytosis in a skeletally mature rat model of osteoarthritis.

AIMS: Our goals in the current experiments were to determine if (a) upregulation of Wnt signaling would induce osteoarthritis changes in stable stifle joints and (b) if downregulation of Wnt signaling in destabilized joints would influence the progression of OA. METHODS: At 37 weeks of age, rats were injected in the stifle joint with a recombinant adeno-associated viral vector containing the Wnt-inhibitor Dkk-1 or a Wnt10b transgene. At 40 weeks of age, rats underwent surgical destabilization of the joint. At 50 weeks of age, stifle joints were submitted for micro-computed tomography and histopathological analysis. RESULTS: Injection of either Wnt10b or Dkk-1 transgenes in stable joints improved bone architectural parameters, but worsened soft tissue integrity. Osteophytosis was decreased by Dkk-1, but unchanged by Wnt10b. Destabilization negatively influenced bone architecture, increased osteophytosis, and decreased soft tissue integrity. Dkk-1 exacerbated the negative effects of destabilization, whereas Wnt10b had little effect on these parameters. Osteophytosis was improved, whereas soft tissue integrity was worsened by both transgenes in destabilized joints. CONCLUSIONS: The Wnt-inhibitor Dkk-1 does not appear to completely inhibit the effects of Wnt signaling on bone remodeling. In vivo upregulation of Wnt10b and its inhibitor, Dkk-1, can produce both parallel or contrasting phenotypic responses depending on the specific parameter measured and the fidelity of the examined joint. These observations elucidate different roles for Wnt signaling in stable versus destabilized joints and may help to explain the conflicting results previously reported for the role of Dkk-1 in joint disease.

Preexisting Neutralizing Antibodies to Adeno-Associated Virus Capsids in Large Animals Other Than Monkeys May Confound In Vivo Gene Therapy Studies.

Adeno-associated virus (AAV) vectors are currently being tested not only in small animal models such as mice but also in large animal models, including pigs, dogs, and horses. Natural exposure to AAV occurs in most of the species used in these studies and potentially elicits a neutralizing humoral immune response to AAV. In this study, we show the prevalence of neutralizing antibodies (NAbs) to several AAV serotypes in these large animals as measured by an in vitro NAb assay and the ability of these NAbs to block AAV transduction in an in vivo mouse model of NAb passive transfer assay. The results of this study clearly show the importance of evaluating large animal models for the presence of AAV NAbs before enrolling them in AAV-mediated gene therapy studies.

Clinical use of computed tomography and surface markers to assist internal fixation within the equine hoof.

OBJECTIVE: To describe clinical use of computed tomography (CT) and hoof surface markers to facilitate internal fixation within the confines of the hoof wall. STUDY DESIGN: Retrospective case series. ANIMALS: Horses (n = 16) that had CT-guided internal fixation of the distal phalanx (DP) or distal sesamoid bone (DSB). METHODS: Drill bit entry point and direction were planned from CT image series performed on hooves with grids of barium paste dots at proposed entry and projected exit sites. Post-implantation CT images were obtained to check screw position and length as well as fracture reduction. Imaging, reduction, and surgical and general anesthesia times were evaluated. Outcome was recorded. RESULTS: Screw position and length were considered near optimal in all horses, with no consequential malposition of bits or screws. Fracture reduction was evident in all cases. Preoperative planning times (at least 2 CT image acquisitions and grid creation) ranged from 10 to 20 minutes. Surgery time ranged from 45 to 90 minutes (mean, 61 minutes) and general anesthesia time ranged from 115 to 220 minutes (mean, 171 minutes). CONCLUSIONS: The combination of CT and surface marker grids allowed accurate positioning of screws in clinical DP and DSB fractures. The technique was simple and rapid. An aiming device is useful for the technique.

Diagnostic and therapeutic arthroscopy in the standing horse.

This article describes diagnostic arthroscopy and arthroscopic management of selected lesions in the standing equine patient. Details on case selection, patient and operating room preparation, and surgical technique are presented. This information will add techniques that avoid general anesthesia to the equine surgeon's armamentarium.

Acid ceramidase maintains the chondrogenic phenotype of expanded primary chondrocytes and improves the chondrogenic differentiation of bone marrow-derived mesenchymal stem cells.

Acid ceramidase is required to maintain the metabolic balance of several important bioactive lipids, including ceramide, sphingosine and sphingosine-1-phosphate. Here we show that addition of recombinant acid ceramidase (rAC) to primary chondrocyte culture media maintained low levels of ceramide and led to elevated sphingosine by 48 hours. Surprisingly, after three weeks of expansion the chondrogenic phenotype of these cells also was markedly improved, as assessed by a combination of histochemical staining (Alcian Blue and Safranin-O), western blotting (e.g., Sox9, aggrecan, collagen 2A1), and/or qPCR. The same effects were evident in rat, equine and human cells, and were observed in monolayer and 3-D cultures. rAC also reduced the number of apoptotic cells in some culture conditions, contributing to overall improved cell quality. In addition to these effects on primary chondrocytes, when rAC was added to freshly harvested rat, equine or feline bone marrow cultures an ~2-fold enrichment of mesenchymal stem cells (MSCs) was observed by one week. rAC also improved the chondrogenic differentiation of MSCs, as revealed by histochemical and immunostaining. These latter effects were synergistic with TGF-beta1. Based on these results we propose that rAC could be used to improve the outcome of cell-based cartilage repair by maintaining the quality of the expanded cells, and also might be useful in vivo to induce endogenous cartilage repair in combination with other techniques. The results also suggest that short-term changes in sphingolipid metabolism may lead to longer-term effects on the chondrogenic phenotype.