Apoptosis of ligamentous cells of the cranial cruciate ligament from stable stifle joints of dogs with partial cranial cruciate ligament rupture

OBJECTIVE: To describe the presence and amount of apoptotic ligamentous cells in different areas of partially ruptured canine cranial cruciate ligaments (prCCLs) and to compare these findings with apoptosis of ligamentous cells in totally ruptured cranial cruciate ligaments (trCCLs). ANIMALS: 20 dogs with prCCLs and 14 dogs with trCCLs. PROCEDURES: Dogs with prCCLs or trCCLs were admitted to the veterinary hospital for stifle joint treatment. Biopsy specimens of the intact area of prCCLs (group A) and the ruptured area of prCCLs (group B) as well as specimens from trCCLs (group C) were harvested during arthroscopy. Caspase-3 and poly (ADP-ribose) polymerase (PARP) detection were used to detect apoptotic ligamentous cells by immunohistochemistry. RESULTS: No difference was found in the degree of synovitis or osteophytosis between prCCLs and trCCLs. No difference was found in degenerative changes in ligaments between groups A and B. A substantial amount of apoptotic cells could be found in > 90% of all stained slides. A correlation (r(s) = 0.71) was found between the number of caspase-3-and PARP-positive cells. No significant difference was found in the amount of apoptotic cells among the 3 groups. No significant correlation could be detected between the degree of synovitis and apoptotic cells or osteophyte production and apoptotic cells. CONCLUSIONS AND CLINICAL RELEVANCE: The lack of difference between the 3 groups indicates that apoptosis could be a factor in the internal disease process leading to CCL rupture and is not primarily a consequence of the acute rupture of the ligament.     

Synovitis in dogs with stable stifle joints and incipient cranial cruciate ligament rupture: a cross-sectional study

Objective: To evaluate stifle joints of dogs for synovitis, before development of joint instability and cranial cruciate ligament rupture (CrCLR). Study Design: Cross‐sectional study. Animals: Dogs (n=16) with CrCLR and stable contralateral stifles; 10 control dogs with intact CrCL. Methods: Arthritis and tibial translation were graded radiographically. Synovitis severity and cruciate pathology were assessed arthroscopically. Presence of inflammatory cells in synovial membrane biopsies was scored histologically. CrCLR stifle pairs and control stifles were compared. Results: Radiographic evidence of arthritis, cranial tibial translation, and arthroscopic synovitis were increased in unstable stifles, when compared with stable contralateral stifles in CrCLR dogs (P<.05). Arthroscopic synovitis in both joints of CrCLR dogs was increased compared with controls, was correlated with radiographic arthritis (S_R=0.71, P<.05), and was present in all stable contralateral stifles. Arthroscopically, 75% of stable stifle joints had CrCL fiber disruption, which correlated with severity of synovitis (S_R=0.56, P<.05). Histologic evidence of synovitis was identified in all CrCLR dogs, but was only significantly correlated with arthroscopic observations in stable stifles (r²=0.57, P<.005). Conclusion: Synovitis is an early feature of the CrCLR arthropathy in dogs before development of joint instability clinically. Severity of synovitis is correlated with radiographic arthritis in joints with minimal to no clinically detectable CrCL damage.     

Observer variability of tibial plateau slope measurement in 40 dogs with cranial cruciate ligament-deficient stifle joints

OBJECTIVE: To determine (1) the inter- and intraobserver variability in measurement of tibial plateau angle (TPA), (2) whether this inter- and intraobserver variability is related to the characteristics of the dog (age, size, and amount of degenerative joint disease [DJD]) and the experience level of the observer, and (3) the extent of any relationship between interobserver variability of TPA and the variability of the observers' selection of the specific cranial and caudal points along the tibial plateau. STUDY DESIGN: Examination of tibial radiographs of 40 dogs clinically affected with a cranial cruciate ligament (CrCL)-deficient stifle joint. METHODS: Eleven different observers, divided into 3 groups based on their level of experience with the tibial plateau leveling osteotomy (TPLO) technique, measured the TPA on all 40 radiographs on 5 different occasions. The degree of DJD present in the stifle joint was independently graded as an overall measure and then again as it specifically related to the cranial and caudal points along the tibial plateau. The total observed variabilities of the TPA were assessed with reference to interobserver differences, intraobserver differences, and among the groups of observers with respect to the different dog characteristics. Finally, the specific points selected on the radiographs were reexamined to determine whether any variability was present in cranial and caudal point selection. RESULTS: The interobserver standard deviation of the TPA measurements for each dog was 0.8 degrees, and the intraobserver standard deviation was 1.5 degrees. The TPA measurements obtained by the 11 observers differed significantly from each other (P <.001); however, there was no significant difference of TPA among the different groups of observers (P =.67). There was no significant correlation observed between either the inter- or intraobserver variability and the dog characteristics. Specific point data and their relationship to the various variables of dog characteristics and inter- and intraobserver TPA variability revealed significant correlations only to the amount of DJD present at the caudal point (P =.001). CONCLUSIONS: Interobserver variation, but no significant group variation, was present. Overall DJD did not appear to be related to the variability in TPA angle measurement. Most of the interobserver variability was attributable to variability in horizontal point selection at both the cranial and caudal points and vertical point selection at the caudal point. It appears that degenerative changes that specifically obscure the points on the tibial plateau, especially at the caudal point, are responsible for most of the interobserver variation. CLINICAL RELEVANCE: The desired postoperative TPA of 5 degrees is dependent on a precise initial measure of TPA preoperatively. This study indicates that there is statistically significant interobserver variability with measurement of TPA, which, therefore, can result in a similar amount of variability with the final tibial plateau slope obtained postoperatively.     

Radiographic assessment of the progression of osteoarthrosis in the contralateral stifle joint of dogs with a ruptured cranial cruciate ligament

The formation and progression of osteoarthrosis in the unaffected contralateral stifle joints of 14 dogs with a unilateral cranial cruciate ligament rupture were monitored radiographically in terms of a global score and the scores for 10 parameters specific for the stifle joint. The dogs were examined initially and six and 12 months later by three observers, and the variability between the observers' scores was also assessed. The score for osteophytes at the tibial attachment site of the ligament was the most reliable parameter, and that for the increase in femoropatellar joint space was the least reliable. In the contralateral stifle joints there were significant increases after six and 12 months in osteophyte formation caudal to the tibial plateau, and in subchondral sclerosis of the tibial plateau and of the long digital extensor muscle groove. These three parameters progressed more regularly during the disease process than the other parameters. The global osteoarthrosis score of the contralateral stifle joint was an important risk factor for sustaining a rupture of the cranial cruciate ligament in that joint during the next six months.     

Surgical stabilisation of the stifle in cranial cruciate ligament injury in cattle

Surgical stabilisation of cranial cruciate ligament injury was performed on 23 stifles in 22 dairy cattle ranging in age from 11 months to 14 years. Duration of time from injury until surgery varied from two days to three years. A row of Lembert sutures was placed in both the lateral and medial retinacular tissues. In addition, an imbrication suture was placed from the lateral femorotibial ligament to the lateral or middle patellar ligament in five cattle. External support was used in 15 cattle in the form of a modified Robert Jones/Thomas-Schroeder splint. Thirteen cattle improved markedly, four recovered completely and three improved moderately. Surgery was unsuccessful on seven cases. When surgery was unsuccessful, moderate to severe lesions of degenerative joint disease were usually present.     

Caudal cruciate ligament damage in dogs with cranial cruciate ligament rupture

Objective: To investigate the incidence of caudal cruciate ligament (CaCL) damage in dogs with cranial cruciate ligament rupture (CCLR). Study Design: Prospective clinical study. Animals: Dogs (n=24) admitted for surgical stabilization of the stifle after CCLR and 8 healthy dogs with intact cranial cruciate ligament (CCL) and CaCL studied as controls. Methods: Preoperative radiographs and stifle joint images (arthrotomy, 6; arthroscopy, 18) were collected from dogs with CCLR. Severity of arthritis, synovitis, CCL damage, and CaCL damage were assessed using numerical rating scales. The CaCL was probed to determine whether minor fraying or a full thickness defect in the ligament was present. Data collected from the study population were compared with the control population of dogs. Results: The CaCL was damaged in 21/24 (88%) of dogs with CCLR; 6/24 (25%) had a full thickness defect in the CaCL. Severity of stifle synovitis and severity of damage to the CaCL were positively correlated (P<.05). Conclusions: The CaCL is damaged in a high percentage of dogs with CCLR. A significant and positive correlation exists between the degree of synovitis present and the extent of CaCL damage. Clinical Relevance: In dogs with CCLR, cruciate ligament pathology typically involves both the CCL and CaCL. As the severity of synovitis and the extent of CaCL damage are related, this observation supports the hypothesis that stifle synovitis may contribute to CCL and CaCL degeneration and subsequent damage.     

COL-3 inhibition of collagen fragmentation in ruptured cranial cruciate ligament explants from dogs with stifle arthritis

Inhibition of collagen fragment generation in canine cranial cruciate ligament (CCL) explant cultures by the matrix metalloprotease inhibitor (6-demethyl)-6-deoxy-4-dedimethylamino tetracycline (COL-3) was studied. Cranial cruciate ligament specimens were collected from dogs with inflammatory stifle arthritis/CCL rupture and dogs with normal stifles. Explant cultures from each CCL specimen included one COL-3 treated explant and a baseline control; explants from 12 ruptured CCLs were prepared in triplicate and a protease inhibitor cocktail positive control was used. Explant supernatants were analyzed for generation of collagen fragments after two days. Treatment of ruptured CCL explants with 10(-4)M COL-3 decreased generation of collagen fragments. The extent of this inhibition was increased in explants treated with a protease inhibitor cocktail. Generation of collagen fragments was increased in ruptured CCLs, when compared with intact CCLs. It is concluded that generation of collagen fragments was increased in pathological ruptured CCL explants. This degradation could be significantly inhibited in vitro by 10(-4)M COL-3.     

Proximal tibial epiphysiodesis to reduce tibial plateau slope in young dogs with cranial cruciate ligament deficient stifle

The purpose of this study was to evaluate the ability of proximal tibial epiphysiodesis to reduce the tibial plateau slope in young dogs with cranial cruciate ligament (CCL) deficient stifles. Of the 14 treated dogs, eight had a bilateral injury, for a total of 22 joints. After physical and radiographical examination and measurement of tibial plateau slope, all of the dogs underwent surgery. Insertion of the screw was placed in the most proximal part of the tibial plateau, in its medio-lateral centre, aiming to the tibial shaft and using a K wire predriven as a guide; correct position of the screw was confirmed with intraoperative fluoroscopy or radiography. In all of the dogs the tibial slope was decreased at the time of physis fusion and the degree of change depended on the age and the breed of the dog at the time of surgery. The minimum change was 4 degrees and the maximum was 24 degrees. There was a statistically significant difference between tibial slope measured before surgery compared to tibial slope measured at the last follow-up visit after surgery. This study shows that the partial proximal tibial fusion in dogs with ACL injuries was effective in reducing the tibial slope during the residual growing time to such an extent to stabilize the joint, provided that the surgery had been carried out when there was still residual growing potential. The technique appeared to be mini-invasive and malalignment complications could be avoided by correct and precise insertion of the screw.     

Expression of immune response genes in the stifle joint of dogs with oligoarthritis and degenerative cranial cruciate ligament rupture

Dysregulation of immune responses within joints plays an important role in development of inflammatory arthritis. We determined expression of a panel of immune response and matrix turnover genes in synovial fluid collected from a group of dogs with stifle oligoarthritis and associated degenerative cranial cruciate ligament (CCL) rupture (n=27). We also studied synovial fluid gene expression in dogs affected with other forms of degenerative arthritis (n=9) and in the stifle joint of healthy dogs with intact CCL (n=14). After collection, synovial cells were pelleted and RNA was isolated. Relative expression of cathepsin K, cathepsin S, tartrate-resistant acid phosphatase (TRAP), matrix metalloproteinase-9 (MMP-9), invariant chain (li), toll-like receptor-2 (TLR-2), and TLR-9 was determined using real-time quantitative RT-PCR. Data were normalized to peripheral blood mononuclear cells (PBMC) as an internal control. Relative expression of cathepsin K, MMP-9, TRAP, and li was increased in the stifle synovial fluid of dogs with oligoarthritis, when compared with the stifles of healthy dogs (P<0.05). In contrast, relative expression of all of the genes-of-interest in synovial fluid from joints affected with other forms of arthritis was not significantly different from the stifles of healthy dogs. TRAP expression was also significantly increased in the stifle joints of dogs with oligoarthritis, when compared to joint expression of TRAP in dogs with other forms of degenerative arthritis (P<0.05). In the dogs with stifle oligoarthritis, expression of both matrix turnover and immune response genes was increased in stifle synovial fluid, when compared with the internal PBMC control, whereas in healthy dogs and dogs with other forms of arthritis, only expression of matrix turnover genes was increased in synovial fluid, when compared with the internal PBMC control (P<0.05). Taken together, these findings suggest that antigen-specific immune responses within the stifle joint may be involved in the pathogenesis of persistent synovitis and associated joint degradation in dogs with oligoarthritis and degenerative CCL rupture.     

Osteochondral lesion and cranial cruciate ligament rupture in an immature dog stifle

An osteochondral flap was present on the medial femoral condyle of a stifle of a six-month-old female rottweiler in which there was also a ruptured cranial cruciate ligament. The lesion was probably osteochondritis dissecans, but the possibilities of it being secondary to joint in-stability or a subchondral fracture, as has been described in people with knee ligamentous injuries, are discussed.     

Histiocytic sarcoma of the stifle in a dog with cranial cruciate ligament failure and TPLO treatment

A 10-year-old spayed female Golden Retriever was admitted with chronic lameness of the right hind limb. A tibial plateau leveling osteotomy (TPLO) had been performed on this leg approximately three years previously. A progressively growing soft tissue mass, affecting the right stifle, previously treated with TPLO was biopsied and found to be a histiocytic sarcoma. Previously proposed links between the development of neoplasia in the stifle region and the presence of chronic synovitis, osteotomy, orthopaedic implants, and specifically the Slocum TPLO plate, are briefly discussed.     

Pathologic changes in grossly normal menisci in dogs with rupture of the cranial cruciate ligament

OBJECTIVE: To determine whether histopathologic changes are detectable in grossly normal medial menisci from dogs with rupture of the cranial cruciate ligament (CCL). DESIGN: Case series. SAMPLE POPULATION: 40 medial menisci from dogs with rupture of the CCL and 20 medial menisci from control dogs without stifle joint disease. PROCEDURE: Data evaluated included age, duration of clinical signs, and whether rupture of the CCL was complete or incomplete. Three groups (n = 20/group) were also compared on the basis of 5 histologic criteria; group-1 menisci appeared grossly normal and were obtained from dogs with naturally occurring rupture of the CCL, group-2 menisci were grossly abnormal and were also obtained from dogs with naturally occurring CCL ruptures, and group-3 menisci were collected at postmortem from dogs without stifle joint disease that were of similar age and weight as dogs in groups 1 and 2. RESULTS: Group-2 menisci were significantly different from group-1 and -3 menisci in all histologic criteria. Group-1 menisci were significantly different from control menisci in only 1 of the 5 histologic criteria (cartilage differentiation). Dogs that were > or =3 years old had significantly more surface cellularity than did dogs that were < 3 years old. A significant difference was not detected between groups 1 and 2 with regard to completeness of rupture. CONCLUSIONS AND CLINICAL RELEVANCE: Histologic changes in meniscal cartilage correlate with gross appearance of the cartilage at time of surgery for rupture of the CCL. On the basis of minimal histologic changes, routine removal of grossly normal menisci does not appear to be warranted.     

Radiographic features of a lateral extracapsular wire suture in the canine cranial cruciate deficient stifle

Radiographs of 74 dogs (84 stifles) presented with a cranial cruciate ligament rupture and surgically treated using a lateral extracapsular wire (LEW) were reviewed. A strand of orthopaedic wire was surgically placed caudally around the lateral fabella and through a predrilled hole in the tibial crest. At six week follow-up, the LEW was broken at least once in 26 of 33 stifles, predominantly in the area of the lateral fabella. In five stifles, the LEW had slipped off the fabella and was displaced distally along the gastrocnemius muscle. Six months after surgery, the LEW was often broken at several sites. LEW migration was rare. Osteolysis and sclerosis at the site of LEW penetration through the tibial crest occurred in about two-thirds of the stifles examined.     

Matrix metalloproteinase activity in stifle synovial fluid of cranial cruciate ligament deficient dogs and effect of postoperative doxycycline treatment

This prospective clinical study investigated the activity of matrix metalloproteinases (MMPs) in stifle synovial fluid (SF) of 13 dogs with acute cranial cruciate ligament (CCL) rupture, and the effect of a postoperative doxycycline treatment. MMP-2, 3, 9 and 13 activities were compared with respect to the time of sampling (preoperatively or 1 month after surgical stabilisation) and the type of postoperative adjuvant treatment (doxycycline or not). No significant activity was detected for both MMP-3 and MMP-13. MMP-2 and MMP-9 activities were found to be significantly highly increased in SF of CCL ruptured stifles compared to control stifles of unaffected dogs. No significant effect from surgical stabilisation and postoperative doxycycline treatment on MMP-2 and MMP-9 activities was found, indicating that doxycycline may not be an appropriate postoperative medical treatment after CCL rupture.