Effects of intra-articular administration of methylprednisolone acetate on normal articular cartilage and on healing of experimentally induced osteochondral defects in horses.

The effects of intra-articular administration of methylprednisolone acetate (MPA) on the healing of full-thickness osteochondral defects and on normal cartilage were evaluated in 8 horses. In group-1 horses (n = 4), a 1-cm-diameter, full-thickness defect was created bilaterally in the articular cartilage on the dorsal distal surface of the radial carpal bone. Cartilage defects were not created in group-2 horses (n = 4). One middle carpal joint was randomly selected in each horse (groups 1 and 2), and treated with an intra-articular injection of 100 mg of MPA, once a week for 4 treatments. Injections began 1 week after surgery in group-1 horses. The contralateral middle carpal joint received intra-articular injections of an equivalent volume of 0.9% sodium chloride solution (SCS), and served as a control. Horses were evaluated for 16 weeks, then were euthanatized, and the middle carpal joints were examined and photographed. Synovial and articular cartilage specimens were obtained for histologic and histochemical evaluation. Gross morphometric evaluation of the healing defects in group-1 horses revealed that 48.6% of the defect in control joints and 0% of the defect in MPA-treated joints was resurfaced with a smooth, white tissue, histologically confirmed as fibrocartilage. This replacement tissue was a firmly attached fibrocartilage in control joints and a thin fibrous tissue in MPA-treated joints. The articular cartilage in joints treated with MPA had morphologic changes, including chondrocyte cluster formation, loss of palisading architecture, and cellular necrosis in both groups of horses.(ABSTRACT TRUNCATED AT 250 WORDS)     

Surgical remodelling of the femoral trochlea: an experimental study.

Surgical deepening of the groove in the femoral trochlea is a technique commonly employed in the correction of patellar dislocation in dogs. In order to discover how this deep articular defect repairs the operation was studied experimentally in young goats. Observations were made on the appearance of the trochlea and the histologic changes that had taken place after approximately six, 12, and 40 weeks. The abraded groove became filled with fibrocartilage with a relatively smooth surface and it articulated freely with the patella. The majority of the replacement tissues arose by differentiation of the granulation tissue which initially filled the groove, but there appeared to have been some contribution made by the hyaline cartilage at the margins of the groove.     

Effects of tissue antigen matching on the healing of fresh cancellous bone allografts in dogs

The radiographic and histologic healing patterns of fresh cancellous bone allografts were compared with those of fresh cancellous autografts in dogs. Two groups of allografts were studied: 1 group of littermate pairs with minor histocompatibility mismatches, and 1 group of unrelated pairs with major histocompatibility mismatches. Pairs were chosen on the basis of preoperative serologic typing and mixed lymphocyte cultural assay of lymphocyte-defined compatibility. Generally, grafts with minor mismatches and autografts achieved bony union in 6 weeks with grafted bone serving as a scaffold for new bone formation. Grafts with major mismatches were resorbed and replaced by fibrocartilage. Healing of osteotomy sites grafted with mismatched bone occurred by ingrowth of host periosteal and endosteal new bone.     

Histochemical study of normal and collapsed tracheas in dogs

Tracheas from 15 toy breed dogs with normal tracheas and 6 dogs with collapsed tracheas were examined histologically and histochemically. Tracheal cartilage was analyzed for chondroitin sulfate by means of alcian blue (CEC method) and for calcium with alizarin red S. Cartilage arcs from dogs with collapsed tracheas had areas that were apparently hypocellular, and some had other areas that appeared like fibrocartilage or fibrous tissue. Histochemically, collapsed tracheal cartilage had less chondroitin sulfate and calcium than did normal tracheal cartilage. Cartilage arcs from the collapsed tracheas were not uniformly affected to the same degree, and parts of a given tracheal arc appeared normal, whereas other parts had an abnormal histologic appearance.     

Orthotopic Osteochondral Transplantation of the Canine Proximal Femur

An 18-week study was conducted to evaluate orthotopic osteochondral transplantation of the proximal femur in the dog. Eighteen dogs were divided into 3 groups of 6 each. The first group received autografts, the second received fresh allografts, and the third received grafts that had been frozen in a bone bank for 24–28 days. The grafts were fixed in position using dynamic compression plates. The grafted limbs were maintained in a sling and thus were nonfunctional and non-weight bearing throughout the 18-week study. Postoperatively the dogs were given oral tetracycline to assess osteo- cyte viability. The dogs were radiographed at 2–week intervals and 1 dog in each group was euthanatized every 3 weeks. The femurs were examined using standard histopathologic and fluorescent labeling techniques. All femoral heads were luxated by the 2nd postoperative week. The bones of all the femoral heads underwent avascular necrosis and degenerative changes were present in the transplanted cartilage by the 6th postoperative week. During the first 18 weeks following transplantation there was little radiographic and histologic difference among the 3 types of grafts regarding the nature and rate of bone healing. Based on the data obtained from techniques utilized in this study, the femoral head, neck, and articular cartilage did not survive, while the femoral diaphysis did survive the transplantation process. Different techniques may alter to some degree the results obtained.     

The Effect of Trans-Stifle External Skeletal Fixation and Hyaluronic Acid Therapy on Articular Cartilage in the Dog

Transarticular external skeletal (TES) fixators were applied unilaterally to the stifle joints of 10 young adult dogs. After 4 weeks, the fixators were removed from all dogs. Two dogs were not allowed a remobilization period, whereas 8 dogs were provided with 4 additional weeks of weight-bearing activity in a kennel run. Four dogs were given high-molecular weight hyaluronic acid by intra-articular injection weekly during the remobilization period. Clinical gait evaluations and range of motion were determined during the remobilization period. Articular cartilage samples from both stifle joints of all dogs were evaluated histologically and histochemically. No significant differences in gait scores or range of motion were noted between treated and untreated dogs. Articular cartilage proteoglycan content was reduced after 4 weeks of trans-stifle external skeletal fixation as determined by loss of alcian blue (AB) histochemical staining. Improved homogeneity of histochemical staining was observed after remobilization. However, remobilization was associated with histological damage to the surface and tangential layers of articular cartilage. Remobilization combined with hyaluronic acid (HA) therapy improved histochemical staining and reduced structural damage to articular cartilage when compared with remobilization alone.     

Functional, radiographic, and histologic assessment of healing of autogenous osteochondral grafts and full-thickness cartilage defects in the talus of dogs.

A circular (5.5 mm diameter) full-thickness cartilage defect was created on the medial ridge of the talus in 12 skeletally mature dogs. In 6 dogs, the articular surface of the lesion was repaired, using an osteochondral graft obtained from the ipsilateral manus. The graft (digit I, first phalanx, distal articular surface and diaphysis) was contoured to obtain a press fit in the drilled talar recipient site. In 6 dogs, the lesion was not treated and healed by fibrous tissue replacement. Functional assessment (lameness, hock range of motion, joint stability, joint crepitus, and mid-femoral muscle circumference) was completed before surgery and at postoperative weeks 2 through 20. Radiographic assessment (periarticular soft tissue width, joint space width, osteophyte formation, and graft incorporation) was completed before surgery and at postoperative weeks 0, 6, 12, and 20. To facilitate histologic assessment, tissues were stained with toluidine blue and H&E. Histologic assessment of the articular surface on the surgically treated talus, ipsilateral tibia, and contralateral talus was completed, using a modification of the Mankin grading system. Subchondral bone was examined to assess graft viability and incorporation. Analysis of the ordinal data was completed, using a Mann-Whitney rank sum test. All dogs were fully weight bearing by postoperative week 7. Dogs without grafts had significantly (P = 0.036) better clinical function at postoperative week 6. Significant difference in functional assessment was not evident at postoperative week 20. Immediate postoperative radiographic assessment revealed significant (P = 0.005) difference between nongrafted and grafted groups. Significant difference was not observed at postoperative week 6, 12, or 20. All grafts appeared radiographically incorporated by postoperative week 12. All grafts restored joint surface congruity, whereas 3 of 6 nongrafted lesions had poor articular congruity. Of 6 grafts, 4 partially retained normal hyaline cartilage, resulting in significantly (P = 0.014) lower Mankin grades. Significant histologic differences between groups were not apparent when the apposing tibia and control talus were examined. Talar reconstruction by use of a phalangeal osteochondral graft is a viable surgical procedure. These data indicate that normal articular and subchondral architecture are more closely approximated by osteochondral reconstruction than by fibrous tissue repair.     

Radiographic and Histologic Changes After Thoracolumbar Disc Curettage

Forty-eight thoracolumbar disc spaces were approached by a dorsolateral muscle-separating technique in six mature dogs. Twenty-four of the discs were fenestrated and curetted, and the changes were followed radiographically and histologically for 16 weeks. Disc space collapse was visible radiographically immediately after the surgical procedure, and those disc spaces remained narrow throughout the observation period. Histologically, there was evidence of chondrocyte activation within 2 weeks in the excavated disc spaces. By 16 weeks, fibrocartilage had filled the void in the curetted disc spaces.     

Use of biochemical markers of osteoarthritis to investigate the potential disease-modifying effect of tibial plateau levelling osteotomy

OBJECTIVES: To evaluate the hypothesis that the concentration of the 1/20/5D4 epitope of keratan sulphate, cartilage oligomeric matrix protein and total sulphated glycosaminoglycans in synovial fluids from dogs with cranial cruciate ligament disease would be affected by tibial plateau levelling osteotomy. In addition, to evaluate the hypothesis that medial meniscal release or meniscal injury would alter the expression of these candidate biomarkers. METHODS: Forty-one dogs with naturally occurring cranial cruciate ligament disease were recruited prospectively. Synovial fluids were collected from the index joint before surgery and six weeks and six months postsurgery. Following tibial plateau levelling osteotomy, synovial fluids were assayed for 1/20/5D4 epitope of keratan sulphate and cartilage oligomeric matrix protein concentration using an inhibition ELISA and for sulphated glycosaminoglycans using a direct dye-binding assay. RESULTS: The sulphated glycosaminoglycans ratio did not change significantly during the study. Medial meniscal injury at entry was associated with lower concentrations of synovial fluid cartilage oligomeric matrix protein (P<0.05, unpaired t test). There was no association between medial meniscal release and the changes in marker concentrations, either from 0 to six weeks or 0 to six months. CLINICAL SIGNIFICANCE: Tibial plateau levelling osteotomy did not significantly alter the expression of the named candidate biomarkers. These findings reflect the limited nature of the arthrotomy or indicate that tibial plateau levelling osteotomy does not influence the progression of osteoarthritis (OA). From these studies, there is no evidence that tibial plateau levelling osteotomy affects cartilage metabolism.     

Nitric oxide metabolite production in the cranial cruciate ligament, synovial membrane, and articular cartilage of dogs with cranial cruciate ligament rupture

OBJECTIVE: To measure concentrations of nitric oxide metabolites (nitrite-nitrate [NOt]) in cartilage, synovial membrane, and cranial cruciate ligament (CCL) in dogs and evaluate associations with osteoarthritis in dogs with CCL rupture. ANIMALS: 46 dogs with CCL rupture and 54 control dogs without joint disease. PROCEDURE: Tissue specimens for histologic examination and explant culture were harvested during surgery in the CCL group or immediately after euthanasia in the control group; NOt concentrations were measured in supernatant of explant cultures and compared among dogs with various degrees of osteoarthritis and between dogs with and without CCL rupture. RESULTS: Osteoarthritic cartilage had significantly higher NOt concentration (1,171.6 nmol/g) than did healthy cartilage (491.0 nmol/g); NOt concentration was associated with severity of macroscopic and microscopic lesions. Synovial membrane NOt concentration did not differ between dogs with and without CCL rupture. Ruptured CCL produced less NOt than did intact ligaments. In control dogs, NOt concentrations were similar for intact ligaments (568.1 nmol/g) and articular cartilage (491.0 nmol/g). Synthesis of NOt was inhibited substantially by coincubation with inhibitors. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that NOt in canine joint tissues originates from the inducible nitric oxide synthase pathway. Nitric oxide metabolite production in cartilage was greater in dogs with osteoarthritis than in healthy dogs and was associated with lesion severity, suggesting that nitric oxide inhibitors may be considered as a treatment for osteoarthritis. The CCL produces substantial concentrations of NOt; the importance of this finding is unknown.     

Repair of full-thickness defects in the femoral trochlea of dogs after trochlear arthroplasty

Trochlear arthroplasty was performed on one stifle in each of 18 Greyhounds; their contralateral pelvic limbs and the pelvic limbs of 2 dogs treated with sham surgery were controls. At 2, 4, 8, and 16 weeks, radiography, glycosaminoglycan histochemical analysis, histologic examination, and scanning electron microscopy of the trochlear surface were done. All dogs used the limb and walked without an observable limp within 2 weeks after surgery. Radiography at euthanasia did not reveal degenerative changes. Well-vascularized, highly cellular, loose fibrous connective tissue was reorganized with time into a dense fibrous connective tissue covering the entire wound. Scanning electron microscopy revealed that the replacement tissue surface was rough, torn, and debris-covered. Differential histologic staining established that change to fibrocartilage had begun deep within the replacement tissue. Histochemical analyses corroborated those observations.     

The Effect of Exercise on the Healing of Articular Cartilage Defects in the Equine Carpus

Arthroscopic surgery was performed on 12 horses (2-4 years of age) to create a 7 x 14 mm full-thickness cartilage defect in one radial carpal bone and in the contralateral third carpal bone. Six horses remained confined to a small paddock and six horses underwent a program of increasing exercise consisting of walking, trotting, and cantering for 13 weeks. All lesions showed evidence of healing at week 6 that progressed to more complete healing at week 13. There was no difference in the amount of repair tissue covering the defect. Histologically, the lesions healed with a combination of fibrous tissue and fibrocartilage. The repair tissue was significantly thicker in the exercised horses but there was no difference in repair quality. It was concluded that radial carpal and third carpal lesions have an equal ability to heal and that early postoperative exercise is not detrimental to the repair tissue within these carpal cartilage defects.     

Influence of interleukin-1beta and hyaluronan on proteoglycan release from equine navicular hyaline cartilage and fibrocartilage

Proteoglycan (PG) release, in response to recombinant human interleukin-1beta (rh-IL-1beta), was measured in cartilage explants obtained from the equine distal sesamoid bone (navicular bone). Fibrocartilage from the surface of the navicular bone apposing the deep digital flexor tendon and hyaline cartilage from the surface of the navicular bone articulating with the middle phalanx were labelled with 35SO4. Hyaline cartilage from the distal metacarpus was used as a control tissue. Following radiolabel incorporation, the three cartilage types were treated with rh-IL-1beta (100 U/mL) in the presence of hyaluronan (0.2, 2, 20, 200 and 2000 microgram/mL). rh-IL-1beta-Induced PG release was measured by scintillation assay of PG-bound radiolabel. Increases in PG release of 94% (P < 0.01), 101% (P < 0.05) and 122% (P < 0.05), in response to rh-IL-1beta, were noted in fibrocartilage, navicular hyaline cartilage and metacarpal hyaline cartilage, respectively. Hyaluronan (0.2 microgram/mL) significantly reduced rh-IL-1beta-induced PG release in metacarpal hyaline cartilage (P < 0.01). In fibrocartilage and navicular hyaline cartilage, hyaluronan did not reduce PG release and at some concentrations appeared to increase PG release, although this was not statistically significant. These experiments show that (i) fibrocartilage and hyaline cartilage of the navicular bone release PGs in response to rh-IL-1beta, and (ii) hyaluronan does not prevent rh-IL-1beta-induced breakdown of navicular bone cartilage.     

Biochemical characterisation of navicular hyaline cartilage,navicular fibrocartilage and the deep digital flexor tendon in horses with navicular disease

The study hypothesis was that navicular disease is a process analogous to degenerative joint disease, which leads to changes in navicular fibrocartilage and in deep digital flexor tendon (DDFT) matrix composition and that the process extends to the adjacent distal interphalangeal joint. The objectives were to compare the biochemical composition of the navicular articular and palmar cartilages from 18 horses with navicular disease with 49 horses with no history of front limb lameness, and to compare navicular fibrocartilage with medial meniscus of the stifle and collateral cartilage of the hoof. Cartilage oligomeric matrix protein (COMP), deoxyribonucleic acid (DNA), total glycosaminoglycan (GAG), metalloproteinases MMP-2 and MMP-9 and water content in tissues were measured. Hyaline cartilage had the highest content of COMP and COMP content in hyaline cartilage and tendon was higher in lame horses than in sound horses (p<0.05). The concentration of MMP-2 amount in hyaline cartilage was higher in lame horses than in sound horses. The MMP-2 amounts were significantly higher in tendons compared to other tissue types. Overall, 79% of the lame horses with lesions had MMP-9 in their tendons and the amount was higher than in sound horses (p<0.05). In horses with navicular disease there were matrix changes in navicular hyaline and fibrocartilage as well as the DDFT with potential implications for the pathogenesis and management of the condition.     

Development of the navicular bone in foetal and young horses, including the arterial supply

A macroscopic, arteriographic and histological study of the development and the arterial anatomy of the navicular bone of 33 foetuses and 55 young horses is described. After 125 days of gestation the blood supply consists of two routes: one situated in the superficial layer of the fibrocartilage and the other similar to the blood supply of the navicular bone of the normal mature horse. After 270 days gestation, the blood vessels in the fibrocartilage gradually regressed and retracted until they have disappeared at six months after birth. At two months after birth the first macroscopic thinning of the fibrocartilage was noticed. From seven months to one year about 45 per cent of the navicular bones showed a slight thinning of the fibrocartilage. A positive correlation was found between radiographic abnormalities (ie enlargement of the nutrient foramina) and the frequency of thinning of the fibrocartilage. Radiographic abnormalities were first recognised 14 days after birth, whereas the arteriogram showed the first changes such as fewer or no arteries entering distally at the distal extremities at 10 weeks after birth. At four weeks after birth the first arterial wall changes were found, ie intimal thickening with or without splitting of the internal elastic membrane. From that age onward, the number of navicular bones with arterial wall changes gradually increased. Starting at five months after birth only 6 to 20 per cent of the arteries in the navicular bones without radiographic abnormalities showed arterial wall changes. However, the navicular bones with radiographic abnormalities showed arterial wall changes in 25 to 80 per cent of the arteries.     

Healing of full-thickness cartilage compared with full-thickness cartilage and subchondral bone defects in the equine third carpal bone.

The effect of lesion depth on the quality of third carpal bone cartilage repair was examined. A 1-cm diameter articular defect penetrating the calcified cartilage in one limb and the subchondral bone plate in the opposite limb was created in the radial facet of the third carpal bones. Clinical and xeroradiographic examinations were performed every 4 weeks until 4 months (3 horses) and 6 months (3 horses) after surgery. The synovial membrane, non-opposing articular surfaces and articular defects were examined grossly, histologically and histochemically. Grossly, deeper defects contained thicker, whiter tissue, but both joints contained generalised degenerative changes. Defects extending through calcified cartilage were filled deeply by fibrocartilage and superficially by fibrous connective tissue. Defects extending through subchondral bone were consistently filled with hyaline-like cartilage in the depths of the lesion, fibrocartilage in the intermediate layer and fibrous connective tissue superficially. The results indicate that subchondral bone is the source of hyaline-like cartilage repair tissue and suggest that quality of healing of cartilage defects may be improved by penetrating the subchondral bone plate. It also appears that the synovitis associated with the procedure must be controlled before the procedure can be advocated for treatment of clinical cases.     

Meniscal Release in Cruciate Ligament Intact Stifles Causes Lameness and Medial Compartment Cartilage Pathology in Dogs 12 Weeks Postoperatively

Objective— To evaluate after 12 weeks the effects of caudal medial meniscal release (MR) in the cranial cruciate ligament-intact canine stifle.
Study Design— Blinded, prospective in vivo study.
Animals— Purpose-bred hound dogs (n=10).
Methods— Either MR (n=5) or a sham (SH) surgery (n=5) was performed via arthroscopy. Orthopedic examination and subjective lameness evaluation were performed in each dog preoperatively and at 4, 8, and 12 weeks after surgery. Twelve weeks postoperatively, ultrasonographic, radiographic, and arthroscopic examinations were performed on the operated stifles. Gross pathology of the articular cartilage, cruciate ligaments, and menisci was assessed. India ink staining of the femoral and tibial articular surfaces was performed to determine the percent area of articular cartilage damage.
Results— At 8 and 12 weeks after surgery, MR dogs were lamer than SH dogs. At 12 weeks, the degree of radiographic OA was significantly higher in MR stifles than in SH stifles. Gross and sonographic meniscal pathology was more severe in MR stifles compared with SH stifles. MR stifles had significantly more severe articular cartilage pathology compared with SH stifles 12 weeks after surgery; pathology was most severe in the medial compartment.
Conclusions— MR alone is associated with articular cartilage loss, further meniscal pathology, degenerative joint disease, and lameness.
Clinical Relevance— Subsequent osteoarthritis and dysfunction of the stifle joint should be considered when making clinical decisions regarding MR in dogs.     

THE SUPINATED MEDIOLATERAL RADIOGRAPH FOR DETECTION OF HUMERAL HEAD OSTEOCHONDROSIS IN THE DOG

Mediolateral and supinated mediolateral radiographs of the shoulder joint were compared in 19 dogs. Twenty shoulders, representing 15 dogs (5 had bilateral lesions), had osteochondrosis of the humeral head. The flattened humeral head and subchondral defect were detectable in both views in all affected shoulders. The lesions were slightly more easily detected in the supinated view. The supinated view more consistently demonstrated the presence of a calcified cartilage flap and therefore, could be useful in determining a course of therapy. In four dogs (8 shoulders) without osteochondrosis and six normal shoulders from affected dogs, there were no instances in which a shoulder appeared normal on one view, but demonstrated a lesion on the other. The supinated view should be obtained in addition to the mediolateral view in dogs with osteochondrosis of the humeral head.     

Cylindrical press-fit osteochondral allografts for resurfacing the equine metatarsophalangeal joint

OBJECTIVE: To investigate the feasibility of resurfacing the equine fetlock joint using cylindrical, orthotopic, press-fit, osteochondral allografts. STUDY DESIGN: Experimental study. ANIMALS: Ten mature, mixed-breed horses. METHODS: Cylindrical, osteochondral grafts (6.5-mm diameter) were harvested aseptically from cadaveric equine metatarsophalangeal joints. Allografts were transplanted into 6 horses; 4 horses were sham operated. The surgical approach involved creation of a bone block at the origin of the medial collateral ligament and luxation of the metatarsophalangeal joint. Grafts were placed into the medial and lateral metatarsal condyles. Radiographs were taken at 8 and 25 weeks, and lameness was evaluated at 25 weeks. Horses were killed at 25 weeks. Analyses included gross evaluation, microradiography, paravital staining, light microscopy, and cartilage biochemistry. RESULTS: No complications occurred that could be attributed to the surgical procedure. Graft congruency with the surrounding articular cartilage was fair to excellent. Two horses were sound at 25 weeks. Most grafts had more than 90% articular cartilage coverage, and histologic and microradiographic analysis revealed good graft incorporation and articular cartilage survival. Sulphated glycosaminoglycan concentration was decreased in grafted tissue. CONCLUSIONS: We attribute the viability of osteochondral allografts in the equine fetlock to adequate congruency, stable graft fixation, and the use of orthotopic tissue. Host response to the allograft bone tissue did not affect cartilage viability. CLINICAL RELEVANCE: Before clinical use, improvements to instrumentation are required that would decrease damage to grafts and minimize technique-associated incongruencies of the articular surface at the time of grafting. Larger grafts would also likely be required to resurface a greater surface area.