Richard D. Coutts

Chondrocyte apoptosis and nitric oxide production during experimentally induced osteoarthritis.

OBJECTIVE Chondrocytes produce nitric oxide (NO) and undergo apoptosis in response to exogenous NO. This study sought to examine the relationship between NO synthesis, chondrocyte apoptosis, and the development of cartilage degradation during experimental osteoarthritis (OA). METHODS OA was induced in rabbits by anterior cruciate ligament transection (ACLT). Knees were harvested after 4 weeks and assessed for OA severity and chondrocyte apoptosis. Conditioned media from cultured cartilage explants were analyzed for nitrite content. Cartilage sections were analyzed by immunohistochemistry for the presence of nitrotyrosine. RESULTS All ACLT knees demonstrated osteoarthritic changes. Conditioned media from ACLT cartilage organ cultures contained higher levels of nitrite as compared with cartilage samples from the nonoperated side or from rabbits that had not received ACLT. Cultures of specific areas of cartilage from ACLT knees showed high levels of NO production in the medial femoral and medial tibial cartilage. Approximately 28.7% of chondrocytes isolated from ACLT cartilage and 6.7% of chondrocytes from cartilage of the nonoperated side underwent apoptosis. In situ staining demonstrated apoptotic cells in the superficial and middle zones of ACLT cartilage. A high number of apoptotic cells was present at the pannus-cartilage junction. In control cartilage, the superficial zone contained a small number of cells in apoptosis. The prevalence of apoptotic cells was significantly correlated with the levels of nitrite production and OA grade. CONCLUSION These observations suggest that, during the early phases of OA, NO production may lead to chondrocyte apoptosis, and that both events contribute to the pathogenesis of cartilage degradation. Inhibitors of NO synthesis and chondrocyte apoptosis may therefore be of therapeutic value after cartilage injury and in patients with OA.

Characterization of a model of osteoarthritis in the rabbit knee

A new computerized method of histomorphometry was used to assess the development of osteoarthritis (OA) in a rabbit model. Three groups of 10 New Zealand White rabbits with closed epiphyses underwent unilateral anterior cruciate ligament transection (ACLT) and contralateral arthrotomy (sham). Groups were killed at 4, 8 and 12 weeks. At the time of death the femoral condyles were assessed grossly following the application of India ink using the following grading scale. Grade 1: intact surface; grade 2: minimal fibrillation; grade 3: overt fibrillation; grade 4: erosion. All histological sections were assessed using a color image analysis system. The mean thickness and area were measured for a defined cartilage region. The root mean square surface roughness (based on deviations from an idealized smooth surface) was calculated to assess the surface profile of the articular cartilage. The results were as follows. After ACLT, no full-thickness ulceration was noted at 4 weeks. Four of the medial femoral condyles at 8 weeks and six at 12 weeks showed full-thickness ulceration of the articular cartilage. The per cent cartilage area and cartilage thickness (ACLT divided by sham) in almost all regions showed decreases with time, indicating progressive erosion. The surface of the ACLT knees was much rougher than that of sham of the knees. These results demonstrate the usefulness of a quantitative methodology using a computerized video analysis system to assess the articular cartilage following ACLT in a rabbit model for the development of OA.

A comparison of cortical bone atrophy secondary to fixation with plates with large differences in bending stiffness

Two internal-fixation plates with large differences in bending stiffness were applied to pairs of intact femora of six adult mongrel dogs to study the osteoporosis induced by rigid fixation. After nine and twelve months of plate application a significant, increased amount of bone atrophy was seen on the rigidly plated side. Biomechanical measurements of specimens from various segments of both femoral diaphyses showed the bones to have similar mechanical properties (as tissue), but different structural properties (as organ). These findings suggest that the osteoporosis due to rigid plate fixation occurs by thinning of the cortex rather than by reduction of the mechanical properties of the osseous tissue.

Osteochondral repair using perichondrial cells : A 1-year study in rabbits

Articular cartilage repair remains a clinical and scientific challenge with increasing interest focused on the transplantation of chondrogenic cells. This study evaluated the repair response during a 1-year period after implantation of allogenic perichondrium cell polylactic acid composite grafts into 3.7 × 5 mm osteochondral defects drilled into the medial femoral condyles of 82 adult New Zealand White rabbits. The repair tissue was evaluated grossly, histologically, histomorphometrically, biochemically, and biomechanically at 6 weeks, 12 weeks, 6 months, and 1 year after implantation. After gross evaluation, cartilaginous material appeared to fill the defect in 70 experimental knees, for an overall repair frequency of 85%. The histomorphometric results and the histologic appearances were variable. None of the specimens were completely normal at 1 year. Only specimens with subchondral bone reformation displayed a definable cartilage appearing surface with chondrocytes surrounded by dense matrix. Subchondral bone reformation was inconsistent, reaching 50% at 1 year. Biochemically, the repair tissue matured during a 1-year period into a hyaline Type II collagen dominant tissue, whereas glycosaminoglycan content remained low at all time periods. The measured compressive properties of the repair tissue at 1 year were not significantly different from those of the contralateral knee that was not surgically treated. The treatment of osteochondral defects in the rabbit knee with allogenic perichondrium cell polylactic acid composite grafts yielded a high percentage of grossly successful repairs that showed inconsistent subchondral bone reformation. These results suggest that healthy subchondral bone is important to articular cartilage repair. They also highlight that a cartilaginous appearing tissue at gross inspection may not represent structurally normal articular cartilage. Continued multidisciplinary studies on the arthroplastic potential of rib perichondrial cells are needed before human studies, which rarely can extend beyond gross assessment of repair tissue appearance can be undertaken.

Rib perichondrial grafts for the repair of full-thickness articular-cartilage defects. A morphological and biochemical study in rabbits.

The purpose of this study was to investigate the use of perichondrial grafts in articular cartilage defects and to characterize the newly formed cartilage. In a rabbit model, rib perichondrium was used to repair full-thickness defects in the femoral condyle. The quality of repair was then evaluated histologically and biochemically at six and twelve weeks after grafting. Unacceptable results were obtained in 50 per cent of the rabbits. These failures were due to condylar fracture in 20 per cent, failure of graft attachment in 20 per cent, and infection in 10 per cent. The technique of grafting must be improved to increase the percentage of successful grafts in which neocartilage with a relatively normal chemical composition fills the articular cartilage defect. Successful grafts proliferate to fill the full-thickness defect with neocartilage, which has biochemical characteristics that are similar to those of hyaline cartilage.

The effects of hyaluronan during the development of osteoarthritis

Ninety-nine mature New Zealand White (NZW) rabbits underwent unilateral anterior cruciate ligament transection (ACLT) and were divided into three groups. The contralateral non-operated knees served as controls. The first group (SA) received intra-articular injections of 0.3 ml hyaluronan (HA: MW; 8 x 10(5)) beginning 4 weeks after ACLT, once a week for 5 weeks. The second group (SV) was injected with vehicle (carrier of HA) in the same fashion as the SA group. The third group (SN) served as a nontreatment group post ACLT. All animals were killed 9 weeks post-surgery and were assessed by gross morphology, histomorphometry and biochemical analysis. Gross morphologic changes on the femoral cartilage in the SA group were less severe than those in the SV and SN groups. Cartilage thickness, cartilage area, and thickness of synovial lining cell layer histomorphometric parameters were measured, showing a positive effect of HA on the preservation of articular cartilage and synovial tissue. Similarly, the cartilage and synovial tissues from knees injected with HA did not demonstrate significant alterations from contralateral controls as measured by biochemical analysis [i.e., water content, pyridinoline concentration, glycosaminoglycan (GAG) content for the cartilage, and DNA concentration for the synovial tissue].

Physiology and therapeutic value of passive joint motion.

Despite the long history of therapeutic experience with different types and amounts of passive joint motion, its effects and the principles of its use remain controversial. Through empiric success, a spectrum of passive motion has evolved for various clinical purposes, including joint diagnosis; correction of joint deformities; mobilization of stiff joints; stimulation of joint healing; neuromuscular re-education; and prevention of immobilization complications (e.g., contracture formation, connective tissue atrophy, relative healing inhibition, and associated stasis abnormalities). However, the potential abuses of passive motion (e.g., causing additional tissue trauma, mobilizing unprotected joints, and stretching the wrong joints or tissues) have created serious doubts as to the value of this therapy and raised important questions concerning the lack of proper definition (e.g., force, direction, speed, and duration) and the unknown margins of safety. Clinical and experimental evidence supports the probable effectiveness of passive joint motion on joint and tissue levels, but without a better quantitative understanding of the mechanisms of action, dose-responsiveness, specific tissue effects, and, most important, their controls. Thus, passive motion will continue to be used suboptimally with inconsistent results. When these clinical and research deficiencies are corrected, passive motion will attain its proper place as a powerful and reliable orthopedic tool.

The effects of rigidity of internal fixation plates on long bone remodeling. A biomechanical and quantitative histological study.

The effect of rigidity of internal fixation plates on long bone remodeling was studied using two types of plates with considerable differences in stiffness. The plated bones were subjected to bioengineering quantitative histological, and cortical thickness studies after 9 and 12 months. The biomechanical results, together with the quantitative histological measurements of the macroscopic architecture, showed that tissue characteristics of the plated bones were similar. However, because of the larger cortical area, the less regidly plated femora can sustain significantly higher loads and energy before failure. Cortical thickness measurements also showed that rigid plate immobilization results in thinning of the cortex of the underlying bone. The experimental results suggest that cortical bone remodels according to functional stress demands, and the osteoporosis secondary to rigid plate protection is consequent to thinking of its cortex.

Pseudoseptic Reactions to Hylan Viscosupplementation: Diagnosis and Treatment

Hyaluronans are used widely in the treatment of osteoarthritis of the knee. Three commercial hyaluronan preparations currently are available in the United States: sodium hyaluronate (Hyalgan), sodium hyaluronate (Supartz), and hylan G-F 20 (Synvisc). Although the sodium hyaluronates are derived naturally, hylan is chemically modified to increase its molecular weight. All three products have been shown to be well tolerated in clinical trials, however, there have been reports in the literature of pseudoseptic reactions, or severe acute inflammatory reactions, after injections with hylan. Our study reviewed the reported incidence of pseudosepsis. The pathogenic mechanisms and clinical treatment of this reaction are presented.

The increasing need for nonoperative treatment of patients with osteoarthritis.

Osteoarthritis affects more patients than almost any other musculoskeletal disorder. The number of patients suffering joint pain and stiffness as a result of this disease will increase rapidly in the next decade. Although operative treatments of patients with osteoarthritis will continue to improve and the number of operative procedures will increase slightly in the next decade, only a small fraction of the patients with osteoarthritis will require operative procedures. The most pressing healthcare need for the majority of patients with osteoarthritis is nonoperative care that helps relieve symptoms and improve function, and in some instances slows progression. In rare instances, the symptoms of osteoarthritis improve spontaneously, but most patients need nonoperative care for decades. Orthopaedists need to improve their ability to provide nonoperative care for patients with osteoarthritis. They should be skilled in the early diagnosis of osteoarthritis and in the use of current common nonoperative treatments including patient education, activity modification, shoe modifications, braces, oral analgesics, oral nonsteroidal antiinflammatory medications, oral dietary supplements, and intraarticular injections. Furthermore, orthopaedists should be prepared to incorporate new nonoperative treatments for patients with osteoarthritis into their practice.