David Amiel

The effect of prolonged physical training on the properties of long bone: a study of Wolff's Law.

UNLABELLED Five one-year-old immature swine were subjected to twelve months of exercise training. Four matched swine with no training served as controls. After they were killed, four-millimeter-wide strips of bone taken from the anterior, medial, posterior, and lateral quadrants of the central femoral diaphysis were subjected to four-point bending tests to failure. It was found that although exercise did not change the mechanical properties of the cortical bone, it resulted in significant increases in the averaged femoral cross-sectional properties: 17 per cent in cortical thickness, 23 per cent in cortical cross-sectional area, and 21 per cent and 27 per cent in maximum and minimum area moments of inertia, respectively. These changes were due primarily to reduction in the diameter of the medullary canal. The analyses of bone composition showed that the bone density and biochemical contents of the control and exercised animals were similar, but the total volume and the dry, ash, and calcium weights of the exercised bone were significantly higher. These combined results suggest that prolonged exercise has a significant effect on the quantity of the bone, but not on its quality. CLINICAL RELEVANCE It has long been recognized that stress deprivation from immobilization in plaster casts results in profound bone atrophy, and it is generally accepted that a minimum level of activity is necessary for homeostasis of bone. These results show that exercise at a level comparable to that prescribed in running fitness programs for humans (65 to 80 per cent of maximum heart rate) can not only maintain homeostasis, but produce actual hypertrophy of bone. This work further suggests the importance of graduated, prolonged, supervised rehabilitation programs in overcoming osteoporotic states.

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.

Articular cartilage transplantation. Clinical results in the knee.

Between December 1983 and August 1991, 55 consecutive patients (55 knees) who underwent articular cartilage transplantation to their damaged knees were enrolled in the study. Average followup was 75 months (range, 11-147 months). Eight-two percent were younger than 45 years of age. Patients were evaluated through an 18-point scale, with 6 points each allocated to pain, range of motion, and function. An excellent knee was pain free, had full range of motion, and permitted unlimited activity. A good knee allowed full time employment and moderate activity. Eleven of 15 (73%) allografts transplanted 10 or more years ago were still good or excellent at the time of last followup. Overall, 45 of 55 (76%) knees that received the transplants were rated good or excellent. Specifically, 36 of 43 (84%) patients with unipolar transplants regained normal use of their resurfaced knee. The results after bipolar resurfacing were less encouraging, with only six of 12 (50%) knees rated good or excellent. The described technique of osteochondral shell allograft resurfacing of the knee capitalize on the different healing potentials of bone and cartilage by transplanting the viable articular cartilage organ in its entirety along with just enough of the underlying bone to allow for graft incorporation through creeping substitution. The results support the use of fresh osteochondral shell allograft transplantation for the treatment of large, full thickness articular cartilage defects to the medial or lateral femoral condyles and to the patella.

Prolonged Storage Effects on the Articular Cartilage of Fresh Human Osteochondral Allografts

BACKGROUND Fresh osteochondral allograft transplantation is a well-established technique for the treatment of cartilage defects of the knee. It is believed that the basic paradigm of the technique is that the transplantation of viable chondrocytes maintains the articular cartilage matrix over time. Allograft tissue is typically transplanted up to forty-two days after the death of the donor, but it is unknown how the conditions and duration of storage affect the properties of fresh human osteochondral allografts. This study examined the quality of human allograft cartilage as a function of storage for a duration of one, seven, fourteen, and twenty-eight days. We hypothesized that chondrocyte viability, chondrocyte metabolic activity, and the biochemical and biomechanical properties of articular cartilage would remain unchanged after storage for twenty-eight days. METHODS Sixty osteochondral plugs were harvested from ten fresh human femoral condyles within forty-eight hours after the death of the donor and were stored in culture medium at 4 degrees C. At one, seven, fourteen, and twenty-eight days after harvest, the osteochondral plugs were analyzed for (1) viability and viable cell density by confocal microscopy, (2) proteoglycan synthesis by quantification of (35)SO(4) incorporation, (3) glycosaminoglycan content, (4) indentation stiffness, (5) compressive modulus and hydraulic permeability by static and dynamic compression testing, and (6) tensile modulus by equilibrium tensile testing. RESULTS Chondrocyte viability and viable cell density remained unchanged after storage for seven and fourteen days (p > 0.7) and then declined at twenty-eight days (p < 0.001). Proteoglycan synthesis remained unchanged at seven days (p > 0.1) and then declined at fourteen days (p < 0.01) and twenty-eight days (p < 0.001). No significant differences were detected in glycosaminoglycan content (p > 0.8), indentation stiffness (p > 0.4), compressive modulus (p > 0.05), permeability (p > 0.3), or equilibrium tensile modulus after storage for twenty-eight days (p > 0.9). CONCLUSIONS These data demonstrate that fresh human osteochondral allograft tissue stored for more than fourteen days undergoes significant decreases in chondrocyte viability, viable cell density, and metabolic activity, with preservation of glycosaminoglycan content and biomechanical properties. The cartilage matrix is preserved during storage for twenty-eight days, but the chondrocytes necessary to maintain the matrix after transplantation decreased over that time-period.

The Importance of Controlled Passive Mobilization on Flexor Tendon Healing: A Biomechanical Study

The effects of controlled passive motion on primary tendon repair were studied using the canine forepaw flexor apparatus as experimental model. The animals were divided into seven groups based on duration(3 to 12 weeks post repair) and mode of immobilization and partial mobilization. The repaired tendons were subjected to biomechanical evaluation of their gliding function and tensile strength characteristics. The results showed positive effects of controlled passive motion on tendon repair. The rate of tendon repair was significantly improved over those animals that were continuously immobilized. At 12 weeks, the repaired flexors from the motion group had regained over one-third of the ultimate tensile load as compared to their contralateral intact controls. Of equal importance is that these repaired tendons maintained good gliding function within the sheath during the repair process. The gliding function of these tendons was also significantly better than those subjected to continuous immobilization.

The Effect of Immobilization on Collagen Turnover in Connective Tissue: A Biochemical-Biomechanical Correlation

Immobilization of the knee joint for 9 weeks results in a reduction of the mechanical properties in the lateral collateral ligament. Specifically, ligament stiffness is reduced in this tissue. No statistical change in collagen mass was detected for the medial collateral ligament (MCL) or patellar tendon. An increase in collagen turnover (synthesis and degradation) was, however, found in the immobilized medial collateral ligament and patellar tendon. It is thus proposed that stiffness reduction is due to a change in the ligament substance itself, rather than a result of tissue atrophy.

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.

Normal ligament properties and ligament healing.

There is still a long way to go to achieve the goals of ligament replacement through modification of normal, intrinsic ligament healing processes. We are learning about the biology of ligamentous tissues, including the problem of regeneration, and that in itself is an all-important first step toward seeking solutions.

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.

Collagen Cross-Linking Alterations in Joint Contractures: Changes in the Reducible Cross-Links in Periarticular Connective Tissue Collagen After Nine Weeks of Immobilization

A significant increase in the NaBH4 reducible intermolecular cross-links in the 9 week immobilized rabbit periarticular connective tissue was found. Dihydroxylysinonorleucine, hydroxylysinonorleucine, and histidinohydroxymerodesmosine were the major cross-links which increased during the period of immobilization. No change in the hydroxlysine/lysine ratio between the immobilized and control periarticular connective tissue collagen was detected during the 9 weeks of enforced immobilization of the rabbit knee. Since the collagen mass in the immobilized periarticular connective tissue does not change, or at most is reduced 10%, we suggest that there is an increase in collagen cross-links expressed both in per unit weight of collagen and on the basis of collagen mass per knee due to the lack of physical stress and motion.