Combined effects of estrogen and progesterone on the anterior cruciate ligament.

Previous studies from the authors' laboratory have established the presence of estrogen and progesterone receptors in the human anterior cruciate ligament. The purpose of the current study was to investigate the combined effects of 1beta-estradiol and progesterone on cell proliferation and procollagen synthesis of the human anterior cruciate ligament fibroblasts. Fibroblast proliferation and procollagen synthesis in response to logarithmic concentrations of 17beta-estradiol (0.0025 ng/mL, 0.025 ng/mL, 0.25 ng/mL) and progesterone (1 ng/mL, 10 ng/mL, 100 ng/mL) were assessed with the measurement of 3H-thymidine incorporation and Types I and III procollagen specific equilibrium radioimmunoassays. On Days 1, 3, and 5 there was a dose dependent decrease in the fibroblast proliferation and procollagen Type I synthesis with increasing estradiol concentrations. The effect was attenuated with increasing progesterone concentrations. Controlling for estrogen levels, a dose dependent increase in fibroblast proliferation and procollagen Type I synthesis was observed with increasing progesterone concentrations. The effect was more pronounced at lower concentrations of estrogen, suggesting estrogen levels were the dominant factor. The effects of estrogen and progesterone became less apparent by Day 7. No significant differences in Type III procollagen synthesis were seen with varying estradiol concentrations at any of the designated times. These early physiologic changes in fibroblast proliferation and Type I procollagen synthesis may provide a biologic explanation for the increased anterior cruciate ligament injury rate observed in female athletes, suggesting the acute cyclical hormonal variations in the female athlete during menstruation predispose her to ligamentous injury.

The effect of regional gene therapy with bone morphogenetic protein-2-producing bone-marrow cells on the repair of segmental femoral defects in rats.

BACKGROUND: Recombinant human bone morphogenetic proteins (rhBMPs) can induce bone formation, but the inability to identify an ideal delivery system limits their clinical application. We used ex vivo adenoviral gene transfer to create BMP-2-producing bone-marrow cells, which allow delivery of the BMP-2 to a specific anatomical site. The autologous BMP-2-producing bone-marrow cells then were used to heal a critical-sized femoral segmental defect in syngeneic rats. METHODS: Femoral defects in five groups of rats were filled with 5 x 10(6) BMP-2-producing bone-marrow cells, created through adenoviral gene transfer (twenty-four femora, Group I); twenty micrograms of rhBMP-2 (sixteen femora, Group II); 5 x 10(6) beta-galactosidase-producing rat-bone-marrow cells, created through adenoviral gene transfer of the lacZ gene (twelve femora, Group III); 5 x 10(6) uninfected rat-bone-marrow cells (ten femora, Group IV); or guanidine hydrochloride-extracted demineralized bone matrix only (ten femora, Group V). Guanidine hydrochloride-extracted demineralized bone matrix served as a substrate in all experimental groups. Specimens that were removed two months postoperatively underwent histological and histomorphometric analysis as well as biomechanical testing. RESULTS: Twenty-two of the twenty-four defects in Group I (BMP-2-producing bone-marrow cells) and all sixteen defects in Group II (rhBMP-2) had healed radiographically at two months postoperatively compared with only one of the thirty-two defects in the three control groups (beta-galactosidase-producing rat-bone-marrow cells, uninfected rat-bone-marrow cells, and guanidine hydrochloride-extracted demineralized bone matrix alone). Histological analysis of the specimens revealed that defects that had received BMP-2-producing bone-marrow cells (Group I) were filled with coarse trabecular bone at two months postoperatively, whereas in those that had received rhBMP-2 (Group II) the bone was thin and lace-like. Defects that had been treated with bone-marrow cells producing beta-galactosidase (Group III), uninfected bone-marrow cells (Group IV), or guanidine hydrochloride-extracted demineralized bone matrix only (Group V) demonstrated little or no bone formation. Histomorphometric analysis revealed a significantly greater total area of bone formation in the defects treated with the BMP-2-producing bone-marrow cells than in those treated with the rhBMP-2 (p = 0.036). Biomechanical testing demonstrated no significant differences, with the numbers available, between the healed femora that had received BMP-2-producing bone-marrow cells and the untreated (control) femora with respect to ultimate torque to failure or energy to failure. CONCLUSIONS: This study demonstrated that BMP-2-producing bone-marrow cells created by means of adenoviral gene transfer produce sufficient protein to heal a segmental femoral defect. We also established the feasibility of ex vivo gene transfer with the use of biologically acute autologous short-term cultures of bone-marrow cells.

Effect of estrogen on cellular metabolism of the human anterior cruciate ligament.

Previous studies from this laboratory have established the presence of estrogen receptors in the human anterior cruciate ligament. The purpose of this study was to investigate the effects of 17 beta-estradiol on cell proliferation and procollagen levels, as an indicator of collagen synthesis, in the human anterior cruciate ligament fibroblasts. Fibroblast proliferation and procollagen synthesis in response to near log concentrations of 17 beta-estradiol (at 0.0029 ng/mL, 0.025 ng/mL, 0.25 ng/mL, 2.5 ng/mL, and 25 ng/mL) were assessed with the measurement of 3H-thymidine incorporation and Types 1 and 3 procollagen specific equilibrium radioimmunoassays. On Days 1 and 3, there was a dose dependent decrease in the proliferation of anterior cruciate ligament fibroblasts with increasing estradiol concentrations. This dose dependent effect of decreased fibroblast proliferation with increasing estradiol concentrations became less apparent at 7, 10, and 14 days. On Days 1 and 3, procollagen synthesis decreased in a dose dependent manner with increasing estradiol concentrations. On Days 7, 10, and 14, this dose dependent effect was attenuated. No significant differences in Type 3 procollagen synthesis by anterior cruciate ligament fibroblasts were observed with varying estradiol concentrations at any of the designated points. These early physiologic changes in fibroblast proliferation and Type I procollagen synthesis may provide a biologic explanation for the increased anterior cruciate ligament injury rate observed in female athletes, suggesting that it is the acute cyclic variations in the female athlete who is menstruating that predisposes her to ligamentous injury.

In situ calibration of miniature sensors implanted into the anterior cruciate ligament part I: strain measurements.

The goals of this study were to (a) evaluate the differential variable reluctance transducer as an instrument for measuring tissue strain in the anteromedial band of the anterior cruciate ligament, (b) develop a series of calibration curves (for simple states of knee loading) from which resultant force in the ligament could be estimated from measured strain levels in the anteromedial band of the ligament, and (c) study the effects of knee flexion angle and mode of applied loading on output from the transducer. Thirteen fresh-frozen cadaveric knee specimens underwent mechanical isolation of a bone cap containing the tibial insertion of the anterior cruciate ligament and attachment of a load cell to measure resultant force in the ligament. The transducer (with barbed prongs) was inserted into the anteromedial band of the anterior cruciate ligament to record local elongation of the instrumented fibers as resultant force was generated in the ligament. A series of calibration curves (anteromedial bundle strain versus resultant force in the anterior cruciate ligament) were determined at selected knee flexion angles as external loads were applied to the knee. During passive knee extension, strain readings did not always follow the pattern of resultant force in the ligament; erratic strain readings were often measured beyond 20 degrees of flexion, where the anteromedial band was slack. For anterior tibial loading, the anteromedial band was a more active contributor to resultant ligament force beyond 45 degrees of flexion and was less active near full extension; mean resultant forces in the range of 150-200 N produced strain levels on the order of 3-4%. The anteromedial band was also active during application of internal tibial torque; mean resultant forces on the order of 180-220 N produced strains on the order of 2%. Resultant forces generated by varus moment were relatively low, and the anteromedial band was not always strained. Mean coefficients of variation for resultant force in the ligament (five repeated measurements) ranged between 0.038 and 0.111. Mean coefficients of variation for five repeated placements of the strain transducer in the same site ranged from 0.209 to 0.342. Insertion and removal of this transducer at the anteromedial band produced observable damage to the ligament. In our study, repeatable measurements were possible only if both prongs of the transducer were sutured to the ligament fibers.

In situ calibration of miniature sensors implanted into the anterior cruciate ligament part II: force probe measurements.

The arthroscopically implantable force probe transducer, which measures the effects of local ligament fiber tension, was inserted into the anteromedial band of the anterior cruciate ligament after measurements with the differential variable reluctance transducer were completed in Part I of this study. The overall goals in Part II remained the same, with additional experiments included to determine the sensitivity of output voltage from the transducer to medial-lateral placement of the device within the anteromedial band and to depth of placement within a given insertion hole. Calibration curves of output voltage from the arthroscopically implantable force probe transducer versus resultant force in the ligament were generated during a separate series of knee-loading experiments identical to those performed in Part I. The output voltage for a given probe placement was highly sensitive to the depth of implantation into the anteromedial band. When the probe was completely buried within the ligament, voltage outputs were often sporadic or absent even though surface fibers had clearly developed tension. When the probe was only partially inserted into the hole, such that the end of the probe was slightly proud to the surface, voltage output was significantly higher as the device measured tension in the superficial fibers. Voltage outputs for proud placement were always significantly higher than corresponding voltages for deep placements for all test conditions. With proud placements, voltage outputs were not sensitive to small deviations in medial-lateral position within the anteromedial band. Mean coefficients of variation for output voltage (four repeated placements of the probe into the same central hole) ranged from 0.156 to 0.359 (deep and proud insertions). Output voltage from the probe generally followed the pattern of resultant force in the ligament during passive knee extension. For anterior tibial loading, the contribution of deep fibers to resultant force did not depend on the knee flexion angle at which the test was conducted; the contribution of superficial fibers was greatest beyond 45 degrees of flexion and least at full extension. The contributions of the anteromedial band to resultant force in the ligament were not significantly different between the three modes of loading (anterior tibial force, internal tibia torque, and varus moment) at either 0 or 10 degrees of flexion; this was true for both superficial and deep fibers. We found it necessary to secure the probe within the insertion site using a suture (for both deep and proud placements) to obtain repeatable readings. Puncturing the anteromedial band clearly produced tissue damage; the insertion hole often produced a permanent plane of cleavage in the anteromedial band. However, this tissue damage did not alter the overall ability of the ligament to generate resultant force.

Regional gene therapy with a BMP-2-producing murine stromal cell line induces heterotopic and orthotopic bone formation in rodents.

The ability to continuously deliver osteoinductive proteins to a specific anatomic site would facilitate the treatment of fracture nonunions and other clinical problems associated with bone loss. We have developed a murine model of regional gene therapy. A bone-marrow stromal cell line infected with an adenovirus expressing recombinant bone morphogenetic protein-2 cDNA secreted biologically active bone morphogenetic protein-2. These bone morphogenetic protein-2-producing cells were able to induce abundant heterotopic bone formation when implanted into the quadriceps muscle of severe combined immune deficient mice and also successfully healed large segmental femoral defects in nude rats. These studies demonstrate that regional gene therapy with continuous delivery of osteoinductive factors to a specific anatomic site can enhance the formation and repair of bone.

Outcome after total hip arthroplasty. Comparison of a traditional disease-specific and a quality-of-life measurement of outcome.

The purpose of this study was to examine the relationship between the Harris Hip Score (HHS), a traditional method of patient assessment of a total hip arthroplasty (THA), and the Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36), a commonly used health-related quality-of-life survey. One hundred forty patients returning for routine clinical follow-up evaluation of a primary THA were asked to fill out the SF-36 quality-of-life survey, as well as questions concerning their perceptions of their THA. The patient's surgeon assessed the THA with the traditional HHS. The correlations between the HHS and the SF-36 domains were highest in the physical component summary scores for male patients of all ages and female patients 65 years of age or older. The correlations were lower for the mental component summary scores of all patients, but particularly in female patients younger than 65. When the SF-36 scores were compared with age and sex-matched population norms, both age and sex were found to be important. Men younger than 65 had scores lower than norms in the physical function domains, but were comparable in the mental health domains. The older men had scores comparable to the norms in all domains. Female patients of all ages, however, had lower scores in the physical function domains. The greatest differences were noted in the female patients younger than 65. The HHS is commonly used to assess disease-specific pain and function in THA patients; however, the results of this study suggest that the SF-36 health survey can capture additional important quality-of-life domains that are influenced by a THA and that these domains are influenced by the age and sex of the patient. The combination of a disease-specific scoring system and a quality-of-life survey would allow a more global assessment of a THA in all patients. Studies evaluating the results of THAs should either assess the results of male and female patients separately when sample size is sufficiently large or use sex as a possible covariate in a multivariate analysis.

Fibroblast growth factor and epidermal growth factor receptors in ligament healing.

Although recent in vivo studies indicate that basic fibroblast growth factor hastens the healing and strength of the medical collateral ligament after injury, in vitro studies with epidermal growth factor and basic fibroblast growth factor have shown increased fibroblast proliferation with the exogenous administration of these growth factors. Using an established spontaneously healing rabbit injury model, the surgical transection of the medial collateral ligament was undertaken in 12 anesthetized male adult rabbits. Immunohistochemical localization using monoclonal antibodies to the basic fibroblast growth factor receptor and epidermal growth factor receptor were used to identify the distribution and relative concentrations of the individual receptors at 3, 7, 14, and 28 days after surgery. Realizing that the trophic effects of basic fibroblast growth factor and epidermal growth factor are determined exclusively by their individual receptors, this study confirms the hypothesis that basic fibroblast growth factor and epidermal growth factor receptor proteins are present and increase during the early stages of medial collateral ligament healing. The understanding of this rabbit ligament injury model has far reaching implications to ligament healing seen in humans. By elucidating the spatial and temporal regulation of the basic fibroblast growth factor and epidermal growth factor receptor proteins, exogenous growth factor therapy, once approved for human use, potentially can be synchronized with maximal protein receptor levels.

Estrogen affects the cellular metabolism of the anterior cruciate ligament. A potential explanation for female athletic injury.

Investigations from this laboratory have established the presence of estrogen receptors in the human anterior cruciate ligament. This study further investigates the effects of 17 beta-estradiol on the cellular proliferation and collagen synthesis of fibroblasts derived from the rabbit anterior cruciate ligament. Fibroblast proliferation and collagen synthesis in response to near log concentrations of 17 beta-estradiol (at 0.0029, 0.025, 0.25, 2.5, and 25 ng/ml) were assessed by measuring [3H]thymidine and [14C]hydroxyproline incorporation, respectively. Collagen synthesis was significantly reduced with increasing local estradiol concentration (P < 0.001). Declining collagen synthesis was first noted at a 17 beta-estradiol concentration of 0.025 ng/ml. Within normal physiologic levels of estrogen (0.025 to 0.25 ng/ml), collagen synthesis was reduced by more than 40% of control, and at pharmacologic levels of 2.5 and 25 ng/ml, by more than 50% of control. A significant reduction of fibroblast proliferation was also observed with increasing estradiol concentrations (P = 0.023). Clinically, alterations in anterior cruciate ligament cellular metabolism caused by estrogen fluctuations may change the composition of the ligament, rendering it more susceptible to injury.

Morphology and matrix composition during early tendon to bone healing.

This study outlines the early morphologic phenomenon of tendon to bone healing in the rabbit model. Twelve skeletally mature, male New Zealand White rabbits received transplantation of the hallucis longus tendon into a 2-mm calcaneal bone tunnel. The morphologic characteristics of the healing tendon to bone interface were evaluated at 1, 2, 4, and 6 weeks after surgery by the use of conventional histology and immunohistochemical localization of collagen Types I, II, and III. Histologic analysis illustrated progressive maturation and reorganization of the tendon to bone interface with subsequent development of tissue collagen fiber continuity between the tendon and bone. Initially, diffuse immunolocalization of all three collagen types was observed within the scar tissue filling the space between the tendon and bone. During a 6-week period, reorganization of the scar tissue into an interface occurred, similar to an indirect insertion. Although a definitive fibrocartilage region did not form, Type II collagen was localized at the remodeling insertion site throughout the first 6 weeks of repair. In addition, Type III collagen fibers, resembling Sharpey's fibers, were noted to span this interface. The characterization of the insertion between tendon and bone is important to the understanding of healing in commonly used orthopaedic grafting procedures, such as anterior cruciate ligament reconstructions.

The efficacy of prophylaxis with low-dose warfarin for prevention of pulmonary embolism following total hip arthroplasty.

The selection of a prophylaxis regimen and its implementation have been influenced considerably by the decreased duration of hospital stays and the pressures of cost containment. The purpose of the present study was to determine the rate of symptomatic pulmonary embolism both before and after discharge, the number of days required to achieve an adequate level of anticoagulation, and the complications associated with the use of low-dose warfarin after total hip arthroplasty. Between 1987 and 1993, 1099 primary and revision total hip arthroplasties were performed in 940 patients who received low-dose warfarin for prophylaxis against thromboembolic disease. The average duration of prophylaxis was fifteen days (range, one to twenty-nine days). The target level of anticoagulation (as indicated by a prothrombin time of fourteen to seventeen seconds) was achieved an average of three days (range, one to sixteen days) after the operation. The level of anticoagulation was lower than the target range at the time of discharge after 257 total hip arthroplasties (23.4 per cent), and the target level was never achieved during the period of hospitalization after fifty-four such procedures (4.9 per cent). Twelve total hip arthroplasties were associated with a symptomatic pulmonary embolism; the over-all prevalence of this complication therefore was 1.1 per cent (95 per cent confidence interval, 0.4 to 1.9 per cent). Four pulmonary emboli were diagnosed before discharge and eight, after discharge. A fatal pulmonary embolism occurred after one procedure (0.1 per cent). Patients who had a history of symptomatic venous thromboembolic disease had a significantly increased risk of symptomatic pulmonary embolism after total hip arthroplasty (p = 0.001, Fisher exact test). A major bleeding episode occurred after thirty-two total hip arthroplasties (2.9 per cent). Patients who had a prothrombin time of more than seventeen seconds had a significantly increased risk of hematoma formation (p = 0.003, chi-square analysis). Prophylaxis with low-dose warfarin is safe and effective for the prevention of pulmonary embolism after total hip arthroplasty.

A biomechanical study of replacement of the posterior cruciate ligament with a graft. Part 1: Isometry, pre-tension of the graft, and anterior-posterior laxity.

UNLABELLED: Twelve fresh-frozen knee specimens from cadavera were subjected to anterior-posterior laxity testing with 200 newtons of force applied to the tibia; testing was performed before and after a femoral load-cell was connected to a mechanically isolated cylindrical cap of subchondral femoral bone containing the femoral origin of the posterior cruciate ligament. The posterior cruciate ligament then was removed, the proximal end of a thin trial isometer wire was attached to one of four points designated on the femur, and displacement of the distal end of the wire relative to the tibia was measured over a 120-degree range of motion. The potted end of a ten-millimeter-wide bone-patellar ligament-bone graft was centered over the femoral origin of the ligament and attached to the femoral load-cell. Isometry measurements were repeated with the wire attached to the bone block of the free end of the graft in the tibial tunnel. Force was recorded at the load-cell (representing force in the intra-articular portion of the graft) as pre-tension was applied, with use of a calibrated spring-scale, to the tibial end of the graft. A laxity-matched pre-tension of the graft was determined such that the anterior-posterior laxity of the reconstructed knee at 90 degrees of flexion was within one millimeter of the laxity that was measured after installation of the load-cell. Anterior-posterior testing was repeated after insertion of the graft at the laxity-matched pre-tension. The least amount of change in the relative displacement of the trial wire over the 120-degree range of flexion occurred when the wire was attached to the proximal point on the femur (a point on the proximal margin of the femoral origin of the posterior cruciate ligament, midway between the anterior and posterior borders of the ligament). The greatest change in the relative displacement was associated with the anterior point (a point on the anterior margin of the femoral origin of the ligament, midway between the proximal and distal borders). The mean relative displacements of the trial wire when it was attached to a point at the center of the femoral origin of the ligament were not significantly different from the corresponding mean displacements of the distal end of the graft when the proximal end of the graft was centered at this point. At 90 degrees of flexion, the force recorded by the load-cell averaged 64 to 74 per cent of the force applied to the tibial end of the graft. The laxity-matched pre-tension of the graft at 90 degrees of flexion (as recorded by the load-cell) ranged from six to 100 newtons (mean and standard deviation, 43.0 +/- 33.4 newtons). With the numbers available, the mean laxities after insertion of the graft were not significantly different, at any angle of flexion, from the corresponding mean values after installation of the load-cell. CLINICAL RELEVANCE: Isometer readings from a trial wire attached to a point on the femur provided an accurate indication of the change in the length of a graft subsequently centered at that point. Anteriorly placed femoral tunnels should be avoided, as the isometer readings indicated increased tension, with flexion of the knee, in a graft placed in this region. The force in the intra-articular portion of the graft was always less than the force applied to the bone block in the tibial tunnel. Therefore, the femoral end of the graft should be tensioned to avoid frictional losses from the severe bend in the graft as it passes over the posterior tibial plateau. With correct pre-tensioning of a graft, normal anterior-posterior laxity at 0 to 90 degrees of flexion can be restored. However, because of the considerable range in the laxity-matched pre-tensions, we recommend that the pre-tension be greater than forty-three newtons for all patients to ensure that normal laxity is restored.

A biomechanical study of replacement of the posterior cruciate ligament with a graft. Part II: Forces in the graft compared with forces in the intact ligament.

UNLABELLED: A femoral load-cell was installed in twelve fresh-frozen knee specimens from cadavera, to measure the resultant force at the femoral origin of the posterior cruciate ligament during a series of tibial-loading tests. The posterior cruciate ligament was removed, and a ten-millimeter-wide bone-patellar ligament-bone graft was inserted. The knee was flexed to 90 degrees, the graft was pre-tensioned to restore the anterior-posterior laxity to that recorded after installation of the load-cell, and the loading tests were repeated. With the tibia locked in neutral rotation and a 200-newton posterior force applied to the tibia, the mean force generated in the intact posterior cruciate ligament ranged from 220 newtons at 90 degrees of flexion to thirty-six newtons at full extension. When the tibia was locked in external rotation during the posterior drawer test, the force was reduced when the knee was flexed 10 to 70 degrees; when the tibia was locked in internal rotation, the mean force was reduced at only 30 and 45 degrees of flexion. The mean forces in the graft were not significantly different, with the numbers available, from the corresponding values for the intact ligament during application of a straight posterior tibial force (neutral tibial rotation), during application of a fifteen-newton-meter flexion or extension moment (hyperflexion or hyperextension), during application of a ten-newton-meter varus or valgus moment, or during application of a ten-newton-meter internal or external tibial torque. With the numbers available, there were no significant differences between the mean tibial rotations associated with the intact posterior cruciate ligament and those associated with the graft at any angle of flexion, without or with applied tibial torque. CLINICAL RELEVANCE: The amount of force generated in the posterior cruciate ligament during the posterior drawer test depends on the angle of flexion at which the test is performed. When the angle of flexion is near 90 degrees, all of the posterior force applied to the tibia is transmitted to the ligament and the force in the ligament is not affected by the position of tibial rotation. When the test is performed at an angle of flexion near 30 degrees and in neutral tibial rotation, other structures (such as the collateral ligaments and the posterior part of the capsule) help to resist the posterior force applied to the tibia. The position of tibial rotation is important when the test is performed with the knee at an angle of flexion near 30 degrees, as secondary structures pre-tensioned by tibial torque act to reduce the amount of force carried by the posterior cruciate ligament even more. With a few minor exceptions, we found that the forces in a graft used to replace the posterior cruciate ligament were approximately the same as those in the intact ligament. Therefore, there appears to be little justification for restricting low-level rehabilitation activities once the fixation of the graft has healed. However, forces in the graft could be quite high during hyperextension and hypertension, as they are in the intact ligament. Thus, bracing in the early postoperative period may be advisable to prevent these motions.

Low-dose warfarin prophylaxis to prevent symptomatic pulmonary embolism after total knee arthroplasty.

Pulmonary embolism poses a risk to patients undergoing total knee arthroplasty. The selection of an appropriate prophylaxis agent and its implementation have been influenced by decreased duration of hospital stay and the pressures of cost containment. The purpose of this study was to determine the inpatient and outpatient pulmonary embolism rates, the number of days required to attain the target level of anticoagulation, and complications associated with the use of a low-dose warfarin prophylaxis protocol after primary and revision total knee arthroplasty. Between 1984 and 1993, there were 815 primary and revision total knee arthroplasties that received low-dose warfarin prophylaxis at our institution. The average time to attainment of the target level of anticoagulation was 3 days. The average duration of warfarin prophylaxis was 12 days. Overall, there were a total of three symptomatic pulmonary embolisms (0.3%; 95% confidence interval, 0.08%-1.1%). There were eight (1%) symptomatic deep vein thromboses (all distal). There were two deaths (0.3%), but neither one was secondary to a pulmonary embolism. Seventeen knees (2.5%) developed a hematoma after surgery, and two of these patients required drainage of the knee. Low-dose warfarin prophylaxis is safe and effective in preventing symptomatic pulmonary embolism after total knee arthroplasty.

Primary immunolocalization of androgen target cells in the human anterior cruciate ligament.

To identify androgen target cells in the human anterior cruciate ligament, immunohistochemical localization of the androgen receptor was performed in 31 specimens of the ligament. All of the specimens were obtained at surgery. Seventeen specimens were from women, and 14 were from men: the average age of the patients was 45.2 years (range: 18-78 years). An immunoperoxidase method using monoclonal antibodies to the androgen receptor was employed to identify androgen target cells in the ligament. Consistent staining of the androgen receptor was demonstrable in six specimens obtained from young men 18-24 years old, and equivocal positive staining was seen in two other specimens from young men. No receptors were demonstrated by this method in any of the specimens from women or older men. Androgen receptors were localized to synoviocytes in the synovium and under the synovial lining, fibroblasts in the ligament stroma, and cells lining the blood-vessel walls of the anterior cruciate ligament. The demonstration of androgen receptors in the cells of the anterior cruciate ligament strongly suggests that male sex hormones may have an effect on the structure and composition of this ligament in young men.

Biomechanical consequences of replacement of the anterior cruciate ligament with a patellar ligament allograft. Part I: insertion of the graft and anterior-posterior testing.

Nineteen fresh-frozen knee specimens from cadavera were tested for anterior-posterior laxity with 200 newtons of force applied to the tibia. A cylindrical cap of subchondral bone containing the tibial insertion of the anterior cruciate ligament was isolated with a coring cutter and was potted in acrylic. A thin wire was connected to the undersurface of the cap, and relative displacement between the cap and the tibia was measured with an isometer as the knee was extended. The cap of bone was connected to a load-cell that recorded force in the intact ligament during anterior-posterior testing with the tibia locked in neutral, internal rotation, and external rotation. The anterior cruciate ligament was then resected, and a femoral tunnel was drilled at the site where the isometer readings from the wire were the same as those obtained for the intact anterior cruciate ligament. A bone-patellar ligament-bone graft was used to reconstruct the anterior cruciate ligament, and the isometer measurements were repeated with the graft in place. The graft was pre-tensioned at 30 degrees of flexion to restore normal anterior-posterior laxity. Anterior-posterior laxity tests were repeated at this level of pre-tension (laxity-matched pre-tension) as well as at a level that was forty-five newtons greater (over-tension). The moment required to extend the knee was measured before and after insertion of the graft at both levels of pre-tension. When the tibia was locked in positions of internal and external rotation, the anterior-posterior laxities and the forces in the anterior cruciate ligament (generated by an anterior force applied to the tibia) were significantly less than the corresponding values with the tibia in neutral rotation at 20, 30, and 45 degrees of flexion (p < or = 0.05). Isometer readings for the intact anterior cruciate ligament indicated that the cap of bone retracted into the joint a mean and standard deviation of 3.1 +/- 0.8 millimeters as the knee was extended from 30 degrees of flexion to full extension. For each specimen, the isometer measurements for the trial wire and for the graft were within 1.5 millimeters of those for the intact anterior cruciate ligament. At laxity-matched pre-tension (mean, 28.2 +/- 16.8 newtons), the mean anterior-posterior laxities of the reconstructed knees were within 1.0 millimeter of the corresponding means for the intact knees between 0 and 45 degrees of flexion. Over-tensioning of the graft by forty-five newtons decreased the anterior-posterior laxity a mean of 1.2 millimeters at 30 degrees of flexion. Over-tensioning of the graft did not change the moment required to bring the knee to full extension.

Biomechanical consequences of replacement of the anterior cruciate ligament with a patellar ligament allograft. Part II: forces in the graft compared with forces in the intact ligament.

Seventeen fresh-frozen knee specimens from cadavera were instrumented with a load-cell attached to a mechanically isolated cylinder of subchondral bone containing the tibial insertion of the anterior cruciate ligament. The forces in the intact anterior cruciate ligament were recorded as the knee was passively extended from 90 degrees of flexion to 5 degrees of hyperextension without and with several constant tibial loads: 100 newtons of anterior tibial force, ten newton-meters of internal and external tibial torque, and ten newton-meters of varus and valgus moment. The anterior cruciate ligament was resected, and a bone-patellar ligament-bone graft was inserted. The knee was flexed to 30 degrees, and the graft was pre-tensioned to restore normal anterior-posterior laxity. The knee-loading experiments were repeated at this level of pre-tension (laxity-matched pre-tension) and at a level that was forty-five newtons greater than the laxity-matched pre-tension (over-tension). During passive extension of the knee, the forces in the graft were always greater than the corresponding forces in the intact anterior cruciate ligament. Over-tensioning of the graft increased the forces in the graft at all angles of flexion. At full extension, the mean force in the anterior cruciate ligament was fifty-six newtons; the mean force in the graft at laxity-matched pre-tension was 168 newtons, and it was 286 newtons in the over-tensioned graft. Greater pre-tensioning may be required when the knee demonstrates apparent tightening of the graft in flexion. The mean forces in the graft generated during all constant loading tests were greater than those for the intact anterior cruciate ligament over the range of flexion. When the graft was over-tensioned, the forces generated by the anterior tibial force and by varus and valgus moment increased but those generated by internal and external tibial torque did not. There was no significant change in the mean tibial rotation as a function of the angle of flexion of the knee after insertion of the graft; normal tibial rotation of the knee during passive extension (the so-called screw home mechanism) was eliminated.

MRI and morphology of the insertion of the patellar tendon after graft harvesting.

We performed MRI on 16 patients who had had reconstruction of the anterior cruciate ligament (ACL) with a mid-third bone-patellar-tendon-bone autograft. Our aim was to assess the tendon and the site of its insertion at an average of seven years after the original operation. In four of these patients biopsies were taken from the donor site when they had revision of their original operation. MRI showed reconstitution of the tendon into the patellar defect with no evidence of bone formation. Six patients had a persistent defect in the patellar tendon itself. Histological examination of the biopsies of the donor site showed an indirect pattern of insertion with absence of the normal fibrocartilage zone. These morphological changes may adversely affect the biomechanical properties of the healed donor site and we suggest that another graft taken from this site may not be suitable for use in a further operation for reconstruction of the ACL.

Primary immunolocalization of estrogen and progesterone target cells in the human anterior cruciate ligament.

To identify estrogen and progesterone target cells in the human anterior cruciate ligament immunohistochemical localization of both estrogen and progesterone receptors was performed in 17 specimens of human anterior cruciate ligament. All ligament specimens were obtained at surgery. Thirteen specimens were from women, and four were from men: the average age was 57 years (range, 18-78 years). Eleven specimens (from nine women and two men) came from total knee replacements for osteoarthritis of the knee: three (from two women and one man), from reconstructions of the anterior cruciate ligament: two (both from women), from medial meniscectomies; and one (from a man), from an amputation secondary to chondrosarcoma of the pelvis. An immunoperoxidase method using monoclonal antibodies to the estrogen and progesterone receptors was employed to identify estrogen and progesterone target cells in the anterior cruciate ligament. Staining of both receptors was demonstrable in 14 specimens and in the remaining three specimens less than 15% of the cells were stained. Both estrogen and progesterone receptors were localized to synoviocytes in the synovial lining, fibroblasts in the anterior cruciate ligament stroma and cells in the blood vessel walls of the ligament. This demonstration of receptors for estrogen and progesterone in the cells of anterior cruciate ligament suggests that female sex hormones may have an effect on its structure and composition.

Effects of combined knee loadings on posterior cruciate ligament force generation.

Resultant forces in the posterior cruciate ligament were measured under paired combinations of posterior tibial force, internal and external tibial torque, and varus and valgus moment. The force generated in the ligament from a straight 100 N posterior tibial force was highly sensitive to the angle of knee flexion. For example, at 90 degrees of flexion the mean resultant force in the posterior cruciate ligament was 112% of the applied posterior tibial force, whereas at 0 degree, only 16% of the applied posterior force was measured in the ligament. When the tibia was preloaded by 10 Nm of external torque, only 9-13% of the 100 N posterior tibial force was transmitted to the posterior cruciate ligament at flexion angles less than 60 degrees; at 90 degrees of flexion, 61% was carried by the ligament. This "off-loading" of the posterior cruciate ligament also occurred when the tibia was preloaded by 10 Nm of internal torque, but only at knee flexion angles between 20 and 40 degrees. The addition of 10 Nm of valgus moment to a knee loaded by a 100 N posterior tibial force increased the mean force in the posterior cruciate ligament at all flexion angles except hyperextension; this represents a common and potentially dangerous loading combination. The addition of 10 Nm of varus moment to a knee loaded by a 100 N posterior tibial force decreased the mean force in the ligament between 10 and 70 degrees of flexion. External tibial torque (alone or combined with varus or valgus moment) was not an important loading mechanism in the posterior cruciate ligament. The application of internal torque plus varus moment at 90 degrees of flexion produced the greatest posterior cruciate ligament forces in our study and represented the only potential injury mechanism that did not involve posterior tibial force.