Estrogen level alters the failure load of the rabbit anterior cruciate ligament.

We hypothesized that increasing the level of circulating serum estrogen would decrease the load at failure for the rabbit anterior cruciate ligament. We developed an animal model in which hormonal manipulations could be correlated with load at failure for the anterior cruciate ligament. Sixteen New Zealand White ovariectomized rabbits were matched and divided into two groups. The eight rabbits that were not treated with the estrogen supplement (Group O) and the eight that were treated with the supplement (Group E) were housed for 30 days. Serum estrogen levels were measured. The knees were stripped of all soft tissue, and the load at failure for the anterior cruciate ligament was measured in a materials testing machine with a displacement rate of 0.5 mm/sec. The load at failure for all 16 specimens in Group E (446 +/- 54 N) (mean +/- SD) was significantly reduced (p = 0.02) compared with that for the nine specimens in Group O (503 +/- 48 N). It is recognized that an increased number of anterior cruciate ligament injuries occurs in female athletes. Although the mechanism responsible for failure of the anterior cruciate ligament in women is yet to be defined, this experiment suggests that estrogen may alter ligament strength.

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.

Combined knee loading states that generate high anterior cruciate ligament forces.

Injuries to the anterior cruciate ligament frequently occur under combined mechanisms of knee loading. This in vitro study was designed to measure levels of ligament force under dual combinations of individual loading states and to determine which combinations generated high force. Resultant force was recorded as the knee was extended passively from 90 degrees of flexion to 5 degrees of hyperextension under constant tibial loadings. The individual loading states were 100 N of anterior tibial force, 10 Nm of varus and valgus moment, and 10 Nm of internal and external tibial torque. Straight anterior tibial force was the most direct loading mechanisms; the mean ligament force was approximately equal to applied anterior tibial force near 30 degrees of flexion and to 150% of applied tibial force at full extension. The addition of internal tibial torque to a knee loaded by anterior tibial force produced dramatic increases of force at full extension and hyperextension. This loading combination produced the highest ligament forces recorded in the study and is the most dangerous in terms of potential injury to the ligament. In direct contrast, the addition of external tibial torque to a knee loaded by anterior tibial force decreased the force dramatically for flexed positions of the knee; at close to 90 degrees of flexion, the anterior cruciate ligament became completely unloaded. The addition of varus moment to a knee loaded by anterior tibial force increased the force in extension and hyperextension, whereas the addition of valgus moment increased the force at flexed positions. These states of combined loading also could present an increased risk for injury. Internal tibial torque is an important loading mechanism of the anterior cruciate ligament for an extended knee. The overall risk of injury to the ligament from varus or valgus moment applied in combination with internal tibial torque is similar to the risk from internal tibial torque alone. External tibial torque was a relatively unimportant mechanism for generating anterior cruciate ligament force.

Temperature dependence of the crossbridge cycle during unloaded shortening and maximum isometric tetanus in frog skeletal muscle.

The primary objective of this study was to determine if the rate limiting step in the crossbridge cycle was the same during maximum rate of shortening and during maintenance of maximum tension in an isometric contraction. To this end the temperature dependence, Q10, of the crossbridge cycle was estimated during unloaded shortening and maximum isometric tetanus. Isolated semitendinosus muscles from the frog were studied at 0 and 10 degrees C. Crossbridge cycling during unloaded shortening was determined from the velocity of unloaded shortening estimated by the slack step technique. Crossbridge cycling during maintained isometric tetanus was determined from the steady rate of energy liberation during the tetanus after allowance for energy liberation due to Ca2+ cycling. The Q10 of the velocity of unloaded shortening was 2.5 and that of the steady rate of energy liberation was 4.6. After correction for the temperature dependence of energy liberation associated with Ca2+ cycling (5.7), the estimated Q10 of the steady rate of energy liberation became 3.9. These estimates of the Q10 of the crossbridge cycle are significantly different. These results support the conclusion that the rate limiting steps during unloaded shortening and maximum isometric force maintenance occur at different steps in the crossbridge cycle. Further the high Q10 of the energy liberation due to Ca2+ cycling may relate to the high concentration of parvalbumin in frog muscle. A second objective of this study was to document in the same muscle the variation of Q10s of mechanical and energetic properties of contraction. Over this temperature range the Q10s ranged from 1.1 to 5.7.

Energetics and mechanics of frog skeletal muscle in hypotonic solution.

Hypotonic solutions are known to potentiate muscle force production and increase actomyosin ATPase activity in solution. As such, both the rate and amount of ATP hydrolysis should increase during contraction. This was tested indirectly by measuring force and energy liberation in Rana pipiens semitendinosus muscles at 0 degrees C in hypotonic solution. Force and the amount and rate of energy liberation increased. This increase is consistent with the interpretation that the rate of ATP hydrolysis is increased in hypotonic solution. Muscles, stretched to beyond myofilament overlap, liberate a substantial fraction of the energy liberated at maximal myofilament overlap. This energy liberation, the activation heat, is thought to reflect the energy utilized to cycle Ca2+. Hypotonic solution decreased the amplitude and the rate of the activation heat, suggesting that the amount and rate of Ca2+ cycled by the sarcoplasmic reticulum is reduced. Thus in hypotonic solution, force production and the rate of ATP hydrolysis by the cross bridges are potentiated despite an apparent decrease in Ca2+ cycling.

Energetics of activation in frog skeletal muscle at sarcomere lengths beyond myofilament overlap.

Experiments were designed to gain information about the effects of extremely long sarcomere lengths (greater than 3.8 microns) on muscle activation. The amount of energy liberated in an isometric twitch by muscles stretched to sarcomere lengths where myofilament overlap is vanishingly small (greater than 3.6 microns) is thought to be an indirect measure of the Ca2+ cycled during contraction. The effects of altering sarcomere length from 3.8 to 4.3 microns on the amount of Ca2+ cycled was measured using twitch energy liberation as an indicator of the Ca2+ cycled. Twitch energy liberation decreased by approximately 20% over this sarcomere length region, suggesting that the amount of Ca2+ released by a single action potential is not altered dramatically when a muscle is stretched to extreme lengths.

Effects of 2-n-butyl-methylenedioxyindene on skeletal muscle mechanics and energetics.

Mechanical and energetic effects of 2-n-butyl-3-dimethylamino-5,6-methylenedioxyindene (2-butyl-MDI) were investigated in isolated frog semitendinosus muscles at 0 degrees C. Previous research on various tissues suggested that this compound functions as an intracellular Ca2+ antagonist. The effects of 2-butyl-MDI (2 X 10(-4) M) with respect to time were progressive and reversible with exposures of 30 min or less. A 30-min exposure to the agent significantly decreased twitch and tetanus force and energy liberation, increased the twitch-to-tetanus ratio, prolonged kinetics of force development, induced a stimulus frequency-dependent tetanic fatigue, and decreased contractile economy (measured as force per unit energy liberation). Energy associated with Ca2+ cycling, activation heat, was depressed by 31 +/- 4%. The significant reduction of activation heat production by 2-butyl-MDI suggests that the quantity of Ca2+ released by the sarcoplasmic reticulum upon stimulation is reduced. However, the complexity of the results summarized above suggests multiple sites and/or modes of action for the agent.