The effects of refreezing on the viscoelastic and tensile properties of ligaments.

Biomechanical testing protocols for ligaments can be extensive and span two or more days. During this time, a specimen may have to undergo more than one cycle of freezing and thawing. Thus, the objective of this study was to evaluate the effects of refreezing on the viscoelastic and tensile properties of ligaments. The femur-medial collateral ligament-tibia complexes (FMTC) from six pairs of rabbit knees were used for this study. Following sacrifice, one leg in each pair was assigned to the fresh group and the FMTC was immediately dissected and prepared for testing. The contralateral knees were fresh-frozen at -20 degrees C for 3 weeks, thawed, dissected and then refrozen for one additional week before being tested as the refrozen group. The cross-sectional area and shape of the medial collateral ligament (MCL) was measured using a laser micrometer system. Stress relaxation and cyclic stress-relaxation tests in uniaxial tension were performed followed by a load to failure test. When the viscoelastic behavior of the MCL was described by the quasi-linear viscoelastic (QLV) theory, no statistically significant differences could be detected for the five constants (A, B, C, tau1, and tau2) between the fresh and refrozen groups (p > or = 0.07) based on our sample size. In addition, the structural properties of the FMTCs and the mechanical properties of the MCLs were also found to be similar between the two groups (p > or = 0.68). These results suggest that careful refreezing of the specimens had little or no effect on the biomechanical properties measured.

A novel methodology to reproduce previously recorded six-degree of freedom kinematics on the same diarthrodial joint.

The objective of this study was to develop a novel method to more accurately reproduce previously recorded 6-DOF kinematics of the tibia with respect to the femur using robotic technology. Furthermore, the effect of performing only a single or multiple registrations and the effect of robot joint configuration were investigated. A single registration consisted of registering the tibia and femur with respect to the robot at full extension and reproducing all kinematics while multiple registrations consisted of registering the bones at each flexion angle and reproducing only the kinematics of the corresponding flexion angle. Kinematics of the knee in response to an anterior (134 N) and combined internal/external (+/-10 N m) and varus/valgus (+/-5 N m) loads were collected at 0 degrees , 15 degrees , 30 degrees , 60 degrees , and 90 degrees of flexion. A six axes, serial-articulated robotic manipulator (PUMA Model 762) was calibrated and the working volume was reduced to improve the robot's accuracy. The effect of the robot joint configuration was determined by performing single and multiple registrations for three selected configurations. For each robot joint configuration, the accuracy in position of the reproduced kinematics improved after multiple registrations (0.7+/-0.3, 1.2+/-0.5, and 0.9+/-0.2 mm, respectively) when compared to only a single registration (1.3+/-0.9, 2.0+/-1.0, and 1.5+/-0.7 mm, respectively) (p<0.05). The accuracy in position of each robot joint configuration was unique as significant differences were detected between each of the configurations. These data demonstrate that the number of registrations and the robot joint configuration both affect the accuracy of the reproduced kinematics. Therefore, when using robotic technology to reproduce previously recorded kinematics, it may be necessary to perform these analyses for each individual robotic system and for each diarthrodial joint, as different joints will require the robot to be placed in different robot joint configurations.

Functional tissue engineering for ligament healing: potential of antisense gene therapy.

Traumatic knee injuries frequently involve the disruption of multiple ligaments, such as a complete tear of the medial collateral ligament (MCL) together with a rupture of the anterior cruciate ligament (ACL) (Miyasaka, K., D. M. Daniel, M. L. Stone, and P. Hirshman. Am. J. Knee Surg. 4:3-8, 1991). Despite the high incidence, clinical management of this type of injury is still debated. Laboratory studies have shown that the ACL and MCL share the responsibility of stabilizing the knee, especially in response to valgus and other rotatory torques as well as anterior tibial loads (Inoue, M., E. McGurk-Burleson, J. M. Hollis, and S. L-Y. Woo. Am. J. Sports Med. 15:15-21, 1987; Kanamori, A., M. Sakane, J. Zeminski, T. W. Rudy, and S. L-Y. Woo. J. Ortho. Sci. 5:567-571, 2000; Ma, C. B., C. D. Papageogiou, R. E. Debski, and S. L. Woo. Acta Orthop. Scand. 71:387-393, 2000; Sakane, M., G. A. Livesay, R. J. Fox, T. W. Rudy, T. J. Runco, and S. L-Y. Woo. Knee Surg. Sports Traumatol. Arthrosc. 7:93-97, 1999). When one structure is deficient, the force in the other increases significantly to compensate. The injured ACL does not heal and requires surgical replacement by tissue grafts. On the other hand, after an isolated MCL tear or in a combined MCL and ACL injury, the MCL can heal spontaneously without surgical intervention and can function well in most cases. Nevertheless, the biomechanical and biochemical properties as well as the histomorphological appearance of the healing MCL are substantially different to those of normal tissue (Bray, R. C., D. J. Butterwick, M. R. Daschak, and J. V. Tyberg. J. Orthop. Res. 14:618-625, 1996; Loitz-Ramage, B. J., C. B. Frank, and N. G. Shrive. Clin. Orthop.:272-280, 1997; Weiss, J. A., S. L-Y. Woo, K. J. Ohland, S. Horibe, and P. O. Newton. J. Orthop. Res. 9:516-528, 1991). In an effort to improve the outcome of injuries to these and other ligaments, therapeutic strategies associated with improving biomechanical, biochemical, and histomorphological properties of ligaments have been investigated in recent years. These therapeutic strategies include growth factor stimulation (Conti, N. A., and L. E. Dahners. Presented at Orthopaedic Research Society, San Francisco, CA; Deie, M., T. Marui, C. R. Allen, K. A. Hildebrand, H. I. Georgescu, et al. Mech. Ageing Dev. 97:121-130, 1997), cell therapy (Menetrey, J., C. Kasemkijwattana, C. S. Day, P. Bosch, F. H. Fu, et al. Tissue Eng. 5:435-442, 1999; Watanabe, N., S. L-Y. Woo, C. Papageorgiou, C. Celechovsky, and S. Takai. Microsc. Res. Tech. 58:39-44, 2002), as well as gene stherapy (Nakamura N., D. A. Hart, R. S. Boorman, Y. Kaneda, N. G. Shrive, et al. J. Orthop. Res. 18:517-523, 2000; Shimomura, T., F. Jia, C. Niyibizi, and S. L-Y. Woo. Connect. Tissue Res.:2003). The knowledge gained by studying these therapeutic strategies could potentially be applied to other ligaments and tendons. In this article, antisense gene therapy to alter gene expression by using antisense oligonucleotides will be examined as a possible solution.

Passive strain distribution in the interosseous ligament of the forearm: implications for injury reconstruction.

PURPOSE: For severe forearm injuries such as an Essex-Lopresti fracture-dislocation, functional reconstruction necessitates repair of the interosseous ligament (IOL) to restore normal load sharing between the radius and ulna. Locating or tensioning such a reconstruction improperly can lead to abnormal load sharing and/or restriction of forearm rotation. The normal IOL strains should indicate the proper location of reconstruction grafts and the proper forearm rotation for tensioning the grafts. The objective of this study was to quantify the passive strain distribution of the IOL of the forearm with passive rotation of the forearm throughout the range of motion. METHODS: The 3-dimensional motions of the radius with respect to the ulna were measured throughout forearm rotation in 10 cadaveric forearms by using an instrumented spatial linkage. From the bone motions and ligament insertion site geometry from dissection and computed tomographic scanning, insertion site motions were determined and used to calculate changes in ligament fiber lengths. RESULTS: The measured strain distribution in the IOL was nonuniform and varied with forearm rotation. The overall magnitude of IOL strain was found to be greatest in supination and smallest in pronation. In supination the strains varied across fibers with strains being greatest in the distal fibers and lowest in the proximal fibers. Strains in neutral rotation were uniform across fibers. Although fibers were generally slack in pronation proximal fibers were less slack than distal fibers. CONCLUSIONS: The results of this study indicate that fiber strains in the IOL vary from proximal to distal and depend on forearm rotation. Our data suggest that to prevent restriction of forearm rotation all grafts should be tensioned in supination, where measured strains were generally highest. Our data also suggest that a 2-bundle IOL reconstruction may be necessary for proper load transfer between the radius and ulna in both supination and pronation.

Distribution of in situ forces in the anterior cruciate ligament in response to rotatory loads.

The anterior cruciate ligament (ACL) can be anatomically divided into anteromedial (AM) and posterolateral (PL) bundles. Current ACL reconstruction techniques focus primarily on reproducing the AM bundle, but are insufficient in response to rotatory loads. The objective of this study was to determine the distribution of in situ force between the two bundles when the knee is subjected to anterior tibial and rotatory loads. Ten cadaveric knees (50+/-10 years) were tested using a robotic/universal force-moment sensor (UFS) testing system. Two external loading conditions were applied: a 134 N anterior tibial load at full knee extension and 15 degrees, 30 degrees, 60 degrees, and 90 degrees of flexion and a combined rotatory load of 10 Nm valgus and 5 Nm internal tibial torque at 15 degrees and 30 degrees of flexion. The resulting 6 degrees of freedom kinematics of the knee and the in situ forces in the ACL and its two bundles were determined. Under an anterior tibial load, the in situ force in the PL bundle was the highest at full extension (67+/-30 N) and decreased with increasing flexion. The in situ force in the AM bundle was lower than in the PL bundle at full extension, but increased with increasing flexion, reaching a maximum (90+/-17 N) at 60 degrees of flexion and then decreasing at 90 degrees. Under a combined rotatory load, the in situ force of the PL bundle was higher at 15 degrees (21+/-11 N) and lower at 30 degrees of flexion (14+/-6 N). The in situ force in the AM bundle was similar at 15 degrees and 30 degrees of knee flexion (30+/-15 vs. 35+/-16 N, respectively). Comparing these two external loading conditions demonstrated the importance of the PL bundle, especially when the knee is near full extension. These findings provide a better understanding of the function of the two bundles of the ACL and could serve as a basis for future considerations of surgical reconstruction in the replacement of the ACL.

Tensile properties of an anterior cruciate ligament graft after bone-patellar tendon-bone press-fit fixation.

Hardware used for fixation of ACL autografts in bone tunnels frequently complicates revision surgery, requiring two-stage procedures when a bone-patellar tendon-bone (B-PT-B) autograft is used for ACL reconstruction. Therefore alternative procedures that eliminate hardware have been advocated. This study compared the mechanical behavior of two fixation procedures: a widely used interference screw (IFS) fixation and a press-fit fixation that is hardware free. Twenty hind limbs from skeletally mature Saanen breed goats were used in this study, ten each in IFS and press-fit groups. After ACL reconstruction the specimens were dissected, leaving a femur-ACL graft-tibia complex (FATC) for uniaxial tensile testing. The tests included a series of three cyclic creep tests (C1-C3) for the evaluation of residual elongation followed by a tensile load to failure test to obtain linear stiffness and ultimate load of the FATCs. Four of ten specimens failed during the cyclic creep test for the press-fit group, compared to one for the IFS group. For the remaining specimens residual elongation following three cyclic creep tests (C1-C3) was 1.7+/-0.5 mm in the press-fit group compared to 1.3+/-0.6 mm in the IFS group, and there was no statistical significant difference between the two fixations. In the load to failure test there was also no statistical significant difference in linear stiffness between the two fixations. However, the ultimate load for the press-fit group (215+/-75 N) was significantly lower than that for the IFS group (328+/-103 N). These results provide the basis for future studies involving the time course of healing of these two procedures using the goat model.