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Surgical Retrieval, Isolation and In vitro Expansion of Human Anterior Cruciate Ligament-derived Cells for Tissue Engineering Applications
Published on: April 30, 2014
The effects of in situ freezing on the anterior cruciate ligament. An experimental study in goats
D W Jackson1, E S Grood, B T Cohn
1Research Department, Southern California Center for Sports Medicine, Long Beach 90806.
Abstract:
We developed an in situ freeze-thaw model designed to simulate an ideally placed and oriented autogenous graft of the anterior cruciate ligament. In this model, the anterior cruciate ligament was exposed, and the femoral insertion, tibial insertion, and body of the anterior cruciate ligament were frozen in situ with specially designed freezing probes. Freeze-thaw cycles were repeated five times. We used the technique in thirty-three mature goats to study the biological and biomechanical outcomes of the devitalized and devascularized anterior cruciate ligament at zero, six, and twenty-six weeks after treatment. Thus, the collagen fibers of the simulated autogenous graft remain in normal anatomical position and the simulated graft is fixed under physiological tension. At twenty-six weeks, no statistically significant differences were noted between treated and contralateral control (untreated) ligaments relative to anterior-posterior translation, maximum force to rupture, stiffness in the linear region of the force-length curve, modulus of elasticity in the linear region, strain to maximum stress, or maximum stress. The only statistically significant difference was an increase in cross-sectional area of the ligament. This increase was 22 and 42 per cent greater than that in the control ligaments at six weeks and six months. At six months, the ligaments in the control group had an average mid-cross-sectional area of 17.7 +/- 1.2 square millimeters and the ligaments in the experimental group, 25.2 +/- 3.1 square millimeters. Changes in the size and density of the collagen fibrils also were demonstrated at six months. These observations are in sharp contrast to our previous studies of replacement of the anterior cruciate ligament, in which an allograft of the ligament or an allograft supplemented with a 3M ligament augmentation device (LAD; 3M, St. Paul, Minnesota) was used. In those studies, an average reduction in maximum strength of 75 per cent for the allografts and 50 per cent for the allografts that had a ligament-augmentation device was found at one year. We concluded that devitalized, devascularized anterior cruciate ligaments do not lose strength if the anatomical position and the orientation of the collagen fibers are not altered.
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