| Objective: 1. This study is trying to simulate the clinical conditions, establishing the rat animal model of traumatic myositis ossificans and evaluate the reliability, stability and practicability of the model, the basis for later related researchers. 2. Study the effect of acupuncture on traumatic ossification muscular and Smad4, the influence of Smad7, and tries to further explore its mechanism.Methods: Experiment 1: choose only 2 months of age 40, healthy adult male SD rats were randomly divided into four groups, each group of 10. Struck method with a fall knee fracture model(fractures), soft tissue contusion model(soft), the knee joint dislocation model is set up by the side pull reverse method(dislocation), respectively after the fracture group, soft injury group, the dislocation group violence massage, rat limb knee joint after each massage with 3 m medical scotch tape and the self-made small splint, rat limb knee fixed. Achilles tendon rupture model set up by using the cut from the broken Achilles tendon method(Achilles). Observe the four groups of the general situation of rat limb. In postoperative 5, 10, the weekend in the X-ray in rats. Experiment 2: selected 80 male SD rats, with a cut from the broken Achilles tendon model is set up, Achilles tendon rupture in rats were randomly divided into two groups, namely the acupuncture group and the control group, each group of 40. Acupuncture group on postoperative day 4 of rat limb acupuncture intervention, limb foot three mile, gb 34, acupuncture creek, kunlun, clockwise turn anti-clockwise twisting three times, flat or flat. Control group fake needle intervention, and simulation of acupuncture, but not with acupuncture, all normal rats. In building after 3, 5, 7, 9, 10 weekend, take eight rats in each group, lateral ankle slice, pathological changes of radiography after Achilles tendon tissue and HE staining observation, Smad4, Smad7 immunohistochemical detection, dynamic change of statistical description. Results: Experiment one:(1) the general observations: 5 weekend, fracture group of rats with knee fracture malunion, soft tissue hyperemia swelling, stiff joints, limited activity, local without and obvious. Soft wound group of rats with knee has no obvious abnormalities, local soft tissue swelling, muscle tension was higher than that of without and obvious. Dislocation group of rats with knee abnormalities, soft tissue swelling, limited activity, ammonites and obvious lumps. Achilles tendon group rats with lateral ankle plantar flexion deformity, swelling, and than plantar flexion back stretch limited functioning, ammonites and Achilles tendon to harden. Weekend, 10 fracture groups of rats with knee fracture malunion, soft tissue swelling, no loss, joint stiffness degree aggravating, 20% local without and obvious lumps in rats. Soft wound group of rats with knee has no obvious abnormalities, local soft tissue swelling, muscle tension increased from the previous, ammonites and obvious lumps. Dislocation group of rats with knee abnormalities, soft tissue swelling is aggravating, limited activity, 10% of the children and obvious. Achilles tendon group rats with lateral ankle plantar flexion deformity, hyperplasia of swelling is aggravating, plantar flexion back stretch limited functioning due to severe, ammonites and Achilles tendon coarsen harden earlier.(2) the weekend 5, fracture, soft injury group, there were no traumatic dislocation of myositis ossificans, Achilles tendon group 60% of traumatic myositis ossificans. Group 20% 10 weekend fractures knee traumatic myositis ossificans, soft wound group of traumatic myositis ossificans does not appear, traumatic myositis ossificans dislocation group 10%. Achilles tendon group rats 100% in traumatic myositis ossificans. Experiment 2:(1) general observation: intraoperative see rat Achilles tendon white, about 2-4 mm wide, 1 mm thick. Postoperative 3 weekend, Achilles tendon end slightly enlargement, disordered arrangement of fibers. 5 after the weekend, enlargement of the Achilles tendon end, disordered arrangement of fibers, the hardening of touch. 7 after the weekend, Achilles tendon end enlargement, disordered arrangement of fibers, the color yellow, the hardening of touch. 9 after weekend, Achilles tendon end obvious enlargement, disordered arrangement of fiber, color is yellow, the hardening of touch. After weekend, 10 Achilles tendon end enlargement, obviously the color yellow, Achilles tendon central part of calcification.(2) acupuncture group and control group of rats after 5 time points total there was a trend toward increased with the extension of time and inducing rate, 5, 7, 9 at the weekend, the same point in time, acupuncture group, the incidence of heterotopic ossification is higher than the control group. Weekend acupuncture group and control group 10 rats in traumatic myositis ossificans.(3) the histological results showed that 3 weeks acupuncture group and the control group were inflammatory cells infiltration, did not see flake homogeneous light blue, 5, 7, 9 at the weekend, at the same time point acupuncture group and ectopic bone, cartilage cells, osteoblasts, mature trabecular bone) the occurrence rate and maturity were higher than the control group, 10 weekend, acupuncture group and the control group were 100% in mature bone tissue, the control check see cartilage, into chondrocytes and cartilage in lacunae.(4) immunohistochemical results: postoperative 3, 5, 7, 9 at the weekend, positive rate of Smad4 acupuncture group were higher than that of control group, both groups in the weekend 5 peak, acupuncture group 7 Smad4 maintain peak over the weekend, but with a pale yellow dyeing ratio increases, the control group 5 Smad4 positive rate began to decline after the weekend, weekend 10 picks up, the positive rate higher than acupuncture group. Acupuncture group of Smad7 weekend is 3 5 lower over the weekend, after rising, 9 a peak over the weekend, after falling, 5 Smad7 reached the lowest at the weekend. Control group 5, and 7 is gradually reduce 3 weekend over the weekend, weekend reaches the lowest 7, after rising. 3, 5, 7, 9 weekend Smad7 acupuncture group were higher than the control group. Conclusion: Experiment one:(1) compared with violent massage method induced heterotopic ossification, Achilles tendon from fault model in the aspects of imaging results more stable and stronger system controlling and repeatability.(2) and BMP, decalcified bone matrix induced ectopic into fragments, Achilles tendon from fault model of heterotopic bone formation mechanisms and clinical secondary to trauma mechanism of heterotopic ossification similarity is higher, and the operation is easy to operation, conveniently. Experiment two:(1) the use of acupuncture method in our study accelerated the process of rat heterotopic ossification Achilles tendon to promote traumatic myositis ossificans rat Achilles tendon.(2) the mechanism may be improved Smad4 protein concentration.(3) acupuncture can improve the inhibitory protein Smad7 protein concentration, but have not on rats in traumatic myositis ossificans have prevention and cure function. |
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