Construction of artificial nerve Schwann cells in rat long nerve defect in the repair of Research_6331

By Dana Perry,2014-10-30 16:06
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Construction of artificial nerve Schwann cells in rat long nerve defect in the repair of Research_6331

    Construction of artificial nerve Schwann cells in rat long nerve defect in the repair of Research

     Abstract The Schwann cells and PLGA composed of tissue-engineered artificial

    nerve repair of peripheral nerve defects. [Methods] Schwann cells were seeded in the

    built-polyglactin 910 fibers in PLGA hollow tubes, built into the neural tissue engineering. To 60 SD rats were randomly divided into 3 groups, each with 20, 20 mm rat sciatic nerve defects in the animal model. A group with cut autologous nerve

    segment sutured, B group of Schwann cells in tissue engineering using artificial neural repair, C group were non-vaccinated Schwann cells in PLGA hollow tube to be

    repaired. 8,12 weeks after the use of neural electrophysiological and histological

    evaluation of separate effects. [Results] In general observation, histological observation, B Group A group of similar performance and recovery. In the nerve

    electrophysiological testing, the weight of calf triceps muscle measured cross-section of

    the bridge material measured the number of nerve fibers in the results of the analysis of group B, but slightly lower than the A group was significantly higher than C group. B rat distal segment bridge after horseradish peroxidase tracer can be seen in the spinal cord anterior horn neurons were labeled. Transmission electron microscopy showed the middle of a large number of objects B group bridging the regeneration of nerve fibers. [Conclusion] The Schwann cells and PLGA tissue engineering of artificial neural repair of rat peripheral nerve defects, you can fix 20 mm long segment of peripheral nerve defects, and get close to the autologous nerve repair in rats results.

     Key words artificial neural nerve repair long segment defect

     Abstract: [Objective] To study the effect of repairing of rat peripheral nerve defect with engineering artificial nerve composed with Schwann cells and poly lactic glycoli aced (PLGA). [Method] Human Schwann cells were vaccinated into PLGA tube with polyglactin 910. Sixty SD rats were divided into 3 groups randomly with 20 rats in each one: group A, self-nerve graft; group B, Schwann cells artificial nerve; group C, pure PLGA. Evaluations of electrophysiological and histological examines were carried out 8 weeks ad 12 weeks after operation. [Result] The outcome of group B showed similar to group A in general and histological study, however it expressed slightly lower in electrophysiological study, muscle weight and nerve fiber quantity. Marked neuron could be found in spinal cord after group B had been traced. Large number of

    regeneration nerve fibers could be seen from electron microscope. [Conclusion] The tissue engineering artificial nerve composed with Schwann cells is able to repair 20 mm defects of peripheral nerve, and it has an approximate treatment effect to the self -

    nerve grafting.

     Key words: artificial nerve; long defect; nerve regeneration

     Artificial neural scaffold material and seeds from the cells, extracellular matrix, as well as the induction and growth-promoting factors such as the organic unity of several

    components, the repair of peripheral nerve injury has important research significance. Schwann cells (Schwann cell) is one of the most important seed cells. The training and purified Schwann cells in the stent with a kind of similar to Bungner orderly

    distribution, and the secretion of a variety of neurotrophic factors to support and guide axonal regeneration is to improve the effectiveness of the key to repairing nerve defects [1,2]. The current domestic and international multi-purpose newborn rats, rabbits and

    other animals, peripheral nerve Schwann cells in primary culture to build artificial neural. In this study, the polyglactin 910 with built-in poly-glycolic acid - poly lactic

    acid (PLGA) scaffolds prepared by catheter, through the inoculation of human

    embryonic Schwann cells in vitro induction culture, build artificial nerves, and bridge in the rat sciatic nerve defect , and autologous nerve graft as control, to study and assess its impact on rat peripheral nerve repair results.

     1 Materials and methods

     1.1 Main reagents, materials and equipment

     polyglactin 910 (polyglycolic acid and polylactic acid copolymer fibers, the United States Johnson & Johnson); PLGA catheter (poly-lactic-glycolic acid copolymer, CAS);

    anti-S-100 monoclonal antibody (Sigma Corporation); 20% fetal bovine serum DMEM (Gibaco companies); rat tail collagen (homemade); 0.1 g / mL of ketamine hydrochloride injection; 20 ~ 40 mg / mL horseradish peroxidase solution; DAB-H2O2

    role of the liquid; 3,3 '- 2 methyl-benzidine (DAB) 5 mg, 0.05 mol / L Tris-HCL buffer

    solution (pH = 7.6) 10 mL, H2O2 (30%) 1μl.

     Induction 3 ~ 4 months babies stillborn 6; 3-month-old SD rats were 32, weighing

    200 ~ 250 g; SXP-1B surgical microscope; Leica freezing microtome; Leica DMIRB

    image analyzer.

     1.2 Preparation of tissue-engineered artificial nerve

     1.2.1 obtain fetal peripheral nerve, carefully peel epineurium, cut 0.5 ~ 1 mm3 pieces, placed in 3.5 cm petri dishes, the use of repeated explant method and

    differential adhesion method to obtain high purity of Schwann cells suspension, cells at about 5 × 104.

     1.2.2 the rat tail collagen-embedded polyglactin 910 fiber (diameter 12 μm, length 2

    cm), placed in petri dishes, each 10 to 12 fibers arranged radial center overlap. France will be using direct explant human embryo Schwann cells were polyglactin 910 fiber. 20% FBS DMEM, 37 ?, 5% CO2 training, spare.

     1.2.3 will be 20 mm length of rat tail collagen pre-coated PLGA catheter inserted

    after Schwann cells. 37 ?, 5% CO2 cultured for 2 weeks, moved to polyglactin 910

    fibers, building nerve conduit for adult workers to continue to foster 2 d, ready to


     1.3 Animal experiment

     To 60 SD rats were randomly divided into 3 groups, each with 20. A group with autologous nerve segment sutured; B group with tissue-engineered artificial nerve

    repair; C group were free of Schwann cells in PLGA hollow tube. Intraperitoneal injection of ketamine 200 mg / kg in anesthetized rats, routine disinfection of shares made after the back of shop towel-hole incision, exposing the sciatic nerve sciatic nerve resection under the micro-scale measurement of 18 mm, resulting from 20 mm of

    natural retraction of the nerve defect. Gastrocnemius from the edge of the distal stump of about 5 mm, with 10 ? of the suture with a needle under a microscope at 10 times the surgery will be on both sides of the bridge material and the outer membrane nerve stump sutured. Closure of the wound, according to the standard laboratory animal feeding for 12 weeks. 8 weeks after surgery in each group 10 rats were randomly taken to observe the first 12 weeks on the remaining 10 rats in each group were observed.

     1.4 Statistical Analysis

     Using SPSS 12.0 software for the experiment to analyze the data in each group, P

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