Objective To explore the effect of tri pterygium glycoside (TG) on the skeletal muscle atrophy and apoptosis after nerve allograft. Methods Twenty Wistar male rats were adopted as donors, weighing 200-250 g, and the sciatic nerves were harvested. Fifty SD male rats were adopted as recipients, weighing 200-250 g. Fifty SD rats were made the models of10 mm right sciatic nerve defect randomly divided into five groups (n=10): group A, group B, group C, group D and group E.groups A and B received fresh nerve allograft, groups C and D received sciatic nerve allograft pretreated with TG, and group E received autograft. The SD rats were given medicine for 5 weeks from the second day after the transplantation: groups A and E were given physiological sal ine, groups B and D TG 5 mg/ (kg·d), and group C TG 2.5 mg/ (kg·d). At 3 and 6 weeks, respectively, after nerve transplantation, general observation was performed; the structure of skeletal muscles was observed by HE staining; the diameter of skeletal muscles was analyzed with Image-Pro Plus v5.2; the ultrastructure of skeletal muscles was observed by TEM; the expressions of Bax and Bcl-2 were detected by immunohistochemical staining; and the apoptosis of skeletal muscles was detected by TUNEL. Results All rats survived to the end of the experiment. In general observation, the skeletal muscles of SD rates atrophied to different degrees 3 weeks after operation. The muscular atrophy in group A was more serious at 6 weeks, and that in the other groups improved. The wet weight, fiber diameter and expression of Bcl-2 in group A were significantly lower than those in groups B, C, D and E (P lt; 0.01)；those in groups B, C and D were lower than those in group E (P lt; 0.05); and there were no significant differences among groups B, C and D (P gt; 0.05). The apoptosis index and expression of Bax in group A were significantly higher than those in groups B, C, D and E (P lt; 0.01)；those in groups B, C and D were higher than in groupE (Plt; 0.05); and there were no significant differences among groups B, C and D (P gt; 0.05). Three weeks after nerve allograft, under the l ight microscope, the muscle fibers became thin; under the TEM, the sarcoplasmic reticulum was expanded. Six weeks after nerve allograft, under the l ight microscope, the gap of the muscle fibers in group A was found to broaden and connective tissue hyperplasia occurred obviously; under the TEM, sarcomere damage, serious silk dissolution and fragmentary Z l ines were seen in group A, but the myofibrils were arranged tidily in the other groups, and the l ight band, dark band and sarcomere were clear. Conclusion TG can decrease the skeletal muscle atrophy and apoptosis after nerve allograft. The donor’s nerve that is pretreated with TG can reduce the dosage of immunosuppressant for the recipient after allograft.
OBJECTIVE: To evaluate the results of limb function and the methods of bone and soft tissue reconstruction of patients treated with allografting. METHODS: From May 1992 to January 1999, 90 patients suffered from bone malignant tumor were treated with allografting in different methods of internal fixations. The average follow-up was 37.5 months. The limb postoperative function, complications related to different surgical methods were compared according to Enneking evaluation system. RESULTS: Skin necrosis, infection, non-union, fracture of allograft were the main complications which affect patients’ limb postoperative functions. Of the 90 fresh-frozen allografting procedures, the final results of operation showed that hip joints and knee joints were better than the shoulder joints. More than 80% of the patients treated with interlocked intramedullary nail and allograft-prosthesis combination led to an over-all result that was excellent and good. Interlocked intermedullary nail was of recommended method of internal fixation. Early exercises of operative limbs could promote function recovery. CONCLUSION: Using of interlocked intramedullary nail and allograft-prosthesis combination are of recommended operation method and can be applied with better results, and early exercises of operative limbs will lead to better functions.
Objective To investigate the rat model of cardiac allograft vasculopathy after heart transplantation in rat abdominal cavity. Methods Forty Wistar rats and 40SDrats were divided into control group and experiment group randomly pair-matching. Rat model ofheterotopic heart transplantation was developed. Low doseCyclosporine A were injected into the abdominal cavity in experiment group, while the control group had not received the Cyclosporine A. Transplant hearts were harvested at two weeks and four weeks post-operatively and changes of coronary artery were observed by light microscope. Results There were no alteration of tunica intima of coronary artery in control group at two weeks and four weeks post-transplantation. Tunica intima of coronary artery increased in thickness at two weeks post-transplantation in experiment group and concentric circular change occurred at four weeks post-transplantation. Lumen of coronary artery constricted transparent and cardiac allograft vasculopathy occurred. Conclusion This animal model is reliable of cardiac allograft vasculopathy.
Objective To explore the biomechanical difference between the different fixations of cortical bone plate allograft. Methods Twenty-seven cadaveric femurs were harvested and were made into the simulated fracture models, which were equally divided into Groups A, B and C. In Group A, the models were fixed with 2 bone plate allografts (110 mm×10 mm×3 mm); in Group B, the models were fixed with 2 struts (110 mm×10 mm×3 mm) and 5 bone screws; in Group C, the models were fixed with 1 strut (110 mm×10 mm×3 mm) and 5 bone screws. The biomechanical tests for the three-piont bending, torsion, and compression were performed. The parameters studied included the values of the displacements in the three-piont bending tests and the compression tests, and the maximum loads during the bending, the compression, and the torsion. Results As for all the stiffness parameters tested, Group A showed the greatest displacements among the threegroups（P＜0.05）, except the compressive stiffness parameter, which was similar to that in Group B. The maximum loads of the three-point bending, the torsion, and the compression in Group A were 1.65±0.34 kN, 554.3±49.34 N, and 7.78±0.82 Nm, respectively; in Group B, they were 1.12±0.37 kN, 428.00±37.40 N,and 3.39±0.22 Nm, respectively; in Group C, they were 0.71±0.46 kN, 218.67±36.53N, and 1.74±0.12 Nm, respectively. Group A had a significantly greater strengththan the other 2 groups(P＜0.05). Conclusion The strength of the cortical bone plate allograft is related to its different fixations. The two cortical bone plate allografts have a greater strength and stiffness than the struts fixed with the bone screws, which can meet the clinical requirement.
OBJECTIVE: To study the effect of basic fibroblast growth factor (bFGF) and hyaluronic acid gel (HAG) combined with freeze-dried bone allograft in repairing segmental bone defect and to explore their mechanism. METHODS: The 15 mm segmental bone/periosteum defects were created on bilateral radius in 50 New Zealand rabbits and were treated with four different kinds of implants on 25 radius respectively (group A: bFGF and HAG combined with freeze-dried bone; group B: bFGF combined with freeze-dried bone; group C: HAG combined with freeze-dried bone; group D: simple freeze-dried bone as a control). The repair of defect was observed radiologically and histologically and were analyzed by radionuclide bone imaging and measurement of calcium contents at different periods. RESULTS: The new bone formation, bone metabolic activity and calcium contents of defects were higher in group A than in group B (P lt; 0.05), and were higher in group B than in groups C and D (P lt; 0.05). There were no significant difference between groups C and D. The bone defects healed in the 8th week in group A, in the 10th week in group B, but did not healed in the 10th week in groups C and D. CONCLUSION: As an osteogenetic factor, bFGF promotes the new bone formation; as a slow-release carrier, HAG enhances the effectiveness of bFGF. The combination of bFGF, HAG and freeze-dried bone allograft can repair the segmental bone defect more effectively.
Objective To investigate the promotion effect of neurotropic reinnervation with chemically extracted acellular nerve allograft. Methods The sciatic nerves of 5 healthy adult SD rats, regardless of gender and weighing 270-300 g, were collected to prepare chemically extracted acellular nerve allograft. Eighteen healthy adult Wistar rats, regardless of genderand weighing 300-320 g, were made the model of left sciatic nerve defect (10 mm) and randomly divided into 2 groups: autograft (control group, n=9) and allograft group (experimental group, n=9). The defects were bridged by acellular nerve allograft in experimental group and by autograft by turning over the proximal and distal ends of the nerve in control group. At 3 months after transplantation, dorsal root ganglion (DRG) resection operation was performed in 6 rats of 2 groups. At 3 weeks after operation, the sural nerves were harvested to calculate the misdirection rate of nerve fibers with pathological staining and compute-assisted image analysis. Cholinesterase staining and carbonic anhydrase staining were performed in the sural nerve of 3 rats that did not undergo DRG resection at 3 months. Results The results of chol inesterase staining and carbonic anhydrase staining showed that experimental group had less brown granules and more black granules than control group. After DRG resection, count of myelinated nerve fiber were 4 257 ± 285 in the experimental group and 4 494 ± 310 in the control group significant (P lt; 0.05). The misdirection rate was 2.27% ± 0.28% and 7.65% ± 0.68% in the experimental group and in the control group respectively, showing significant difference (P lt; 0.05). Conclusion Chemically extracted acellular nerve allograft can not only act as a scaffold in the period of nerve defects repair, but markedly enhance the effects of neurotropic reinnervation.
Objective To investigate the research advance in repair of the peripheral nerve defect with an acellular nerve allograft. Methods The recent related literature was extensively and comprehensively reviewed. The methods and the effects of the allografts with acellular nerves were analyzed. Results The immunogenicity of the allograft was more significantly relieved by the chemical treatment than by the physicaltreatment. The effect of the chemical treatment on the axon regeneration was better than that of the physical treatment. Conclusion Because of the limitation of the host Schwann cell translation in the longsegment acellular nerve allografts, the effect of Schwann cells is not satisfactory and regeneration of the nerve is limited. So, the recellularized treatment with some related measures can enhance the host Schwann cell translation so that this problem can be solved.
Objective To study the effect of vascular endothelial cell growth factor (VEGF) on repair of bone defect with cortical bone allograft. Methods Forty five New Zealand white rabbits, weighted 2.5-3.0 kg, were made bone defect model of 1.5 cm in length in the bilateral radii and then were randomly divided into 3groups. The defect was repaired with only cortical bone allograft in the control group, with the cortical bone allograft and local injection of human recombinantVEGF in the experimental group, and with the cortical bone allograft and abdominal injection of VEGF PAb3 in the antagonist group. Roentgenography, immunohistochemical staining and tetracycline labelling were carried out to evaluate the reparative results 1, 3, 5, 8 and 16 weeks after operation. Results Immunohistochemical staining results showed that a great deal of blood vessels formed in the experimental group, and the number of blood vessels increased gradually with the time and reached the highest value at the 8th week. Tetracyclinelabelling showed the same result.The best results in callus formation, ossification rate and count of microvascular density were shown in the experimental group, while those in the control group were significantly better than those in the antagonist group (Plt;0.05),but there was no significant difference between the experimental group and the control group at the 8th week and the 16th week (Pgt;0.05). Conclusion VEGF can accelerates the bone formation and angiogenesis in the bone allografts, thus it can promote the repair of bone defects.
In order to reduce the immunogenicity of parathyroid allografts and induce immunotolerance, we depleted Ⅰa＋ donor cells of rat parathyroid allografts by anti-Ⅰa monoclonal antibody plus complement, transplanted the treated glands underneath the capsule of the recipient kindey,and observed the median survival time (MST) of the allografts. Our results showed that the MST of the treated group were 60 days, compared with control group (MST:14 days), Ｐ＜0.01. This results indicate that rat parathyroid allografts survival can be prolonged dramatically by depletion of Ⅰa＋ donor cells.
Objective To study an effect of the peripheral nerve allograft with subcutaneous preservation at different times on the sciatic nerve regenerationin rats. Methods Fifty-five Wistar rats were used in this experiment, which were randomly divided into the following 5 groups: the experimental groups (Groups A, B, C, 10 rats), the control group (Group D, 10 rats), and the donorgroup (Group E, 15 rats). In the experimental groups, a 15-mm segment of the sciatic nerve harvested from the donors was separately inserted into the subcutaneous compartment on the left thigh after the 1week (Group A), 2-week (Group B), and 3week (Group C) preservation; the segment of the sciatic nerve in the subcutaneous compartment was removed and transplanted into a 10-mm defect of theright sciatic nerve, which was made immediately. In Group D, a 10-mm sciatic nerve defect was made and immediately repaired in situ on the right thigh. The function of the sciatic nerve was evaluated by the sciatic functional index (SFI) at 2, 4, 6, 8, 10 and 12 weeks after operation. The histological and electrophysiological examinations were performed at 12 weeks after operation. Results After operation, SFI decreased gradually at 12 weeks afteroperation, SFI inGroups A and D was at the minimal level and had a significant difference compared with that in Groups B and C (Plt;0.05).There was no significant difference between Group A and Group D. A large number of the myelinated nerve fibers and a small number of the unmyelinated nerve fibers were regenerated in Groups A and D. The number and the structure of the regenerated nerve were similar to the normal ones. The number and the size of the regenerated axon had a significant difference compared with those in Groups B and C (Plt;0.05). There was no significant difference between Group A and Group D. The conduction velocity and the latent period of the motor nerve had significant differences between Groups A and D and Groups B and C (Plt;0.05), and there was no significant difference betweenGroupA and Group D. Conclusion The nerve allograft with a 1-weeksubcutaneous preservation can promote nerve regeneration better.