OBJECTIVE To explore the effect of basic fibroblast growth factor (bFGF) combined with autogenous vein graft conduit on peripheral nerve regeneration. METHODS Fifty four New Zealand rabbits were divided into three groups. The main trunk of sciatic nerve of rabbit in one side was severed and bridged by autogenous vein. 0.2 ml bFGF solution (4,000 U/ml) was intravenously injected to the vein graft conduit as group A, the same amount of saline solution as group B, and no solution injection as group C. Microscopic examination, axon video analysis and nerve conduct velocity were performed at the 10th, 30th, and 100th day after operation. RESULTS The nerve fibers were grown into vein graft conduit in all groups at 30th after operation, they were more and regular in group A than that of group B and C, and the axon regeneration rate in group A was more than that of group B and C. CONCLUSION bFGF combined with autogenous vein graft conduit can markedly promote nerve regeneration.
Basing on the experimental results, 48 nerve defects (with the length of 3-4 cm in 21 cases, 4.1-5cm in 25 cases and 6cm in 2 cases) were repaired clinically by using vaseularized nerve sheath canal with living Schwann s cells, 87.5 percent of them obtained good results. The advantages were: (1) The neural sheath had rich blood supply with resultant less scar from its healing; (2) The living Schwann s cells would secrete somatomedin to promote the reproduction of neural tissues; and (3) The useless neurofib...
Motor endplate is the structure connected between the nerve terminal and muscle fibre and plays a very important role in conducting nerve impulses to the target, therefore, systemical study of the sequential changes of the motor end-plate upon denervation is quite important.Ninety New Zealand rabbits were divided into nine groups from two weeks to nine months after denervation. Acetycholinesterase(AchE) was analyzed quantitatively to study the sequential changes of the motorendplates of tibialis anterior muscle. The results showed that there was no significant reduction of AchE at theend if one month after denervation, whereas a sharp reduction of AchE afterwards. AchE could not be stained after five month denervation.
OBJECTIVE: To investigate the characteristics and the pathologic classification of electrical-injury nerve using somatosensory evoked potential(SEP) technique. METHODS: SEP were detected and evaluated in 12 cases with electrical-injury nerve during operation, electrical stimulation was commenced from distal side of nerve where the structure of nerve looks normal under operating microscope, up to proximal side until evoking out a stable SEP predeterminate virtual value. Pathological examination and the following functional evaluation were compared with the values of SEP. RESULTS: At the site of nerve looking normal under operating microscope, perineurium appears normal or slightly thicken. But there are obvious fibrosis and fibrotic proliferation between fascicular and intrafascicular. Vessel plexus is not seen. At SEP stabilizely evoked site, nervous construction is normal, there are visible interfascicular vessel plexus and connective tissue appears loose. Comparing SEP values with pathological section, amplitude and latency of SEP is positively correlative with the quality of nerve. Eight cases repaired with SEP technique to select the anastomosis site for nerve transplantation were followed up, two-point discrimination reached grade III (America hand surgery association criterion) within 62.5% cases. CONCLUSION: SEP technique is valuable method for functional evaluation of electrical- injury nerve which has a complicated pathology. The pathology of electrical-injury nerve can be classified into 4 types, type A: fibrosis of nerve; type B: nerve looking normal under operation microscope, perineurium appears thicken, and there are obvious fibrosis and fibrotic proliferation between fascicular and intrafascicular, vessel plexus is rarely to see; type C: nerve looks normal, lymphocyte infiltration exists and it is obvious that there are many physalis-like, retrogressive construction in the section; type D: nervous construction is normal, there are visible interfascicular vessel plexus, and connective tissue appears loose, SEP always can be stably evoked.
ObjectiveTo review the research progress of peripheral nerve mismatch regeneration, and to provide reference for its related basic research and clinical treatment.MethodsThe pathophysiology of peripheral nerve after injury, several main factors affecting the mismatch regeneration of peripheral nerve, and the fate of axon after mismatch regeneration were summarized by referring to the relevant literature at home and abroad in recent years.ResultsDistal pathways and target organs can selectively affect the mismatch regeneration of peripheral nerves; different phenotypes of Schwann cells have different effects on the mismatch regeneration of peripheral nerves; studying the mechanism of action of exosomes from different Schwann cells on different types of axons can provide a new direction for solving the mismatch regeneration of peripheral nerves.ConclusionPeripheral nerve mismatch regeneration is affected by various factors. However, the specific mechanism and characteristics of these factors remain to be further studied.
OBJECTIVE: To explore the mechanism of tissue specificity of neurotropism in peripheral nerve regeneration, we investigated the biological characteristics of the nerve regeneration conditioned fluids(NRCF) on motoneuron of SD rats cultured in vitro. METHODS: Silicon chambers were sutured respectively to the distal stumps of motorial branch of femoral nerve and saphenous nerve to collect NRCF, namely MD-NRCF and SD-NRCF. The rats cortex motoneuron were divided into 4 groups and cocultured with MD-NRCF, SD-NRCF, b-FGF and serum-free medium respectively. The cultured cells were photoed under phase-contrast microscope, their longest neurites and cell-body areas were measured by cell image processing computer system. MTT automated colorimetric microassay was also adopted to quantify the activation of cultured motoneurons in each group. RESULTS: Cells of MD-NRCF group had longer neurites than those of the other three groups, and their activation was also superior to those of the other groups. CONCLUSION: The results suggest that MD-NRCF has more significantly neurite-promoting and neurobiological effects on motoneuron than SD-NRCF and b-FGF.
ObjectiveTo study the effect of the loaded concentration gradient of nerve growth factor (NGF) immobilized conduit on rat peripheral nerve defect repair. MethodsThe peripheral nerve conduits made of poly (ε-caprolactone)-block-poly (L-lactide-co-ε-caprolactone) were prepared with uniform loads or concentration gradient loads by combining differential absorption of NGF/silk fibroin (SF) coating, and the gradient of NGF was immobilized in the nerve conduits. ELISA method was used to exam the NGF release for 12 weeks in vitro. Twenty-four male Sprague Dawley rats (weighing, 220-250 g) were selected to establish the right sciatic nerve defect model (14 mm in length) and randomly divided into 4 groups according to repair methods. The transected nerve was bridged by a blank conduit without NGF in group A, by a conduit containing uniform loads of NGF in group B, by a conduit concentration gradient loads of NGF in group C, and by the autogenous nerve segment in group D. The gross observation, electrophysiological examination, histological observation, and transmission electron microscope observation were carried out to assess the nerve regeneration at 12 weeks after surgery. ResultsThe cumulative release amount of NGF was (14.2±1.4) ng/mg and (13.7±1.3) ng/mg in gradient of NGF loaded conduits and uniform NGF loaded conduits respectively at 12 weeks, showing no significant difference (t=0.564, P=0.570). All the animals survived to completion of the experiment; plantar ulcers occurred at 4 days, which healed at 12 weeks; groups C and D were better than groups A and B in ulcerative healing. At 12 weeks after surgery, the compound muscle action potential of group A was significantly lower than that of groups B, C, and D (P<0.05), and group B was significantly lower than groups C and D (P<0.05), but no significant difference was found between groups C and D (P>0.05). The axon density of group C was significantly higher that of groups A, B, and D (P<0.05); group D was significantly higher than groups A, B, and C, and group C was significantly higher than groups A and B in the axon number, axon diameter, and area of muscle fiber (P<0.05); the thickness of myelin sheath of groups C and D was significantly larger than that of groups A and B (P<0.05), but no significant difference was found between groups C and D (P>0.05). ConclusionGradient of NGF loaded nerve condnits for rat sciatic nerve defect has similar results to autogenous nerve, with a good bridge, which can promote the sciatic nerve regeneration, improve the myelinization of the regenerating nerve, and accelerate the function reconstruction of the regenerating nerve.
To find new technique for repair of peripheral nerve defect, the nerve elongation repair technique was adopted. Two cases with nerve defect were treated by this method. One was a 12 year old male, the defect length of right radial nerve was 7.2 cm at the elbow. The other one was a 28 year old male, the defect length of left ulnar nerve the was 5 cm at elbow. In this method, the nerve was elongated by slow stretch from distal and proximal end of the ruptured nerve. After a few days, the nerve was repaired by direct suture. After operation, the function of nerves were recovered in 119 days and 114 days respectively. Follow-up for 5 years, the function of the effected limbs were recovered to the normal side. It was concluded that: (1) the peripheral never can be elongated by slow stretch; (2) to stretch the nerve end in a rubber tube can prevent adhesion and connective tissue blocking; (3) strength and supporting point of stretching should be designed carefully.
OBJECTIVE To investigate the effects of targeted muscular injection of ciliary neurotrophic factor (CNTF) on the regeneration of injured peripheral nerves. METHODS The left sciatic nerves of 80 Sprague-Dawley rats were excised to form 6 mm defect and the two ends were bridged by silicone tubes, they were randomly divided into two groups, CNTF group and normal saline (NS) group. The CNTF group was given recombinant human CNTF, 1 mg/kg every other day for 30 days, and the NS group was given equal quantity of normal saline as NS group. The sciatic nerve functional index (SFI), electrophysiological assessment, morphometric analysis of axons, and choleratoxin horseradish peroxidase (CB-HRP) retrograde-labelling were measured postoperatively. RESULTS The SFI, electrophysiological parameters (nerve conduction velocity, latency and amplitude of compound muscle action potentials), myelinated axons counts, mean axons diameters and myelin sheath thickness, number of CB-HRP labelled ventral horn motor neurons of spinal cord were significantly higher in CNTF group than that of NS group. CONCLUSION Targeted muscular injection of CNTF can promote the regeneration of peripheral nerve and improve the nerve functional recovery.
Objective To investigate the outcome of repairing the peripheral nerve defects with the tissue engineered nerve constructed by Schwann cells and fibrin glue. Methods Wallerian degenerated sciatic nerve were harvested from the 4-week-old New Zealand rabbits for culture of Schwann cells. The Schwann cells were then separated, amplified and purified, and then were identified by the S-100 protein immunochemical staining. The cultured Schwann cells (1×106/ml) were mixed with fibrin glue to form the Schwann cell-fibrin glue compound, which was observed by the inverted phase contrastmicroscope. The compound filled some silicone tubes (Group A) and biomembrane (Group B) to fabricate the tissue engineered nerves with a purpose of repairing the 10-mm defects in the New Zealand rabbit tibia nerves. The autologous nerve grafting was performed in Group C. The electrophysiological examination and the histomorphological analysis were performed at 10 weeks after the transplantation. Results All the rabbits survived through the experiment. In Group A, all the rabbits developed an ulcer in the soles of their left feet at 3-4weeks after the transplantation, while less ulceration developed in Groups B and C. At 10 weeks after the transplantation, the electrophysiological examination was performed, the elective stimulation failed to pass through the nerve grafts, and no composed muscular action potential was found in all the rabbits in Group A; the elective stimulation could pass through all the nerve grafts in Groups B and C, and could evoke the composed muscular action potential; the composed muscular action potential and the nerve conduct velocity in the two groups were 4.21±0.82 mV and 3.40±5.40 m/s vs. 4.80±1.15 mV and 36.55±6.43 m/s(Pgt;0.05). In Group A, no regrown axon was found in the nerve grafts, but neuromawas found to have formed in the both ends of the silicon tube. In Groups B and C, there was no obvious neuroma formation but regrown axons could be found to have regenerated. The histomorphological analysis on the regrown axons showed thatthere was no statistically significant difference between Groups B and C. Conclusion The tissue engineered nerve fabricated with Schwann cells, fibrin glue, and biomembrane can promote the nerve regeneration, and its reparative effect is similar to that of the autologous nerves; therefore, the future of its clinical practice is brilliant.