Objective To observe the revascularization process of chemically extracted acellular allogeneous nerve graft in repairing rat sciatic nerve defect. Methods Eighty adult male SD rats were selected. The sciatic nerve trunks from ischial tuberosity to the ramus of tibiofibular nerve of 16 SD rats were obtained and were prepared into acellular nerve stents by chemical reagent. Sixty-four SD rats were used to prepare the models of sciatic nerve defect (1.0 cm) and thereafter were randomized into two groups (n=32): experimental group in which acellular allogeneous nerve grafts were adopted and control group in which orthotopic transplantation of autologous nerve grafts were adopted. Postoperatively, the general conditions of all rats were observed, and the gross and ALP staining observation were conducted at 5, 7, 10, 14, 21, 28 days and 2, 3 months, respectively. Results All the incisions were healed by first intention. Trail ing status and toe’s dysfunction in extension happened to the right hindl imb of rats in two groups and were improved 6 weeks after operation. General observation showed that the grafts of two groups connected well to the nerves, with appearances similar to that of normal nerve. ALP staining demonstrated that the experimental group had no ingrowth of microvessel but the control group had ingrowth of microvessel 5 days after operation; the experimental group had ingrowth of microvessel but both groups had no microvessel 7 days after operation; few longitudinal microvessel throughout the grafts were observed in both groups 10, 14 and 21 days after operation; no obvious difference in capillary network of grafts was observed between two groups 28 days after operation; and the microvascular architecture of grafts in both groups were similar to that of normal nerve 2 and 3 months after operation. Conclusion When the chemically extracted allogeneous nerve graft is adopted to repair the peripheral nerve defect, new blood microvessels can grow into grafts timely and effectively.
Objective To know the possibility of nerveregeneration after artery sleeve anastomosis and end-to-side suture Methods Seventy-five SD rats were divided into 5 groups. First, the distal end ofsevered peroneal nerve was sutured end-to -side with artery sleeve anastomosis withnormal nerve tibial trunk in groups A, B, C and D. Second, the tibial epineurium at the suture site was not removed in group A; the epineurium at the suturesite was removed(windowing) in group B; the distal end of pre-injured peroneal nerve was sutured after 14 days and windowing was done in group C; and the neural growth factor was injected into artery sleeve and windowing was done in group D. While the distal end of severed peroneal nerve was sutured end to side directly with normal nerve tibial trunk and windowing was done in group E. The histological observation was made and the number of nerve fibers was recorded after 4, 8 and 12 weeks of operation.Results After 4 weeks, there existed the regeneration of axons and myeline sheaths in groups C, D, E, and no nerve fiber regeneration was seen in group A. After 8 weeks, the regenerating nerve fibers were significantly more in groups C, D and E than in group B and ingroup E than groups C and D(Plt;0.05). After 12 weeks, the regenerating nervefibers were significantly more in groups C，D and E than in group B(Plt;0.05).Conclusion End-to-side coaptation with artery sleeve anastomosis is a new valuable method in repair of peripheral nerve injuries.
Objective To study the mRNA expressions of protein kinase C(PKC) and nerve growth factor (NGF) in rat sciatic nerve and the number ofaxons after phorbol-12-myristate-13-acetate (PMA) was injected into silicone chamber. Methods Forty-two SD adult rats were divided into six groups depending on the time of injury (1 day, 3 days, 1 week, 2 weeks, 3 weeks and 4 weeks). A 0.5 cm nerve was cut in doublerat sciatic nerves and “T” type silicone chamber was sutured. PMA at the concentration of 1×10-9mol/L was injected discontinuously into the right side of Ttype silicone chamber(PMA group) and saline was injected into the left side(control group). Nucleic acid in situ hybridization histochemistry technique and thecomputer imagine analysis were employed to detect dynamic changes of PKC mRNA and NGF mRNA in rat sciatic nerves. The number of axons was measured. Results The expressions of PKC mRNA and NGF mRNA increased after injury, and the expressions of PKC mRNA and NGF mRNA reached the peak 2 weeks and3 weeks after injury respectively in control group. The expressions of PKC mRNA and NGF mRNA in PMA group were significantly increased than those in control group 2,3 and 4 weeks after injury(Plt;0.01).The number of axons in PMA group significantly increased than that in control group(Plt;0.01). Conclusion PKC involved inthe expression of NGF mRNA and nerve regeneration after injury. During the regenerated course, PMA can promote the expression of NGF mRNA and the number of axons after injury.
OBJECTIVE: To explore the possibility to bridge peripheral nerve defects by xenogeneic acellular nerve basal lamina scaffolds. METHODS: Thirty SD rats were randomly divided into 5 groups; in each group, the left sciatic nerves were bridged respectively by predegenerated or fresh xenogeneic acellular nerve basal lamina scaffolds, autogenous nerve grafting, fresh xenogeneic nerve grafting or without bridging. Two kinds of acellular nerve basal lamina scaffolds, extracted by 3% Triton X-100 and 4% deoxycholate sodium from either fresh rabbit tibial nerves or predegenerated ones for 2 weeks, were transplanted to bridge 15 mm rat sciatic nerve gaps. Six months after the grafting, the recovery of function was evaluated by gait analysis, pinch test, morphological and morphometric analysis. RESULTS: The sciatic nerve function indexes (SFI) were -30.7% +/- 6.8% in rats treated with xenogeneic acellular nerve, -36.2% +/- 9.7% with xenogeneic predegenerated acellular nerve, and -33.9% +/- 11.3% with autograft respectively (P gt; 0.05). The number of regenerative myelinated axons, diameter of myelinated fibers and thickness of myelin sheath in acellular xenograft were satisfactory when compared with that in autograft. Regenerated microfascicles distributed in the center of degenerated and acellular nerve group. The regenerated nerve fibers had normal morphological and structural characters under transmission electron microscope. The number and diameter of myelinated fibers in degenerated accellular nerve group was similar to that of autograft group (P gt; 0.05). Whereas the thickness of myelin sheath in degenerated accellular nerve group was significantly less than that of autograft group (P lt; 0.05). CONCLUSION: The above results indicate that xenogeneic acellular nerve basal lamina scaffolds extracted by chemical procedure can be successfully used to repair nerve defects without any immunosuppressants.
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.
A 0.6cm segment of right common peroneal nerve was resected in 60 SpragueDawley rats. The nerve defects were bridged by adhering the epineurium with autogenous nerve, vein, skeletal muscle, tendon and silastic tube. According to the kinds of the grafts used, the rats were divided into 5 groups. In 6 and 12 weeks after operation, the effect was assessed by motor nerve conduction velocity, weight of the anterior tibial muscle, number of distal axons and histological examination. It was demonstrated that the result from autogenous nerve graft was superior to other grafts in all aspects and that of the vein graft was better thanthe other three. The characteristics of the nerve regeneration and the process of maturation in different types of the grafts were discussed. The related microenvironment which caused the difference was also discussed.
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.
ObjectiveTo summarize the applications of Schwann cells (SCs), stem cells, and genetically modified cells (GMCs) in repair of peripheral nerve defects. MethodsThe literature of original experimental study and clinical research related with SCs, stem cells, and GMCs was reviewed and analyzed. ResultsSCs play a key role in repair of peripheral nerve defects; the stem cells can be induced to differentiate into SCs, which can be implanted into nerve conduits to promote the repair of peripheral nerve defect; genetically modified technology can enhance the function of SCs and different stem cells, which has been regarded as a new option for tissue engineered nerve. ConclusionAlthough great progress has been made in tissue engineered nerve recently, mostly limited to the experimental stage. The research of seed cells in application of tissue engineered nerve need be studied deeply.
ObjectiveTo summarize the research progress of autologous vein nerve conduit for the repair of peripheral nerve defect. MethodsThe recent domestic and foreign literature concerning autologous vein nerve conduit for repair of peripheral nerve defect was analyzed and summarized. ResultsA large number of basic researches and clinical applications show that the effect of autologous venous nerve conduit is close to that of autologous nerve transplantation in repairing short nerve defect, especially the compound nerve conduit has a variety of autologous nerve tissue, cells, and growth factors, etc. ConclusionAutologous vein nerve conduit for repair of non-nerve defect can be a good supplement of autologous nerve graft, improvement of autologous venous catheter to repair peripheral nerve defect is the research direction in the future.
ObjectiveTo investigate the effect of electrospun chitosan/polylactic acid (ch/PLA) nerve conduit for repairing peripheral nerve defect in rats. MethodsNerve conducts loaded with ch/PLA was made by the way of electrospun. The mechanical property, hydrophility, biocompatibility were tested, and the scanning electron microscope was used to observe the ultrastructure. The same experiments were also performed on pure PLA nerve conducts as a comparison. Then, 54 Sprague Dawley rats were divided into 3 groups randomly, 18 rats in each group. Firstly, the 10 mm defects in the right sciatic nerves were made in the rats and were respectively repaired with ch/PLA (group A), autografts (group B), and no implant (group C). At 4, 8, and 12 weeks after operation, general observations, sciatic functional index (SFI), electrophysiological evaluation, wet weight of gastrocnemius and soleus muscles, histological examination, immunohistological analysis, and transmission electron microscopy were performed to evaluate the effects. ResultsCompared with pure PLA nerve conducts, the addition of chitosan could improve the mechanical property, hydrophility, biocompatibility, and ultrastructure of the nerve conducts. At 4 weeks postoperatively, the regenerated nerve bridged the nerve defect in group A. The SFI improved gradually in both group A and group B, showing no significant difference (P>0.05). Compound muscle action potentials and nerve conduction velocity could be detected in both group A and group B at 8 and 12 weeks after operation, and significant improvements were shown in both groups (P<0.05). The wet weight and myocyte cross section of gastrocnemius and soleus muscles showed no significant difference between group A and group B (P>0.05), but there was significant difference when compared with group C (P<0.05) at 12 weeks postoperatively. Immunohistological analysis revealed that S-100 positive Schwann cells migrated in both group A and group B, and axon also regenerated by immunohistological staining for growth associated protein 43 and neurofilaments 160. Transmission electron microscopy showed no significant difference in the diameter of nerve fiber between group A and group B (P>0.05), but the thickness of myelin sheath in group A was significantly larger than that in group B (P<0.05). ConclusionThe electrospun ch/PLA nerve conduits can effectively promote the peripheral nerve regeneration, and may promise an alternative to nerve autograft for repairing peripheral nerve defect.