Nerve Graft Group Research Articles

Autogenous venous graft with one-stage prepared Schwann cells as a conduit for repair of long segmental nerve defects.

The use of autogenous venous graft with intraluminal injection of Schwann cells to enhance nerve regeneration of long segmental nerve defects was evaluated in a rabbit tibial nerve-repair model. Schwann cells were isolated from the excised rabbit tibial nerve by using the polylysine differential adhesion method. The cultured cells were identified by immunocytochemical labeling for S-100 protein. Tibial nerve defects in 4-cm segments were created in 24 animals, which were then divided into three groups. In Group 1, the tibial nerve defect was repaired with interposition vein graft alone; in Group 2, the nerve defect was repaired with a vein graft with intraluminal injection of Schwann-cell suspension; in Group 3, the nerve defect was repaired by autogenous nerve graft alone. At 2 months postoperatively, electrophysiologic evaluation showed that an evoked muscle action potential was recorded for the animals in Group 2, with vein grafting plus Schwann cells, and for those in Group 3, with autogenous nerve grafting, but not for those in Group 1, where vein grafting alone was used. The average motor nerve conduction velocity in the group with vein grafting and Schwann cells was 3.4 +/- 1.5 m/sec, which was slower than the nerve grafting group (7.8 +/- 1.8 m/sec). Histologic analysis confirmed there was formation of new nerve fascicles with myelination in the vein graft filled with Schwann cells. No nerve regrowth was found in the vein grafts without Schwann cells. These results suggested that isolated Schwann cells are able to survive in a vein graft, and that the vein graft with intraluminal seeded Schwann cells could be an alternative for repairing injured nerves with long gaps.

Brain-Derived Neurotrophic Factor–Enriched Collagen Tubule as a Substitute for Autologous Nerve Grafts

Autologous nerve interposition grafts are frequently harvested by head and neck surgeons. The sacrifice of these donor nerves guarantees some degree of morbidity, including sensory loss, additional incision sites with associated potential complications, and prolonged operative time. An alternative to autologous nerve grafting is, therefore, desirable. To determine if a collagen tubule (CT) filled with either a plain collagen gel or a brain-derived neurotrophic factor (BDNF)-enriched collagen gel could be used to achieve functional and histologic outcomes equivalent to an autologous nerve graft in bridging a 15-mm nerve gap in the rabbit facial nerve. A prospective, randomized, blinded animal study with a control group. Thirty rabbit facial nerves were resected (15-mm segments) to create nerve gaps. The gaps were bridged using 1 of 3 methods, assigned randomly: a reversed facial nerve (control), a collagen gel-filled CT, or a BDNF-enriched collagen gel-filled CT. The animals were evaluated after 6 weeks in a blinded fashion for functional nerve recovery, axon count, and axonal diameter. There were no significant differences between the autologous nerve graft group, the collagen gel-filled CT group, or the BDNF-enriched collagen gel-filled CT group (n = 10 for each group) for functional nerve recovery (P =.94). The mean axon count and the mean axonal diameter were highest in the BDNF-enriched collagen gel-filled CT group, but these differences failed to reach statistical significance (P =.18 and.96, respectively). Collagen tubules filled with BDNF-enriched collagen gel appear to be at least as good as autologous nerve grafts for bridging short facial nerve gaps. Larger experimental studies are warranted to determine if clinical trials are justified.

Peripheral nerve lengthening by controlled isolated distraction: a new animal model

We have developed a simple and effective animal model to study the distraction neurogenesis utilizing the sciatic nerve-lengthening technique in rats. The model allows macroscopic, physiological, and histological evaluation of the distraction site. Fourteen adult Harlan Sprague Dawley rats (300–350 g) were used in this study. A 10 mm segment of the right sciatic nerve of each animal in the nerve-lengthening group was resected. Gradual nerve lengthening was performed by advancing the proximal nerve stump at a rate of 1 mm/day. The proximal stump neuroma was then resected and a direct nerve anastomosis was performed. On the left side a standard autogenous nerve-grafting procedure was performed with a 10 mm segment of sciatic nerve used as an in situ nerve graft. Three months after the second surgery, the sciatic nerves were exposed and investigated by gross observation and EMG followed by histological processing and tissue analysis. Neomicrovascularization was observed surrounding the sciatic nerve anastomosis in all five specimens of the nerve-lengthening group as compared to the more white-colored scar tissue that was observed in the nerve-grafting group. The EMG results were similar for both groups. Histological studies of the lengthened nerves showed axon morphology equivalent to the grafted nerves. This study demonstrated a clear evidence of the successful nerve regeneration within a segmental nerve gap by nerve lengthening.

Morphometric study of regeneration through vascularized nerve graft in a rabbit sciatic nerve model.

Adequate vascularization is essential for a successful nerve graft. Theoretically, immediate vascularization will minimize fibroblast infiltration and support axonal regeneration. In this study, histomorphologic and morphometric studies were carried out to determine whether vascularized grafts are beneficial, in terms of axonal regeneration. In a rabbit model, 4-cm segments of sciatic nerve were obtained and placed as a nonvascularized graft on one side, and as a pedicled vascularized graft fed by the inferior gluteal vessel on the contralateral side. Histomorphologically, the distribution of myelinated nerve fibers and Schwann cells was evaluated after toluidine blue staining, at 2-, 3-, and 4-month intervals. The following results were obtained. 1) Myelinated nerve fibers were more abundant in the proximal, middle, and distal segments of the vascularized nerve group at 2 and 3 months. 2) The average nerve-fiber diameter was greater in the vascularized nerve graft group at 2, 3, and 4 months (2 to 10 microm). 3) Schwann cells were more abundant in the proximal, middle, and distal segments of the vascularized nerve graft group at all time points. Based on the above findings, the immediate restoration of circulation in the vascularized nerve graft can be accountable for the increased number of surviving Schwann cells, the rapid clearing of axons, and myelin-sheath changes that occur during Wallerian degeneration, thus enabling "morphologically" optimal regeneration.

Functional and structural effects of GM-1 ganglioside treatment on peripheral nerve grafting in the rat.

Peripheral nerve regeneration after traumatic injury and standard repair with a nerve autograft is usually incomplete. This study tested the influence of graft vascularity and pharmacological intervention with GM-1 ganglioside on nerve regeneration in a rat sciatic nerve model. Controls included an unoperated contralateral side and sham-operated groups either with or without the GM-1. During the 5 months of recovery, locomotion was tested by the sciatic function index (SFI). At killing, anesthetized animals were prepared for nerve conduction velocity (NCV) studies, followed by the wet weight of the gastrocnemius muscle (expression of atrophy), toe-chewing (expression of lesion severity and sensory loss), and histological examination of the nerve segments. The SFI showed a slight but significant recovery for both the vascular and avascular groups (34% at 20 weeks), but when GM-1 ganglioside treatment was included, the SFI was poor throughout (20-33%). The average NCV of the graft groups without GM-1 was 46% to 57% of the normal nerve (52.7 m/s), whereas for the groups treated with GM-1, it was 63% to 64% of normal; treatment of the non-vascular graft group significantly improved recovery. A uniformly poor recovery from muscle atrophy was seen for all nerve graft groups (62-67%) compared with normal controls. The mean number of toes per foot chewed was 1.9 and 2.4 in graft groups without GM-1 treatment and 0.9 and 1.3 in graft groups treated with GM-1. This treatment significantly reduced both the extent and the number of animals exhibiting autotomy. The qualitative microscopic appearance of the distal nerve segment in all surgical groups was similar. We conclude that the systemic addition of GM-1 ganglioside enhances only some aspects of regeneration in grafted nerves, possibly with a preferential effect on sensory nerve regeneration and functional recovery.

USE OF THE TURNOVER EPINEURIAL SHEATH TUBE FOR REPAIR OF PERIPHERAL NERVE GAPS

Autologous nerve grafting is the conventional technique for bridging nerve gaps, despite its various disadvantages. In this study, the authors investigated the effects of the turnover epineurial sheath tube (TEST) as an alternative to nerve grafting for the repair of nerve gaps, using a rat sciatic nerve model in four groups: Group 1 (n = 5): sham control; Group 2 (n = 10): segmental nerve resection + no repair; Group 3 (n = 10): segmental nerve resection + nerve grafting; Group 4 (n = 15): segmental nerve resection + TEST. Functional recovery was evaluated by walking-track analysis. The sciatic nerves and gastrocnemius muscles were harvested for histologic and quantitative histomorphometric evaluation at 12 weeks. Sciatic functional indices and histomorphometric analyses revealed statistically significant differences between the sham control and the three experimental groups (p < 0.001). The difference between the TEST group and the nerve graft group was not significant; however functional recovery was significantly improved in these two groups, compared to the non-repaired group (p < 0.05). The authors suggest that the TEST provides a suitable conduit between two stumps, eliminates donor-site morbidity, reduces the number of suture sites, fibrosis, and operating time, and might be an alternative to nerve grafting for nerve gap repair.

Rat tibial nerve regeneration after postoperative administration of cis-diaminedichloroplatinum.

We investigated the results of postoperative administration of cis-diaminedichloroplatinum on nerve regeneration in rats. Forty-two Lewis rats were divided into two groups, receiving end-to-end suture or nerve grafting. Subgroups of the suture group included control, a one-time dose of cis-diaminedichloroplatinum (3 mg/kg), and four doses of the same amount of cis-diaminedichloroplatinum. Subgroups in the nerve grafting group included a control and a four-dose group. Functional recovery was measured by gait analysis using the tibial function index. Biopsies of nerve were taken distal to the suture site for histologic evaluation 20 weeks postoperatively. Tibial function index at 4, 6, and 20 postoperative weeks in the four-dose group was significantly inferior to that of the control group in the suture group. Tibial function index was inferior in the four-dose group until 10 weeks postoperatively compared with control in the nerve grafting group; however, there was no significant difference. The percentage of neural tissue in the one-dose suture group was significantly greater than in the four-dose suture group. These results suggest that postoperative cis-diaminedichloroplatinum administration may have a detrimental effect on nerve regeneration.

Axonal regrowth through a collagen guidance channel bridging spinal cord to the avulsed C6 roots: Functional recovery in primates with brachial plexus injury

Intraspinal implantation of a collagen guidance channel (CGC) to promote axon regeneration was investigated in marmosets with brachial plexus injury. After avulsion of the right C5, C6 and C7 spinal roots, a CGC containing (group B) or not (group A) a nerve segment, or a nerve graft (group C), was ventro-laterally implanted into the cord to bridge the ventral horn and the avulsed C6 roots. No spinal cord dysfunction was observed following surgery. Two months later, the postoperative flaccid paralysis of the lesioned arm improved. In five months, a normal electromyogram of the affected biceps muscle was recorded in all repaired animals. Motor evoked potentials were obtained with a mean amplitude of 13.37 ± 13.66 μV in group A, 13.21 ± 5.16 μV in group B and 37.14 ± 35.16 μV in group C. The force of biceps muscle contraction was 27.33 ± 20.03 g (group A), 24.33 ± 17.03 g (group B) and 37.38 ± 21.70 g (group C). Retrograde tracing by horseradish peroxidase showed labelled motoneurons ipsilaterally located in the C5 and C6 ventral horn, nearby the implantation site. The mean labelled neurons was 32.33 ± 21.13, 219.33 ± 176.29 and 64.33 ± 23.54 in group A, B and C respectively. Histological analysis presented numerous myelinated and unmyelinated regenerating axons in the implant of these animals. Statistical analysis did not show significant difference among the three repaired groups. Our results indicate that spinal neurons can regenerate through a CGC to avulsed nerve roots and induce motor recovery in primates. J. Neurosci. Res. 51:723–734, 1998. © 1998 Wiley-Liss, Inc.

Axonal regrowth through a collagen guidance channel bridging spinal cord to the avulsed C6 roots: functional recovery in primates with brachial plexus injury.

Intraspinal implantation of a collagen guidance channel (CGC) to promote axon regeneration was investigated in marmosets with brachial plexus injury. After avulsion of the right C5, C6 and C7 spinal roots, a CGC containing (group B) or not (group A) a nerve segment, or a nerve graft (group C), was ventro-laterally implanted into the cord to bridge the ventral horn and the avulsed C6 roots. No spinal cord dysfunction was observed following surgery. Two months later, the postoperative flaccid paralysis of the lesioned arm improved. In five months, a normal electromyogram of the affected biceps muscle was recorded in all repaired animals. Motor evoked potentials were obtained with a mean amplitude of 13.37 +/- 13.66 microV in group A, 13.21 +/- 5.16 microV in group B and 37.14 +/- 35.16 microV in group C. The force of biceps muscle contraction was 27.33 +/- 20.03 g (group A), 24.33 +/- 17.03 g (group B) and 37.38 +/- 21.70 g (group C). Retrograde tracing by horseradish peroxidase showed labelled motoneurons ipsilaterally located in the C5 and C6 ventral horn, nearby the implantation site. The mean labelled neurons was 32.33 +/- 21.13, 219.33 +/- 176.29 and 64.33 +/- 23.54 in group A, B and C respectively. Histological analysis presented numerous myelinated and unmyelinated regenerating axons in the implant of these animals. Statistical analysis did not show significant difference among the three repaired groups. Our results indicate that spinal neurons can regenerate through a CGC to avulsed nerve roots and induce motor recovery in primates.

Stimulated jitter measurement in the assessment of recovery after different methods of peripheral nerve repair

The recording of stimulated jitter offers a quantitative method for following the recovery of neuromuscular function after peripheral nerve repair. In groups of rats, electrophysiological recording of jitter was carried out on control animals and on animals 90 days after sciatic nerve division and subsequent repair with either direct end-to-end suture (NS), nerve graft (NG) or freeze thawed muscle graft (FTMG). It was found that values for jitter were highest in the FTMG group. The NS and NG groups demonstrated statistically similar jitter values when compared with each other and with the normal. It was concluded that the speed of nerve regeneration is slower in the FTMG group, at least initially, and that 90 days after sciatic nerve repair the FMTG group had an increase in the number of immature neuromuscular junctions when compared with the NS or NG groups. Jitter measurement would appear to offer a means of detecting small differences in nerve regeneration. The value of this in future developments in nerve repair is discussed.

Nerve regeneration in diabetic rats

This study evaluated the capacity of diabetic rats to recover the ability to walk after nerve repair or nerve graft of the posterior tibial nerve at thigh level. Functional recovery of the posterior tibial nerve was evaluated by walking track analysis during regeneration in streptozotocin-induced diabetic rats. Surgical procedures were performed 8 weeks after induction of diabetes. The nerve repair was epineurial. The nerve graft was a 1.5 cm segment orthotopically replaced. There was no significant difference in functional recovery between normal and diabetic rats for both the nerve repair and nerve graft groups at 6, 12, and 24 weeks after nerve reconstruction. It is concluded that the presence of diabetes is not a contraindication for nerve reconstruction. © 1998 Wiley-Liss, Inc. MICROSURGERY 18:9–11, 1998.

Nerve regeneration in diabetic rats.

This study evaluated the capacity of diabetic rats to recover the ability to walk after nerve repair or nerve graft of the posterior tibial nerve at thigh level. Functional recovery of the posterior tibial nerve was evaluated by walking track analysis during regeneration in streptozotocin-induced diabetic rats. Surgical procedures were performed 8 weeks after induction of diabetes. The nerve repair was epineurial. The nerve graft was a 1.5 cm segment orthotopically replaced. There was no significant difference in functional recovery between normal and diabetic rats for both the nerve repair and nerve graft groups at 6, 12, and 24 weeks after nerve reconstruction. It is concluded that the presence of diabetes is not a contraindication for nerve reconstruction.

Improvement in nerve regeneration by monoclonal antibodies to ICAM-1 and LFA-1 in allogeneic mice.

We examined whether giving monoclonal antibodies (MoAb) to intercellular adhesion molecule 1 (ICAM-1) and leucocyte function associated antigen (LFA-1), which have important roles in the initial stage of rejection after allografts, can improve nerve regeneration in allogeneic mice. Fresh sciatic nerves were grafted using BALB/c mice as donors and C3H/He mice as recipients. Nerve regeneration at six weeks was significantly better in the mice given MoAbs at one and five days than in those given none (n = 5 in each group), although nerve regeneration even in the five-day group was significantly inferior to that in the syngeneic nerve graft group (n = 5). The survival time of the nerve donor skin graft at 12 weeks was not prolonged by treatment with MoAbs, indicating a failure to induce immunological tolerance. However, at 10 months after nerve grafting there were fewer Mac-1, Lyt-1, and Thy-1 positive cells in the five-day group and they showed less immunoreactivity than the untreated group. We conclude that giving MoAbs could effectively improve nerve regeneration in grafted allogeneic nerve segments, although it did not induce immunological tolerance.

Functional assessment of tibial-nerve recovery in the cat using gait analysis: preliminary study.

The purpose of this study was to investigate gait-pattern changes after complete tibial nerve lesion in the cat, and to observe whether nerve repair could reverse some of the changes. In six cats, a 5-cm segment of the tibial nerve was transected. The nerve gap was then repaired with nerve autograft in three animals and was unrepaired in three as controls. The walking patterns of the cats were videotaped, and the hip, knee, ankle, and metatarsophalangeal joint angles were measured at the beginnings of the F, E1, E2, and E3 phases of the step cycle. Two weeks after surgery, abnormal gait patterns were observed, and four gait parameters (E3.Hip, E3.Ankle, E3.M-P, and F.Ankle) were found to be statistically significantly different from normal. Six months after surgery, the nerve-graft group had gait-parameter values approaching normal, while the control group showed no measurable improvement. Correspondingly, electrophysiologic testing revealed considerable nerve regeneration in the nerve-graft group but not in the control group. It was concluded that these gait parameters can be used as valid functional indices to evaluate the degree of tibial nerve recovery in the cat model.

The Role of Cultured Schwann Cell Grafts in the Repair of Gaps within the Peripheral Nervous System of Primates

With recent advances in cell culture techniques it is possible to isolate human SCs from adult peripheral nerves, expand and purify their number in cell culture, and construct a cellular prosthesis from the cultured cells. The current study was designed to ascertain whether these techniques could be used to repair nonhuman primate nerve injuries. In 12 adult femalecynomologousmonkeys, the musculocutaneous (msk) nerve was divided and prevented from regenerating and the brachioradialis nerve (brach) was exposed bilaterally (n=24 nerves) and injured so that a 15-mm gap existed within the nerve. The brach nerves were either repaired with sural nerve autografts (n=6), guidance channels which contained monkey SCs (120×106cells/ml;n=6), or guidance channels without SCs (n=6). The remaining brach nerves (n=6) had either no injury or an injury to the nerve without a repair. Autologous expanded primate SCs were increased in number at least 10-fold over a 2-week period at which time the SC purity exceeded 99.9%. Monkeys in each group, including the control group, regained some degree of elbow flexion after 3 months despite sectioning both the mask nerve and the brach nerve; therefore, we were unable to determine simply on clinical grounds which repair was the most effective in promoting functional recovery. Brach nerves repaired with sural nerve grafts were superior to both the channels which contained SCs and empty channels in regards to the number of myelinated axons proximal, within, and distal to the repair site (P<0.05). Electrophysiologic results closely paralleled the histologic data with evidence of reinnervation of the brachioradialis muscle based on the compound muscle action potential in both sural nerve graft and monkey SC channel repair groups.

Ankle stance angle: a functional index for the evaluation of sciatic nerve recovery after complete transection.

This study attempted to develop a motor functional index, ankle stance angle (ASA), to assess rat sciatic nerve regeneration subsequent to autografting. ASA, 50 degrees in normal rats, is the ankle joint angle at the mid-stance phase of the gait cycle. In a nerve graft group, a 1-cm segment of the right sciatic nerve was transected and then repaired with nerve autograft. In an ungrafted group, the nerve gap was left unrepaired. ASA measured 4 months after surgery was statistically significantly larger in the nerve graft group (36 degrees) than in the ungrafted group (22 degrees). The results suggest that ASA is more sensitive than sciatic function index in detecting functional recovery after a complete sciatic nerve lesion. ASA also showed a significant correlation with the passive range of ankle joint motion and gastrocnemius muscle weight. The study concluded that ASA is a reliable index for assessment of regeneration of rat sciatic nerve after a complete lesion. The intra-rater reliability (r = 0.97 and 0.90) and inter-rater reliability (r = 0.85) tests performed support the conclusions.

Sciatic nerve regeneration in the rat after frozen muscle grafting: A comparative study using somatosensory evoked potentials

In an experimental study, somatosensory evoked potentials were used to evaluate sciatic nerve regeneration in the rat after 12 mm-long conventional nerve grafting, vascularized nerve grafting and frozen muscle grafting. This experimental method was found to be technically easy and highly reproducible. No statistical difference was found between the three groups concerning amplitude of the negative electrical wave recorded at the cortex level after distal stimulation. Conduction velocities were found to be significantly higher in both the vascularized nerve group and the frozen muscle group, compared with the conventional nerve grafting group. The frozen muscle grafting technique is valuable as it gives good experimental results, is easy to carry out and causes minimal damage to the donor site.

Experimental study on nerve regeneration through the basement membrane tubes of the nerve, muscle, and artery

We evaluated neurotization after transplantation with lyophilized nerves, muscles, and arteries, and examined the possibility of practical application of long bridging grafts. Grafts of 10 mm and 25 mm of lyophilized nerves, muscles, and arteries harvested from Fisher rats were transplanted to the sciatic nerves of recipient Lewis rats. The histological changes undergone by short grafts were observed at weekly intervals. The sham-operated and isograft groups were used to compare the results of long grafts. In both the nerve and muscle-graft group, regenerated axons grew out through the residual basement membrane tube. But in the muscle graft group, phagocytosis of myofibril debris took longer than that of degenerated axons. No statistical differences were found between results of TSI, induced EMG, and quantitative analysis of myelinated axons in the nerve and muscle graft groups. No neurotization was noted in the long artery graft. In long grafts, laminin found on the basement membrane may not be sufficient to accelerate neurotization, and arteries should not be used for tubulization.

Arterial bridging for repair of peripheral nerve gap: a comparative study.

A new experimental model was designed in which the regeneration of rat femoral nerve across a 8 mm excised gap was investigated after insertion of the distal and proximal stumps into the anatomically and functionally intact femoral artery (AIAB). This model was compared with groups of free artery and autologous nerve grafting. After a period of 12 weeks, a histological and electrophysiological analysis was carried out, which demonstrated that the AIAB and autologous nerve grafting group had a significantly higher percentage of regeneration compared with the free artery-graft group. The nerve regeneration and intraneural vascular reconstruction that occurred within AIAB group were more successful than those that occurred in the artery and nerve-grafting groups.

Nerve Regeneration in a Bony Bed

In a rabbit model, regeneration of the intratemporal facial nerve was studied as vascularized and nonvascularized nerve grafts. Fifteen New Zealand white rabbits weighing 4 to 4.5 kg were used. A 1-cm gap was created in the intratemporal facial nerve. In the vascularized nerve graft group (group 1), the facial nerve was repaired with a vascularized median nerve graft (n = 6), while in the nonvascularized nerve graft group (group 2), the median nerve was used as a nonvascularized graft (n = 6). In group 3, the facial nerve gap was left unrepaired (n = 3). At 3 months postoperatively, electrophysiologic testing, morphologic nerve study, and morphometric muscle analysis were performed. Although the mean numerical values of axonal counting, nerve conduction, and morphometric muscle study results were slightly better in the vascularized nerve graft group, the differences between the two grafted groups were not statistically significant (p > 0.05). Morphometric nerve analysis, i.e., axon diameter and myelin sheath thickness, showed significant differences in nerve regeneration between the two groups (p < 0.05). These results suggest that a bony bed is not optimal for neovascularization of a nerve graft. We conclude that vascularized nerve grafts are superior to nonvascularized nerve grafts when nerve regeneration occurs in a bony recipient bed. This might be of clinical importance in the repair of facial nerve lesions within the temporal bone.