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Colchicine treatment of the sciatic nerve reduces neurogenic extravasation, but does not affect nociceptive thresholds or collateral sprouting in neuropathic or normal rats.

The effect of topical colchicine treatment of the sciatic nerve on sciatic and saphenous nociceptive thresholds and neurogenic extra-vasation was investigated in normal and neuropathic rats. After a pilot investigation using several different concentrations of colchicine it was determined that treating the sciatic nerve with 5 mM colchicine did not usually affect the heat nociceptive threshold over the sciatic innervated plantar surface of the hindpaw. Mechanical nociception and motor function were also unchanged. Electrical stimulation of the sciatic nerve after intravenous injection of Evans blue dye causes extravasation of the dye in the cutaneous distribution of the nerve. The area and quantity of sciatic extravasation were measured 3 weeks after treating the sciatic nerve with colchicine. This treatment results in a marked loss of neurogenic extravasation, but there were no changes in the sciatic and saphenous mediated heat and mechanical nociceptive thresholds. The area of saphenous nociceptive innervation was mapped using pinch responses and saphenous neurogenic extravasation acutely after sciatic section. There was no change in the cutaneous distribution of saphenous nociceptive fibers when measured 3 weeks after the sciatic colchicine treatment. Some rats had their sciatic nerves transected immediately after colchicine treatment (5 and 50 mM) and the saphenous nociceptive thresholds and autotomy scores were followed postoperatively. Colchicine pretreatment of the sciatic nerve has no effect on the development of hyperalgesia or autotomy. Colchicine blocks axonal transport in peripheral nerve, including the orthograde transport of tachykinins, which probably explains its ability to induce prolonged reductions in sciatic neurogenic extravasation at concentrations that spare C-fiber nociceptor function. Sciatic nerve colchicine treatment does not trigger nociceptive fiber collateral sprouting from the adjacent saphenous nerve, nor does it influence the development of hyperalgesia and autotomy behavior after sciatic transection.

Afferent Pathways↗

Interleukin-1 regulates synthesis of nerve growth factor in non-neuronal cells of rat sciatic nerve.

The Schwann cells and fibroblast-like cells of the intact sciatic nerve of adult rats synthesize very little nerve growth factor (NGF). After lesion, however, there is a dramatic increase in the amounts of both NGF-mRNA and NGF protein synthesized by the sciatic non-neuronal cells. This local increase in NGF synthesis partially replaces the interrupted NGF supply from the periphery to the NGF-responsive sensory and sympathetic neurons, whose axons run within the sciatic nerve. Macrophages, known to invade the site of nerve lesion during wallerian degeneration, are important in the regulation of NGF synthesis. Here we demonstrate that the effect of macrophages on NGF-mRNA levels in cultured explants of sciatic nerve can be mimicked by conditioned media of activated macrophages, and that interleukin-1 is the responsible agent.

Animals↗

Effects of hyperbaric oxygen on skin blood flow and tissue morphology following sciatic nerve constriction.

BACKGROUND: Constriction of the sciatic nerve by loose ligation produces an inflammatory neuropathic injury. This represents an animal model for peripheral mononeuropathy. Oxygen-derived free radicals are suspected to play an important role in the pathogenesis of ischemia/reperfusion injury, leading to neurogenic inflammation. Hyperbaric oxygen (HBO) has been used anecdotally to treat clinically similar conditions in humans, but specific effects on the animal model have not been well studied. OBJECTIVE: This study in a rat model examined the effects of hyperbaric oxygen on skin blood flow and tissue morphology by light and electron microscopy following sciatic nerve constriction. DESIGN: A scientific investigation in a rat model. METHODS: In this study, the neuropathic injury was established by loose ligation of the rat sciatic nerve. The animals were divided into three groups, sham (S, n=8), ligation but no treatment (LN, n=8) and ligation and treatment with hyperbaric oxygen (LT, n=8). The treatment group (n=8) received hyperbaric oxygen treatment immediately following the injury and daily for four additional days at the same time interval. One hundred percent O2 at 3 atmospheres absolute pressure (66 feet sea water) was administered for two hours. The hindpaws of the rats were observed by light microscopy, electron microscopy, laser Doppler flowmetry (LDF), and clinically for the presence of edema. RESULTS: Untreated animals demonstrated marked tissue edema following sciatic constriction, whereas animals that received hyperbaric oxygen had minimal to no edema. The sham group demonstrated normal histology. The group not treated with hyperbaric oxygen demonstrated swollen mitochondria (2-3 times), with loss of cellular integrity, multiple vacuole formation in both nerve and muscle tissue, widened sarcomeres in muscle, and degenerative changes in the nerve myelin sheaths. The group treated with hyperbaric oxygen demonstrated preservation of cellular structure including mitochondrial integrity, no vacuole formation, and maintenance of normal, easily identifiable nerve structure. The sham group had no change of skin blood flow. Skin blood flow of LT group was decreased immediately after ligation (p<0.05) and recovered to baseline level before ligation on Day 5 after four hyperbaric oxygen treatments. Skin blood flow of LN group was decreased immediately after ligation (p <0.01) and did not recover (p <0.01). CONCLUSION: This study evaluated tissue changes after nerve injury caused by loose ligation of the sciatic nerve in rats. Hyperbaric oxygen treatment following sciatic nerve injury reduced tissue edema, improved skin blood flow, and preserved muscle and neuronal ultrastructural integrity.

Journal Article↗

A new anterior approach to the sciatic nerve block.

BACKGROUND: Although several anterior approaches to sciatic nerve block have been described, they are used infrequently. The authors describe a new anterior approach that allows access to the sciatic nerve with the patient in the supine position. METHOD: Sciatic nerve blocks were performed in 22 patients. A line was drawn between the inferior border of the anterosuperior iliac spine and the superior angle of the pubic symphysis tubercle. Next, a perpendicular line bisecting the initial line was drawn and extended 8 cm caudad. The needle was inserted perpendicularly to the skin, and the sciatic nerve was identified at a depth of 10.5 cm (9.5-13.5 cm; median and range) using a nerve stimulator and a 15-cm b-beveled insulated needle. After appropriate localization, either 30 ml mepivacaine, 1.5% (group 1 = knee arthroscopy; n = 16), or 15 ml mepivacaine, 1.5%, plus 15 ml ropivacaine, 0.75%, (group 2 = other procedures; n = 6) was injected. RESULTS: Appropriate landmarks were determined within 1.3 min (0.5-2.0 min). The sciatic nerve was identified in all patients within 2.5 min (1.2-5 min), starting from the beginning of the appropriate landmark determination to the stimulation of its common peroneal nerve component in 13 cases and its tibial nerve component in 9 cases. A complete sensory block in the distribution of both the common peroneal nerve component and the tibial nerve component was obtained within 15 min (5-30 min). A shorter onset was observed in patients who received mepivacaine alone compared with those who received a mixture of mepivacaine plus ropivacaine (10 min [5-25 min] vs. 20 min [10-30 min]; P < 0.05). Recovery time was 4.6 h (2.5-5.5 h) after mepivacaine administration. The addition of ropivacaine produced a block of a much longer duration 13.8 h (5.2-23.6 h); P < 0.05. No complications were observed. CONCLUSIONS: This approach represents an easy and reliable anterior technique for performing sciatic nerve blocks.

Adult↗

Effects of machined/turned, TiO2-blasted and sandblasted/acid-etched titanium oral implant surfaces on nerve conduction: a study on isolated rat sciatic nerves.

The purpose of this study was to explore the clinical relevance of the effects of machined/turned, TiO(2)-blasted and sandblasted/acid-etched titanium oral implant surfaces on nerve conduction. Isolated rat sciatic nerves were placed between two suction electrodes in a pyrex bath containing a tyrode solution. Evoked compound action potentials (cAPs) of the nerves were recorded before and after contact with the implants. The mandibular incisors of randomly selected animals were extracted and changes in cAP amplitudes were used as controls. The differences in final cAP values of Astra Tech implants and rat natural teeth were insignificant (P < 0.05), whereas the differences between other groups were significant (P < 0.05). Machined/turned-surface implants did not cause any change in cAPs. A slight decrease in cAPs was observed for TiO(2)-blasted and sandblasted/acid-etched implants, and the natural teeth. The reductions of cAPs in latter groups were not 50% after an application time of 300 min. The cAP changes of nerves contacting TiO(2)-blasted and sandblasted/acid-etched oral implants fall within physiologic limits in vitro. Machined/turned, TiO(2)-blasted, and sandblasted/acid-etched titanium implant surfaces do not lead to irreversible neurotoxic effects.

Action Potentials↗

The differential expression of N-cadherin in vascularized and nonvascularized nerve grafts: a study in a rat sciatic nerve model.

The authors investigated N-cadherin expression in vascularized and nonvascularized nerve grafts using a rat sciatic nerve model. The vascularized and nonvascularized nerve grafts were elevated, and then both nerves were sutured to the original site. At various periods after the operation, the entire graft was removed. N-cadherin expression was detected via Western blot analysis and histochemical staining using anti-N-cadherin antibody. In both grafts, the level of N-cadherin expression increased after the operation, but during postoperative weeks 6, 9, and 12, the level in the vascularized graft was significantly (p = 0.00023, 0.0018, and 0.0010, respectively) higher than that in the nonvascularized graft. At postoperative week 14, the level of both grafts returned to the control level. Histochemical findings showed that N-cadherin was expressed around the regenerated axons in both grafts, and the degree of regeneration was greater in the vascularized graft than in the nonvascularized graft. These results demonstrate that the expression of N-cadherin increases during the process of axonal regeneration in both grafts, and that the degree of the expression is augmented by vascularization of the nerve graft.

Animals↗

Methods for the experimental functional assessment of rat sciatic nerve regeneration.

In experimental peripheral nerve studies, the rat sciatic nerve model is widely used to examine functional changes after different surgical repairs or pharmacological treatments, following nerve injury. The number and diversity of tests which have been used to assess functional recovery after experimental interventions often makes it difficult to recommend any particular indicator of nerve regeneration. Functional assessment after sciatic nerve lesion has long been focused on walking track analysis, therefore, this article describes in more detail the method to obtain and measure the walking tracks in order to calculate the sciatic functional index (SFI). However, it is important to note that the validity of the SFI has been questioned by several researchers. In addition, the present review includes other traditional tests described in the experimental peripheral nerve literature regarding the rate of return of motor function and sensation, such as the extensor postural thrust (EPT), nociceptive function, and the gastrocnemius-soleus weight parameters. In the last decade, several authors have designed a series of sensitive quantitative methods to assess the recovery of hind limb locomotor function using computerized rat gait analysis. This study aims to review kinematic measures that can be gathered with this technology, including calculation of sciatic functional index, gait-stance duration, ankle kinematics and toe out angle (TOA). A combination of tests, each examining particular components of recovered sensorimotor function is recommended for an overall assessment of rat sciatic nerve regeneration.

Animals↗

Injury-induced reduction of acidic fibroblast growth factor levels in the distal parts of rat sciatic nerve.

Acidic fibroblast growth factor (aFGF) level in sciatic nerve after lesioning was measured by enzyme immunoassay to determine if aFGF functions as a neurotrophic factor like nerve growth factor (NGF). Whereas the NGF level increased in distal segments, the aFGF level there decreased after transection or crushing and recovered to the original level by 10 weeks after crushing. The amount of aFGF mRNA in the sciatic nerve was extremely low to supply the high level of protein found in the sciatic nerve. Sympathetic ganglia, dorsal root ganglia, and spinal cord, which contain neuronal cell bodies extending their axons into the sciatic nerve, showed a greater or similar level of aFGF as sciatic nerve. These results imply that aFGF is synthesized in neuronal cell bodies and distributed anterogradely into their axons. Difference of injury-induced changes in levels between aFGF and NGF suggests distinct mechanisms of the effects elicited from these factors on regeneration of the sciatic nerve.

Animals↗

Vasoactive intestinal polypeptide (VIP) increases in the spinal cord after peripheral axotomy of the sciatic nerve originate from primary afferent neurons.

Following sciatic nerve axotomy, vasoactive intestinal polypeptide (VIP) immunoreactivity increases dramatically in the central terminal areas of the nerve whereas other primary afferent neuropeptides are depleted. The contribution of the peripheral nerve to VIP increases in the spinal cord was investigated by performing sciatic nerve section alone, dorsal rhizotomy of the lumbar roots, axotomy and rhizotomy in combination or section of other peripheral nerves terminating in the same segments as the sciatic nerve. VIP, and for comparison, substance P (SP), cholecystokinin (CCK), somatostatin (SOM), were localized in the lumbar spinal cord and corresponding sensory ganglia using unlabeled antibody immunohistochemistry. After sciatic nerve section, SP, CCK and SOM were depleted in the lumbar dorsal horn whereas VIP increased. After rhizotomy alone all neuropeptide staining including VIP was depleted; axotomy followed by rhizotomy produced the same result. Axotomy of other peripheral nerves terminating in the lumbar cord increased the area of neuropeptide depletion but correspondingly increased the area of VIP staining. A large proportion of small and medium diameter dorsal root ganglion cells were stained for VIP after nerve section or axotomy but not after rhizotomy alone. A radical change in neuropeptide metabolism of dorsal root ganglion cells occurs after peripheral axotomy, in the form of a marked increase in VIP synthesis. An intact dorsal root is necessary for increases in VIP in the spinal cord indicating the primary afferent origin of the response.

Animals↗

Assessment of effects of pinealectomy and exogenous melatonin administration on rat sciatic nerve suture repair: an electrophysiological, electron microscopic, and immunohistochemical study.

BACKGROUND: Collagen scar formation at the cut end of a nerve, an important problem in clinical practice for neurosurgeons in peripheral nerve surgery, obstructs sprouting of axons into appropriate distal fascicles, and thereby limits nerve regeneration. Researchers attempt to control collagen accumulation in the formation of neuroma by various physical and chemical methods, but these have yielded only limited functional success. This is the first experimental study investigating the effects of melatonin (MLT) on nerve repair and neuronal regeneration in rat sciatic nerve suture repair. METHODS: The hypothesis that exogenous MLT administration may inhibit the formation of neuroma in peripheral nerve surgery was investigated in rat sciatic nerve model. In this study, a total of 80 rats were used for control groups (Groups Ia, Ib, IIa, and IId), MLT group (Group Ic), surgical pinealectomy (Px) groups (Groups IIb and IIc), and group of MLT treatment following Px procedure (Group IIe). All animals underwent a surgical intervention consisting of bilateral sciatic nerve section and primary suture repair. At 8 weeks after repair, the animals were killed following completion of recording of nerve action potentials (NAPs). Then, unilateral sciatic nerve specimens including the suture repair region were carefully removed and the excised segments were processed for electron microscopy examination. Afterwards, contralateral sciatic nerve specimens from two animals from each group were removed and stained for immunohistochemical analysis. RESULTS: Results of morphometric analysis revealed that Px procedure caused an elevation of collagen content of the sciatic nerve and macroscopic neuroma formation, and that there was a statistically significant reduction in collagen content of the same region in pinealectomized animals treated with MLT (p<0.001). Accordingly, electrophysiological findings demonstrated that the stimulus intensities required to excite a NAP response were increased in surgical Px group, but the presence of a reduced threshold response was found in the group treated with MLT following Px procedure (p<0.01). Immunohistochemical staining for Type I collagen and Type III collagen was markedly more intense in the epineurium of animals after Px. Virtually no or only weak staining was observed in animals in control groups and the MLT treatment group. Results of immunohistochemical analysis revealed that surgical Px procedure caused a strong immunoreactivity for Type I collagen and Type III collagen in all connective tissue planes of the nerve, especially in the epineurium, and there was a statistically significant reduction in immunoreactivity of the repair region in animals receiving MLT treatment after Px procedure (p<0.001). CONCLUSION: This study demonstrates that exogenous MLT administration significantly inhibits collagen accumulation in the formation of neuroma in the suture repair site and thereby improves nerve regeneration. From a clinical standpoint, the positive effect of MLT administration on neuroma formation and nerve regeneration seems a particularly attractive treatment option. Therefore, we believe that nerve repair with addition of MLT may be a worthwhile option in addition to other treatment modalities in case of MLT deficiency, such as aging. However, further experimental and clinical studies using functional analysis warranted to confirm this result in future.

Action Potentials↗

Selective increase of tumour necrosis factor-alpha in injured and spared myelinated primary afferents after chronic constrictive injury of rat sciatic nerve.

Chronic constriction of the sciatic nerve, leading to a hyperalgesic state, results in a partial lesion wherein some axons are injured and others remain intact. Here we sought to characterize reactive changes which occur in DRG cell bodies of injured and uninjured axons projecting to skin and muscle. Using immunohistochemistry combined with flurorogold and fluororuby retrograde labelling to define DRG cell bodies associated with injured and uninjured axons, we analysed the DRG immunoreactivity (IR) for tumour necrosis factor-alpha (TNF), interleukin-10 (IL-10), the sensory neuron-specific channel vanilloid receptor 1 (VR1), isolectin B4 (IB4) and calcitonin-gene-related peptide (CGRP) 4 days after a unilateral chronic constriction injury (CCI) of the rat sciatic nerve. TNF IR was predominantly localized in neuronal DRG cells. In DRG with an intact nerve, TNF IR was present in 45%, IL-10 IR in 46%, VR1 IR in 44%, IB4 IR in 51% and CGRP IR in 40% of all neuronal profiles. Four days after CCI, TNF IR was increased in medium-sized neurons, whereas IR for IL-10, VR1 and IB4, predominantly present in small neurons, was reduced. Importantly, not only injured but also adjacent spared neurons contributed markedly to increased TNF IR. Neurons projecting to both muscle and skin displayed upregulated TNF IR after CCI. TNF in medium-sized neurons colocalized with neurofilament and trkB, but not with IB4, trkA or RET, suggesting a selective phenotypic switch in presumably low-threshold myelinated primary afferents. Spared myelinated fibres with intact sensory functions but upregulated TNF expression may contribute to behavioural changes observed after nerve injury.

Animals↗

Anterior sciatic nerve block--new landmarks and clinical experience.

BACKGROUND: Anterior sciatic nerve blocks can be complicated by several problems. Pain can be caused by bony contacts and, in obese patients, identification of the landmarks is frequently difficult. METHODS: In a first step, 100 normal anterior-posterior pelvic X-rays were analyzed. The landmarks of the classical anterior approach were drawn on these X-rays and assessed for their sufficiency. Then, in a prospective case study, 200 consecutive patients undergoing total knee replacement were investigated. These patients received femoral and sciatic nerve catheters for postoperative pain management. Using modified anatomical landmarks, sciatic nerve catheters were inserted 5 cm distal from the insertion site of the femoral nerve block perpendicularly in the midline of the lower extremity. This midline connected the insertion site of the femoral nerve catheter to the midpoint between the medial and lateral epicondyle. Correct catheter positioning was verified by magnetic resonance imaging (MRI) in six patients. RESULTS: Evaluation of pelvic X-rays showed that puncture following the classical landmarks pointed in 51% at the lesser trochanter, in 5% medial to the lesser trochanter and in 42% directly at the femur. In the latter patients, location of the sciatic nerve would have been difficult or even impossible. Using our modified anterior approach, the sciatic nerve could be blocked in 196 patients (98%). In nine patients (4.5%) blockade of the posterior femoral cutaneous nerve failed. Vascular puncture happened in 10 (5%) and bony contact in 35 patients (17.5%). Median puncturing depth was 9.5 (7.5-14) cm. Correct sciatic nerve catheter positioning was verified in all patients who underwent MRI. CONCLUSION: Our landmarks for locating the sciatic nerve help to avoid bony contacts and thereby reduce pain during puncture. Our method reliably enabled catheter placement.

Aged↗

[Morphological changes after permanent nerve block by freezing and ethanol injection of the sciatic nerve of the rabbit (author's transl)].

Permanent nerve blocks by intraneurally injected alcohol are often complicated by alcohol-neuritis. Encouraging clinical experiences with permanent blocks by freezing raises the question whether morphological differences between the nerve lesions could explain the difference in their side effects. On 30 rabbits both sciatic nerves were blocked after surgical preparation. The one by intraneural injection of 0,5 ml of 96% Ethanol, the other by freezing with a cryoprobe. The resulting degeneration and the beginning of recovery of the nerves was followed by histological evaluations of the sciatic nerves during the first 28 days after the blockade. The nerve lesions of both types of blockade were complete. That produced by the cryoprobe was limited to the small area of local freezing, whereas the alcohol-block produced the same type of nerve degeneration but with a wide-spread extension reaching the sacral plexus. We discuss whether this slight morphological difference might be sufficient to explain the higher complication rate of alcohol blocks.

Animals↗

Cyclic sciatica related to an extrapelvic endometriosis of the sciatic nerve: new concepts in surgical therapy.

Sciatic pain caused by endometriosis of the sciatic nerve is an uncommon clinical finding and seems to have been verified histologically in only a few cases. Patients complain of typical signs and symptoms of common sciatica that are cyclic in nature. Suggested compression of lumbar root or sciatic nerve or its plexus could be confirmed by electromyography, computed tomography, or magnetic resonance imaging, and by prompt response to hormonal suppression of ovarian function with regression of the radiologic findings. Patients often have required radical surgery with total hysterectomy and bilateral salpingo-oophorectomy. However, conservative surgery with excision of the endometriosis from the nerve can be successful in selected patients who wish to preserve reproductive function. We report a case of sciatic nerve involvement explored by magnetic resonance imaging, with endometriosis in contact with the nerve in the right sciatic trunk.

Adult↗

In vivo incorporation of [3H]palmitic acid into PO protein, the major intrinsic protein of rat sciatic nerve myelin.

Separation of rat sciatic nerve myelin proteins by sodium dodecyl sulfate-slab gel electrophoresis 18 h after injection of [3H]palmitic acid into the nerve demonstrated acylation of the PO protein. When sciatic nerve myelin labeled with [3H]palmitic acid was extracted with acidified chloroform/methanol or chloroform/methanol, the radioactivity associated with PO was retained. These results provided evidence that the radioactivity derived from [3H]palmitic acid in PO protein was not due to labeled phospholipid bound to PO protein by strong physical interaction. Furthermore, the radioactivity associated with purified preparations of PO remained after dialysis and re-electrophoresis, providing additional evidence that [3H]palmitic acid was firmly bound to PO. Treatment of myelin proteins with hydroxylamine at pH 6.6 released most of the radioactivity, indicating that [3H]palmitic acid was covalently bound by ester linkage to PO. Cleavage of purified acylated PO with methanolic NaOH released 85% of the protein-bound radioactivity. Gas-liquid chromatography of the fatty acids released from PO showed labeling of methyl esters of palmitate, stearate, and oleate (56, 16, and 5%, respectively). In addition, a small unlabeled peak with retention time identical with methyl linoleate was also observed. Intraneural injection of a mixture of [3H]palmitic acid and [14C]fucose or [3H]palmitic acid and [35S]sulfate in vivo followed by disc gel electrophoresis and determination of incorporated radioactivity clearly showed that PO was glycosylated, sulfated, and acylated. The potential significance of fatty acids linked to PO is discussed.

Acylation↗

The use of a muscle graft to repair a segmentary nerve defect. An experimental study using the sciatic nerve of rats as model.

The use of a devitalized skeletal muscle graft and conventional nerve graft to repair a 5mm long segmentary sciatic nerve lesion was studied in rats by means of functional, morphometric and spinal cord motor neuron cell response evaluation. Thirty-four rats were used and divided into four groups according to the procedure: (1) sham operation; (2) conventional nerve grafting; (3) muscle grafting; (4) unrepaired lesion. The sciatic functional index (SFI) was evaluated every fortnight up to the 105th postoperative day by measuring three parameters in the rats' footprint. The animals of Groups 2 and 3 presented initial complete functional loss, followed by slow but steady recovery, with final similar SFIs. The histologic and morphometric studies showed an increased small diameter/thin myelin sheath nerve fiber density distally to the lesion site for both types of graft. An increased population of motor neurons was observed in the anterior horn of the lumbar spinal cord segment with both types of grafts, but not in the control groups. The SFI, histologic and morphometric data did not differ significantly between the two types of graft, thus indicating a similar behavior. The authors conclude that a 5mm long skeletal muscle graft works as well as a conventional nerve graft.

Animals↗