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The presence of macrophages and the induction of c-jun protein and proliferation of non-neuronal cells were studied following implantation of silicone tubes with different diameters (i.e. 0.8 or 1.6 mm) around the rat sciatic nerve. Three days after implantation, numerous EDI and ED2 positive macrophages could be observed around the nerve beneath the 1.6 mm tubes. Some EDI and ED2 positive macrophages were also present in the endoneurium. In contrast, there were numerous EDI and ED2 positive macrophages in the endoneurium beneath the tube and distally in nerves surrounded by the 0.8 mm tube. In these experiments, there was also a massive induction of c-jun protein and DNA synthesis in non-neuronal cells, as visualised by c-jun and BrdU antibodies respectively (e.g. a response similar to that observed after a crush lesion). Such activated cells, albeit few, were also present in the endoneurium beneath the tube of nerves with a 1.6 mm tube, but not distal to the tube in the endoneurium. At 7 days, the responses were somewhat amplified but essentially the same as at 3 days. The results showed that the large diameter implants, which do not cause axonal damage, as does the small diameter tube, but result in conditioning of the nerve [4], induced invasion of macrophages around the nerve and activation of some cells in the endoneurium beneath the tube. We suggest that cell activation is caused by factors released from macrophages and that endoneurial cell activation is important for the conditioning of the nerve by the silicone tube implant.

To test the hypothesis that cognitive capacity is correlated with the outcome of functional sensibility after nerve repair, 19 patients were evaluated 2 to 5 years after median or ulnar nerve repair at the distal forearm level. The sensory evaluation included tests for functional sensibility as well as assessments addressing perception thresholds for touch/pressure and vibration. Psychometric tests for cognitive capacity were also carried out. Multiple regression analysis, correcting for the effect of age and the ability to perceive touch/vibration, was used to investigate the relationship between functional sensibility and cognitive capacity, and to determine which of the tested central nervous factors had the greatest influence on the outcome of recovery of functional sensibility. On a ranking list of such factors verbal learning and visuo-spatial logic capacity were the most important ones, indicating significant correlations with functional sensibility. It is concluded that cognitive capacity factors may play an important role for the functional outcome following nerve repair and that variations in such factors may help to explain the variability in the outcome of nerve repair.

To clarify whether long-term impaired glucose tolerance (IGT) is associated with dysfunction of peripheral and autonomic nerves, age-matched men with IGT and diabetes mellitus were followed prospectively for 12-15 years, when peripheral and autonomic nerve function was assessed. The patients comprised four subgroups: (1) 51 IGT subjects (duration of IGT at least 12-15 years); (2) 35 diabetic patients, with IGT 12-15 years ago, who later developed diabetes; (3) 34 diabetic patients, duration of diabetes at least 12-15 years; and (4) 62 age-matched non-diabetic control subjects. Mean age of the whole study population was 61 +/- 2 years (mean +/- SD), not different in the four groups. Peripheral nerve function tests included nerve conduction velocities, amplitudes, distal latencies, F-reflexes, and sensory perception thresholds for heat, cold, and vibration. Autonomic nerve function tests included the heart rate reaction during deep breathing (expiration to inspiration ratio) and to tilt (acceleration and brake indices). Despite 12-15 years of IGT, peripheral nerve function did not differ between IGT and control subjects, whereas autonomic nerve function deviated; an abnormal expiration to inspiration ratio (a sign of vagal nerve dysfunction) was significantly more common (15/51 versus 5/62; p < 0.01) in IGT than in control subjects. Diabetic patients (groups 2 and 3) showed lower conduction velocities (in general 2-4 m s-1 lower) than IGT and control subjects in all tested nerves. In conclusion, diabetes but not IGT, is associated with peripheral nerve dysfunction.

The biochemical and morphological responses of the rabbit vagus nerve to local application of colchicine and to nerve crush were investigated. Fourteen days after the cervical vagus nerve had been crushed or subjected to local application of colchicine for 2 h, nodose ganglia of anaesthetized rabbits were either injected with [35S]methionine or [3H]leucine for studies of slow and fast axonal transport, respectively, or prepared for light microscopical examination. The radio-labelled proteins of the faster of the two slow transport groups (SCb; 25-30 mm day-1) were separated by one- or two-dimensional polyacrylamide gel electrophoresis and both radio-labelled tubulin and actin were quantified by densitometry from resulting fluorographs of gels. A relative increase in radio-labelled tubulin was found in SCb in the crushed and colchicine-treated nerves; this increase persisted for up to 50 days after nerve crush. Morphological changes in nerve cell bodies induced by colchicine were similar, but smaller in magnitude than those in crushed nerves. It is concluded that a temporary arrest of axonal transport produced by colchicine can lead to a redistribution of tubulin transport comparable with that found in regenerating nerve.

The silicone chamber model for nerve regeneration is suitable to test the effects of exogenous agents or surgical manipulations on nerve regeneration. The total 16-day regeneration period used in this model makes it possible to analyze the effects of certain manipulations on the sequential advancement of the individual cellular components (circumferential perineurial-like cells, vessels, Schwann cells, axons, and myelin) into the chamber fibrin matrix. In the present study we compared the effects on cellular migration of a 7 day delayed chamber repair vs. chamber repair immediately after transection (control chambers) of the rat sciatic nerve. Regeneration was evaluated with light and electron microscopic techniques. Chambers implanted after a delay of 7 days had a statistically significant more advanced migration of vessels, Schwann cells, and axons from the proximal nerve stump and also a significantly increased vascular density as compared to control chambers. We conclude that a 7 day delayed nerve repair stimulates nerve regeneration in this specific silicone chamber model.

In the present study the inflammatory response was quantitatively evaluated during peripheral nerve regeneration. The fluid from silicone nerve regeneration chambers, inserted in rats, was collected during the early period of regeneration of transected sciatic nerves (6 h-7 d) and analysed with respect to inflammatory cells and mediators (leukotriene B4, LTB4, and interleukin-1 alpha, IL-1 alpha). Leucocytes were detected during the entire period (up to 7 d after implantation). The highest concentration was detected after 24 h. PMNG (polymorphonuclear granulocyte) was the predominant cell type in the chamber fluid during the initial 5d of regeneration. Analysis of the concentration of LTB4 demonstrated two peaks (at 24 h and 5 d). The IL-1 alpha concentration displayed an early and relatively smaller peak after 24 h and a second and much larger peak after 7 d, concomitant with an increase of the number of mononuclear cells.

Effects of acute compression for 2 hours around the sciatic nerve trunk at 30 or 80 mmHg on the regeneration potential in rat sciatic nerves were studied. Sham compression or mobilization was performed contralaterally. A week later a crush injury was inflicted proximal to the compressed segment. After another 3 or 6 days the length of axonal outgrowth was measured, using the pinch test technique. We found that compression at either level caused an increased length of axonal outgrowth compared to the mobilized or sham-compressed nerves. The results show that an acute compression at low pressures does have a conditioning lesion effect on peripheral nerves.

Axonal transport and morphological changes were studied in the rabbit vagus nerve after the nerves had been subjected to compression at either 0, 50 or 200 mmHg for two hours. Slow axonally transported proteins, tubulin and actin, were radiolabelled with 35S-methionine two, seven or 14 days after the injury and the distribution of radiolabelled tubulin and actin within component b of slow transport was measured three days later by densitometric analysis of fluorographs of polyacrylamide gel. No significant differences were found in the distribution of tubulin two (50 and 200 mmHg) or seven (200 mmHg) days after injury, but at 14 days (200 mmHg) there was significantly increased radiolabelling of tubulin relative to actin in the nerve 60 to 70 mm from the nodose ganglion. Morphometric measurements of the nerve cell bodies two days after the compression injury at 200 mmHg revealed no significant changes. Previous work has shown that morphological changes, similar to those found after axotomy, were present in nerve cell bodies seven days after a compression injury. This, taken together with the present results, indicates that compression can induce both morphological and biochemical changes in the neurone. The altered axonal transport of tubulin associated with nerve injury follows a slower time course and does not precede the morphological changes. The findings may be of relevance when discussing the double crush syndrome.

Matrix formation within silicone tubes of different geometries implanted between the stumps of the transected rat sciatic nerve was studied. A matrix, composed of longitudinally oriented fibrin strands containing fibronectin, was formed within one day. The matrix then increased in size. The matrix contained macrophages and other inflammatory cells. Matrix size could be increased if the diameter of the tube was increased or if holes were made in the implanted tube. In contrast, matrix diameter decreased if the tube length was increased or if circulation was compromised in the inserts. The results suggest that the size, orientation and cellular components of the matrix have profound effects on the regenerative response of the transected nerve.

The formation of a regenerate between sciatic nerve segments or stumps inserted into Y-tunnelled silicone chambers was studied under conditions where regenerating axons were prevented from entering the chamber. This was accomplished by using an isolated segment of the nerve as a proximal insert. After one week, a cellular regenerate spanned the proximal and distal inserts. The size of the regenerate increased if circulation was preserved in the distal inserts. At four weeks, a perineurium-like sheath surrounded the regenerate and longitudinally oriented Schwann cell columns could be observed throughout the regenerate. A similar 'pseudo-nerve' formed towards a piece of distally inserted tendon. Thus, the information required for the formation of a nerve-like structure is inherent to the non-neuronal cells entering the chamber. Schwann cells, in contrast to regenerating axons, do not exhibit preferential growth towards nervous tissue.

The effects of controlled vibrations of defined frequency (80 Hz), acceleration (32 m/s2 root mean square), and duration (5 hours daily, 2 or 5 days) induced to the hind limb of rats on the regeneration potential in the sciatic nerve after a test crush lesion were determined. Exposure to vibration induced a marked and significant increase in outgrowth length of axons from the crush injury as evaluated after 3 and 6 days with the pinch reflex test. This effect was still observed 1 month but not 3 months after exposure to vibration. Even such a short duration of vibration exposure as 2 days induced an increased length of outgrowth. Such a conditioning effect may be due to local changes in the environment of the axons or to changes in the nerve cell bodies in the dorsal root ganglion. The results indicate that an alarm reaction exists in the nerve at a time point where no structural changes are observed in the nerve. By inducing such a conditioning lesion to nerve tissue, vibration represents a trauma corresponding to a crush lesion or transection of the nerve.

Specificity of muscle reinnervation and the recovery of muscle contractility were studied after repair of the transected rat sciatic nerve. Six different techniques were compared: epineurial suture, perineural suture, whole nerve graft, interfascicular grafts, skeletal muscle bridge and tubulization. Muscle tetanic force and specificity of reinnervation were evaluated 12 weeks after nerve repair. Recovery of tetanic force was superior after repair with epineurial sutures. There was no statistical significance between the other methods in respect of tetanic force. The specificity of muscle reinnervation was best after tubulization, repair with interfascicular grafts and perineurial suture.

The hind paws of rats were subjected to vibration at a frequency of 80 Hz., an acceleration of 32 m./s.2 rms (i.e. ah.w approximately 6.3 m./s.2 rms) for five hours daily during five consecutive days. Morphological, histochemical and immunohistochemical analyses of the soleus, extensor digitorum longus and the plantar muscles in the vibrated limb and the contralateral control limb were performed. No changes were seen in the soleus or extensor digitorum longus muscles but different degrees of degeneration of the muscle fibres were seen in the plantar muscle sections as well as signs of regeneration. No changes were observed in the contralateral unexposed limb. It is concluded that it is not only nervous tissue but also muscle tissue that can be affected by vibration. The changes seem to be confined to muscles close to the vibration exciter.

The basic pathophysiology of an acute and chronic nerve compression lesion is complex. Compression of a peripheral nerve induces marked changes in intraneural microcirculation and nerve fiber structure, impairment of axonal transport, and alterations in vascular permeability, with edema formation and deterioration of nerve function. The peripheral nerves of subjects with underlying neuropathies are more susceptible to compression injury.

The perception thresholds for vibration stimuli of the hand at seven frequencies (8-500 Hz) were evaluated in 300 patients referred to the Department of Hand Surgery in Malmö during the years 1985-1990 for various types of neuropathy (carpal tunnel syndrome, vibration-induced neuropathy, cervical rhizopathy, brachialgia, ulnar neuropathy, radial tunnel syndrome, and polyneuropathy). A sensibility index was calculated by dividing the integrated area under the obtained vibrogram curve of each object tested (areaT) by that of the corresponding area under a superimposed and age matched reference curve (areaR). Sensibility index of less than 0.8 was regarded as the cut off value. The index and little fingers bilaterally were tested to reflect median and ulnar nerve function. There were considerable variations in patterns of pathology among the various groups of patients. In patients with unilateral carpal tunnel syndrome only 10 patients (23%) had abnormalities limited to one recording, while 21 patients (47%) showed abnormalities in all four recordings. This indicates that an isolated carpal tunnel syndrome may reflect more generalised disease of the peripheral nervous system.

The projection of regenerating axons and the specificity of motor reinnervation were studied after repair of the transected rat sciatic nerve with Y-tunnelled silicone chambers. This geometry was used experimentally to face either the proximal tibial or peroneal fascicle with two distal fascicular options usually the distal peroneal and tibial fascicle. A 4 mm gap separated the proximal and distal fascicles. Four weeks after the repair, preferential motor reinnervation could be demonstrated and there were always more axons projecting towards the distal homonymous fascicle. In contrast, if the distal stumps were disconnected from the target no fascicle specific projection of axons was observed. This was true even if segments from the median and ulnar nerve were used to replace either the distal tibial or peroneal segments. It appeared as though the size and not the type of fascicle determined the number of attracted axons. The results suggest that there is no fascicle specific guidance of regenerating nerve fibers.

The effects of application of a chromic catgut suture (conditioning lesion) placed close to the sciatic or tibial nerves on regeneration of the sciatic nerve after a crush lesion (test lesion), that had been induced after an appropriate conditioning interval (two or four weeks) were assessed. The catgut suture induced an inflammatory reaction around the nerve during the four weeks after application of the catgut suture (conditioning interval) but no signs of degeneration of nerve fibres were seen. There was a significant increase in length of outgrowth of sensory nerve fibres as measured by the pinch reflex test when the test lesion was applied after two and four weeks' exposure to the catgut suture. The rate of regeneration was increased by about 11% when the catgut suture had been applied for four weeks before the test lesion was made. The findings indicate that an inflammatory reaction around a peripheral nerve may act as a conditioning lesion, thereby stimulating regeneration of the nerve.

To find out if chronic nerve compression could affect the peripheral nerve by acting as a "conditioning lesion", silicone tubes (internal diameter 0.8-1.6 mm) were placed round the sciatic or tibial nerves of adult rats for different time periods. After the period of compression the tubes were removed and a test crush lesion was made on the sciatic nerve in the thigh. Regeneration distances of the sensory nerve fibres were analysed by the pinch reflex test after a further three or six days. Compression of sciatic or tibial nerves induced a significant increase in outgrowth length compared with nerves subjected to mobilisation and crush lesion alone. All the tubes tested had a conditioning effect, but only tubes with a small internal diameter seemed to induce nerve fibre degeneration as assessed by immunocytochemistry. The results showed that chronic compression induced a regenerative response in the peripheral nerve. The mechanism by which this was accomplished remains obscure but it may involve both the neurons and the non-neuronal cells.

The medial collateral ligaments in the knee joints of 15 rabbits were replaced with expanded polytetrafluoroethylene (E-PTFE). The joints were not immobilised postoperatively, and their structure and function were evaluated after 12 months. Range of motion was equal in all groups. Stability of the knee joints and breaking strength of the ligaments were similar in the group in which the ligament had been reconstructed and in the sham operated control group (n = 8). The group in which the ligament had been severed and left to heal by formation of scar tissue (n = 7) had significantly reduced joint stability and the breaking strength of the medial collateral ligament was significantly less. The E-PTFE ligaments were well incorporated into the surrounding tissues. There were no adverse tissue reactions and the articular cartilage looked normal both macroscopically and microscopically. These results indicate that E-PTFE might be a useful substitute for damaged ligaments, at least in smaller joints like metacarpophalangeal and interphalangeal joints.

Reinnervation of the gastrocnemius and anterior tibial muscles was assessed by measurements of tetanic force after repair of sciatic nerves with Y-shaped or X-shaped silicone tubes in rats. The transsected proximal stump of either the tibial or the peroneal fascicle was introduced into the opening of a Y-shaped silicone tube, or both fascicles were introduced into an X-shaped tube. The distal tibial and peroneal fascicles were inserted into the distal outlets of the tubes leaving a gap of 4 mm between proximal and distal stumps. In the X-shaped tubes the proximal inserts were placed opposite or adjacent to their respective distal parts. Sixteen weeks later reinnervation was evaluated by measurements of tetanic force of the gastrocnemius and anterior tibial muscles after electrical stimulation of the fascicles. There was preferential reinnervation in both types of tubes. In Y-shaped tubes about 90% of the tetanic force could be recorded from both muscles after stimulation of the peroneal and tibial fascicles, respectively. Recovery was lower in the X-shaped tubes, amounting to about 75%. Contractions evoked by misrouted fibres were similar (roughly 40%) in both models. We conclude that motor axons preferentially, but not exclusively, selected a path to reinnervate their original target muscle.

The microanatomy of the neuron and the peripheral nerve, which is a composite tissue, should be considered when discussing the pathophysiology of nerve compression injuries. Acute and chronic compression of peripheral nerve can induce changes in intraneural microcirculation and nerve fiber structure, increase vascular permeability with subsequent edema formation, and impair anterograde and retrograde axonal transport, which all contribute to the clinical symptoms and deterioration of nerve function. Morphologic and functional changes in the nerve cell bodies can be observed after compression which may be due to inhibition of axonal transport. The latter findings might be a theoretical explanation for double crush syndromes. Diabetes mellitus may confer on the peripheral nerve an increased susceptibility to compression injuries. Clinical stages of compression syndromes, such as carpal tunnel syndrome, may be related to specific pathophysiologic events occurring in the nerve.

To investigate the importance of nutritive environment in the healing of reconstructed ligaments, the morphology of sutured free patellar ligaments in synovial fluid of the contralateral knee joint, without re-establishment of microcirculation, was studied in rabbits. After two weeks (n = 6) and one (n = 6) and three (n = 5) months, the sutured ligament was removed from the joint and fibroblasts were seen growing into the suture gap. The ligaments were enveloped in a capsule of fibroblasts, which was more developed in those ligaments that were studied three months after operation. The collagen of the piece of ligament was disorganised at all time points, and tensile strength (n = 6) was low compared with non-sutured patellar ligament. Some ligaments disappeared or were partly attached to the synovial membrane and revascularised. The findings indicate that diffusion of nutrients may be important for the survival and healing of reconstructed ligaments, as the ligaments can heal in synovial fluid without re-establishment of the microcirculation.

The ulnar nerve of a 21-year old man was repaired at the wrist by a silicone chamber technique 10 days after a traumatic transection. A 3 mm gap was left between the nerve ends inside the chamber. At follow-up three years later, motor and sensory recovery was excellent. At exploration at that time a macroscopically normal nerve was found in the tube.

Gore-Tex (expanded Polytetrafluoroethylene, E-PTFE) was used to replace the distal pulleys on the proximal phalanges of 20 rabbits. Morphology and function of the reconstructed pulleys were evaluated at 12-20 weeks. The breaking strength of the E-PTFE pulley equalled that of a normal pulley. Range of motion, tendon excursion, and force of flexion were no different from those of normal pulleys. No adhesions between flexor tendons and synovial tendon sheaths or the E-PTFE pulleys could be detected. No adverse tissue reactions were seen. Fibroblast-like cells from the surrounding tissues had grown into the membrane. In other experiments, where the pulleys had been either removed or detached, local fibrosis and adhesions were seen, and the deep flexor tendons had ruptured. The results of the present study indicate that E-PTFE can be used as a biosynthetic replacement for damaged pulleys.

To study the effects of drying on regeneration, crush lesions were made on the sciatic nerve of 38 rats and the injured area was then air-dried for various periods of time (0-60 min). The regeneration distance of sensory nerve fibres was measured by the pinch reflex test three or six days later. Regeneration was significantly impaired by drying the nerve for 30 or 60 min, but shorter periods (10 min) also seemed to affect axonal outgrowth. Regeneration distances were increased in nerves that were subjected to drying and then allowed to recover for a week (conditioning interval) before the nerve was crushed. The results show that drying of a peripheral nerve is detrimental to nerve regeneration, and we suggest that nerves should not be exposed to air for more than a few minutes during operation without irrigation.

Non-myelinated C-fibre responses during sympathetic trunk stimulation were studied in rabbit common peroneal nerve 2 weeks after the nerve had been subjected to compression at 400 mmHg for 30 min. Our previous studies have demonstrated that during sympathetic trunk stimulation the compound action potential of uninjured somatic C-fibres is characterized by a reduced amplitude and an increased latency. In the present study, nerve compression changed the C-fibre response to sympathetic stimulation. Three out of eight nerves reacted to nerve compression by increased C-fibre compound action potential amplitude in response to sympathetic stimulation. In three other rabbits with compressed nerves the C-fibre action potential amplitude was unchanged, and in the remaining two rabbits the action potential amplitude was decreased during sympathetic stimulation. The action potential latency increased in all tested compressed C-fibres. The phenomenon of increased C-fibre amplitude during sympathetic activation has not been observed in uninjured nerves. As in uninjured nerves, noradrenaline infusion produced an increased C-fibre action potential amplitude and latency in six animals. Sympathetic stimulation did not affect the A-fibre response. These results indicate that sympathetic activity influences the conduction properties in C-fibres of somatic origin and that the response can be changed after a nerve injury. The findings may be of importance for the understanding of pain aggravation in different types of nerve injuries during increased sympathetic activity.

The hind leg of adult rats was exposed to vibrations (82 Hz; amplitude peak-to-peak 0.21 mm) for 4 hours during 5 consecutive days. Light and electron microscopic examination of the plantar and sciatic nerves were done immediately after the exposure period or after a 2- or 4-week recovery period. Light microscopic examination did not reveal any distinct signs of injury. However, ultrastructurally unmyelinated fibers in the plantar nerves showed distinct changes, with deranged axoplasmic structure and/or accumulation of smooth endoplasmatic reticulum. These changes were to a large extent reversible in 2 weeks and appeared normalized after a 4-week recovery period. No ultrastructural changes could be observed in the sciatic nerve. However, when the sciatic nerve was crushed after 5 days of vibration exposure, axonal outgrowth was increased 23% as compared with controls. These findings confirm that vibration induces nerve fiber damage, in this experimental model expressed as a "conditioning effect" contributing to increased regeneration potential of the corresponding neurones.

In all types of peripheral nerve injury, it is important to realize that the lesion affects one extended cell, the neurone, which extends from the central nervous system down to the target tissue in the extremity. Compression of a peripheral nerve can disturb the intraneural transport (axonal transport) of a large variety of substances. This may be followed by morphological and biochemical changes in the nerve cell body. These central changes may effect the axon as a whole and confer on the nerve an increased susceptibility to trauma. Studies concerning the reaction of neurones to compression, relevant when discussing the double crush syndrome, are reviewed.

The mechanisms influencing regeneration of peripheral nerves are incompletely known, but growth factors are supposed to play a key role. In the present study, we demonstrate, with the aid of immunohistochemical methods, that somatomedin C (Sm-C/insulin-like growth factor I/IGF-I) rapidly increased from low to high concentrations, reaching peak values in 2 weeks, in regenerating sciatic nerves of adult rats. In addition, IGF-I was demonstrated extracellularly, never observed in the control nerves. Reactive Schwann cells appeared to be the major source for IGF-synthesis. Higher concentrations were seen in tubulated nerves as compared to sutured ones. It is proposed that IGF-I exerts important growth supporting effects on regenerating peripheral nerves.

Effects of graded compression on nerve function were analyzed in order to evaluate the relative importance of pressure level and duration of compression for functional deterioration. The pressure was applied by means of a small inflatable cuff. The effects of two pressure levels, i.e., 80 mm Hg applied for 2 hr or 400 mm Hg applied for 15 min, were studied in rabbit tibial nerves. The lower pressure tested, which is known to induce ischemia of the compressed nerve segment, also causes some degree of mechanical deformation of the nerve trunk, which leads to incomplete recovery following pressure release. The duration of compression is of importance for the degree of nerve injury even at the higher pressure level tested.

The hind limbs of anaesthetized rats were exposed to vibration trauma (81 Hz; amplitude peak to peak 0.50 mm) for 4 hours during 2 consecutive days. The animals were examined in groups of 4 immediately after the last exposure, and after 1, 2, 3, 5, 7, 10, 14 and 28 days. The Achilles tendons and the tendons of the anterior tibialis muscles were sampled and processed to demonstrate IGF-I immunoreactivity. In the normal Achilles tendon and in the tendon of the anterior tibial muscle, slight IGF-I immunoreactivity was seen in many of the long slender fibroblasts between the collagen bundles. A strong increase in the IGF-I immunoreactivity appeared in the anterior tibialis muscle tendon 3 days after the last vibration exposure. In addition, the tendon fibroblasts became hypertrophic. A similar but less striking increase in IGF-I immunoreactivity appeared in the Achilles tendon. The peak intensity and frequency of stained cells were achieved after 7 days for both tendons. The intensity then levelled off, and was normalized after 28 days. It is concluded that acute exposure to vibrations induces reactive changes in fibroblasts in tendons, which may reflect a change to a more active synthesising state, as a response to the vibration trauma. The transiently altered expression of IGF-I immunoreactivity forms a link in a chain of events regulating the functional activity level of fibroblasts in response to a trauma.

Regeneration of severed peripheral nerves is unfortunately often incomplete, due to loss of nerve fibers and neuroma formation. A new approach is presented with the intention of improving the conditions for nerve repair. In the first of the two stages, a pseudosynovial tube is formed around a silicone rubber rod, surrounded by a stainless steel spiral, which was placed in the backs of rats. This tube, in the second stage, is used as a free "tube graft" to bridge gaps of about 10-12 mm lengths in the severed sciatic nerve. The tube was kept open by the metal spiral. Regenerating nerve fibers with their sprouts grew into the initially open space in the tube. A new nerve trunk was formed, comprised of closely packed myelinated and unmyelinated axons, organized into fascicles. Demonstration by electron microscopy and by EMG recording of reinnervation of foot muscles supported successful long-term results. The fascicles were delimited by perineurial and epineurial sheaths and, furthermore, showed signs of maturation. It was also demonstrated that the nerve-fiber regeneration ceased after a few weeks if there was no distal nerve inserted into the tube. The importance of optimizing the interaction between local factors and regenerating nerve fibers for reestablishment of functionally valuable motor units is discussed.

Compound action potentials of both myelinated (A) and non-myelinated (C) fibres in the common peroneal nerve of rabbits were studied during and after acute, graded compression of the nerve at 200 or 400 mmHg applied for 2 h or during ischaemia created by nitrogen inhalation or aortic occlusion. Compression of the nerve at 200 mmHg blocked the AI component (large myelinated fibres) after about 23 min, while compression at 400 mmHg shortened this time to 11 min. The A2 component (thinner myelinated fibres) had a lower conduction velocity and a higher resistance to compression. There was just a slight decrease in conduction velocity of the non-myelinated fibres when the nerves were compressed at 200 mmHg for 2 h. However, compression at 400 mmHg for 2 h induced a marked deterioration of amplitude and conduction velocity of the C-fibres. There was an incomplete restitution of function of A- and C-fibres during 2 h of recovery. The thinner myelinated fibres were more susceptible to deprivation of oxygen than the thicker ones, while non-myelinated fibres differed in response according to method of ischaemia induction. It is concluded that non-myelinated fibres are very resistant to compression and a very high pressure (greater than 400 mmHg) is needed to affect these fibres.

Gore-tex chambers were used to bridge a 6 mm gap between the proximal and distal nerve stumps of a rat sciatic nerve. The wall structure of these chambers is characterized by "nodes" interconnected by smaller fibrils. Chambers with internodal distances of 5, 10 and 30 microns were used. Some 30 microns chambers were coated from the outside with Gore-tex (0.2 micron internodal distance) and others were coated from the inside. Regeneration after 12 weeks, as evidenced by muscle action potential recordings and light microscopy, was successful regardless of what type of chamber had been used. The organization of the nerve structure varied among different chamber types. A well organized coaxial nerve structure with myelinated axons was observed if inner-coated chambers were used. In chambers that were not coated or in outer-coated chambers tissue completely filled the chambers, and myelinated axons were arranged in mini-fascicles surrounded by loose connective tissue.

Morphological studies have indicated that proximal nerve ends of transected rat sciatic nerves regenerating into preformed mesothelial chambers show a different organization as compared to neuromas developed in contact with a muscle fascia. We have studied the physiological properties of nerve fibres arising from these types of preparations with reference to ongoing activity, response to mechanical stimulation and noradrenaline sensitivity. The study included also fibres arising from ligated and encapsulated neuromas. Fibres with ongoing activity arising from the neuroma could be found from neuromas in contact with a muscle fascia and also from ligated and encapsulated neuromas. This ongoing activity was enhanced by mechanical stimulation and i.v. infusion of noradrenaline. In contrast, fibres arising from proximal nerve ends in mesothelial chambers did not show ongoing activity. These silent fibres responded dynamically to light mechanical stimulation. Noradrenaline did not induce ongoing activity in these fibres.

In 1944 Weiss & Taylor presented experimental evidence against "neurotropism" in nerve regeneration. We used a silicone Y-chamber system to repeat some of those experiments. The proximal stump of transected rat sciatic nerve was introduced into the proximal inlet of the Y. One of the distal outlets was left empty, plugged or occupied by a tendon graft, the other outlet being occupied by a nerve graft. Analysis after 4 and 12 weeks showed in all cases a preferential or exclusive axonal growth towards the nerve piece. The results, indicating the existence of "tissue specificity", are contradictory to the results reported by Weiss & Taylor (30). The findings are discussed with respect to possible influence of humoral, cellular and molecular factors associated with the distal nerve stump as well as the matrix, formed between both nerve segments.

Peripheral neuropathy represents a well-known complication from long-term exposure to vibration. In the present study an experimental model is presented with the purpose of analyzing the formation of intraneural edema following vibration exposure. Vibration (82 Hz, peak-to-peak amplitude 0.21 mm) was induced in the hind limb of rats by the use of vibrating electric motors during 4 h/d for 5 d. Tracer techniques (with albumin Evans blue and horseradish peroxidase) were used to study the permeability of intraneural microvessels after the vibration exposure on day 5. It was found that the vibration trauma in this model induced epineurial edema in the sciatic nerve. It is hypothesized that the formation of intraneural edema may be an important pathophysiological factor in the occurrence of vibration-induced neuropathy.

Preformed, autologous mesothelial chambers were utilized to study axonal growth following selective predegeneration of the distal nerve stump and/or preconditioning of the proximal nerve stump. The left and/or right sciatic nerve of rats was exposed and transected in the thigh. Two weeks after transection, the left proximal nerve stump was cross-anastomosed with the right distal nerve stump by using a mesothelial chamber leaving a 15-mm gap between the two nerve stumps. Previous studies have shown that axonal overgrowth normally does not occur over this gap distance to the distal stump. Three months after cross-anastomosing, regeneration across the 15-mm gap was evaluated by muscle action potential recordings and light microscopical examination. In experiments in which a distal nerve stump was selectively degenerated and the proximal segment was freshly cut, axons had bridged the 15-mm gap in six of seven rats. When a proximal preconditioned nerve stump was matched with a freshly cut distal stump, axonal overgrowth occurred in only 4 of 10 experiments. In experiments including a proximal preconditioned nerve stump and a distal predegenerated stump, axons bridged the gap in 6 of 8 experiments. We concluded that a priming lesion, including manipulation with proximal and/or distal stump, enhances axonal growth in mesothelial chambers.

An experimental model is presented for studying nerve regeneration over gaps of various lengths between the both ends of a severed nerve. After transferring left and right sciatic nerves of rat to the back, the gap between the two nerve ends could be bridged by a preformed, tube-shaped mesothelial chamber of a desired length. When the gap length was 10 mm or less, a well developed nerve structure was generated in the chamber between the nerve ends, and axons from the left sciatic nerve reinnervated muscles in the right limb via the right sciatic nerve. When the gap length was extended to 15 mm or more no such regeneration occurred. When no distal nerve end was introduced into the chamber, there was a limited growth into this chamber over only 5-6 mm. This "limited growth phenomenon" is discussed with respect to a lack of "trophic" or cellular support from a distal nerve segment. It is also proposed that the termination of growth, seen under these circumstances, may suggest a new principle for avoiding the development of neuromas after nerve transections.

An experimental model is presented for studying axonal growth after experimental hyperthyroidism and hypothyroidism. The left sciatic nerve of the rat was transected and transposed to the back. The proximal nerve stump was inserted into a 50-mm-long mesothelial chamber leaving the distal end of the chamber open. Different groups of young adult rats were given daily injections of thyroxine (10 micrograms/100 g body weight) or the goitrogen, thiamazol, in the drinking water (0.125 g/liter) for 12 weeks. Thyroxine treatment increased significantly the extent of axonal outgrowth from the proximal nerve stump compared with untreated rats. Experimental hypothyroidism (thiamazol treatment), evidenced by a retarded body growth, did not affect the extent of axonal outgrowth. In other experiments the left proximal nerve stump was cross-anastomosed with the right distal nerve stump. The two nerve stumps were bridged with a mesothelial chamber leaving a 15-mm gap. This gap distance is known from our previous studies to inhibit axonal overgrowth to the distal nerve stump. As evidenced by histological evaluation, in three of six thyroxine-treated rats, axons had bridged the 15-mm gap. We conclude that experimentally induced hyperthyroidism enhances axonal growth in mesothelial chambers.

Effects of compression at low pressures on slow and fast axonal transport was investigated in rabbit vagus nerve. Proteins in the sensory fibres were radiolabelled by injection of [3H]leucine or [35S]methionine into the nodose ganglion. A small compression chamber and/or ligatures were applied around the cervical part of the vagus nerve for 8 h, at an appropriate time for the subsequent analysis of the effects of compression on both slow and fast transport of radiolabelled proteins. In normal nerves there were two waves of slowly transported proteins with rates of about 12-15 and 25-30 mm/day, respectively. SDS-polyacrylamide gel electrophoresis was used and confirmed that the main proteins which accumulated proximal to the ligatures had a molecular weight of 54 000-56 000. Neither compression of the nerve at 20 mm Hg nor sham-compression induced any statistically significant accumulation of slowly transported proteins at the site of compression. A higher pressure, i.e. 30 mm Hg, induced a marked but incomplete accumulation of slowly transported proteins. Fast transport was partially inhibited in some, but not all, nerves, when 20 mm Hg was applied for 8 h, in contrast to the lack of effect found previously with the same pressure applied for only 2 h. Despite these slight differences, the results indicate that both slow and fast transport are impaired by low pressure levels of around 20-30 mm Hg, which are comparable with those found in human compression neuropathies. The impaired provision of cytoskeletal elements to the distal axon may be of significance in the pathophysiology of nerve entrapment syndromes.

We have compared the anatomic and functional regeneration of a transected sciatic nerve following regrowth from its proximal stump through either preformed empty mesothelial chambers or autologous nerve grafts bridging a 10 mm gap. Within the mesothelial chambers an organized multifascicular nerve trunk forms between the proximal and distal stumps. After 3 months, distal segment cross sections from the mesothelial chamber and nerve graft groups did not differ with respect to axonal density or distribution of axonal diameters. Mean conduction velocities across the gaps were also similar, although the nerve graft group had a wider distribution of velocities. Little or no regeneration was evident when the gap between the nerve stumps was left empty. These results suggest that if the regrowing proximal stump is in an appropriate environment, it can form a well organized and oriented nerve trunk. In the mesothelial chambers, the regenerating nerve is surrounded by a loose cellular stroma and a small amount of interstitial fluid, which was found to contain trophic activity for cultured rodent sensory neurons. Such factors may also support nerve regeneration in vivo.

An experimental model is described in which collagenase in different concentrations and volumes were applied epidurally or intrathecally in the rabbit lumbar spine. This made it possible to study the tissue effects in a situation similar to that of collagenase leaking from a disc or accidental epidural or intrathecal injection at chemonucleolysis. Epidurally applied collagenase, in higher concentration, caused a local thinning of the dura. This effect was reduced at lower concentrations and volumes. Intrathecally injected collagenase, even in small amounts, caused intrathecal hemorrhage and acute paraplegia in the hind limbs. Therefore, in the clinical situation, intrathecal injection of collagenase must be avoided.

Hind legs of adult rats were exposed to vibrations (81 Hz; amplitude 0.50 mm peak to peak) for 4 h during two consecutive days. The sciatic, tibial and plantar nerves were isolated and processed for immunohistochemical demonstration of IGF-I (insulin-like growth factor I; somatomedin C) immunoreactivity at different time intervals after the vibration exposure. In sham-exposed rats the axons in peripheral nerves showed no or faint IGF-I immunoreactivity while most Schwann cells were negative. Exposure of the hind legs to vibrations induced increased IGF-I immunoreactivity in the Schwann cells, demonstrable at the end of the exposure period and reaching maximal intensity 2-3 days after vibration exposure. Several distended axons similarly showed increased staining. The IGF-I immunoreactivity decreased after 7-10 days to almost the level in the control nerves. The most extensive changes were observed in the plantar nerves. The tibial nerves similarly expressed strongly increased IGF-I immunoreactivity in their Schwann cells. The sciatic nerve showed, however, only slightly to moderately increased staining. Cells in the epineurium of the plantar and, to a limited extent, of the tibial nerves expressed concomitantly increased IGF-I immunoreactivity. We conclude that the transiently increased IGF-I immunoreactivity in peripheral nerves reflects reactive changes caused by vibrations and most prominently expressed by the Schwann cells.

The hypothesis that nerves in diabetes mellitus exhibit an increased susceptibility to compression was experimentally tested. Inhibition of fast axonal transport was induced by local compression in sciatic nerves of rats with streptozotocin-induced diabetes mellitus. Fast anterograde axonal transport was measured after application of 3H-leucine to the motor neurone cell bodies in the spinal cord. The sciatic nerve was subjected to local, graded compression in vivo by a small compression chamber. The amount of accumulation of proteins was quantified by calculation of a transport block ratio. Compression at 30 mm Hg for 3 h induced a significantly greater (p less than 0.05) accumulation of axonally transported proteins at the site of compression in nerves of diabetic animals (transport block ratio: 1.01 +/- 0.35; n = 7) than in nerves of controls (0.67 +/- 0.16; n = 7). Accumulation was significantly higher in ligature experiments of both control (1.34 +/- 0.44; n = 8; p less than 0.01) and diabetic animals (1.45 +/- 0.30; n = 8; p less than 0.05), indicating that the block of transport in compressed nerves was incomplete. Neither sham compressed diabetic (0.50 +/- 0.09; n = 6) nor control (0.49 +/- 0.11; n = 6) nerves showed any block of axonal transport. The possible causes of the increased inhibition of fast axonal transport in diabetic rats are discussed. The results indicate that diabetes may lead to an increased susceptibility of peripheral nerves to compression.

The minimal pressure for impairment of fast anterograde axonal transport was determined in rabbit vagus nerve. Proteins, transported by fast anterograde axonal transport, were labeled by a microinjection of [3H]leucine into the nodose ganglion, and a small compression chamber was applied around the cervical vagus nerve. In this way the nerve was subjected to acute, graded compression. Compression at 20 mm Hg for 2 h as well as sham compression did not induce accumulation of axonally transported proteins at the level of compression. However, a pressure of 30 mm Hg for 2 h induced a block of axonal transport at the site of compression. The causes of the axonal transport block are discussed as well as the minimal pressure level in relation to pressures found in clinical nerve compression lesions.

Effects of experimental compression at different pressures on retrograde axonal transport were studied in rabbit vagus nerve. Proteins in the sensory neurones were radiolabelled by injection of [3H]leucine into the nodose ganglion. Sixteen hours after labelling, a small compression chamber and/or ligatures were applied around the cervical part of the vagus nerve for 8 h. Compression of the vagus nerve at 20, 30 and 200 mm Hg pressure induced a graded inhibition of both retrograde and anterograde transport of the radiolabelled proteins. Neither retrograde nor anterograde transport was affected by the presence of the non-inflated chamber. The results indicate that compression at pressures similar to those found in human carpal tunnel syndrome can block retrograde axonal transport. The consequences of inhibition of retrograde and anterograde axonal transport for the metabolism in the nerve cell bodies are discussed.