A laminin graft replaces neurorrhaphy in the restorative surgery of the rat sciatic nerve.

We investigated the role of laminin in functional recovery of a peripheral nerve injury using electrophysiological and behavioral approaches on the rat sciatic nerve in vivo. These studies were complemented by neurofilament protein immunocytochemistry on the sciatic nerve 20 days after an operation, in which an 8-mm piece of the nerve was removed and replaced by a graft of laminin, its neurite outgrowth-promoting peptide, a control peptide, collagen, or by resuturing of the removed piece of the nerve. Electrophysiological measurements of muscle strength 4 months after the sciatic nerve transection showed that a laminin graft was as effective as neurorrhaphy in supporting functional recovery of an injured peripheral nerve. A laminin graft also significantly reduced autotomy in the operated animals. Immunocytochemistry confirmed that both a laminin graft and resuturing supported growth of the 200-kDa neurofilament-positive axons into the distal stump of the nerve within 20 days of operation. A graft with a neurite outgrowth-promoting peptide of the B2 chain of laminin supported similar axon growth, whereas another peptide graft also derived from laminin or a collagen graft did not support axon growth. All grafts allowed Schwann cell growth into the distal stumps of the nerves, but neurites accompanied them only in the regeneration-supporting grafts and in the resutured nerves. The Schwann cells of the regenerating nerves expressed high levels of the neurite outgrowth-promoting domain of the B2 chain of laminin, whereas the Schwann cells of the degenerating nerves failed to express this domain in the distal stumps of the degenerating nerves. These results provide the first in vivo evidence for the functional role of laminin in peripheral nerve regeneration. As the neurite outgrowth-promoting domain of the B2 chain of laminin is as efficient as laminin or resuturing in supporting a short-term recovery of an injured sciatic nerve, this area may be a regeneration-promoting domain of this glycoprotein. More importantly, as grafting significantly reduces post-traumatic pain behavior in the operated animals, the laminin graft surgery may provide a useful method for clinical restoration of the injured peripheral nerves.

Response characteristics of tooth pulp-driven postsynaptic neurons in the spinal trigeminal subnucleus oralis of the cat.

Neuronal activity in the spinal trigeminal subnucleus oralis in response to electrical tooth stimulation was recorded in the anaesthetized cat in order to compare the electrophysiological characteristics of the oralis neurons with those of subnucleus caudalis and interpolaris neurons recorded in previous studies. The most sensitive oralis neurons had lower thresholds and shorter latencies than the most sensitive caudalis and interpolaris neurons. The thresholds of the oralis neurons were lower and their strength-duration curves flatter than those depicting liminal dental pain in man but similar to those depicting liminal jaw reflexes in the cat. Noxious conditioning stimulus elevated the threshold of only 1 of 10 neurons tested. The converging input from the skin and oral mucosa was from low-threshold mechanoreceptors. The results indicate that the response properties of the subnucleus oralis neurons differ significantly from those of other spinal subnuclei. Human pain thresholds cannot be explained by the liminal response properties of oralis neurons. These neurons might be important in the mediation of liminal reflex events evoked by dental stimuli.

Role of intradental A- and C-type nerve fibres in dental pain mechanisms.

The arousal of the two components of pain (the first rapid or sharp pain and the second dull pain) are considered to be related to activation of A delta- and C-type nociceptive primary afferents, respectively. The same dichotomy of pain sensations may also exist in teeth, although due to the short distance between the site of stimulation and the brain the two sensations might not be as clearly separated as in stimulation of, for example, the extremities. The sensations evoked by stimulation of human teeth vary according to the type of the stimuli applied. Low-intensity electrical stimulation is able to induce non-painful (prepain) sensations. At high current intensities pain is evoked. Drilling, probing and air-drying of exposed dentin induce only pain. Most studies also indicate that thermal stimulation only induces painful sensations. The quality of dental pain can vary. Typically, dentinal stimulation of teeth with healthy pulps induces sharp pain. On the other hand intense heat stimulation can result in dull pain which radiates to a wider area of the face and jaws. This component of the stimulus-induced pain seems to share some characteristics of toothache associated with painful pulpitis. Single fibre recordings of intradental nerve activity in experimental animals have shown that in addition to A-fibres a considerable number of C-type primary afferents innervate the dental pulp. This is in accordance with the results of neuroanatomical studies, which indicate that 70-80% of pulpal axons in human, monkey, dog, and cat teeth are unmyelinated. Intradental A- and C-fibre groups seem to be functionally different and can be activated separately by certain external stimuli. Comparison of the response characteristics of the pulp nerve fibres and the sensations induced from human teeth indicate that: 1) A-fibres are responsible for the sensitivity of dentine and thus for the mediation of the sharp pain induced by dentinal stimulation, 2) Prepain sensations induced by electrical stimulation result from activation of the lowest threshold A-fibres some of which can be classified as A beta-fibres according to their conduction velocities. Comparison of the responses of the A beta- and A delta-fibres indicate that they belong to the same functional group, 3) Intradental C-fibres are activated only if the external stimuli reach the pulp proper. Their activation may contribute to the dull pain induced by intense thermal stimulation of the tooth and to that associated with pulpal inflammation.

Effect of perivascular sympathectomy on distal adrenergic innervation in the hands of monkeys.

Perivascular sympathectomy has been thought to cause distal adrenergic denervation. We performed perivascular sympathectomy for a distance of 1 cm. on two common digital arteries in the right hands of two anaesthetised Macaca arctoides monkeys. Four days later, samples were taken for glyoxylic acid-induced fluorescence examination of the operated and opposite control hand. The distal adrenergic nerves were morphologically normal in appearance after the perivascular sympathectomy. The operation should perhaps be called adventitectomy rather than perivascular sympathectomy and its positive effects may be due to the loss of adventitial support for the vasospastic arteries rather than adrenergic denervation.

Reflex responses in the digastric and tongue muscles to stimulation of intradental nerves in the cat.

Electrical stimulation of tooth pulp nerves induces the digastric jaw-opening reflex in the cat, apparently due to activation of intradental A-fibres; C-fibres do not seem to be involved. In fact, reflex responses to activation of pulpal C-fibres have not been studied. In the present experiments on anesthetized cats we recorded EMG reflex responses of the digastric and tongue muscles to stimulation of the intact tooth crown, exposed dentine, and the pulp. We used stimuli that selectively activate either A- or C-fibres of the pulp. Slow heating of the tooth and application of capsaicin into the pulp, both procedures known to excite only C-fibres in the pulp, evoked licking movements of the tongue and prolonged EMG-responses in the tongue and, less consistently, digastric muscles. Similar muscle responses were elicited by high intensity electrical current pulses applied to the tooth. At low current intensities only short-duration digastric activation (jaw-opening reflex) was induced. Similar digastric jaw-opening was also evoked by drilling and air-drying of dentine, both stimuli able to activate only intradental A-fibres. These results indicate that activation of both A- and C-type pulp nerve fibres can induce reflectory muscle activation, and further, they support the concept of afferent intradental C-fibre innervation.

Effect of naloxone on tooth pulp-evoked jaw-opening reflex in the barbiturate-anaesthetized cat.

The threshold of the tooth pulp-evoked jaw-opening reflex was uninfluenced by administration of 0.5 or 5.0 mg kg-1, i.v. naloxone, a specific opioid antagonist, in the barbiturate-anaesthetized cat. Furthermore, facilitatory or inhibitory interactions between two successive tooth pulp-evoked jaw reflex responses were not influenced by naloxone. It is concluded that naloxone-sensitive opioid receptor-mediated mechanisms do not contribute to the modulation of tooth pulp-evoked reflexes by conditioning dental stimuli. Also they do not exert a tonic inhibition on the sensory or motor part of the tooth pulp-driven reflexes. Experiments performed for comparison showed that the non-nociceptive polysynaptic reflex discharge in the flexo-motor neurons of the limb was not influenced by naloxone either.

Response characteristics of tooth pulp-driven postsynaptic neurons in the spinal trigeminal subnucleus interpolaris of the cat: comparison with primary afferent fiber, subnucleus caudalis, reflex, and sensory responses.

Tooth pulp-evoked single neuron responses were recorded in the spinal trigeminal subnucleus interpolaris of the cat. The thresholds to monopolar electric pulses of varying duration (0.2-20 ms) were determined using a constant current stimulator. The thresholds were comparable with those of primary afferent A-fibers, although the most sensitive primary afferent fibers have lower thresholds. The thresholds and latencies showed that none of the interpolaris neurons received their input solely from intradental C-fibers. The most sensitive subnucleus interpolaris neurons had lower thresholds than the respective subnucleus caudalis neurons studied in our previous work. The thresholds and strength-duration curves of the most sensitive interpolaris neurons and of the tooth pulp-elicited jaw-opening reflex are nearly similar, although the jaw reflex can be elicited at an intensity which is slightly lower than that needed to activate the most sensitive interpolaris neurons of the present sample. The most sensitive interpolaris neurons were activated at current intensities that were below the intensity needed to produce liminal dental pain in man, and the strength-duration curves of these neurons were flatter than the curve depicting liminal dental pain sensation in man. The relationship between stimulus intensity and response magnitude could be well described by power functions, the median exponent of which was 1.251. A conditioning stimulation of the tooth pulp at low intensity produced a short (less than 25 ms) enhancement of the response to the following test stimulus, whereas a high intensity conditioning stimulus produced a longer (greater than 40 ms) suppression of the response to the following stimulus. The threshold of 33% of the neurons was elevated during a noxious tail pinch, and this elevation was not reversed by naloxone, an opioid antagonist. The results indicate that in the trigeminal subnucleus interpolaris there are tooth pulp-driven neurons with an input from intradental A-fibers and that a considerable temporal summation of impulses from primary afferent fibers is needed to activate most of them. Human dental pain thresholds cannot be explained by the liminal response properties of the most sensitive interpolaris neurons, but they may be important in the mediation of near-threshold reflex events. It is possible, however, that the high-threshold interpolaris neurons may have a role in the mediation of sensory responses.

Human pain thresholds after the application of lypressin, a vasopressin analogue.

Recent animal studies indicate that vasopressin has analgetic properties. The aim of this study was to find out if lypressin, a vasopressin analogue, produces analgesia in man. The effect of i.n. lypressin (5 and 10 I.U.) on experimental pain was tested in healthy humans. The lower dose proved high enough to produce a significant antidiuretic effect. Lypressin did not have any marked analgetic effect at these doses either on ischaemic, cutaneous thermal, or dental pain. The results indicate that lypressin cannot be used for pain relief in man at doses low enough not to produce a hazardous water retention.

Spinal block, after dantrolene pretreatment, for resection of a thigh muscle herniation in a young malignant hyperthermia susceptible man.

We describe a young man who experienced malignant hyperpyrexia, probably triggered by suxamethonium and/or enflurane during his second operation for an epigastric hernia. His malignant hyperthermia susceptibility was later verified using the caffeine/halothane contracture test in vitro. Subsequently, a tumorous mass, consisting of herniated and hypertrophied muscle grew in his thigh, and was resected under spinal anaesthesia. Whereas dantrolene (2.5 mg/kg i.v.) pretreatment produced impaired swallowing, the subsequent high spinal block, in addition, resulted in laboured breathing. It is stressed that respiratory power should be monitored when patients pretreated with dantrolene are given spinal anaesthesia. The muscular symptoms and test results in the patient's relatives are also discussed.

Tooth pulp-evoked activity in the spinal trigeminal nucleus caudalis of cat: comparison to primary afferent fiber, reflex, and sensory responses.

Tooth pulp-evoked single-neuron responses were recorded in the spinal trigeminal nucleus caudalis of the cat. The thresholds to monopolar electric pulses of various durations (0.2 to 20 ms) were determined using a constant current stimulator. With stimulus pulse durations of 10 to 20 ms, the thresholds were comparable with those of primary afferent A-fibers, although the most sensitive primary afferent fibers had lower thresholds. Primary afferent C-fibers had higher thresholds than the postsynaptic neurons studied. The threshold for the tooth pulp-elicited jaw-opening response was obtained at a lower stimulus intensity than the liminal response in most postsynaptic neurons of this study. The threshold rise of the postsynaptic trigeminal neurons with decreasing stimulus pulse duration (from 5 to 0.2 ms) was much steeper than that of primary afferent A-fibers or jaw-opening response. The strength-duration curves for tooth pulp-elicited pain sensations in man resemble those of spinal trigeminal neurons. Sixty-two percent of the units had a threshold elevation during a noxious pinch of the tail. The results indicate that the activation of postsynaptic trigeminal neurons requires a considerable temporal summation of primary afferent impulses. The jaw reflex thresholds cannot be explained by the properties of the neurons in the subnucleus caudalis of the trigeminal tract. The results support the concept that dental pain is based on the activation of spinal trigeminal nucleus caudalis neurons receiving their input from intradental A-fibers.

Pain and stress: correlation of stress hormone release to pain modulation in man.

The contribution of stress-induced and opioid-dependent mechanisms to the modulation of experimental pain was studied in man under different conditions. The contribution of these mechanisms to the possible attenuation of acute cardiac pain in human patients was also studied. According to the present series of investigations, stress-induced mechanisms might be involved in the modulation of pain caused by physical exercise but not by concurrent subacute pain or transcutaneous nerve stimulation. The lack of any negative correlation between the pain intensity and the release of stress hormones indicates that stress mechanisms do not attenuate acute ischaemic pain of the cardiac origin. The use of an opioid-antagonist, naloxone, and the measurement of plasma levels of beta-endorphin did not reveal any contribution of endogenous opinoids to pain modulation in the current study.

Effects of repeated bupivacaine administration on sciatic nerve and surrounding muscle tissue in rats.

The effects of repeated administration of 0.5% bupivacaine or saline into the sciatic notch of rats were studied by light microscopy, electron microscopy and a neurophysiological technique. Very severe myositis, including local necrosis, developed in six of 12 rats treated twice daily with 1 ml bupivacaine for either 3 or 7 days. A 3-h infusion of 1.5 ml 0.5% bupivacaine resulted in minor injury to muscle tissue. A marked degree of disruption and vacuolization of myelin sheaths was evident in nerves exposed to bupivacaine for 3 days. Lymphocyte accumulation was confined to the area surrounding the nervous tissue in 7 of 10 of the preparations from rats treated for 3 days or by a 3-h infusion. No histological changes were detected in nerve and muscle tissue from the opposite extremity exposed to saline. After a recovery period of 3 weeks, no differences in the nerve or muscle histology were seen between samples from bupivacaine- or saline-treated animals. The amplitude of the compound action potential of sciatic nerves was, however, significantly lower after bupivacaine treatment (7 days, 1 ml twice daily). Thus, impaired function may continue despite the lack of histological intraneural injury.

Elevation of dental pain threshold induced in man by physical exercise is not reversed by cyproheptadine-mediated suppression of growth hormone release.

The effect of cyproheptadine on growth hormone (GH) secretion and dental pain threshold elevation during physical exercise was studied in healthy human subjects. Different levels of exercise (200-300 W) were produced by a cycle-ergometer. Dental pain thresholds were tested with a constant current pulp tester. In all 6 subjects dental pain thresholds and the heart rate were increased with increasing work load. Cyproheptadine did not have any significant effect on dental pain threshold elevations, although it suppressed the exercise-induced GH release. The results indicate that the exercise-induced dental pain threshold elevation is not based on GH-related stress mechanisms, since cyproheptadine did not reverse the pain threshold elevation.

Tooth pulp-evoked jaw-opening reflex in the cat: evidence for central facilitation induced by noxious discharge in the intradental nerve fibers.

The tooth pulp-evoked jaw-opening reflex was studied in the barbiturate-anesthetized cat. At liminal intensity of the stimulus, a stable short-latency response was obtained in the digastricus and in the tongue. At a higher stimulus intensity, there occasionally appeared to be a prolonged discharge of variable duration in the digastricus, and a second period of activity in the tongue after a silent period. The threshold intensity for these late discharges was supraliminal for the intradental A-fibers and subliminal for intradental C-fibers. Noxious conditioning stimulation of a tooth led to a temporary decrease of the threshold for the jaw-opening reflex elicited from a contralateral or adjacent tooth; only conditioning stimulation at an intensity producing a marked arousal reaction was effective in this respect. Infiltration of the tooth apex with epinephrine produced a local elevation of the threshold for the tooth pulp-evoked jaw-opening reflex. Distant noxious conditioning stimulation (tail pinch) did not influence the jaw-opening threshold. The results indicated that based on some central mechanisms, conditioning noxious stimulation of a tooth can produce a facilitation of the jaw-opening reflex.

Liminal and supraliminal response characteristics of mechanoreceptive neurons in the cuneate nucleus of cat.

The response characteristics of mechanoreceptive neurons (RA, SA, and PC) innervating the foot pad of cat were determined in the cuneate nucleus. The mechanical stimuli were single sinusoidal pulses of varying frequency (20, 60, 150, and 240 Hz), and vibratory trains of varying frequency (80 and 240 Hz) and duration (50, 100, and 400 ms). Thresholds and stimulus-response functions were determined with single pulses. Absolute thresholds (1 impulse/train), tuning thresholds (1 impulse/cycle), and atonal intervals (the range between absolute and tuning thresholds) were determined with vibratory stimulus trains. When tested with single pulses the thresholds resembled those of primary afferents in all unit populations. The stimulus-response function of PC units but not of all RA units were comparable to those of primary afferents. Noxious conditioning stimulation did not influence the thresholds of cuneate mechanoreceptors in the tested sample (N = 6). Mostly PC units were tested with vibratory trains. Absolute thresholds were not dependent on stimulus duration, which is a consistent finding with peripheral units. In contrast to peripheral units, the tuning thresholds in most PC units were elevated with increasing stimulus duration. The variability in the range of atonal intervals was much larger than in the periphery. Thus, it seems that both the type of the tactile signal and the type of the studied mechanoreceptive neuron are critical parameters in determining whether the response characteristics of neurons in the cuneate and in the periphery are identical or not.

Response variability to repeated mechanical stimulation of the skin in the dorsal column system.

The response variability in the primary afferent fibers and in the post-synaptic neurons of- the cuneatus was determined using repetitive mechanical stimulation of the skin in cat. The response variability was larger in the post-synaptic neurons. Of the two studied mechanoreceptor types, the RA (rapidly adapting) units had significantly more variability at the cuneate level than the PC (Pacinian) units, but at the peripheral level no such difference was found. The results suggest that the PC units transmit signals more securely through the cuneatus than the RA units.

Modification of dental pain and cutaneous thermal sensitivity by physical exercise in man.

The effect of physical exercise on dental pain thresholds, the release of pituitary stress hormones and thermal sensitivity of skin was tested in healthy human subjects. Different levels of exercise (100-300 W) at different pedal frequencies were produced by a cycle ergometer. Thermal limen (the interval between warm and cool thresholds) determined from glabrous hand, hairy forearm and leg was used as a parameter of thermal sensitivity. In all subjects the heart rate and blood pressure were increased with increasing work load. Dental pain thresholds were elevated at high work loads with a concomitant activation of pituitary stress hormone (especially growth hormone) release. Thermal limens at all 3 sites were increased work load, too, independent of the pedal frequency. The increase of thermal limen was most marked in the leg and least in the glabrous hand. The results indicate that physical exercise produces a non-segmental, load-dependent decrease of pain and thermal sensitivity with a concomitant activation of pituitary stress mechanisms. The magnitude of modification varies with skin region. Activation of inhibitory mechanisms at spinal levels via muscle and proprioceptive afferents, in a way suggested by the gate control theory of pain mechanisms, seems to have only a minor, if any, contribution to the present findings, since a higher pedal frequency did not produce a more marked decrease of sensitivity.

Effect of tourniquet-induced ischemia on magnitude scaling of tactile stimuli applied to the human hand.

The effect of tourniquet-induced ischemia was tested on magnitude scaling of supraliminal tactile stimuli (single-cycle sine waves) applied to the human hand contra- or ipsilaterally to the cuff. Magnitude estimations were not influenced on the contralateral side. In the ischemic hand there was a reduction of magnitude estimates beginning 12 min. after inflation of the cuff, independent of the stimulus intensity. The results indicate that peripheral noxious stimulation does not produce a nonsegmental reduction of tactile sensitivity. Moreover, it is suggested that the thickest afferent mechanoreceptive fibers (A-beta) have a major role in the mediation of tactile signals both at liminal and supraliminal levels.

The influence of exercise on dental pain thresholds and the release of stress hormones.

Different levels of exercise (50-200 W) were produced by a bicycle ergometer. In all six subjects the heart rate and blood pressure were increased with increasing work load. Dental pain thresholds tended to increase with increasing work load, too. Plasma ACTH levels were above the normal range during the whole experiment in all subjects, whereas plasma cortisol and prolactin levels were elevated only in one subject. Growth hormone levels had a tendency to elevation at 200 W. There was no correlation between the release of cortisol, prolactin or ACTH and the dental pain threshold elevation. However, there was significant correlation between the release of growth hormone and the dental pain threshold elevation. The results indicate that physical exercise at submaximal work loads is enough to produce a pain threshold elevation in some subjects, with a minor coactivation of stress mechanisms.

The function of intradental nerves in relation to the sensations induced by dental stimulation.

Stimulation of intradental nerves has been widely used in pain research as a method for selective activation of pain pathways. It is believed that the only sensation experienced by human subjects in response to activation of pulp nerves is that of pain. However, this concept is not strictly correct. With electrical stimulation at threshold level or near to it a sensation which is not necessarily painful ("prepain") is experienced. When the stimulus intensity is increased suprathreshold, the sensation tends to change to a painful and unpleasant one. The changes in sensations are probably caused by activation of intradental nerve units with different thresholds and conduction velocities. In cats the fastest conducting pulp nerve fibres have the lowest thresholds and slowly conducting units are activated at much higher current levels. In most experiments on human teeth using natural stimuli like hot and cold the only sensation experienced has been pain. It seems also difficult for the subjects to find any difference between different stimuli. Correspondingly, in animal experiments it has been shown that different stimuli applied to dentine are capable of activating the same intradental nerve units probably with a common mechanism (hydrodynamic). However, some recent studies indicate that sensation of cold could be induced by stimulating human teeth.