Clinical benefit of botulinum toxin for treatment of persistent TMD-related myofascial pain: A randomized, placebo-controlled, cross-over trial.

BACKGROUND: Injections of botulinum toxin type A (BoNT-A) have been proposed as an additional treatment modality for patients suffering chronic temporomandibular disorder (TMD)-related myofascial pain (MFP). BoNT-A impairs muscle function, along with its analgesic effect, and a minimal effective dose should be used. The objective of this randomized placebo-controlled crossover study was to evaluate the clinical benefit of a moderate dose (50 U) of BoNT-A. METHODS: Sixty-six subjects were randomized into two groups, one which received BoNT-A first and a second which received a saline solution (SS) first. Follow-ups were performed 2, 11, and 16 weeks after the injections. Diagnostic criteria for temporomandibular disorders (DC/TMD) diagnostic algorithms were used to evaluate characteristic pain intensity (CPI) and pain-related disability based on the Graded Chronic Pain Scale (GCPS). Electromyographic and bite force were also evaluated. RESULTS: The within-group analysis showed a significant improvement in pain intensity and pain-related disability after BoNT-A (p < 0.001, p = 0.005, p = 0.011) and SS (p = 0.003, p = 0.005, p = 0.046) injections up to week 16. The between-group analysis of pain-related variables revealed no differences between groups at any time. Nonetheless, BoNT-A, but not SS, caused a significant decline in muscle performance. The number needed to treat (NNT) regarding a clinically significant pain reduction (≥30%) was 6.3, 57.0, and 19.0 at 2, 11, and 16-week follow-ups favoring BoNT-A. CONCLUSIONS: Injections of 50 U of BoNT-A might improve MFP symptoms, but the specific effect of the drug on pain compared to the placebo is not obvious.

Insights into the genetic architecture of morphological traits in two passerine bird species.

Knowledge about the underlying genetic architecture of phenotypic traits is needed to understand and predict evolutionary dynamics. The number of causal loci, magnitude of the effects and location in the genome are, however, still largely unknown. Here, we use genome-wide single-nucleotide polymorphism (SNP) data from two large-scale data sets on house sparrows and collared flycatchers to examine the genetic architecture of different morphological traits (tarsus length, wing length, body mass, bill depth, bill length, total and visible badge size and white wing patches). Genomic heritabilities were estimated using relatedness calculated from SNPs. The proportion of variance captured by the SNPs (SNP-based heritability) was lower in house sparrows compared with collared flycatchers, as expected given marker density (6348 SNPs in house sparrows versus 38 689 SNPs in collared flycatchers). Indeed, after downsampling to similar SNP density and sample size, this estimate was no longer markedly different between species. Chromosome-partitioning analyses demonstrated that the proportion of variance explained by each chromosome was significantly positively related to the chromosome size for some traits and, generally, that larger chromosomes tended to explain proportionally more variation than smaller chromosomes. Finally, we found two genome-wide significant associations with very small-effect sizes. One SNP on chromosome 20 was associated with bill length in house sparrows and explained 1.2% of phenotypic variation (VP), and one SNP on chromosome 4 was associated with tarsus length in collared flycatchers (3% of VP). Although we cannot exclude the possibility of undetected large-effect loci, our results indicate a polygenic basis for morphological traits.

Diagnostic criteria for temporomandibular disorders (DC/TMD): interexaminer reliability of the Finnish version of Axis I clinical diagnoses.

Recently, updated diagnostic criteria for temporomandibular disorders (DC/TMD) were published to assess TMD in a standardised way in clinical and research settings. The DC/TMD protocol has been translated into Finnish using specific cultural equivalency procedures. To assess the interexaminer reliability using the Finnish translations of the DC/TMD-FIN Axis I clinical diagnostic assessment instruments. Reliability assessment data were collected during a 1-day DC/TMD Examiner Training Course at the University of Turku, Finland, in collaboration with the International DC/TMD Training and Calibration Center in Malmö University. Clinical TMD examinations according to the Finnish pre-final version of the DC/TMD Axis I assessment protocol were performed by four experienced TMD specialists on altogether 16 models. Kappa coefficient, overall percentage agreement (%A) as well as positive (PA) and negative (NA) agreements were used to define the reliability. Myofascial pain with referral, headache attributed to TMD and disc displacement (DD) without reduction without limited opening showed excellent kappa values (range 0·87-1·00). Fair-to-good reliability was observed for diagnoses of myalgia (k = 0·67), arthralgia (k = 0·71) and DD with reduction (k = 0·64). The PA was high for all pain-related diagnoses and DD without reduction without limited opening (medians ≥83%), and acceptable for DD with reduction (median 67%). The NA was high (medians ≥87%) for all DC/TMD diagnoses, except for myalgia which showed acceptable NA (median 75%). The %A was high for all assessed diagnoses (medians >85%). The findings of this study showed DC/TMD-FIN Axis I to demonstrate sufficiently high reliability for pain-related TMD diagnoses.

Long-term use of occlusal appliance has impact on sleep structure.

Acrylic occlusal appliances (OAs) have been used for temporomandibular disorders and sleep bruxism, but the effects of the treatment are still insufficiently evaluated. Two all-night polysomnographic recordings were made in a sleep laboratory on 14 bruxists (9 females and 5 males with mean age of 27·5 years). The measurement included basic polysomnography with additional masseter muscle electromyogram and movement recording (static charge-sensitive bed method) using randomisation. The base night recording was followed by the second study night after 8 weeks regular use of OA. The OA was made on the occlusal surface of the teeth of the upper jaw, and it was used at night time during the study period. With the OA, rapid eye movement sleep changed from 23·3% to 19·6% (P = 0·078), and slow wave sleep increased significantly from 10·2% to 14·7% (P = 0·039). Masseter contraction (MC) episodes occurred with similar frequency (9·7 vs. 10·5 episodes per hour, P = 0·272). The intensity of the rhythmic MC bursts within an episode decreased from 5·5 to 4·4 (P = 0·027). The groups were post hoc divided into responders and non-responders using a 20% change in MC episode per hour as a cut-off point. The results indicated that 43% of bruxists increased activity (negative responders), while 36% decreased (positive responders), and in 21%, there was no change in the level. It is concluded that OA does not have significant feedback inhibition on masseter muscle motor activity during sleep. However, OA may increase slow wave sleep.

Intron sequences of arginine kinase in an intertidal snail suggest an ecotype-specific selective sweep and a gene duplication.

Many species with restricted gene flow repeatedly respond similarly to local selection pressures. To fully understand the genetic mechanisms behind this process, the phylogeographic history of the species (inferred from neutral markers) as well as the loci under selection need to be known. Here we sequenced an intron in the arginine kinase gene (Ark), which shows strong clinal variation between two locally adapted ecotypes of the flat periwinkle, Littorina fabalis. The 'small-sheltered' ecotype was almost fixed for one haplotype, H1, in populations on both sides of the North Sea, unlike the 'large-moderately exposed ecotype', which segregated for ten different haplotypes. This contrasts with neutral markers, where the two ecotypes are equally variable. H1 could have been driven to high frequency in an ancestral population and then repeatedly spread to sheltered habitats due to local selection pressures with the colonization of both sides of the North Sea, after the last glacial maximum (~18 000 years ago). An alternative explanation is that a positively selected mutation, in or linked to Ark, arose after the range expansion and secondarily spread through sheltered populations throughout the distribution range, causing this ecotype to evolve in a concerted fashion. Also, we were able to sequence up to four haplotypes consistently from some individuals, suggesting a gene duplication in Ark.

Complete lack of mitochondrial divergence between two species of NE Atlantic marine intertidal gastropods.

Some mitochondrial introgression is common between closely related species, but distinct species rarely show substantial introgression in their entire distribution range. In this study, however, we report a complete lack of mitochondrial divergence between two sympatric species of flat periwinkles (Littorina fabalis and Littorina obtusata) which, based on previous allozyme studies, diverged approximately 1 Ma. We re-examined their species status using both morphology (morphometric analysis) and neutral genetic markers (microsatellites) and our results confirmed that these species are well separated. Despite this, the two species shared all common cytochrome-b haplotypes throughout their NE Atlantic distribution and no deep split between typical L. fabalis and L. obtusata haplotypes could be found. We suggest that incomplete lineage sorting explains most of the lack of mitochondrial divergence between these species. However, coalescent-based analyses and the sympatric sharing of unique haplotypes suggest that introgressive hybridization also has occurred.

Postural and dynamic masseter and anterior temporalis muscle EMG repeatability in serial assessments.

Electromyographic (EMG) assessment has been used as a non-invasive tool to objectively assess muscle function, although with controversial research and clinical potential. The aim of this study was to assess within-, inter-subject and between-day repeatability of serial EMG recordings. The study sample included 10 asymptomatic subjects with no history of temporomandibular disorders or muscle parafunctions. Bilateral masseter and anterior temporalis muscle EMG parameters were assessed in two standardized serial recordings (day 1 to day 2) using a portable EMG equipment (ME 6000 recorder, Mega Electronics, Kuopio, Finland). The functional tasks included postural/resting activities as pre- and post-recording series of 30 s each and jaw opening/closing, intercuspal and maximal voluntary clenching activities of 5 s, repeated three times. The assessed EMG parameters included the mean amplitude, s.d. and error. In addition, the power spectrum EMG parameter assessment included the median power frequencies and the averaged EMG spectrum data values. The results of the intraclass correlation coefficient analysis indicated reliability for nearly all of the intercuspal and all clenching EMG amplitude and power spectrum parameters. This was complemented by the repeated measures anova and post hoc analyses that indicated non-significant differences between day 1 and 2 in task- and muscle-related analyses. Most variability was noted in postural and some in opening/closing tasks. In conclusion this study assessed the reliability, repeatability and limitations of postural and various dynamic masseter and temporalis EMG recordings for serial assessment.

Cortical representation of experimental tooth pain in humans.

Cortical processing of electrically induced pain from the tooth pulp was studied in healthy volunteers with fMRI. In a first experiment, cortical representation of tooth pain was compared with that of painful mechanical stimulation to the hand. The contralateral S1 cortex was activated during painful mechanical stimulation of the hand, whereas tooth pain lead to bilateral activation of S1. The S2 and insular region were bilaterally activated by both stimuli. In S2, the center of gravity of the activation during painful mechanical stimulation was more medial/posterior compared to tooth pain. In the insular region, tooth pain induced a stronger activation of the anterior and medial parts. The posterior part of the anterior cingulate gyrus was more strongly activated by painful stimulation of the hand. Differential activations were also found in motor and frontal areas including the orbital frontal cortex where tooth pain lead to greater activations. In a second experiment, we compared the effect of weak with strong tooth pain. A significantly greater activation by more painful tooth stimuli was found in most of those areas in which tooth pain had induced more activation than hand pain. In the medial frontal and right superior frontal gyri, we found an inverse relationship between pain intensity and BOLD contrast. We concluded that tooth pain activates a cortical network which is in several respects different from that activated by painful mechanical stimulation of the hand, not only in the somatotopically organized somatosensory areas but also in parts of the 'medial' pain projection system.

Painful tooth stimulation elevates matrix metalloproteinase-8 levels locally in human gingival crevicular fluid.

Recent studies have demonstrated that pulpal pain can induce neurogenic inflammatory reactions in gingiva and the expression of pro-inflammatory neuropeptides in gingival crevicular fluid (GCF). Neuropeptides co-ordinate the activity of immuno-effector cells and may influence the secretion of matrix metalloproteinase (MMP)-8, the major tissue-destructive protease in GCF. With this background, we studied whether experimental pulpal pain can trigger changes in GCF MMP-8 levels. The molecular forms of MMP-8 in the GCF of stimulated and non-stimulated teeth were analyzed by Western immunoblot, and MMP-8 levels by quantitative immunofluorometric assay. Painful stimulation of the upper incisor provoked significant elevations in GCF MMP-8 levels of the stimulated tooth. Western immunoblot revealed elevations in both neutrophil- and mesenchymal-type MMP-8 isoforms. At the same time, the GCF MMP-8 levels of the non-stimulated teeth were not changed. Analysis of these data indicated that pulpal pain can induce local elevations in MMP-8 levels in GCF.

The importance of stimulus site and intensity in differences of pain-induced vascular reflexes in human orofacial regions.

Studies in anaesthetized animals have indicated that noxious stimulation may produce marked blood flow changes in various orofacial structures, but the influence of painful stimulation on the blood flow regulation of the orofacial area of humans has been studied only to a limited extent. The purpose of this investigation was to study whether there are differences in temporal and spatial patterns of pain-induced vasoactive reflexes between various orofacial regions and hand in healthy human volunteers. Dynamic changes in blood flow in various orofacial regions elicited by painful stimulation of the tooth and finger were measured by means of Laser Doppler imaging (LDI) and computer-assisted infrared thermography (IRT). Blood flow of the finger was recorded by laser Doppler flowmetry (LDF) and plethysmography (PLET). During both stimulus paradigms there was a transient elevation in heart rate (HR) and blood pressure (BP). At the same time there was a significant blood flow decrease in the finger (LDF, PLET) and in the nose (LDI, IRT). In contrast to tooth stimulation, finger stimulation caused a more marked blood flow reduction in the finger. Only high intensity tooth stimulation, but not finger stimulation, caused a long-lasting vasodilatation both in lower and upper lip. The blood flow changes in the lips were not correlated with changes in systemic blood pressure or heart rate. In the cheek, there were no marked flow changes during either finger or tooth stimulation. These data indicate that painful tooth (regional) stimulation, but not finger (remote) stimulation, can induce a long-lasting vasodilatation in parts of orofacial tissues which cannot be explained by changes in central cardiovascular parameters. This tooth-stimulation-induced blood flow increase supports the hypothesis of a special vasodilator reflex mechanism in the orofacial area. Furthermore, tooth-stimulation-induced vasoconstriction in the nose and dilatation in the lips indicate that separate vasoactive reflex mechanisms may exist for different orofacial regions.

The influence of naloxone on dental pain threshold elevation produced by peripheral conditioning stimulation at high frequency.

The phasic dental pain threshold elevation produced by high-frequency transcutaneous electrical nerve stimulation (TENS) in healthy humans was not abolished by naloxone. A prolonged (20 min) high-frequency TENS did not cause any tonic pain threshold elevation. The results indicate that opioid-dependent systems have only a minor if any contribution to alleviation of experimental pain during high-frequency TENS.

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.

Dexamethasone attenuates exercise-induced dental analgesia in man.

The effect of dexamethasone on exercise-induced adrenocorticotropin (ACTH) secretion and dental analgesia was studied in healthy human subjects. Different levels of exercise (100-200 W) were produced by a cycle ergometer. Dental pain thresholds were tested with a constant current stimulator. Dental pain thresholds were elevated with increasing work loads, and the elevation was still significant 30 min after the end of the exercise. Dexamethasone produced a significant reversal of exercise-induced pain threshold elevations concomitantly with the suppression of exercise-induced ACTH release. The results suggest that the corticotropin releasing factor-ACTH axis is involved in the exercise-induced analgesia.

Ischemic pain nonsegmentally produces a predominant reduction of pain and thermal sensitivity in man: a selective role for endogenous opioids.

Ischemic pain was produced by a blood pressure cuff placed to the arm of healthy human subjects for 15 min which produced a mean pain score of 59% (visual analogue scale). Ischemia induced a significant dental pain threshold elevation (mean 67%) and 2 mg of naloxone did not reduce it. Thermal sensitivity of the upper lip had a tendency to reduction during ischemia and 2 mg of naloxone reduced this effect. Tactile thresholds in the forehead or in the contralateral arm were not markedly elevated. Neither ACTH nor prolactin level in the plasma was related to the dental pain threshold elevation during ischemia. The findings of the present study suggest that ischemic pain nonsegmentally produces a predominant inhibition of responses to thin afferents. Endogenous opioids may markedly contribute to the reduction of thermal sensitivity induced by ischemia, but their contribution to dental pain threshold elevations seems to be less important. Stress or other adenohypophyseal mechanisms involving the release of ACTH or prolactin do not explain the effects of ischemia found in the present study.

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.

Blood flow increase in the human lip after high-intensity tooth stimulation is not based on cholinergic mechanisms.

The purpose of this study was to investigate whether parasympathetic cholinergic pathways are involved in the regulation of orofacial blood flow. The effect of atropin (1 mg, iv.) on blood flow responses in the lower lip, nose and hand during painful tooth stimulation was studied in healthy human subjects (n=8). In all subjects, tooth stimulation caused a long lasting vasodilatation in the lower lip. During stimulation there was a transient elevation in heart rate (HR) and blood pressure (BP) concomitantly with a blood flow decrease in the finger and nose. With atropin, the pain-induced HR and BP elevations and blood flow reductions in the nose and finger were decreased. However, the pain-induced blood flow increase in the lip was not changed. This study indicates that the tooth stimulation-induced vasodilatation in the lip is not based on parasympathetic cholinergic mechanisms.

Differences between tooth stimulation and capsaicin-induced neurogenic vasodilatation in human gingiva.

Animal experiments have shown that the application of capsaicin to oral mucosa leads to a neurogenic inflammation associated with blood flow elevations in gingivomucosal tissues. In this investigation, we measured the tooth stimulation and capsaicin-evoked blood flow responses in maxillary gingiva in humans to study whether axon-reflex-mediated vasodilatation crosses the midline of the maxilla. The vasoactive reactions were mapped by laser Doppler imaging. Unilateral stimulation of alveolar mucosa and attached gingiva by capsaicin evoked a distinct neurogenic vasodilatation in ipsilateral gingiva, which rapidly attenuated at the midline. Capsaicin stimulation of alveolar mucosa provoked clear inflammatory reactions. In contrast to capsaicin stimuli, tooth stimulation produced symmetrical vasodilatations bilaterally in the gingiva. The ipsilateral responses were significantly smaller during tooth stimulation than during capsaicin stimuli. Analysis of these data suggests that capsaicin-induced inflammatory reactions in gingivomucosal tissues do not cross the midline in the anterior maxilla. The enhanced reaction found during stimulation of alveolar mucosa indicates that alveolar mucosa is more sensitive to chemical irritants than attached gingiva.

Differential effects of noxious conditioning stimulation of the cheek by capsaicin on human sensory and inhibitory masseter reflex responses evoked by tooth pulp stimulation.

In this study, we investigated whether selective activation of nociceptive primary afferent fibers by capsaicin would induce modulations on tooth-pulp-evoked sensory or inhibitory masseter reflex responses in healthy human subjects. The contribution of central N-methyl-D-aspartate (NMDA) receptor mechanisms in capsaicin-induced effects on sensory or reflex responses was evaluated by dextromethorphan, an NMDA-receptor antagonist. The inhibitory masseter reflex was evoked by electrical stimulation (constant current, single pulses) of the upper incisor while the subject was biting at 10% of his maximal force. The sensation of the tooth pulp stimulation was evaluated by visual analogue scale (VAS). The magnitude, duration, and the the latency of the reflex were determined by bite force measurements. The inhibitor masseter reflex could be induced by non-painful tooth pulp stimulation, and the inhibition was enhanced as a function of increasing stimulus intensity. Capsaicin (1%) applied topically to the skin of the cheek produced a spontaneous burning pain sensation. During capsaicin treatment, the VAS ratings for the sensation induced by tooth pulp stimulation were significantly reduced, whereas no significant changes were found in the tooth-pulp-induced masseter reflex responses. Double-blind treatment with dextromethorphan at a dose of 100 mg (= the highest does without side-effects) had no effect on sensory or reflex responses. These data indicate that noxious stimulation of the facial skin by capsaicin induces differential effects on tooth-pulp-evoked sensory and inhibitory masseter reflex responses: Sensory responses are strongly attenuated, while masseter reflex responses are not significantly changed. Dextromethorphan at a clinically applicable dose does not influence tooth-pulp-evoked sensory or reflex responses or their modulation by capsaicin. Furthermore, the lack of modulation of the masseter reflex response by capsaicin differs from the capsaicin-induced enhancement of a nocifensive limb flexion reflex described earlier.

Masticatory force and function in patients with hemispheric brain infarction and hemiplegia.

Recent functional animal studies have reported that the motor control of masticatory muscle function is bilaterally guided by both hemispheres, which may fundamentally differ from the cortical control of limb muscle function. In this study, we investigated whether unilateral cortical brain infarction induces different impairments in masticatory and upper limb motor performance. Evidence of the importance of both hemispheres in controlling masticatory movements would be greater if the masticatory function were shown to be unimpaired in patients with severe hemiplegia. The masticatory function of 16 patients with severe hemiparesis caused by brain infarction in the region of the middle cerebral artery was studied by means of interview, clinical examination, and bite-force measurements. Finger-thumb grip-force measurements and clinical examination of the upper limbs were also performed for evaluation of the effect of infarction on upper limb motor function. Localization of the infarction was confirmed with computer tomography and magnetic resonance imaging. The Scandinavian Stroke Scale demonstrated that each patient had a major unilateral cortical infarction which had caused a marked handicap with a serious impairment of upper limb function on the contralateral side. The clinical examination revealed no major signs of temporomandibular disorders, and the masticatory muscles, when examined by palpation, contracted symmetrically. None of the patients with unilateral brain infarction showed any differences in bite forces between the healthy and paralyzed sides. These results indicate that, in hemiparetic patients, great differences may exist between the motor performances of the masticatory and upper limb muscles. The present investigation clinically illustrates the importance of both hemispheres in the control of masticatory function and movements.

Blood flow increase in the orofacial area of humans induced by painful stimulation.

The purpose of this study was to investigate if painful stimulation produces blood flow changes in the tooth pulp and the facial skin in humans. Also, we attempted to find out if the possible blood flow changes induced by painful stimulation could be explained by central sympathetic and parasympathetic reflex mechanisms, by an antidromic activation of nociceptive axons (axon reflex), or by a change in central cardiovascular parameters. Laser Doppler flowmeter was used to assess the blood flow changes. Electrical tooth pulp stimulation at painful intensities induced a blood flow increase in the ipsilateral lip adjacent to the stimulus site, and vice versa. Nonpainful stimulation had no effects. Painful thermal stimulation of the upper lip also produced an increase in the blood flow of the ipsilateral upper incisor. The blood flow changes in the lip produced by dental stimulation were not correlated with changes in systemic blood pressure or heart rate. Painful electrical stimulation of the hand did not induce any changes in the pulpal blood flow, whereas painful dental stimulation produced a blood flow decrease in the finger but no change in the contralateral lip or cheek. In monkey experiments a regional block of the central conduction of the inferior alveolar nerve at the level of the mandibular foramen produced varying results: the blood flow increase in the lower incisor produced by noxious thermal stimulation of the ipsilateral lower lip was not abolished in two experiments but was abolished in other two experiments. It is concluded that painful stimulation can induce significant increases in the blood flow of the orofacial regions in humans. This increase is predominantly restricted to the region adjacent to the stimulus site and cannot be explained by changes in the central cardiovascular parameters. Central neuronal reflex mechanisms and an axon reflex may both underlie these blood flow increases.