Volume 4, Issue 2

Clear reporting of research is crucial to the scientific process. Poorly designed and reported studies are damaging not only to the efforts of individual researchers, but also to science as a whole. Standardised reporting methods, such as those already established for reporting randomised clinical trials, have led to improved study design and facilitated the processes of clinical systematic review and meta-analysis. Such standards were lacking in the pre-clinical field until the development of the ARRIVE (Animal Research: Reporting In Vivo Experiments) guidelines. These were prompted following a survey which highlighted a widespread lack of robust and consistent reporting of pre-clinical in vivo research, with reports frequently omitting basic information required for study replication and quality assessment. The resulting twenty item checklist in ARRIVE covers all aspects of experimental design with particular emphasis on bias reduction and methodological transparency. Influential publishers and research funders have already adopted ARRIVE. Further dissemination and acknowledgement of the importance of these guidelines is vital to their widespread implementation. Conclusions and implications Wide implementation of the ARRIVE guidelines for reporting of in vivo preclinical research, especially pain research, are essential for a much needed increased transparency and quality in publishing such research. ARRIVE will also positively influence improvements in experimental design and quality, assist the conduct of accurate replication studies of important new findings and facilitate meta-analyses of preclinical research.

Background In recent years, the prescription of serotonin-noradrenalin reuptake inhibitors (SNRIs) for treatment of fibromyalgia (FM) has increased with reports of their efficacy. The SNRI milnacipran is approved by the U.S. Food and Drug Administration (FDA) for treatment of FM, yet, the mechanisms by which milnacipran reduces FM symptoms are unknown. A large number of neuroimaging studies have demonstrated altered brain function in patients with FM but the effect of milnacipran on central pain processing has not been investigated. The primary objective of this study was to assess the effect of milnacipran on sensitivity to pressure-evoked pain in FM. Secondary objectives were to assess the effect of milnacipran on cerebral processing of pressure-evoked pain using fMRI and the tolerability and safety of milnacipran 200 mg/day in FM. Methods 92 patients were randomized to either 13-weeks milnacipran treatment (200 mg/day) or placebo in this double-blind, placebo-controlled multicenter clinical trial. Psychophysical measures and functional MRI (fMRI) assessments were performed before and after treatment using a computer-controlled pressure-pain stimulator. Here, we present the results of several a priori defined statistical analyses. Results Milnacipran-treated patients displayed a trend toward lower pressure-pain sensitivity after treatment, compared to placebo, and the difference was greater at higher pain intensities. A single group fMRI analysis of milnacipran-treated patients indicated increased pain-evoked brain activity in the caudatus nucleus, anterior insula and amygdala after treatment, compared to before treatment; regions implicated in pain inhibitory processes. A 2 × 2 repeated measures fMRI analysis, comparing milnacipran and placebo, before and after treatment, showed that milnacipran-treated patients had greater pain-evoked activity in the precuneus/posterior cingulate cortex after treatment; a region previously implicated in intrinsic brain function and FM pathology. This finding was only significant when uncorrected for multiple comparisons. The safety analysis revealed that patients from both treatment groups had treatment-emergent adverse events where nausea was the most common complaint, reported by 43.5% of placebo patients and 71.7% of milnacipran-treated patients. Patients on milnacipran were more likely to discontinue treatment because of side effects. Conclusions Our results provide preliminary indications of increased pain inhibitory responses in milnacipran-treated FM patients, compared to placebo. The psychophysical assessments did not reach statistical significance but reveal a trend toward higher pressure-pain tolerance after treatment with milnacipran, compared to placebo, especially for higher pain intensities. Our fMRI analyses point toward increased activation of the precuneus/posterior cingulum in patients treated with milnacipran, however results were not corrected for multiple comparisons. The precuneus/posterior cingulum is a key region of the default mode network and has previously been associated with abnormal function in FM. Future studies may further explore activity within the default mode network as a potential biomarker for abnormal central pain processing. Implications The present study provides novel insights for future studies where functional neuroimaging may be used to elucidate the central mechanisms of common pharmacological treatments for chronic pain. Furthermore, our results point toward a potential mechanism for pain normalization in response to milnacipran, involving regions of the default mode network although this finding needs to be replicated in future studies.

Background and aims Preclinical data suggest that the chemokine receptor 2 (CCR2) is involved in the pathophysiology of neuropathic pain through modulation of neuronal excitability, synaptic transmission and activation of spinal cord microglia. CCR2-antagonists have shown to be effective in preclinical models of neuropathic pain. The aim of this study was to evaluate the analgesic efficacy, safety and tolerability of a novel CCR2-antagonist, AZD2423, in patients with painful diabetic neuropathy (PDN). Methods This was a double-blind, randomized, parallel-group, multi-center study in patients with symmetric distal sensory polyneuropathy due to type 1 or 2 diabetes and duration of neuropathic pain between 3 months and 5 years. Concomitant treatment with neuropathic pain medications (e.g. anticonvulsants, tricyclic antidepressants, serotonin-noradrenaline uptake inhibitors, opioids, topical lidocaine or capsaicin) was not allowed. 134 patients with PDN were equally randomized to 28 days oral administration of 20 mg AZD2423,150 mg AZD2423, or placebo. The primary efficacy variable was the change of average pain score from 5-days baseline to the last 5 days of treatment, measured with numerical rating scale (NRS, 0–10). The secondary efficacy measures included NRS worst pain scores, patient global impression of change, pain interference on sleep and activity, and neuropathic pain symptom inventory (NPSI). Results The change of NRS average pain score was not significantly different between treatment groups (AZD2423 20mg: –1.50; AZD2423 150 mg: –1.35; placebo: –1.61). The NPSI total score and three out of five subscores (evoked pain, pressing/deep pain and paresthesia/dysesthesia) tended to be reduced more by AZD2423 150 mg than by placebo. No other secondary efficacy variables differed between treatment groups. The frequency and type of adverse events for AZD2423 were similar to placebo. The achieved plasma levels of AZD2423 in the two dose groups were in line with predictions from pharmacokinetic data previously obtained in healthy volunteers. Dose-dependent increase of plasma levels of the ligand of CCR2 (CCL2; chemokine ligand 2) and decrease of the mean levels of monocytes (–27% by AZD2423 150 mg) suggested that the administrated doses of AZD2423 interacted with the CCR2 target. Conclusion The CCR2-antagonist AZD2423 showed no analgesic efficacy in PDN based on NRS average pain scores and global and functional pain outcome measures. The NPSI data suggested possible effects on certain sensory components of pain. There were no major safety or tolerability concerns. Implications Treatment with a CCR2-antagonist does not have a clinically important analgesic effect in an overall PDN population.

Aabstract Introduction Chronic musculoskeletal pain represents a significant health problem among adults in Norway. The prevalence of chronic pain is reported to be 35-53% in cross sectional studies of both genders. For many years, it has been a common opinion among medical doctors that chronic pain may indeed reduce a person’s quality of life, but not affect life expectancy. However, over the previous two decades, reports about mortality and cause of death in individuals with chronic pain have been published. So far, several studies conclude that there is an increased mortality in patients with chronic pain, but it is not clear what causes this. Increased occurrences of cardio-vascular death or cancer death have been reported in some studies, but not verified in other studies. Aims of the study The aims of this study were to estimate the mortality rate in females with different extent of pain, to identify potential risk factors for death and to investigate if the causes of death differ according to prior reported pain. Methods This is a prospective population-based study of all women between 20 and 50 years registered in Arendal, Norway, in 1989 ( N = 2498 individuals). At follow-up in 2007, 2261 living females were retraced, 89 had died. All subjects received a questionnaire containing questions about chronic pain (pain ≥ 3 months duration in muscles, joints, back or the whole body) as well as 13 sub-questions about pain-modulating factors, non-specific health complaints and sleep problems, by mail in 1990, 1995 and 2007. Only subjects who answered the questionnaire in 1990 were included in the analyses. Of the deceased, 71 had answered the questionnaire in 1990. A multivariate model for cox regression analysis was used in order to clarify if chronic pain, sleep problems, feeling anxious, frightened or nervous and number of unspecific health were risk factors for death. The causes of death of 87 of the deceased individuals were obtained by linking the ID-number with the Norwegian Cause of Death Registry. Results The ratio of deceased responders was 2% (14/870) among those with no pain versus 5% (57/1168) among those with chronic pain at baseline. When separating into chronic regional pain and chronic widespread pain, the mortality rate was respectively 4% and 8% in the different groups. Age adjusted hazard ratio for mortality rate in individuals with initially chronic pain was [HR 2.5 (CI 1.4–4.5)] compared to pain free individuals. In the multivariate analysis, having chronic pain [HR 2.1 (1.1–4.2)] and feeling anxious, frightened or nervous [HR 3.2 (1.8–5.6)] were associated with increased risk of death. There was no difference in death from cardiovascular disease or malignancies between the groups of pain free individuals vs. the group of individuals with chronic pain. Conclusion The mortality rate was significantly higher for individuals with chronic pain compared to pain free individuals, adjusted for age. In addition, feeling anxious, frightened or nervous were risk factors for death. There was an increase in all-cause mortality.

Background and objectives Unmyelinated C-fibres comprise the largest group of somatic afferents and have demonstrated a crucial role not only in the perception of high-threshold mechanically, thermally or chemically induced pain, but also in non-harmful low-threshold mechanical stimuli [1,2]. The objective of our study was to characterize differential sensitivity changes of C-fibre related subclasses of high-threshold and low-threshold polymodal nociceptors and low-threshold mechanoreceptors to the local anaesthetic (LA) mepivacaine during nerve block of the purely sensory lateral femoral cutaneous nerve (LFCN) in human. We assumed a diverse response of different classes of afferents to the two different concentrations of the LA mepivacaine (Scandicaine). Methods In a double-blind randomized experimental setting, an ultrasound-guided nerve block of the LFCN was performed in 10 healthy male subjects, each with two different concentrations of mepivacaine (0.5 and 1%). Responsiveness of afferent nerve fibres to different noxious and non-noxious stimuli was tested by Quantitative Sensory Testing (QST) 30, 180, and 300 min after nerve block. Both LA concentrations of mepivacaine were compared for time course of the areas of anaesthesia for the tested sensory modalities. Results Initial extension of anaesthetic areas at 30 min did not differ between both LA concentrations. At 180 min only the anaesthetic areas to nociceptive stimuli were reduced at the site of lower mepivacaine injection (260mN: 204mm 2 (18; 244; median difference and 95% confidence interval; p < 0.05), heat: 276mm 2 (3; 305)). In contrast, no significant differences were found between the two concentration when non-nociceptive stimuli were used (100mN: 187mm 2 (4; 240), p >0.05, brush: 159mm 2 (–59; 202)). Conclusion Equal initial sizes of anaesthesia areas for all sensory modalities can be explained by supramaximal perineural LA molecule concentration in both administered mepivacaine dosages. Upon washout of the LA nociceptive function is restored faster as compared to non-nociceptive sensation and higher concentration of the LA are required to maintain the analgesia. Quantitative sensory testing is able to detect different susceptibility of low threshold mechanosensors and subtypes of nociceptive C-fibres to mepivacaine. Using painful mechanical, heat and electrical stimulation different classes of nociceptors will be activated. The analgesic areas to electrical stimulation were particularly small; one might therefore hypothesize that the proposed protocol allows to also differentiate mechano-insensitive (“silent”) and mechanosensitive (“polymodal”) nociceptors. Implications QST is a non-invasive method to functionally examine sensory modalities and their pharmacological modulation in humans. The method is sufficiently sensitive to differentiate the analgesic properties of mepivacaine at 0.5 and 1% and might also be adequate to different classes of nociceptors. Further development of nociceptive stimuli including supra-threshold encoding characteristics will enable to investigate peripheral analgesic effects more specifically and thus might help to design new analgesics with preferential effect on high frequency discharge of nociceptors.

Aabstract Background and purpose Women exhibit higher prevalence of most painful disorders. Several explanations have been proposed for this discrepancy, one being that endogenous pain modulatory pathways, which affect incoming nociceptive signals, act differently in men and women. A less efficient pain inhibitory system has been proposed as a contributing factor to explain why women exhibit higher prevalence of most painful disorders. The present study determined whether muscle pain, induced experimentally by electrical stimulation, is inhibited by a painful heat stimulus. This conditioned pain modulation (CPM) paradigm was used to determine whether women show signs of reduced inhibition compared to men. Methods Forty self-reported healthy individuals (20 female, 20 male) participated in a cross-over design with painful and non-painful heat as a conditioning stimulus. Test stimuli were painful intramuscular electrical stimulation of the tibialis anterior muscle at two intensities; low (1.1 × pain threshold) and high (1.6 × pain threshold). Painful conditioning was contact heat (45–49 ° C) to the contralateral forearm. Nonpainful conditioning was contact heat at 35 °C. Ten test stimuli were delivered in three blocks (before, during and after conditioning) in two sessions (painful and non-painful conditioning). The women were tested during days 12-14 of the menstrual cycle. This interval corresponds to the ovulatory phase of the menstrual cycle, the interval during which women are reported to show the largest inhibitory effects. Results Test stimuli were rated significantly lower during painful conditioning, compared with before conditioning. This was found for both low and high test stimulus intensities. Anonspecific attenuation was seen during non-painful conditioning for the low test stimulus intensity. Test stimuli were rated significantly lower also 3 min after conditioning, compared with before conditioning. The inhibitory effects were not different between men and women. Similar findings were obtained also if six non-CPM-responders (subjects rating test stimuli higher during conditioning than before conditioning) were excluded. Conclusions and implications The present findings indicate that painful contact heat inhibits electrically induced muscle pain and that inhibition was not different between men and women, when women were tested in the interval 12-14 days after their last menstruation. Some inhibition of muscle pain was seen during non-painful conditioning, indicating that nonspecific inhibitory effects were triggered. Also the nonspecific inhibitory effects were similar in men and women.

ABSTRACT Background Peripheral and central sensitisation is prominent in knee osteoarthritis (KOA) and could be important for the reduced efficacy in some cases after as well surgery as pharmacological interventions. Although sensitisation is important in KOA it is not known to what degree it contributes to the overall clinical pain problem. The aim was therefore to investigate how much a combination of quantitative pain measures assessing various pain mechanisms (local and spreading hyperalgesia, temporal and spatial summation, descending inhibition) could predict peak pain intensity in patients with KOA. Methods While resting in a comfortable recumbent position the pressure pain thresholds (PPT) in the peripatellar region (eight locations) and at the tibialis anterior muscle (TA) were assessed by handheld pressure algometry, computer-controlled pressure algometry and cuff-algometry in the affected leg of 17 KOA patients without pain or sensory dysfunctions in other regions than the knee. Cuff-algometry was used to detect spatial pain summation of the lower leg. Temporal pain summation was assessed by repeated pressure stimulation on the TA muscle. The conditioning pain modulation (CPM) was evaluated by conditioning tonic arm pain and by PPT from the peripatellar region. The participants rated their peak pain intensity in the previous 24 h using on a 10 cm visual analogue scale. Results A multiple-regression model based on TA pressure pain sensitivity (spreading sensitisation) and temporal pain summation on the lower leg accounted for 55% of the variance in peak pain intensity experienced by the patients (P=0.001). Significant correlations ( P < 0.05) were found between PPTs assessed by handheld pressure algometry in the peripatellar region and at TA ( R = 0.94), PPTs assessed by computer-controlled pressure algometry and handheld pressure algometry in the peripatellar region ( R = 0.71), PPTs assessed by computer-controlled pressure algometry in the peripatellar region and handheld pressure algometry at TA ( R = 0.71) and temporal summation at the knee and at TA ( R = 0.73). Conclusion Based on the multiple regression model 55% variance of the perceived maximal pain intensity in painful KOA could be explained by the quantitative experimental pain measures reflecting central pain mechanisms (spreading sensitisation, temporal summation). The lack of other correlations between the methods used in assessing pain mechanisms in this study highlights the importance of applying different tests and different pain modalities when assessing the sensitised pain system as different methods add complementary information. Implications Clinical pain intensity can be explained by influences of different central pain mechanisms in KOA. This has implications for pain management in KOA where treatment addressing central pain components may be more important than previously acknowledged.

Aabstract Background and purpose Central hypersensitivity, defined as an increased excitability of the central nervous system, is considered as the main factor behind facilitation of central pain processes and is probably a very important factor in the induction and maintenance of chronic pain. Widespread hyposensitivity is less studied than hypersensitivity states, but recent work indicates that hypoesthesia may be present in chronic non-neuropathic pain conditions and could have negative prognostic value. Electrical pain and reflex thresholds are well established measures of central pain sensitivity in human pain research. One potential application of these assessments in clinical practice is the detection of central hyper- or hyposensitivity in individual patients. In order to identify these disturbances in the central pain processing of individual patients, knowledge of reference values is essential. We computed percentile normative values of nociceptive withdrawal reflex (NWR) and pain thresholds to different electrical stimulation paradigms. The aim was to provide reference values for the assessment of widespread central hyper- and hyposensitivity in individual patients. Methods 300 pain-free subjects (150 males and 150 females, 18-80 years) were studied. Pain and reflex thresholds to single and repeated electrical stimulation (applied on the innervations area of the sural nerve), and the area of reflex receptive fields (RRF) were recorded. The RRF area was defined as the skin area of the sole of the foot from which a NWR could be evoked in the tibialis anterior muscle, expressed as proportion of the foot sole. For the threshold assessments, quantile regressions were performed to compute critical normative values for widespread central hypersensitivity (5th, 10th and 25th percentiles) and hyposensitivity (75th, 90th and 95th percentiles). For the RRF the opposite applied, computing normative values for widespread central hypersensitivity as 75th, 90th and 95th percentiles, and normative values for hyposensitivity as 5th, 10th and 25th percentiles. The following covariates were included in the regression analyses: gender, age, height, weight, body mass index, body side of testing, depression (Beck Depression Inventory), anxiety (State-Trait-Anxiety-Inventory), catastrophizing (Catastrophizing Scale of the Coping Strategies Questionnaire) and Short-Form 36. Results Age had a significant impact on the subjective pain threshold to single electrical stimuli. The reflex threshold to single electrical stimuli was lower on the dominant compared to the non-dominant side. Therefore, the percentiles for single stimulus pain threshold have been stratified by age and the percentiles for single stimulus reflex threshold by body side (dominant vs. non-dominant). Critical normative values of all tests were computed for widespread central hypersensitivity (5th, 10th and 25th percentiles) and hyposensitivity (75th, 90th and 95th percentiles). The values are provided in the table of the manuscript. Conclusions and implications The computed estimates of critical normative values for the electrical pain test can be used in clinical practice for the assessment of widespread central hypersensitivity and hyposensitivity in individual patients, and in future research on mechanism-based treatment of chronic pain.