Jeppe Nørgaard Poulsen

A human surrogate model of itch utilizing the TRPA1 agonist trans-cinnamaldehyde

The thermoreceptive transient receptor potential ankyrin 1 (TRPA1) is important in the transmission of itch, and its agonist trans-cinnamaldehyde has occasionally been reported to be a pruritogen in humans. However, no studies have accurately quantified the capabilities of trans-cinnamaldehyde to induce itch and related dysesthetic sensations. The present study examined alterations in somatosensory and vasomotor parameters in response to topical trans-cinnamaldehyde 5% and vehicle (ethanol) in 24 healthy subjects. During the study the following parameters were recorded: itch area and intensity, hyperknesis, alloknesis, neurogenic flare, skin blood flow and temperature. Trans-cinnamaldehyde evoked moderate itch sensation, flare, hyperknesis and alloknesis (p < 0.001). Blood flow and skin temperature were elevated in the area of trans-cinnamaldehyde application (p < 0.001). Significant positive correlations were found between blood flow and skin temperature, itch area and blood flow, and itch area and skin temperature. Topical trans-cinnamaldehyde proved feasible as a human itch model with applicability in studying itch mechanisms or anti-pruritic drug profiling.

Botulinum neurotoxin type A modulates vesicular release of glutamate from satellite glial cells

This study investigated the presence of cell membrane docking proteins synaptosomal‐associated protein, 25 and 23 kD (SNAP‐25 and SNAP‐23) in satellite glial cells (SGCs) of rat trigeminal ganglion; whether cultured SGCs would release glutamate in a time‐ and calcium‐dependent manner following calcium‐ionophore ionomycin stimulation; and if botulinum neurotoxin type A (BoNTA), in a dose‐dependent manner, could block or decrease vesicular release of glutamate. SGCs were isolated from the trigeminal ganglia (TG) of adult Wistar rats and cultured for 7 days. The presence of SNAPs in TG sections and isolated SGCs were investigated using immunohistochemistry and immunocytochemistry, respectively. SGCs were stimulated with ionomycin (5 μM for 4, 8, 12 and 30 min.) to release glutamate. SGCs were then pre‐incubated with BoNTA (24 hrs with 0.1, 1, 10 and 100 pM) to investigate if BoNTA could potentially block ionomycin‐stimulated glutamate release. Glutamate concentrations were measured by ELISA. SNAP‐25 and SNAP‐23 were present in SGCs in TG sections and in cultured SGCs. Ionomycin significantly increased glutamate release from cultured SGCs 30 min. following the treatment (P < 0.001). BoNTA (100 pM) significantly decreased glutamate release (P < 0.01). Results from this study demonstrated that SGCs, when stimulated with ionomycin, released glutamate that was inhibited by BoNTA, possibly through cleavage of SNAP‐25 and/or SNAP‐23. These novel findings demonstrate the existence of vesicular glutamate release from SGCs, which could potentially play a role in the trigeminal sensory transmission. In addition, interaction of BoNTA with non‐neuronal cells at the level of TG suggests a potential analgesic mechanism of action of BoNTA.

Investigating the expression of metabotropic glutamate receptors in trigeminal ganglion neurons and satellite glial cells: implications for craniofacial pain

Abstract Context/objective: Previous studies have demonstrated that various subtypes of the metabotropic glutamate receptors (mGluRs) are expressed in the dorsal root ganglion (DRG) of the peripheral nervous system (PNS), implicating that glutamate potentially contributes to sensory transmission through these receptors. While mGluR expression has been investigated largely in the DRG, the present study focused on mGluR expression on neurons and satellite glial cells (SGCs) of the trigeminal ganglion (TG). Materials and methods: To address the presence of mGluRs in rat TG neurons and their corresponding SGCs, the trigeminal ganglia from six adult male Wistar rats were isolated and immunohistochemistry and immunocytochemistry were performed. The expression of mGluR1α-, mGluR2/3- and mGluR8 on TG neurons and SGCs was investigated in tissue slices and isolated cells. Results: 35.1 ± 6.0% of the TG neurons were positive for mGluR1α, whereas 39.9 ± 7.7% and 55.5 ± 6.3% were positive for mGluR2/3 and mGluR8, respectively. Immunoreactive neurons expressing mGluRs were mainly medium- to large sized, with a smaller population of small-sized neurons showing immunoreactivity. The SGCs showed immunoreactivity toward mGluR1α and mGluR8, but not mGluR2/3, both in the tissue and in isolated cells. Conclusions: Findings from the present study showed that trigeminal neurons express mGluR1α, mGluR2/3 and mGluR8, while SGCs only express mGluR1α and mGluR8. This novel evidence may advance investigations on a possible role of mGluRs in relation to trigeminal pain transmission within the craniofacial region.

Cold and L-menthol-induced sensitization in healthy volunteers--a cold hypersensitivity analogue to the heat/capsaicin model.

Abstract Topical high-concentration L-menthol is the only established human experimental pain model to study mechanisms underlying cold hyperalgesia. We aimed at investigating the combinatorial effect of cold stimuli and topical L-menthol on cold pain and secondary mechanical hyperalgesia. Analogue to the heat–capsaicin model on skin sensitization, we proposed that cold/menthol enhances or prolong L-menthol-evoked sensitization. Topical 40% L-menthol or vehicle was applied (20 minutes) on the volar forearms of 20 healthy females and males (age, 28.7 ± 0.6 years). Cold stimulation of 5°C for 5 minutes was then applied to the treated area 3 times with 40-minute intervals. Cold detection threshold and pain, mechanical hyperalgesia (pinprick), static and dynamic mechanical allodynia (von Frey and brush), skin blood flow (laser speckle), and temperature (thermocamera) were assessed. Cold detection threshold and cold pain threshold (CPT) increased after L-menthol and remained high after the cold rekindling cycles (P < 0.001). L-menthol evoked secondary hyperalgesia to pinprick (P < 0.001) particularly in females (P < 0.05) and also induced secondary allodynia to von Frey and brush (P < 0.001). Application of cold stimuli kept these areas enlarged with a higher response in females to brush after the third cold cycle (P < 0.05). Skin blood flow increased after L-menthol (P < 0.001) and stayed stable after cold cycles. Repeated application of cold on skin treated by L-menthol facilitated and prolonged L-menthol–induced cold pain and hyperalgesia. This model may prove beneficial for testing analgesic compounds when a sufficient duration of time is needed to see drug effects on CPT or mechanical hypersensitivity.

Oxaliplatin enhances gap junction-mediated coupling in cell cultures of mouse trigeminal ganglia.

Communications between satellite glial cells and neighboring neurons within sensory ganglia may contribute to neuropathic and inflammatory pain. To elucidate the role of satellite glial cells in chemotherapy-induced pain, we examined the effects of oxaliplatin on the gap junction-mediated coupling between these cells. We also examined whether the gap junction blocker, carbenoxolone, can reverse the coupling. Primary cultures of mice trigeminal ganglia, 24-48h after cell isolation, were used. Satellite glial cells were injected with Lucifer yellow in the presence or absence of oxaliplatin (60 μM). In addition, the effect of carbenoxolone (100 μM) on coupling, and the expression of connexin 43 proteins were evaluated. Dye coupling between adjacent satellite glial cells was significantly increased (2.3-fold, P<0.05) following a 2h incubation with oxaliplatin. Adding carbenoxolone to the oxaliplatin-treated cultures reversed oxaliplatin-evoked coupling to baseline (P<0.05). Immunostaining showed no difference between expression of connexin 43 in control and oxaliplatin-treated cultures. Our findings indicated that oxaliplatin-increased gap junction-mediated coupling between satellite glial cells in primary cultures of mouse trigeminal ganglia, and carbenoxolone reversed this effect. Hence, it is proposed that increased gap junction-mediated coupling was seen between satellite glial cells in TG. This observation together with our previous data obtained from a behavioral study suggests that this phenomenon might contribute to chemotherapy-induced nociception following oxaliplatin treatment.

Simulated airplane headache: a proxy towards identification of underlying mechanisms

BackgroundAirplane Headache (AH) occurs during flights and often appears as an intense, short lasting headache during take-off or landing. Reports are limited on pathological mechanisms underlying the occurrence of this headache. Proper diagnosis and treatments would benefit from identification of potential pathways involved in AH pathogenesis. This study aimed at providing a simulated airplane headache condition as a proxy towards identification of its underlying mechanisms.MethodsFourteen participants including 7 volunteers suffering from AH and 7 healthy matched controls were recruited after meeting the diagnostic and safety criteria based on an approved study protocol. Simulation of AH was achieved by entering a pressure chamber with similar characteristics of an airplane flight. Selected potential biomarkers including salivary prostaglandin E2 (PGE2), cortisol, facial thermo-images, blood pressure, pulse, and saturation pulse oxygen (SPO) were defined and values were collected before, during and after flight simulation in the pressure chamber. Salivary samples were analyzed with ELISA techniques, while data analysis and statistical tests were handled with SPSS version 22.0.ResultsAll participants in the AH-group experienced a headache attack similar to AH experience during flight. The non-AH-group did not experience any headaches. Our data showed that the values for PGE2, cortisol and SPO were significantly different in the AH-group in comparison with the non-AH-group during the flight simulation in the pressure chamber.ConclusionThe pressure chamber proved useful not only to provoke AH-like attack but also to study potential biomarkers for AH in this study. PGE2, and cortisol levels together with SPO presented dysregulation during the simulated AH-attack in affected individuals compared with healthy controls. Based on these findings we propose to use pressure chamber as a model to induce AH, and thus assess new potential biomarkers for AH in future studies.

Individuals with dark eyes and hair exhibit higher pain sensitivity

Abstract Emerging evidence suggests that some phenotypic features, such as eye or hair colour, might predict pain. We investigated if light and dark eye and hair colour would influence pain in 60 healthy subjects divided in groups of 15 according to their eye–hair colour and gender. Pressure pain thresholds (PPTs), cold pressor test (CPT), and quality of the perceived pain were assessed. Findings indicated that dark pigmentation phenotype is more sensitive in response to CPT.

Sleep deprivation sensitizes human craniofacial muscles

Abstract It is unknown whether and how sleep deprivation influences craniofacial muscle sensitivity in healthy humans. We investigated whether total sleep deprivation (TSD) and one night of recovery sleep (RS) can alter mechanical pain sensitivity in temporal and masseter muscles. Fifteen healthy volunteers participated in three consecutive sessions. Pressure pain thresholds were measured on the temporal and masseter muscles. Both temporal and masseter muscles became sensitized after 24 h of TSD. RS reversed the muscle sensitization.

Helmet-induced headache among Danish military personnel:

Aims: External compression headache is defined as a headache caused by an external physical compression applied on the head. It affects about 4% of the general population; however, certain populations (e.g. construction workers and military personnel) with particular needs of headwear or helmet are at higher risk of developing this type of headache. External compression headache is poorly studied in relation to specific populations. This study aimed to investigate the prevalence and pattern of helmet-induced external compression headache among Danish military personnel of the Northern Jutland region in Denmark. Methods: Data acquisition was based on a custom-made questionnaire delivered to volunteers who used helmets in the Danish military service and who agreed to participate in this study. The military of the Northern Jutland region of Denmark facilitated recruitment of the participants. The questionnaires were delivered on paper and the collected (anonymous) answers (total 279) were used for further analysis. Results: About 30% of the study participants reported headache in relation to wearing a military helmet. Headache was defined as a pressing pain predominantly in the front of the head with an average intensity of 4 on a visual analogue scale of 0 (no pain) to 10 (worst pain imaginable). It was also found that helmets with different designs influenced both the occurrence of headache and its characteristics. Conclusions: This study is the first to demonstrate the prevalence and pattern of compression headache among military personnel in North Jutland, Denmark. The findings of this study call for further attention to helmet-induced external compression headache and strategies to minimize the burden.

Effect of cocoa on endorphin levels and craniofacial muscle sensitivity in healthy individuals

Abstract Aims Migraine headache is a recurrent incapacitating neurovascular disorder. A combination of events occurring within the trigeminovascular system has been suggested to underlie migraine pathogenesis, including release of inflammatory neuropeptides with subsequent effects on meningeal vessels and sensory transmission. Increased craniofacial muscle tenderness is also present in patients with migraine and a lower β-endorphin concentration has been observed in these patients. Several external factors have been identified, which can modulate migraine. One such factor is cocoa, though controversies still exist whether this substance exerts pro-algesic or analgesic effects. The present study evaluated effects of pure cocoa on a mechanically-induced headache model in healthy individuals. Serum β-endorphin concentrations were also measured to identify if any changes occurred in response to cocoa consumption. Methods Healthy volunteers (8 men and 7 women, average age: 22±1.8 years) participated in a crossover study (approval number: N-20160015) consisting of two sessions with consumption of water or cocoa. A mechanical headband (custom-made, Aalborg University) was utilized to induce a moderate headache with pain rated as 4 on a Visual Analogue Scale (VAS0–10). In each session Pressure Pain Threshold (PPT) was measured by a hand held algometer at temporalis muscles and blood samples were collected to assess β-endorphin levels by ELISA. ANOVA analysis and independent two-sample t-tests were performed for comparisons. P < 0.05 was considered as significant. Results Consumption of cocoa compared to water did not change the pressure sensitivity in the craniofacial region without mechanical headband. No changes occurred in PPT with the mechanical headband either.Regardless of substance consumption (water/cocoa), endorphin levels remained unchanged. Conclusions Findings demonstrated that pure cocoa in the applied concentration and within the timeline of this study did not seem to exert any analgesic or pro-algesic effect on the mechanical sensitivity of craniofacial muscles or β-endorphin levels.