Silvia Lo Vecchio

A dose-response study of topical allyl-isothiocyanate (mustard oil) as human surrogate model of pain, hyperalgesia, and neurogenic inflammation

Abstract Despite being a ubiquitous animal pain model, the natural TRPA1-agonist allyl isothiocyanate (AITC, also known as “mustard oil”) has only been sparsely investigated as a potential human surrogate model of pain, sensitization, and neurogenic inflammation. Its dose–response as an algogenic, sensitizing irritant remains to be elucidated in human skin. Three concentrations of AITC (10%, 50%, and 90%) and vehicle (paraffin) were applied for 5 minutes to 3 × 3 cm areas on the volar forearms in 14 healthy volunteers, and evoked pain intensity (visual analog scale 0-100 mm) and pain quality were assessed. In addition, a comprehensive battery of quantitative sensory tests was conducted, including assessment of mechanical and thermal sensitivity. Neurogenic inflammation was quantified using full-field laser perfusion imaging. Erythema and hyperpigmentation were assessed before, immediately after, and ≈64 hours after AITC exposure. AITC induced significant dose-dependent, moderate-to-severe spontaneous burning pain, mechanical and heat hyperalgesia, and dynamic mechanical allodynia (P < 0.05). No significant differences in induced pain hypersensitivity were observed between the 50% and 90% AITC concentrations. Acute and prolonged inflammation was evoked by all concentrations, and assessments by full-field laser perfusion imaging demonstrated a significant dose-dependent increase with a ceiling effect from 50% to 90%. Topical AITC application produces pain and somatosensory sensitization in a dose-dependent manner with optimal concentrations recommended to be >10% and ⩽50%. The model is translatable to humans and could be useful in pharmacological proof-of-concept studies of TRPA1-antagonists, analgesics, and anti-inflammatory compounds or for exploratory clinical purposes, eg, loss- or gain-of-function in peripheral neuropathies.

The Effect of Combined Skin and Deep Tissue Inflammatory Pain Models

OBJECTIVE Cutaneous hyperalgesia is prominent in the ultraviolet-B (UVB) model of inflammatory pain. This study investigated possible interactions between cutaneous and deep tissues hyperalgesia. METHODS A total of 16 healthy volunteers participated in the study. Skin inflammation was induced unilaterally by UVB irradiation (three times of the individual minimal erythema dose) in square-shaped areas on the upper-trapezius and lower-back. Moderate delayed onset muscle soreness (DOMS) was induced bilaterally in the low-back by eccentric exercise. Cutaneous blood flow, mechanical thresholds, pressure pain thresholds (PPTs), temporal summation to repetitive pressure stimulation, and stimulus-response functions (SR curve) relating graded pressure stimulations and pain intensity were measured within and outside the irradiated areas, before and 24 hours after irradiation and eccentric muscle exercise. RESULTS Compared with baseline (P < 0.05): the assessments 24 hours after irradiation demonstrated: 1) increased superficial blood flow inside and outside the irradiated areas and in the DOMS site; 2) reduced mechanical thresholds within the irradiated areas; 3) left-shifted SR curve function within and outside the irradiated areas; and 4) facilitated temporal summation of pain inside the irradiated areas and in the DOMS site. There was no significant influence of muscle hyperalgesia on skin measures in normal or UVB-inflamed skin. CONCLUSIONS Moderate degrees of muscle sensitization could not facilitate UVB-induced cutaneous mechanical sensitivity, whereas UVB-induced neurogenic inflammation is enhanced when the DOMS is present.

Topography of itch: evidence of distinct coding for pruriception in the trigeminal nerve

Introduction: Little is known about the topographical distribution of pruriception (in particular for nonhistaminergic itch), although conditions with chronic itch frequently occur in distinct anatomic and often bilateral patterns. This study aimed to investigate regional differences in the sensitivity to itch stimuli by assessing the intensity of itch, pain, and cutaneous neurogenic flare evoked by histamine and cowhage in different anatomic regions in 20 healthy volunteers. Methods: Itch was induced by 1% histamine applied with a prick lancet or by insertion of 25±5 cowhage spicules in 4 regions: volar/dorsal forearm, lower back, and chin. The duration and intensity of itch and pain following each pruritic stimulus were measured by a continuous visual analogue scale (VAS0-100). Sensitivity to touch-evoked itch was assessed by von Frey filaments and cutaneous flare was quantified by full-field laser perfusion imaging. Results: Peak itch intensity was lower at the chin (19.4±3.6) compared with other areas (mean of 3 locations; 41.3±4.4), independently of whether histamine or cowhage was applied (P<0.01). Baseline sensitivity to touch-evoked itch was higher on the chin (P<0.01), but here hyperknesis did not develop in contrast to other areas (P<0.05). Cutaneous flare was more intense but had a smaller dispersion at the chin, compared with other areas (P<0.01). Discussion: In conclusion, sensitivity to histaminergic and non-histaminergic itch diverges considerably between body regions. Lower density of pruriceptive CMH and CMI-neurons or distinct neuronal substrates for itch in the mandibular part of the trigeminal area may explain the observed reduced itch and vasomotor responses.

Hyperalgesia and allodynia to superficial and deep-tissue mechanical stimulation within and outside of the UVB irradiated area in human skin

Abstract Background and aims The ultraviolet-B (UVB) inflammatory model is a well-established model of inflammatory pain. This study investigated whether UVB-induced cutaneous inflammation would enhance pain responses from the underlying deep somatic areas. Methods Skin inflammation was induced, in 24 healthy volunteers, by UVB irradiation (three times of the individual minimal erythema UVB dose) in square-shaped areas on the forearm and lower back. Assessments of cutaneous blood flow, pin-prick thresholds, pressure pain thresholds and tolerance, stimulus–response functions relating graded pressure stimulations and pain intensity (visual analogue scale, VAS) were performed within and outside the irradiated area. Results Twenty-four hours after UVB irradiation, a significant increase in superficial blood flow in the irradiated skin area was demonstrated compared with baseline (P < 0.01) indicating that inflammation was induced. Compared with baseline, UVB irradiation significantly reduced the pin-prick thresholds, pressure pain thresholds and tolerance within and outside of the irradiated area (P < 0.05). The stimulus–response function was left-shifted compared with baseline both within and outside the irradiated area (P < 0.05) with a more pronounced left-shift within the irradiated area (P < 0.01). Application of topical anaesthesia 24h after irradiation in 5 subjects, both within and outside the irradiated area, could only increase the pin-prick thresholds outside the irradiated area. Conclusion The UVB irradiation of the skin not only provokes cutaneous primary and secondary hyper-algesia but also causes hyperalgesia to blunt pressure stimulations 24h after the UVB exposure. Implications The presented UVB model can be used as a translational model from animals into healthy subjects. This model can potentially be used to screen drug candidates with anti-inflammatory properties in early stages of drug development.

Hyperalgesia in human skin and deep-tissues inside and outside of a UVB irradiated area

Abstract Background/aims The ultraviolet B (UVB) inflammatory pain model is often used to induce a steady hyperalgesic area in human skin. UVB causes a well-described erythema, developing maximal response within about 24 h. The aim of the present study was to investigate if cutaneous UVB irradiation can influence both superficial and deep-tissue mechanical pain thresholds in the site of irradiation and in the surrounding area. Methods An area of 3 cm × 4 cm, located on the low back of 16 healthy volunteers, was irradiated by UVB (Medlight, Germany; 3xMED: Minimal Erythema Dose). The degree of inflammation was detected by measuring superficial blood flow before and after irradiation, inside and outside the stimulated area. Applying quantitative sensory assessments, mechanical pain threshold changes were detected one day after irradiation, within and outside of the irradiated area. Sensitivity to cutaneous mechanical stimuli was assessed using pin prick and deep-tissue pressure pain thresholds were evaluated on 12 spots (4 within and 8 outside, 1.5 cm distant from the irradiated area) by a computer-controlled pressure algometer (Aalborg University, Denmark; 1.0 cm2 flat probe, 0.5 cm2 flat probe and a V-shaped probe with a contact surface of 0.03 cm2). Results 24 h after exposure, the irradiated skin showed clear erythema with a boundary matching the irradiated area and a statistically significant increase in cutaneous blood flow (P < 0.001) compared with baseline assessment. Cutaneous pin prick pain thresholds and deep-tissue pressure pain thresholds (all probes) were significantly decreased inside and outside the irradiated area (P < 0.05). Conclusions Cutaneous UVB irradiation reduces mechanical pain thresholds to pin-prick and pressure stimulation which may indicate allodynic responses in both the skin and in deep-tissues. Expansion of the responses to areas outside the irradiated zone confirmed the presence of secondary hyperalgesia to mechanical stimuli.

UVB- and NGF-induced cutaneous sensitization in humans selectively augments cowhage- and histamine-induced pain and evokes mechanical hyperknesis

Exaggerated itch responses to pruritic chemical provocations and mechanical stimuli are evident in patients with chronic itch, for example, in atopic dermatitis. Currently used human models of itch do not account for such itch sensitization features, and the mechanisms underlying clinical itch sensitization are unknown. This study utilized two established human models of cutaneous nociceptive sensitization to explore how pre‐established inflammatory hyperalgesia (ultraviolet‐B‐irradiation; “UVB”) and non‐inflammatory neurotrophic pain sensitization (nerve growth factor; “NGF”) alter sensitivity to chemical and mechanically evoked itch. Twenty healthy volunteers participated in the UVB experiment. Six volar forearm areas (2 cm diameter) were UVB irradiated with ≤2 × minimal erythemal dose, and two non‐irradiated areas were used as controls. Sixteen healthy volunteers participated in the NGF experiment and had 2 μg intradermally injected (4 × 50 μL in 2 cm diameter areas) into both volar forearms. Isotonic saline was applied as control. Pain sensitivity measurements (mechanical and heat pain thresholds) were conducted to validate the models. Subsequently, itch was evoked using histamine and cowhage spicules in the sensitized skin areas, and itch/pain was rated using visual analogue scales. Mechanical hyperknesis (increased itch to punctuate stimuli) was probed with von Frey filaments before/after each itch provocation. Both UVB‐ and NGF models induced robust primary mechanical hyperalgesia (P < .01) and hyperknesis (P < .05). Neither of the models augmented itch in response to chemical itch provocations but significant increases specifically for pain ratings were observed for both histamine and cowhage (P < .05). This suggests that these models are of limited value as proxies for itch sensitization to pruritogens observed, e.g., in inflammatory dermatoses.