archives of oral biology 52 (2007) 149–153

available at www.sciencedirect.com

journal homepage: www.intl.elsevierhealth.com/journals/arob

Effects of capsaicin and menthol on oral thermal

sensory thresholds

A. Kalantzis, P.P. Robinson, A.R. Loescher *

University of Sheffield, UK

article info abstract

Article history: Objective: To determine the long-term effect of capsaicin and short-term effect of menthol
Accepted 8 September 2006 on oral thermal thresholds.
Design: The thresholds for cold detection (CDT), warm detection (WDT), cold pain (CPT) and
Keywords: warm pain (WPT) were determined in 11 regular chilli-eaters (capsaicin group) and 11
Oral control subjects that were closely matched for age, gender and ethnicity. The effect of
Quantitative thermal sensory menthol was determined by asking all 22 participants to suck a lozenge containing 0.52%
thresholds menthol for 5 min.
Capsaicin Results: An ANOVA revealed a significant difference between the capsaicin and control
Menthol groups (P = 0.014), with the greatest difference in the WDT (capsaicin group
4.7  2.7 [S.D.] 8C; control group 2.3  2.2 8C). Immediately after sucking a menthol lozenge
there was a significant rise in the CDT (2.2  1.1 8C to 5.9  6.2 8C; P < 0.01) and WDT
(3.6  2.7 8C to 7.6  4.4 8C; P < 0.001).
Conclusions: The consumption of foods containing capsaicin and menthol significantly
alters thermal sensory thresholds in the oral cavity. Dietary habits should therefore be
taken into account when intra-oral thermal thresholds are determined.
# 2006 Elsevier Ltd. All rights reserved.

1. Introduction and a few thinly myelinated Ad fibres.4 Capsaicin applied

The accurate determination of thermal sensory thresholds,
and therefore the function of thinly myelinated Ad and non-
myelinated C fibres, has become increasingly routine in the
assessment of patients with neuropathy. Although many
studies have now reported thermal sensory thresholds within
the oral cavity (for example1–3), the influence of the diet on oral
thermal thresholds has not previously been considered.
Capsaicin is the main pungent ingredient in ‘‘hot’’ chilli
peppers and despite the burning pain we experience when
eating ‘‘hot’’ dishes, it is for most people an enjoyable
experience. The capsaicin-sensitive receptor has been identi-
fied as the TRPV1 (transient receptor potential vanilloid type 1)
and is located on a subpopulation of non-myelinated C fibres
topically to the skin causes a burning pain and mechanical
hyperalgesia5,6 but the repeated topical application of capsaicin
to skin over a period of time causes the epidermal nerve fibres to
degenerate and the warm thermal thresholds to diminish.6–8
The oral cavity is richly innervated by TRPV1 immunoreactive
nerve fibres,9,10 which are stimulated when we eat ‘‘hot’’ dishes
containing chilli. The first aim of this study was to determine if
the regular consumption of capsaicin in ‘‘hot’’ foods could alter
oral thermal thresholds in man.
A low concentration of menthol and eucalyptus are
frequently added to sweets, cigarettes and even toothpaste
because of their cooling and freshening effect, whilst higher
concentrations may cause burning, irritation or even pain.11 In
addition to the expected enhancement of cold sensation12

* Corresponding author at: Department of Oral Surgery, School of Clinical Dentistry, University of Sheffield, Claremont Crescent, Sheffield
S10 2 TA, UK. Tel.: +44 114 2717849; fax: +44 114 2717863.
E-mail address: a.loescher@sheffield.ac.uk (A.R. Loescher).
0003–9969/$ – see front matter # 2006 Elsevier Ltd. All rights reserved.
doi:10.1016/j.archoralbio.2006.09.001
150 archives of oral biology 52 (2007) 149–153

menthol has also been shown to enhance warmth perception Table 1 – Ingredients of Hall’s menthol lozenges
in the lip13 forearm14 and tongue.15 Menthol directly stimu- Menthol (0.52%, w/w)
lates the TRPM8 receptor (transient receptor potential mela- Eucalyptus oil (0.26%, w/w)
statin type 8), which like TRPV1 is expressed in small diameter Sugar (2.018 g sucrose)
Glucose syrup (0.308 g glucose)
primary sensory neurones. TRPM8 is also activated by
Water
temperatures of 25–28 8C and has been localised within the E270 (lactic acid), E325 (sodium lactate), E330 (citric acid), E332
trigeminal ganglion and on the fungiform papillae of the (potassium citrate)
tongue.16 Like capsaicin, menthol not only activates the cold Peppermint flavour, cooling flavours
specific Ad fibres but also a population of C fibres. The cooling
and soothing effects of lozenges containing only a small
amount of menthol are well recognised. The second aim of 2.3. Statistical analysis
this study was to determine if the small quantity of menthol in
a lozenge could alter oral thermal thresholds. Statistical analysis was undertaken initially using an ANOVA
with a random effect to take account of repeated measure-
ments on any individual subject and fixed factors for modality
2. Methods (CDT, WDT, etc.), groups (controls or capsaicin) and modality
diet interactions. A significant difference revealed by the
2.1. Subjects ANOVA (P-values < 0.05 were regarded as significant) was
investigated by either an independent-sample t-test (capsaicin
Twenty-two healthy Caucasian female volunteers (age range and control groups) or a paired t-test (pre- and post-menthol)
20–28) participated in the study and were divided into two using a Bonferroni correction.
groups. The capsaicin group (11 participants) who ate chilli
(capsaicin) at least three times each week (average 3.7) and the
control group (11 participants) who ate chilli less than twice a 3. Results
month. The project had been reviewed by the South Yorkshire
Research Ethics Committee (SSREC/02/96). 3.1. Capsaicin

2.2. Quantitative thermal sensory analysis The means and standard deviations of the threshold for the
CDT, WDT, CPT and WPT are shown in Figs. 1 and 2. The
The investigations were undertaken in a quiet room, free from ANOVA revealed a significant difference (P = 0.014) between
distractions (temperature 18–20 8C) with the subjects sitting in the control and capsaicin groups for at least one modality and
a comfortable armchair. Thermal thresholds were measured the greatest difference was in the warm detection thresholds
using the Medoc thermal sensory analyser II (TSA) (Medoc, (capsaicin 4.7  2.7 8C, controls 2.3  2.2 8C). However, because
Israel) and a 16 mm  16 mm Peltier thermode. The TSA was of the use of a multiple test correction factor, post hoc analysis
controlled by computer and the progress of the test was using t-tests indicated that this difference (P = 0.04) did not
visualised on a screen seen by the researcher but not the achieve significance (using a Bonferroni correction a P-
participant. The thermode was held by the subject with light value < 0.0125 is required for significance).
pressure on the right side of the dorsal surface of the tongue.
The thresholds were measured from a baseline of 32 8C, using 3.2. Menthol
the method of limits. A temperature change of 1.0 8C s 1 was
used for cold and warm detection thresholds (CDT and WDT) Comparisons of the mean values of CDT, WDT, CPT and WPT
and 1.5 8C s 1 for cold and warm pain thresholds (CPT and before and after treatment of the mouth with menthol are
WPT). Subjects were instructed to press a button, which shown in Figs. 3 and 4. The ANOVA revealed a significant
stopped the stimulus, immediately they felt the thermode
turning cold (CDT) or warm (WDT), or when the cold sensation
became uncomfortable or unpleasant (CPT) or the warm
sensation became uncomfortable or unpleasant (WPT). Ther-
mal thresholds were always determined in the following
order: CDT, WDT, CPT and lastly WPT. For each of these
thermal sensory modalities five consecutive stimuli were
applied with a variable inter-stimulus interval of 4–6 s, and the
median value recorded. Each thermal threshold was then
expressed as the change in temperature from the baseline of
32 8C required to elicit a response.
All 22 subjects were asked to suck a lozenge (Halls Extra
Strong menthols, Cadbury Trebor Basset, Birmingham, UK,
Table 1) containing 0.52% menthol for 5 min immediately Fig. 1 – A bar chart showing the CDT and WDT (WS.D.) in
before repeating the measurement of the thermal thresholds both the control and the capsaicin groups. The thresholds
(CDT, WDT, CPT and WPT) on the right dorsal surface of the are expressed as the change in temperature from the
tongue. baseline (32 8C) that was required to elicit a response.
 archives of oral biology 52 (2007) 149–153
Fig. 2 – A bar chart showing the CPT and WPT (WS.D.) in
both the control and the capsaicin groups. The thresholds
are expressed as the change in temperature from the
baseline (32 8C) that was required to elicit a response.
difference pre- and post-menthol for at least one modality
(P = 0.017) and post hoc analysis using paired t-tests showed
that after treatment with menthol CDT increased by 3.71 8C on
average (P < 0.01) and WDT increased by 4.05 8C (P < 0.001).
There was no significant difference between the effect of
menthol on the control and capsaicin groups.
Fig. 3 – A bar chart showing the CDT and WDT (WS.D.)
before and immediately after sucking a lozenge containing
0.52% menthol. The thresholds are expressed as the
change in temperature from the baseline (32 8C) that was
required to elicit a response. The asterisks indicate a
significant change in the detection thresholds post-
menthol, *P < 0.01; **P < 0.001.
Fig. 4 – A bar chart showing the CPT and WPT (WS.D.) before
and immediately after sucking a lozenge containing 0.52%
menthol. The thresholds are expressed as the change in
temperature from the baseline (32 8C) that was required to
elicit a response.
151
4. Discussion
The initial application of a low dose of topical capsaicin to the
skin or mucosa activates nociceptive afferents causing a
burning sensation and an associated increased sensitivity to
both noxious and innocuous stimuli. Capsaicin mediates its
effect through the TRPV1 receptor which is preferentially
expressed on non-myelinated C fibres17,18 and small diameter
myelinated (Ad) fibres.19 The majority of nerve fibres within
the epidermis are nociceptors20,21 and the repeated topical
application of capsaicin to the skin causes the degeneration of
small diameter afferent fibres.22 Khalili et al.23 reported that
the repeated topical application of capsaicin cream on the
volar aspect of the forearm produced a 76% decrease in the
number of epidermal nerve fibres within 24 h of starting
treatment and a near complete disappearance after 3 days.
These effects of topical capsaicin are the basis for its
therapeutic use in a number of chronic pain, itch and
hypersensitivity states.
In the present study we have shown that frequently eating
‘‘hot’’ foods containing capsaicin alters the thermal detection
thresholds in the oral cavity; with the biggest difference being
in the detection of warm stimuli. These results are not
unexpected if the types of nerve fibres mediating each thermal
modality and the effects of capsaicin on the nerves expressing
TRPV1 are considered. Previous studies in animals and man
have shown that cold detection is evoked by activity in Ad
fibres; warm detection by activity in C fibres and heat and cold
pain by activity in both Ad and C fibres.24 The TRPV1 is located
on a subpopulation of non-myelinated C fibres and a few
thinly myelinated Ad fibres.4 Higher warm detection thresh-
olds in the capsaicin group would therefore be expected, and
are in keeping with reports by Malmberg et al.25 and Simone
and Ochoa6 on studies in the skin. Alternatively it is possible
that individuals with an increased sensitivity (or reduced
threshold) to capsaicin find eating ‘‘hot’’ foods unpleasant or
painful, causing them to consume less capsaicin in their diet;
such individuals would be likely to be found in our control
group.
TRPV1 is normally considered to detect noxious heat with a
threshold of 43 8C26 but TRPV1 knockout animals are still able
to respond to noxious heat.27 This indicates that a there must
be a pathway other than TRPV1 for the detection of noxious
heat. A possible candidate for noxious heat sensation is the
recently cloned TRPV2 receptor, which is activated by high
temperatures (52 8C) and is expressed in medium to large
diameter cells (with myelinated axons) in the dorsal root
ganglion.28 We suggest that in addition to some TRPV1 fibres
that may also have persisted, the presence of a non-TRPV1
pathway, has enabled participants to detect heat pain in the
present study.
Unlike the present study on oral mucosa, the topical
application of capsaicin to the skin has been shown to alter
warm pain thresholds6,22,23,29 and cold pain thresholds.22
Although we found an increase in these values, the difference
was not statistically significant. One possible explanation is
the difference in the timing, concentration and duration of the
capsaicin application. Simone and Ochoa6 clearly demon-
strated that capsaicin initially has an excitatory effect on heat
pain but this is subsequently followed by a longer lasting
152 archives of oral biology 52 (2007) 149–153
inhibitory effect. Malmberg et al.25 reported that the applica-
tion of a single high dose of capsaicin could cause a similar
pattern of epidermal nerve fibre degeneration to that of a low
dose given repeatedly over many weeks. Alternatively, the
difference between the results may reflect subtle changes in
the methodology used in each experiment. In the present
study the ‘method of levels’30 was used so that the stimulus
was gradually increased until it was detected by the subject,
whereas in other studies15,23 subjects have been asked to ask
to rate the intensity of a stimulus. The size of the thermode
used will also influence the results and Khalili et al.23
recommend the use of a small thermode to detect early small
fibre neuropathies. A large thermode applied to richly
innervated tissue, such as the oral mucosa, will cause
temporal and spatial summation and subtle changes will
not be detected. Finally, in the present study the exact
chemical contents of the ‘‘hot’’ food ingested by the
participants was not known. It is probable that ingredients
other than capsaicin, such as allicin (see below) may have
affected the outcome.
The lozenges used in this study not only contained menthol
(0.52%) but also eucalyptus oil (0.26%) and a number of other
‘cooling agents’ (not named by the manufacturers). Although
we primarily intended to study the effects of menthol,
Behrendt et al.31 reported that many other chemicals includ-
ing eucalyptol and hydroxycitronellal (citrus odorant) also
activate TRPM8. Another cold receptor, TRPA1 (also known as
ANKTM1), has been identified with an activation temperature
of 17 8C, close to the threshold for nociceptors. Although
TRPA1 is not activated by menthol it is activated by other
cooling agents such as allicin (the active ingredient in fresh
garlic) and pungent isothiocyanates.32 With time it is likely
that yet more cold receptors will be identified. The effect of
multiple agents, acting through a number of different
receptors, is far more complex than was previously envisaged.
In the same way that mechanosensitive receptors adapt to
prolonged force it has been shown that thermoreceptive
neurones within the dorsal root ganglion adapt to thermal and
menthol stimuli.33 The adaptation of cold thermoreceptors
that occurs during the sucking of a menthol sweet may
account for the raised thermal cold detection thresholds
reported in this study.
The action of menthol to increase the warm detection
thresholds was an unexpected finding although Green13,15
reported that after the topical application of menthol (0.2–2%)
to the tongue the intensity of a warmth stimulus was reduced.
A possible explanation is the finding that some TRPM8 positive
neurones also express TRPV1.34,35 Interestingly, in 1996 Cliff
and Green36 applied menthol and capsaicin to the tongue and
concluded that both menthol (TRPM8) and capsaicin (TRPV1)-
sensitive fibres had a common sensory pathway but different
excitatory mechanisms. Lundy and Contreras37 recorded from
thermally sensitive lingual fibres supplying the anterior
tongue in the rat and demonstrated that a weak concentration
of menthol and warm stimuli decreased neural activity, whilst
a strong concentration of menthol and cold stimuli had the
opposite effect and increased neural discharge.
Yosipovitch et al.38 reported that the application of 10%
menthol (in ethanol) to the skin of the forearm did not alter
either detection or pain thermal sensory thresholds; whereas
Namer et al.39 found that the application of a 40% solution of
menthol caused cold allodynia. This effect of menthol is
dependent upon both the concentration used13,36 and the
duration of the application.40 In the present study the menthol
lozenge was completely dissolved (sucked over a period of
approximately 5 min) before the thermal sensory tests were
undertaken. The test stimuli were then always applied in the
same order: cold detection, warm detection, cold pain and
lastly heat pain. The total time taken to complete this testing
was approximately 10 min. As the effect of menthol on the oral
mucosal receptors may have only lasted for a short period of
time, it may not have persisted for the full duration of the
sensory testing. This may partly explain why the detection
thresholds were raised in the present study but the pain
thresholds remained unaltered.
This study has clearly demonstrated that differences in
dietary habits can influence thermal sensory thresholds and
therefore must be taken into account when intra-oral sensory
testing is undertaken. Many of the patients who currently
undergo intra-oral thermal sensory testing have previously
sustained unilateral nerve injuries and are therefore able to
serve as their own controls by comparing the injured side with
the uninjured contralateral side. However, if intra-oral
thermal thresholds are being compared between different
subsets of the population, special care must be taken to ensure
that dietary habits are matched.
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