Protective Effect of Astaxanthin on Vocal Fold Injury

and Inflammation Due to Vocal Loading: A Clinical Trial

*Mami Kaneko, *Yo Kishimoto, *Ryo Suzuki, *Yoshitaka Kawai, *Ichiro Tateya, and †Shigeru Hirano, *†Kyoto, Japan

Summary: Objectives. Professional voice users, such as singers and teachers, are at greater risk of developing vocal
fold injury from excessive use of voice; thus, protection of the vocal fold is essential. One of the most important factors
that aggravates injury is the production of reactive oxygen species at the wound site. The purpose of the current study
was to assess the effect of astaxanthin, a strong antioxidant, on the protection of the vocal fold from injury and in-
flammation due to vocal loading.
Study Design. This study is an institutional review board-approved human clinical trial.
Methods. Ten male subjects underwent a 60-minute vocal loading session and received vocal assessments prior to,
immediately after, and 30 minutes postvocal loading (AST(−) status). All subjects were then prescribed 24 mg/day of
astaxanthin for 28 days, after which they received the same vocal task and assessments (AST(+) status). Phonatory
parameters were compared between both groups.
Results. Aerodynamic assessment, acoustic analysis, and GRBAS scale (grade, roughness, breathiness, asthenia, and
strain) were significantly worse in the AST(−) status immediately after vocal loading, but improved by 30 minutes after
loading. In contrast, none of the phonatory parameters in the AST(+) status were statistically worse, even when mea-
sured immediately after vocal loading. No allergic responses or adverse effects were observed after administration of
astaxanthin.
Conclusions. The current results suggest that astaxanthin can protect the vocal fold from injury and inflammation
caused by vocal loading possibly through the regulation of oxidative stress.
Key Words: Astaxanthin–Vocal fold–Vocal loading–Reactive oxygen species–Clinical trial.

INTRODUCTION reported that a transient episode of raised-intensity phonation

Excessive use or abuse of the voice can lead to symptoms of hoarse-
ness, weak voice, sore throat, and dysphonia. It has been suggested
that professional voice users, such as teachers, singers, sales pro-
fessionals, or Buddhist priests, are more at risk of developing vocal
disorders than others.1 In general, conservative treatment methods
such as voice therapy—mainly consisting of voice rest, vocal
hygiene, and medical management—are used as the first line of
treatment. However, voice rest is often difficult in these occupa-
tions and adequate voice therapy cannot be performed, making
treatment difficult.2 Thus, the development of more useful and sci-
entifically validated treatments is greatly needed.
Kojima et al3 quantified the extent of epithelial surface damage
within the vocal fold following vibration exposure using an in
vivo rabbit phonation model. They applied varying phonation
times (30, 60, or 120 minutes) and magnitudes (control, modal
intensity phonation, or raised intensity phonation) of the vibra-
tion exposure. Scanning and transmission electron microscopic
examinations revealed a significant reduction in microprojec-
tion density, microprojection height, and depth of the epithelial
surface, with increasing time and phonation magnitude. These
results suggest an increased risk of epithelial surface damage
with excessive and prolonged vibration exposure. Swanson4 also
Accepted for publication June 28, 2016.
Financial disclosure: None.
Conflict of interest: None.
From the *Department of Otolaryngology-Head and Neck Surgery, Graduate School of
Medicine, Kyoto University, Kyoto, Japan; and the †Department of Otolaryngology Head
& Neck Surgery, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Address correspondence and reprint requests to Shigeru Hirano, Department of
Otolaryngology Head & Neck Surgery, Kyoto Prefectural University of Medicine,
Kamigyo-ku, Kyoto 602-8566, Japan. E-mail: hirano@koto.kpu-m.ac.jp
Journal of Voice, Vol. 31, No. 3, pp. 352–358
0892-1997
© 2017 The Voice Foundation. Published by Elsevier Inc. All rights reserved.
http://dx.doi.org/10.1016/j.jvoice.2016.06.017
caused a significant increase in the expression of vocal fold in-
flammatory messenger RNA (mRNA), including interleukin (IL)-
1β, COX-2, and TGFβ1, compared with modal phonation and
sham control in an in vivo rabbit phonation model.
Reactive oxygen species (ROS) are normal byproducts of cel-
lular metabolic processes, including the mitochondrial respiratory
chain.5 Although ROS is essential for various signaling pathways,6
overexposure to ROS, referred to as oxidative stress, is a harmful
process that can be a significant source of damage to cellular
components, including lipids, proteins, and DNA. ROS over-
exposure can also contribute to cellular aging and malignant
transformation.7
A large amount of ROS is produced by inflammatory cells via
nicotinamide adenine dinucleotide phosphate oxidase, which plays
a central role in the defense against wound infection.8 However,
prolonged oxidative and inflammatory stress following skin wound-
ing is thought to be associated with excessive scarring.9
Mizuta et al10 suggested that a large amount of ROS is pro-
duced during the early phase of vocal fold wound healing, until
postinjury day 3, and this period may be crucial for regulating
ROS levels. They also suggested that inflammatory cells may
contribute to ROS generation. Mizuta et al11 also investigated
the effect of astaxanthin on the regulation of oxidative stress and
scarring during vocal fold wound healing. Their results sug-
gested that astaxanthin has the potential to prevent vocal fold
scarring by regulating oxidative stress during the early phase of
healing.
Astaxanthin is a xanthophyll carotenoid found in various mi-
croorganisms and marine animals.7,12–14 Astaxanthin has a strong
antioxidant effect as a potent quencher of singlet oxygen, and
many studies have shown favorable effects of astaxanthin on the
regulation of oxidative stress.7,13,15–20 Several studies have also
Mami Kaneko et al Clinical Trial of Astaxanthin for Vocal Loading
reported on the administration of astaxanthin for human health
purposes and have confirmed its antioxidant activity and safety.21,22
However, there are no clinical studies that assessed the effect
of astaxanthin on vocal fold injury or inflammation. The purpose
of the current clinical study was to assess the effects of astaxanthin
on the protection of the vocal fold from injury and inflamma-
tion due to vocal loading.
METHODS
Subjects
Ten non-singer male subjects participated in the study: the age
range was 23–30 years old and the mean age was 24.2 years.
All subjects were in good overall health based on self-reports
and had no history of major systemic diseases or laryngeal surgery.
Any vocal fold lesions or functional problems were excluded
by laryngoscopic examination. In addition, participants who used
any medications that might influence voice (eg, diuretics, de-
congestants, and antihistamines)23 or with a history of a smoking
habit were excluded.
Procedures
The general experimental paradigm is shown in Figure 1.
Day 0
All subjects first received vocal assessments, as described in detail
below. Subjects then underwent a 60-minute vocal loading session
with reading a Japanese novel. During loading, subjects alter-
nated between 15 minutes of loud phonation with 5 minutes of
rest, for a total of three cycles over the 60-minute loading period.24
Immediately after the 60-minute vocal loading session, sub-
jects received the same vocal assessments as performed prior
to loading. Subjects then rested for 30 minutes. Subjects were
allowed to drink water during the break period after the vocal
load task and after completion of loading, but were not allowed
to talk during the break. After 30 minutes of rest, subjects re-
ceived the vocal assessments again. After all vocal assessments
and vocal loading were completed, subjects received guidance
to take 24 mg/day of astaxanthin for 28 days and to refrain from
vocal abuse for 1 month. We referred to the data obtained prior
to taking astaxanthin as the “AST(−) status”.
FIGURE 1. General experimental paradigm.
353
Days 7–34
All subjects took 24 mg/day of astaxanthin. We confirmed the
condition of their health and compliance with taking astaxanthin
via phone interviews every week after they started treatment.
Day 35
All subjects received the same vocal assessments and vocal
loading that were performed on Day 0. Data obtained after the
subjects took astaxanthin are referred to as the “AST(+) status”.
During vocal loading, a sound level meter (NL-42; RION Co.,
Tokyo, Japan) was positioned at a constant distance of 25 cm
from the subject’s mouth so that the experimenter could monitor
relative intensity levels during loading. The examiner moni-
tored the meter nearly constantly and cued subjects to maintain
a target intensity range of 75–90 dB during phonatory loading.24
All procedures were approved by the institutional review boards
of Kyoto University.
Assessment
Vocal outcomes were evaluated prior to, immediately after, and
30 minutes postvocal loading. The following assessments were
performed: GRBAS scale (grade, roughness, breathiness, as-
thenia, and strain), aerodynamic assessment, acoustic analysis,
intensity, pitch range, stroboscopic examinations, and the Vocal
Fatigue Index (VFI). Stroboscopic examination was performed
using a digital video stroboscopy system with a 70° rigid en-
doscope, Model 9295 (KayPENTAX, Lincoln Park, NJ). Two
raters blinded to AST(+) and AST(−) status assessed all the stro-
boscopic images of the larynx at the same time.
Aerodynamic assessments, which included maximum pho-
nation time (MPT), mean flow rate (MFR), phonation threshold
pressure (PTP), intensity, and pitch range, were examined with
a phonation analyzer at the patient’s normal pitch and loud-
ness (PA-500; Nagashima Co., Osaka, Japan). Acoustic analyses
evaluated jitter and shimmer using the Multidimensional Voice
Program (Model 5105, KayPENTAX).
The GRBAS scale is an anchored perceptual analysis. GRBAS
was independently evaluated by two trained raters, a laryngologist
and a speech language pathologist. This scale was first devel-
oped by the Japanese Society of Logopedics and Phoniatrics and
has become popular worldwide.25 The GRBAS scale is scored
from 0 to 3 in which 0 = within normal limits, 1 = slight, 2 = mod-
erate, and 3 = severe. The ratings on the five subscales (G, R,
B, A, and S) were summed, and two raters blinded to AST(+)
and AST(−) status assessed all GRBAS scales at the same time.
The mean rating score between the two raters was calculated.
Inter-rater reliability was evaluated using Spearman’s rank cor-
relation coefficient, and the results showed significant correlation
between the two, with a correlation coefficient of r = 0.8
(P < 0.001).
The VFI is a standardized tool that can identify individuals
with probable vocal fatigue with good reliability, validity, sen-
sitivity, and specificity.26
Statistical test
Statistical analysis was performed using commercially avail-
able software (Excel Statistics 2012; Social Survey Research
354 Journal of Voice, Vol. 31, No. 3, 2017

TABLE 1.
Preloading Baseline for Each Parameter (Mean Value + Standard Deviation)
Prebaseline
AST(−) AST(+)
Mean (±SD) Mean (±SD) Significance (P Value)
MPT (s) 24.1 (±5.88) 24 (±7.78) 0.97
MFR (mL/s) 157.72 (±45.58) 170.55 (±41.09) 0.52
PTP (mmH2O) 6.89 (±1.27) 6.8 (±1.97) 0.91
Jitter (%) 0.42 (±0.18) 0.34 (±0.256) 0.43
Shimmer (%) 1.93 (±0.86) 1.96 (±0.97) 0.95
Intensity (dB) 79.84 (±4.19) 81.17 (±2.93) 0.42
Pitch range (Hz) 259.2 (±182.11) 275.4 (±175.37) 0.84
GRBAS 5 (±0) 5 (±0) –
VFI (points) 4.3 (±5.42) 3.3 (±4.4) 0.66
AST(+), AST(+) status; AST(−), AST(−) status.
Abbreviations: GRBAS, grade, roughness, breathiness, asthenia, and strain; MFR, mean flow rate; MPT, maximum phonation time; PTP, phonation thresh-
old pressure; SD, standard deviation; VFI, Vocal Fatigue Index.

Information Co., Ltd., Tokyo, Japan). For the preloading base-
line of each parameter, an unpaired t test was performed (Table 1).
Statistical tests using data obtained prior to, immediately after,
and 30 minutes postvocal loading were performed for each pa-
rameter. Significant differences were reported at an alpha level
of 0.05. All reported P values were two sided. A P value of less
than 0.05 was considered significant. Statistical analysis was per-
formed using the Wilcoxon signed-rank test.
RESULTS
Figure 2 and Table 2 show the mean value with standard devi-
ation for each parameter prior to, immediately after, and 30
minutes postvocal loading for each status. We regarded each time
point as “pre,” “post,” or “30 minutes post,” respectively. No al-
lergic responses or adverse effects were noted in any case.
Aerodynamic assessment
MPT was significantly shortened in the AST(−) status postloading
(P = 0.018), but there was no statistically significant change in
the AST(+) status (P = 0.1). MFR significantly increased in the
AST(−) status postloading (P = 0.037), but there was no statis-
tically significant change in the AST(+) status (P = 0.8). PTP
significantly increased in the AST(−) status postloading com-
pared with the AST(+) status (P = 0.028).
Acoustic analysis
Jitter significantly increased in the AST(−) status postloading
(P = 0.037), but there was no statistically significant change in
the AST(+) status (P = 0.39). Jitter also significantly increased
in the AST(−) status 30 minutes postloading compared with the
AST(+) status (P = 0.036). Shimmer significantly increased in
the AST(−) status postloading (P = 0.005), but there was no sta-
tistically significant change in the AST(+) status (P = 0.2).
Shimmer also significantly increased in the AST(−) status
postloading compared with the AST(+) status (P = 0.017).
Intensity and pitch range
Intensity and pitch range did not statistically change in either
status at any time point.
Auditory-perceptual ratings
Scores on the GRBAS scale significantly worsened in the AST(−)
status postloading (P = 0.005), but there was no statistically sig-
nificant change in the AST(+) status (P = 0.18). GRBAS scale
also significantly worsened in the AST(−) status postloading com-
pared with the AST(+) status (P = 0.008).
Self-ratings
VFI significantly worsened in the AST(−) status post and 30
minutes postloading (P = 0.005, P = 0.008, respectively), and VFI
significantly worsened in the AST(+) status postloading
(P = 0.008), but there was no statistically significant change in
the AST(+) status 30 minutes postloading (P = 0.18). VFI also
significantly worsened in the AST(−) status post and 30 minutes
postloading compared with the AST(+) status (P = 0.008,
P = 0.011, respectively).
Stroboscopic examination
In four subjects, the bilateral vocal fold mucosa appeared slightly
inflamed in the AST(−) status postloading, but there were no
changes in the AST(+) status.
DISCUSSION
Vocal fold scarring can occur following injury, inflammation,
or surgical intervention. Vocal fold scarring leads to the disrup-
tion of the layered structure of the lamina propria, which is
essential for optimal vibration.27–29 Once the vocal fold is scarred,
severe dysphonia can occur. Although many therapeutic strat-
egies have been evaluated for vocal fold scarring, a consistent
treatment has yet to be developed. Therefore, the prevention of
scarring is an important therapeutic target.11
Mami Kaneko et al Clinical Trial of Astaxanthin for Vocal Loading 355

FIGURE 2. Phonatory outcomes between the AST(−) and AST(+) status pre, post, and 30 minutes postvocal loading. *P < 0.05, **P < 0.01.
MPT, MFR, jitter, shimmer, and GRBAS score significantly worsened in the AST(−) status postloading, whereas there was no significant change
in the AST(+) status. VFI significantly worsened in the AST(−) status 30 minutes postloading, but there was no statistically significant change in
the AST(+) status. PTP and shimmer significantly worsened in the AST(−) status postloading compared with the AST(+) status. VFI significantly
worsened in the AST(−) status post and 30 minutes postloading compared with the AST(+) status. GRBAS, grade, roughness, breathiness, asthe-
nia, and strain; MFR, mean flow rate; MPT, maximum phonation time; VFI, Vocal Fatigue Index.
 356
TABLE 2.
Phonatory Outcome
AST(−) AST(+) AST(−), AST(+)
Mean (±SD) 95% CI P Value Mean (±SD) 95% CI P Value P Value
MPT (s)
Pre 24.1 (±5.88) 20.46–27.74 Pre, post: P = 0.018* 24 (±7.78) 19.18–28.82 Pre, post: P = 0.1 –
Post 19.8 (±7.97) 14.86–24.74 Pre, post 30: P = 0.64 20.6 (±7.89) 15.71–25.49 Pre, post 30: P = 0.17 P = 0.65
30-min post 23.2 (±8.74) 17.78–28.62 22.3 (±6.26) 18.42–26.18 P = 0.68
MFR (mL/s)
Pre 157.7 (±45.6) 129.47–185.97 Pre, post: P = 0.037* 170.6 (±41.1) 145.08–196.02 Pre, post: P = 0.8 –
Post 185.6 (±68.3) 143.23–227.91 Pre, post 30: P = 0.44 167.9 (±40.2) 142.97–192.77 Pre, post 30: P = 0.51 P = 0.39
30-min post 172.9 (±59.2) 136.19–209.55 165 (±41.9) 139.02–190.98 P = 0.88
PTP (mmH2O)
Pre 6.89 (±1.27) 6.1–7.67 Pre, post: P = 0.06 6.8 (±1.97) 5.58–8.02 Pre, post: P = 0.11 –
Post 7.57 (±1.9) 6.39–8.74 Pre, post 30: P = 0.65 6.32 (±2.19) 4.96–7.67 Pre, post 30: P = 0.76 P = 0.028*
30-min post 6.79 (±1.84) 5.65–7.94 6.79 (±1.69) 5.74–7.83 P = 0.8
Jitter (%)
Pre 0.42 (±0.17) 0.31–0.53 Pre, post: P = 0.037* 0.34 (±0.26) 0.18–0.5 Pre, post: P = 0.39 –
Post 0.69 (±0.43) 0.42–0.95 Pre, post 30: P = 0.33 0.72 (±1.33) 0.1–1.54 Pre, post 30: P = 0.37 P = 0.07
30-min post 0.49 (±0.28) 0.32–0.66 0.27 (±0.14) 0.18–0.35 P = 0.036*
Shimmer (%)
Pre 1.93 (±0.86) 1.4–2.46 Pre, post: P = 0.005** 1.96 (±0.97) 1.35–2.56 Pre, post: P = 0.2 –
Post 3.5 (±1.43) 2.61–4.38 Pre, post 30: P = 0.33 1.75 (±0.86) 1.22–2.29 Pre, post 30: P = 0.26 P = 0.017*
30-min post 1.86 (±0.39) 1.62–2.1 1.72 (±0.7) 1.28–2.16 P = 0.8
Intensity (dB)
Pre 79.8 (±4.19) 77.25–82.43 Pre, post: P = 0.51 81.2 (±2.93) 79.36–82.98 Pre, post: P = 0.51 –
Post 80.4 (±3.44) 78.32–82.59 Pre, post 30: P = 0.88 80.8 (±3.44) 78.69–82.95 Pre, post 30: P = 0.26 P = 0.8
30-min post 80.2 (±2.64) 78.55–81.81 80.6 (±3.04) 78.94–82.7 P = 0.41
Pitch range (Hz)
Pre 259.5 (±182.1) 146.63–372.37 Pre, post: P = 0.28 275.4 (±182.1) 166.71–384.09 Pre, post: P = 0.88 –
Post 238 (±65.4) 135.46–340.54 Pre, post 30: P = 0.88 290.3 (±165.4) 172.95–407.65 Pre, post 30: P = 0.41 P = 0.58

Journal of Voice, Vol. 31, No. 3, 2017

30-min post 280.8 (±165.3) 178.38–383.22 300.5 (±199.7) 166.51–424.29 P = 0.72
GRBAS
Pre 5 (±0.00) – Pre, post: P = 0.005** 5 (±0.00) – Pre, post: P = 0.18 –
Post 7.4 (±0.84) 6.88–7.92 Pre, post 30: P = 0.18 5.4 (±0.84) 4.88–5.92 Pre, post 30: P = 0.18 P = 0.008**
30-min post 5.4 (±0.84) 4.88–5.92 5 (±0.00) – P = 0.18
VFI (points)
Pre 4.3 (±5.42) 0.94–7.66 Pre, post: P = 0.005** 3.3 (±4.4) 0.57–6.03 Pre, post: P = 0.008** –
Post 21.8 (±15.1) 12.45–31.15 Pre, post 30: 12.2 (±8.75) 6.77–17.63 Pre, post 30: P = 0.09 P = 0.008**
30-min post 14.5 (±9.48) 8.63–20.37 P = 0.008** 6.6 (±6.5) 2.57–10.63 P = 0.011*
AST(+), AST (+) status; AST(−), AST(−) status.
*P < 0.05, **P < 0.01. Abbreviations: CI, confidence interval; GRBAS, grade, roughness, breathiness, asthenia, and strain; MFR, mean flow rate; MPT, maximum phonation time; PTP, phonation threshold
pressure; SD, standard deviation; VFI, Vocal Fatigue Index.
Mami Kaneko et al Clinical Trial of Astaxanthin for Vocal Loading
Mizuta et al11 suggested the potential of astaxanthin to reduce
oxidative stress during the early phase of vocal fold wound healing
and to prevent vocal fold scarring in a rat model. They showed10
that following injury, the expression of 4-hydroxy-2-nonenal (4-
HNE) significantly increased until postinjury day 3. Astaxanthin
treatment, however, significantly reduced the expression of 4-HNE
compared with sham treatment, suggesting that astaxanthin regu-
lates oxidative stress. Moreover, the sham-treated group showed
a severe decrease in hyaluronic acid (HA) deposition and con-
traction of the lamina propria compared with the normal group.
In contrast, the astaxanthin-treated group showed a significant
increase in HA deposition and attenuation of the lamina propria
contraction compared with the sham-treated group.
Many studies have reported using astaxanthin as an antioxi-
dant. Mitochondrial dysfunction induced by oxidative stress is
a critical factor in many diseases such as cancer and lifestyle-
related diseases. Astaxanthin is considered to be one of the best
carotenoids for protecting cells, lipids, and membrane lipopro-
teins against oxidative damage.14 Wolf et al15 reported that
astaxanthin decreases endogenous mitochondrial production of
ROS and protects mitochondria against the reduction in mem-
brane function that typically occurs under oxidative stress.
Furthermore, several animal studies have demonstrated the pro-
tective effect of astaxanthin in an ischemia-reperfusion myocardial
model.16–18 The consumption of astaxanthin can also prevent or
reduce the risk of various human disorders, such as decreasing
inflammation,21 reducing cholesterol,22 improving asthenopia,30
inhibiting the progress of age-related macular degeneration and
vision loss,31 and improving semen quality in infertile men.32 Thus,
astaxanthin is a potential therapeutic agent for various diseases
involving oxidative stress in both animals and humans.
Several studies have reported on the deleterious effects of vocal
loading. Enflo et al examined the effects of loud, prolonged vowel
vocalization (at least 80 dB for 20 minutes) on collision thresh-
old pressure (CTP) and PTP for singer and non-singer voices,
and showed that CTP and PTP increased significantly after vocal
loading in non-singer subjects.33 Chang and Karnell reported that
PTP returned to baseline levels within 1 hour after a 2-hour oral
reading task.34 Based on these studies, the current study was de-
signed with an assessment at 30 minutes postloading because
vocal loading was 1 hour.
Several studies have demonstrated that vocal loading causes vocal
fold inflammation in animal and human trials. Abbott et al24 as-
sessed the expression of inflammatory cytokines after 1 hour of
vocal loading, and found increased concentrations of IL-1β, tumor
necrosis factor-α, and matrix metalloproteinase-8 in the laryn-
geal mucus after loading. The increased levels of these mediators
corresponded with the clinical appearance of vocal fold injury.35
In the current study, vocal loading was thought to cause vocal fold
inflammation, which resulted in the worsening of vocal function.
A weak point of the current study was the lack of placebo
control. A placebo control arm should have been helpful in this
study. As there was no knowledge about the effects of anti-
oxidant on the maintenance of the vocal fold during vocal load,
we performed this study as a pilot study to explore the possi-
ble effects of astaxanthin. We also tried to avoid subjects’ intent
to reduce their effort on vocal load during the treatment round
357
by monitoring the intensity of the voice and the amount of reading
during vocal loading, which might contribute to making this study
justifiable. The results of the current clinical trial were encour-
aging and suggest the potential ability of astaxanthin to protect
the vocal fold from injury and inflammation due to vocal loading.
All cases showed anti-inflammatory effects, indicated by sig-
nificant improvements in aerodynamics, acoustic function,
auditory-perceptual ratings, and VFI. During the follow-up period,
no adverse effects were reported. Future studies will be needed
to examine the effects of astaxanthin on professional voice users
such as singers.
Although the sample size was relatively small, the current ex-
ploratory study is the first to examine the effects of astaxanthin
on the protection of the vocal fold from injury and inflamma-
tion due to vocal loading, and the results are encouraging when
it comes to the use of astaxanthin in the maintenance of the vocal
fold in frequent voice users.
CONCLUSIONS
The current study demonstrated that astaxanthin significantly
reduced functional voice deterioration after vocal loading. These
results suggest that astaxanthin is a potent therapeutic agent for
the protection of the vocal fold from injury and inflammation
caused by excessive vocal use.
Acknowledgment
We appreciate Astareal Co., Tokyo, for providing the product
of astaxanthin (Astareal ACT).
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