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Noni Seed Oil Topical Safety, Efficacy, and Potential Mechanisms of Action

Authors:
  • Juve International LLC
Journal of Cosmetics, Dermatological Sciences and Applications, 2012, 2, 74-78
http://dx.doi.org/10.4236/jcdsa.2012.22017 Published Online June 2012 (http://www.SciRP.org/journal/jcdsa)
Noni Seed Oil Topical Safety, Efficacy, and Potential
Mechanisms of Action*
Afa K. Palu#, Brett J. West, C. Jarakae Jensen
Research and Development Department, Morinda Inc., American Fork, USA.
Email: #afap@tni.com
Received February 7th, 2012; revised March 13th, 2012; accepted March 30th, 2012
ABSTRACT
Background: Noni fruit and leaf juices were used to treat acne in Polynesian folk medicine. However, the potential
benefit of noni seed oil (NSO) for human skin health is not as well known. Objective: The topical safety of NSO was
evaluated in a repeat insult patch test with 49 adult volunteers. Comedogenicity was evaluated in 23 adolescent volun-
teers for four weeks. Anti-inflammatory activity was determined in-vitro with COX-2 and 5-LOX enzyme inhibition
assays. Results: No reactions to NSO were observed in any of the volunteers. NSO also reduced the number of open
and closed comedones in the comedogenicity test. NSO inhibited COX-2 and 5-LOX enzymes in a concentration de-
pendent manner, but with COX-2 inhibition being more pronounced than 5-LOX. Conclusion: Noni seed oil is safe for
topical use and is non-comedogenic. It also has potential anti-inflammatory activity, being a dual inhibitor of COX-2
and 5-LOX enzymes. These results indicate the potential utility of NSO for skin care applications, as well as warrant
further investigation of efficacy in 5-LOX and COX-2 related pathophysiological conditions.
Keywords: Morinda citrifolia; Noni Seed Oil; COX-2; 5-LOX; Comedones; Safety
1. Introduction
Morinda citrifolia L. (family Rubiaceae), commonly known
as noni, has been used in Polynesia for food and medi-
cine for thousands of years [1]. The medicinal usages of
noni fruit juice, flowers, leaves, bark, and roots are well-
known among the natives of Polynesia, but less is known
about the potential health benefits of noni seeds and oil
extracted from them [2]. Benthall reported that noni
seeds were consumed as a food by the Burmese Indians
[3]. Polynesians also ate noni seeds, but perhaps, to a
much lesser degree than the Burmese Indians. Therefore,
this use among Pacific Islanders is not well documented
in the scientific literature. But older generations of Ton-
gan were known to pick ripe noni fruit and eat them,
while removing the seeds and roasting them over an open
fire before consumption. This practice was not widely
observed in Tonga but was likely limited to elderly mem-
bers of specific villages. One of the Tongan traditional
healers from the village of Ha’alalo concluded that one
has to acquire the taste of roasted noni seeds to enjoy
them on a regular basis [4].
Noni seeds were considered a by-product of the noni
fruit juice industry until a process for extracting the seed
oil was recently developed [5]. Noni fruits contain 200 -
250 seeds [6]. However, many thousands of noni seeds
are required to produce one fluid ounce of oil. With a 4%
- 6% yield and a cost of more than $400 US dollars per
gallon, noni seed oil (NSO) is a precious resource. As
such, NSO production is cost prohibitive with small scale
production. But when linked to a large-scale noni fruit-
juice operation, it is more economically feasible.
We report two human clinical trials that demonstrate
the safety of topical use of NSO on human skin and its
effect on comedogenicity, with potential mechanisms of
action on acne. This is the first report of NSO inhibitory
effects on COX-2 and 5-LOX enzymes, especially as it
relates to its potential mechanism of action against acne,
as well as the formal evaluation of safety and come-
dogenicity.
2. Materials and Methods
2.1. Ethics Reviews
The human study protocols were reviewed by an institu-
tional ethics review committee. Signed informed consent
was obtained from all volunteers prior to initiation of the
studies, which were conducted in accordance with the
Ethical Principles for Medical Research Involving Hu-
man Subjects as outlined in the Declaration of Helsinki
*Conflict of interest: All authors are employees of Morinda Inc., largest
manufacturer of noni seed oil in the world.
#Corresponding author.
Copyright © 2012 SciRes. JCDSA
Noni Seed Oil Topical Safety, Efficacy, and Potential Mechanisms of Action 75
and in OCR HIPAA Privacy.
2.2. Noni Seed Oil Preparation
Noni seeds were collected from discarded material from
the fruit finishing process on the island of Tahiti. Several
batches of seeds were washed and dried. Samples of the
dried seeds were retained for compositional analysis.
Following drying, the seeds were cracked and ground to
2 - 20 mm particle size. The flaked seeds were then
pressed in a screw press to expel the oil. For greater re-
covery, the remaining seed cake was mixed with a food-
grade nonpolar extraction solvent at a maximum tem-
perature of 79˚C for 30 min. The extract was collected
and the solvent removed by flash evaporation. The ex-
tracted oil was then combined with the pressed oil to
produce crude noni seed oil [5].
2.3. Repeat Insult Patch Test
The repeat insult patch test is a maximization test that
has been used for more than 40 years to evaluate contact
allergens in human subjects [7]. The inclusion criteria for
the repeat insult patch test were healthy adult volunteers
with no skin conditions and limited skin pigmentation
which would not interfere with reaction readings. Those
excluded were persons with known sensitivities to cos-
metics, pregnant or lactating, or had used medication that
might prevent or limit skin reactions, such as antihista-
mines and corticosteroids, within 30 days prior to the test.
NSO was evaluated in 46 females and 3 males (ages:
46.59 ± 15.57). During the induction phase, sodium
lauryl sulfate (SLS) served as an adjuvant and was mixed
with NSO. Occlusive application of 25 µL of the mixture
to the back was accomplished with a Finn Chamber and
held in place for 48 h with Scanpor tape. For each volun-
teer, application of the mixture occurred 5 times at the
same site on the back, with 48 h rest between applica-
tions. After the induction phase, the skin of the volun-
teers was not exposed for at least 7 d. Following this rest
period, the skin of the volunteers was exposed to NSO,
without SLS, by occlusive application for 48 h at a site
different from the one used during induction. For 72 h
following application, subjects were evaluated for possi-
ble delayed reaction, including redness, itching, and irri-
tation. The skin of the volunteers was scored for reac-
tions according to the International Contact Dermatitis
Research Group and under the supervision of a consult-
ing dermatologist [8]. Reactions were scored on a scale
of 0 (no reaction) to 4 (erythema, induration, and bullae).
2.4. Comedogenicity Test
The effects of NSO on comedone counts was evaluated
in 23 (8 males, 15 females) healthy adolescent Caucasian
subjects (ages: 15.6 ± 1.4 yr) in an open label human
clinical trial. Subjects were instructed to apply equal
amounts of oil to the same 2 cm by 2 cm site on the left
side of the forehead daily for four weeks. Application
sites were visually examined by a trained technician at
the initiation of the test period, under 1.5-2X magnifica-
tion, as well as four weeks later. The number of come-
dones was recorded at each examination. Additionally, a
follicular biopsy was used to quantify the open and
closed comedones before and after the trial [9]. The bi-
opsy was performed by applying a quick-setting cyanoa-
crylate polymer to the same site where the oil was ap-
plied. After the polymer film had set, it was gently re-
moved from the skin, thereby extracting the contents of
sebaceous follicles. The contents of the film were then
examined microscopically for the presence of keratotic
plugs, with total numbers being recorded. To document
and confirm the visual examinations and follicular biop-
sies, the application areas were photographed digitally at
a distance of 25 cm. The resultant photographic magnifi-
cation was 1.5×. The total number of open and closed
comedones for each subject, before and after application
of NSO, was determined from these measurements.
2.5. Noni Seed Oil Effects on COX-2
NSO was evaluated for its inhibitory effect on COX-2
enzymes, according to established protocols [10,11]. Briefly,
COX-2, from human recombinant insect Sf21 cells, was
combined with a mixture of 0.5 and 1 mg/mL NSO in
1% DMSO, and a 0.3 µM arachidonic acid-substrate.
The mixture was then incubated in buffer [100 mM Tris-
HCl, pH 7.71, 1 mM glutathione, 1 µM hematin, and 500
µM phenols] for 5 min at 37˚C. Rofecoxib was used as a
positive control. Prostaglandin E2 (PGE2) was quantified
using enzyme immunoassay (EIA) and then reported as
percent inhibition. Assays were run in duplicate.
2.6. Noni Seed Oil Effects on 5-LOX
NSO was also evaluated for its inhibitory effects on 5-
lipoxygenase (5-LOX) enzymes following established
protocols [12,13]. Briefly, 5-LOX enzymes, obtained
from human PBML cells, were added to 0.5 and 1 mg/mL
concentrations of NSO in 1% DMSO. Arachidonic acid
was added to the mixture as a substrate and incubated in
HBSS buffer for 15 min at 37˚C. Nordihydroguaiaretic
acid (NDGA) was used as a positive control. Leukotriene
B4 (LTB4) was quantified using EIA, and results were
reported as percent inhibition (Table 1). Assays were run
in duplicate.
2.7. Data Analysis
Descriptive statistics, such as the median and mean ±
Copyright © 2012 SciRes. JCDSA
Noni Seed Oil Topical Safety, Efficacy, and Potential Mechanisms of Action
76
standard deviation (SD), were calculated. Student’s t-test,
two-tailed and unequal variance, was used to comparepre
and post-trial means in both trials.
3. Results
3.1. Repeat Insult Patch Test
Previously, no evidence of acute oral toxicity was ob-
served for NSO, and it was non-genotoxic in the Salmo-
nella typhimurium reverse mutation assay (Ames test) or
the in vitro mammalian chromosomal aberration assay
[14]. During the challenge phase of the repeat-insult
patch test of NSO, there were no reactions in any of the
49 volunteers. These results indicate that noni seed oil
presents very little risk in causing any allergic contact
dermatitis.
3.2. Comedogenic Test
In adolescent subjects, daily application of NSO for four
weeks did not increase comedone frequency or the num-
ber of acne comedones. But it did decrease the number of
open and close comedones by the end of the trial (Table
2).
3.3. Noni Seed Oil Effects on COX-2 Enzymes
NSO, in concentrations of 0.5 and 1 mg/mL, inhibited
COX-2 enzymes by 85% and 84%, respectively.
3.4. Noni Seed Oil Effects on 5-LOX Enzymes
NSO, in concentrations of 0.5 and 1 mg/mL, inhibited
5-LOX enzymes by 49% and 40%, respectively.
4. Discussion
The current study showed that noni seed oil inhibits
Table 1. Mean percent inhibition of noni seed oil (NSO) on
COX-2 and 5-LOX enzymes.
NSO Concentration (mg/mL) % Inhibition
COX-2 5-LOX
0.5 85 49
1.0 84 40
Table 2. Pre and post trial comedone counts in a 4 week
trial of noni seed oil daily application to the forehead of
adolescent volunteers.
Open Comedone Counts Closed Comedone Counts
Baseline End of Study Baseline End of Study
Mean 3.60 2.43 4.36 3.04
Median 2 1 4 3
Std. Dev. 6.96 3.68 3.23 2.18
COX-2 and 5-LOX enzymes. However, the inhibition of
COX-2 was more pronounced than that of the 5-LOX
enzymes. Further, NSO reduced the number of open and
close comedones in healthy human subjects after 4 weeks
of daily application.
Even though the inhibitory effects on 5-LOX enzymes
was not pronounced compared to COX-2, 5-LOX en-
zymes are activated in many diseases and set in motion
inflammatory responses that may lead to, among other
thing, acne [15,16]. Therefore, it is reasonable to assume
that moderate inhibition of 5-LOX enzymes, in conjunc-
tion with COX-2 inhibitory activity, will ameliorate acne
and inflammatory skin conditions.
We report for the first time the potential anti-inflam-
matory effect on NSO through dual inhibition effects of
COX-2 and 5-LOX. Belch and Hill [17] demonstrated that
fatty acids, such as a linoleic acid derivative, may inhibit
PGE2 synthesis by COX enzymes via substrate competi-
tion. Such an effect may explain, at least in part, the in-
hibition of COX-2 enzymes by NSO. The possibility of
the inhibitory activity being due to linoleic acid is sup-
ported by analyses published by West and colleagues [14]
which showed that linoleic acid accounted for 89.9 ±
19.2 g/kg of dried noni seeds and 594.5 ± 60.5 g/kg of
the crude seed oil. These quantities make NSO a rich
source of linoleic acid. Linoleic acid reduces the size of
follicular casts and micro-comedones by as much as 25%
[18]. This further demonstrates the active role of the fatty
acids in NSO in the anti-inflammatory and anti-come-
dogenic activities observed in our study.
Dual inhibition of COX-2 and 5-LOX enzymes is ef-
fective for more than just the treatment of pain and in-
flammation [19-21]. The current results are consistent
with previous data from our laboratory which demon-
strated that noni fruit juice is a selective inhibitor of
COX-2 enzymes [22]. We also demonstrated that noni
fruit juice was a dual inhibitor of COX-2 and 5-LOX
enzymes [23]. This dual inhibition may explain the pain-
reducing and anti-inflammatory effect of noni juice, as
reported by healers and consumers. But this is a novel
discovery for noni seed oil.
Some plant-derived oils have the potential to induce
comedone formation [24,25]. Therefore, new vegetable
oils must be examined for their comedogenic potential
prior to any topical use by humans. In our studies, NSO
reduced open and closed comedone counts, indicating an
absence of comedogenicity. This finding suggests that
NSO has little or no potential for clogging up pores of
the skin when used as a cosmetic ingredient. Analyses by
West and colleagues [14] showed that NSO contains li-
noleic and oleic acids, which are considered healthy fatty
acids. In fact, oleic fatty acid can be used in cooking of
foods [26-28] and for formulations of various cosmetics
[29-33].
Copyright © 2012 SciRes. JCDSA
Noni Seed Oil Topical Safety, Efficacy, and Potential Mechanisms of Action 77
5. Conclusion
The data support the potential for noni seed oil to reduce
inflammation via its dual inhibition of COX-2 and 5-
LOX enzymes. The fact that it reduced open and close
comedones and did not cause any reaction in the repeat
insult patch test indicates that it is a safe raw material for
cosmetic use.
6. Acknowledgements
Financial support for this project was provided by
Morinda Bioactives. Gratitude is also expressed to Silivia
L. T. Palu from ‘Uiha, Ha’apai and Viliami Palepale
from Lehi, Utah for their contribution on traditional us-
age of noni and noni seed consumption.
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Noni juice is a globally popular health beverage originating in the tropics. Traditional Tahitian healers believe the noni plant to be useful for a wide range of maladies, and noni juice consumers throughout the world have similar perceptions. Nevertheless, human clinical trials are necessary for a precise understanding of what the health benefits of noni juice are. A review of published human intervention studies suggests that noni juice may provide protection against tobacco smoke-induced DNA damage, blood lipid and homocysteine elevation as well as systemic inflammation. Human intervention studies also indicate that noni juice may improve joint health, increase physical endurance, increase immune activity, inhibit glycation of proteins, aid weight management, help maintain bone health in women, help maintain normal blood pressure, and improve gum health. Further, these studies point to notable antioxidant activity in noni juice, more so than other fruit juices which served as trial placebos. It is this antioxidant effect and its interaction with the immune system and inflammation pathways that may account for many of the observed health benefits of noni juice. However, the existing evidence does have some limitations as far as its general application to noni juice products; all the peer-reviewed human interventions studies to date have involved only one source of French Polynesian noni juice. Geographical factors and variations in processing methods are known to produce commercial noni juice products with divergent phytochemical and nutrient compositions. Therefore, other sources of noni products may have different toxicological and pharmacological profiles.
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Noni (Morinda citrifolia L.) is native to the Polynesian and recognized in the tropical and subtropical countries as a sustainable crop with feasible commercial applications. It has been reported that the interest in developing noni plant as a novel source of bioactive compounds are increasing by the day. This review describes the safety, nutritional values, and the properties of noni seed oil (NSO) with potential industrial uses. In particular, the bioactive compounds, anti-nutrients, antioxidant activity, and IC50 values of noni seed and the chemical composition of NSO are also described. NSO has high contents of polyunsaturated fatty acids, total phytosterols and tocopherols that could be better choices for patients with high cholesterol and cardiovascular diseases. Extracts of noni seed have been shown to possess bioactive compounds that exhibit antioxidant, anti-mutagenic, anti-tumor, anti-inflammatory, anti-allergic, anti-viral, anti-fungal, anti-microbial, and anti-carcinogenic properties. Bioactive compound-rich noni fruit seed could be a potential source of functional foods. Moreover, noni seeds could be a valuable new source of vegetable oil because of its nutritional properties and non-toxic nature along with the increasing supply of seeds as by-products from noni juice industry. Comprehensive studies are needed on NSO to explore more potential product development. Moreover, further study is needed on the development of nutraceutical food products from noni seed by-products.
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