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J 43
C © ORIGINAL ARTICLES
SPECIAL TOPIC
Anti-Inammatory Activities of Colloidal Oatmeal
(Avena sativa) Contribute to the Eectiveness of Oats in
Treatment of Itch Associated With Dry, Irritated Skin
Kurt A. Reynertson PhD, Michelle Garay MS, Judith Nebus MBA, Suhyoun Chon PhD, Simarna Kaur PhD,
Khalid Mahmood PhD, Menas Kizoulis BA, Michael D. Southall PhD
Background: Oat (Avena sativa) in colloidal form is a centuries-old topical treatment for a variety of skin conditions, including skin rash-
es, erythema, burns, itch, and eczema; however, few studies have investigated the exact mechanism of action for the anti-inammatory
activity of colloidal oatmeal.
Methods: Four extracts of colloidal oatmeal were made with various solvents and tested in anti-inammatory and antioxidant assays. In
addition, an investigator blind study was performed with twenty-nine healthy female subjects who exhibited bilateral mild to moderate
itch with moderate to severe dry skin on their lower legs. Subjects were treated with a colloidal oatmeal skin protectant lotion.
Results: Extracts of colloidal oatmeal diminished pro-inammatory cytokines in vitro and the colloidal oat skin protectant lotion showed
signicant clinical improvements in skin dryness, scaling, roughness, and itch intensity.
Conclusions: These results demonstrate that colloidal oat extracts exhibit direct anti-oxidant and anti-inammatory activities, which
may provide the mechanisms for observed dermatological benets while using the colloidal oatmeal skin protectant lotion.
J Drugs Dermatol. 2015;14(1):43-48.
ABSTRACT
INTRODUCTION
Oats (Avena sativa) have been cultivated since the
Bronze Age, and have been used in traditional medi-
cine for centuries. As a topical treatment, colloidal
oatmeal has emollient and anti-inammatory properties, and
is commonly used for skin rashes, erythema, burns, itch, and
eczema.1-3 Historically, investigations into the phytochemical
constituents of oat have focused primarily on their value as a
food. For example, β-glucan is the “soluble ber” that makes
oats a heart-healthy food. β-glucans have also been used as
scaffolds for the growth of bioarticial skin, and are known to
assist in wound healing, response to injury and infection, and
have a great water retention capacity.4
Colloidal oatmeal is the nely ground whole oat kernel or groat,
and is an active natural ingredient covered by the FDA OTC Skin
Protectant monograph in the US. The oat grain is ground and
processed until no more than 3% of the total particles exceed
150 µm and no more than 20% exceeds 75 µm.5 The compo-
sition of colloidal oatmeal largely consists of starch (65-85%),
protein (15-20%), lipids (3-11%), ber (5%) and β-glucans (5%).3,6
Oat lipids are primarily composed of triglycerides, along with po-
lar lipids and unsaturated free fatty acids. Oat triglycerides are
rich in omega-3 linoleic and omega-6 linolenic acids and essential
fatty acids7 which are necessary for normal mammalian health
and important for skin barrier function.8-10 In addition, oat lipids
contain important mammalian cell membrane components, such
as phospholipids, glycolipids, and sterols. Lipid oxidation protec-
tion is supplied by mixed tocopherols (vitamin E) and tocotrienols.
Colloidal oatmeal is also a rich source of phenolic antioxidants
and saponins. Avenanthramides, nitrogen-containing pheno-
lic compounds specic to oats, are potent antioxidants and
anti-inammatory agents that have been previously shown to
inhibit NF-κB and IL-8 release in a dose dependent manner.3,11-13
Saponins are glycosylated metabolites which help to protect
oat plants from disease,14 and which can also help create stable
emulsions when colloidal oatmeal is used in a formulation.
Despite a rich history of traditional use, there remain some gaps
in the understanding of the exact mechanisms that give col-
loidal oatmeal its clinical benets; we conducted a series of in
vitro experiments and a clinical study to help identify the mecha-
nism of action for the clinical benet of colloidal oatmeal. We
made four extracts of colloidal oatmeal with organic and aque-
ous solvents to concentrate constituents based on compound
polarity, and subjected them to antioxidant and anti-inamma-
tory assays. In addition, an investigator-blind clinical study was
conducted to evaluate the efcacy of a colloidal oatmeal skin
protectant lotion in alleviating extra dry, itchy skin.
Healthy female subjects with bilateral itch and moderate to se-
vere dry skin on their lower legs were enrolled in a 2-week study.
Subjects applied a set amount of the lotion to the lower leg
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44
K. A. Reynertson, M. Garay, J. Nebus, et al.
UV-Induced Reactive Oxygen Species Production
in Keratinocytes
Primary normal human keratinocytes were incubated for 30min
with 5 μM of the hydrogen peroxide-sensitive uorescent
probe 5-(and-6)-chloromethyl-2′,7′-dichlorodihydro-uorescein
diacetate, acetyl ester (CM-H2DCFDA; Invitrogen Carlsbad, CA).
After incubation, cells were washed 2X with PBS to remove
excess probe and treated with extracts at indicated concen-
trations; the plate was then exposed to UV (1000W-Oriel solar
simulator equipped with a 1mm Schott WG 320 lter; UV dose
applied 4.2kJ/m2 as measured at 360nm). The plate was read
immediately post-UV exposure on a uorescent plate reader
set at wavelengths 485nm excitation/530 nm emission to de-
tect basal peroxide formation. Statistical signicance (P< 0.05)
was determined by 2-tailed t-test.
Gene Expression
Primary human keratinocytes were treated for 24 hours in the
presences of 1.2 mM CaCl2. RNA was extracted using Qiagen
RNeasy kit with DNase I digestion (Valencia, CA). Reverse
transcription was performed using High Capacity cDNA kit
(Life Technologies). 40 to 60 ng of cDNA samples were used in
a QPCR reaction to measure IL-8. Taqman gene expression as-
say was purchased from Life Technologies (Grand Island, NY).
QPCR reaction were performed using ABI 7500 fast amplier.
All gene expression data were normalized by reference genes,
polymerase (RNA) II polypeptide A (POLR2A). Statistical sig-
nicance (P< 0.05) was determined by one-way ANOVA.
Clinical Study Design
The study was an investigator-blind design conducted in
Colorado, USA. Subjects applied a colloidal oatmeal skin
protectant lotion (Skin Relief 24Hr Moisturizing Lotion, Avee-
no®, Skillman, NJ) to the lower leg area twice a day for the
two-week study period. Benets of the colloidal oatmeal skin
protectant lotion were established by clinical evaluations, di-
aries, self-assessments, TEWL, corneometer, and imaging at
day 0, followed by determinations at days 1, 7, and 14. Writ-
ten informed consent was obtained from each subject prior
to enrollment in the study. The protocol and informed con-
sent agreement for this study were reviewed and approved
by an Institutional Review Board (IRB).
area twice a day. Benets of the lotion were assessed by clinical
evaluations, instrumental measures and self-assessments. Itch
intensity was monitored daily through patient diaries.
This clinical study showed that the colloidal oatmeal skin pro-
tectant lotion was effective at relieving itch and alleviating
moderate to severe dry skin while improving stratum corne-
um function in patients with itchy, extra dry skin. In addition,
extracts of colloidal oat demonstrate anti-inammatory and an-
tioxidant activity, which may account for some of the observed
clinical benets.
MATERIALS AND METHODS
Primary human keratinocytes were obtained from Lonza Walk-
ersville, Inc. (Walkersville, MD) and Promocell (Heidelberg,
Germany). Human recombinant TNF-α was procured from Pep-
rotech Inc. (Rocky Hill, NJ).
Preparation of Extracts of Colloidal Oatmeal
Four extracts of colloidal oatmeal were prepared using HPLC-
grade hexanes, aqueous acetone, aqueous methanol, and
water to generate extracts enriched in phytochemicals based
on polarity. Approximately 10 g of colloidal oat was suspend-
ed in 100 mL of solvent and placed on a shaker to maintain
the slurry in constant motion. After 24 hours, the suspensions
were centrifuged, ltered, and concentrated on a rotary evap-
orator under reduced pressure at a temperature not exceeding
40 °C. The extracts were then transferred to a tarred container,
dried under nitrogen, and lyophilized to remove all traces of
solvent. For cell culture experiments, stock solutions of each
extract were dissolved in DMSO (50 mg/mL) and diluted into
media so that DMSO concentrations less than 0.01%.
TNF-α induced IL-8 Release
Primary normal human keratinocytes were treated with test
samples in DMSO for 2 hours before exposure to solar ultra-
violet light (1000W-Oriel solar simulator equipped with a 1-mm
Schott WG 320 lter; UV dose applied: 70 kJ/m2 as measured
at 360 nm). After 24 hours, supernatants were analyzed for IL-8
cytokine release using commercially available kits (Millipore
Corp., Billerica, MA).Statistical signicance (P< 0.05) was de-
termined by 2-tailed t-test.
NF-κB Luciferase Promoter Assay in Keratinocytes
Primary normal human keratinocytes were transiently transfect-
ed with 0.25 ug/well total DNA containing pNF-κB-Luc reporter
plasmid (Stratagene, La Jolla, CA) and the internal control Renilla
luciferase reporter (pRL-TK; Promega Corporation, Madison, WI)
using Lipofectamine 2000 transfection reagent (Invitrogen Cor-
poration, Carlsbad, CA). At 24 hours post transfection, cells were
pre-treated with extracts for 2 hours, followed by treatment with
100 ng/ml TNF-α (Peprotech Inc., Rocky Hill, NJ) for 24 hours. Sta-
tistical signicance (P< 0.05) was determined by 2-tailed t-test.
oat extracts contain components
that not only protect oat lipids from
spoilage, they also form a synergistic
mixture that enhances oat oil’s capacity
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45
K. A. Reynertson, M. Garay, J. Nebus, et al.
change from baseline and incidence of positive responders
were calculated for each time point.
RESULTS
Preparations of Extracts
The hexane extract of colloidal oatmeal (HCO) generated an
oily residue (3.8% yield); the aqueous acetone extract of col-
loidal oatmeal (ACO) generated a sticky amorphous powder
(2.6% yield); the aqueous methanol extract of colloidal oatmeal
(MCO) generated a dry amorphous powder (2.5% yield); the
water extract of colloidal oatmeal (WCO) generated a white
powder (0.7% yield). An approximate qualitative composition
of each extract based on the nature of extraction processes is
presented in Table 1 along with a summary of bioactivities.
Gene Expression
The effect of the colloidal oat extracts on the basal (unstimu-
lated) gene expression of IL-8 by human keratinocytes were
investigated. MCO, ACO, and HCO oat extracts (50 µg/mL)
signicantly decrease the mRNA transcript levels of IL-8 (P<
0.001, P< 0.001, and P< 0.01, respectively) in unstimulated ke-
ratinocytes (Figure 1A).
TNF-α induced IL-8 release
Treatment with TNF-α induced a significant increase in
interleukin-8 (IL-8) production by human keratinocytes.
Pre-treatment with MCO or WCO oat extracts significantly
inhibited the expression of IL-8 (P= 0.004 and P= 0.0002, re-
spectively) (Figure 1B).
NF-κB Luciferase Promoter Assay in Keratinocytes
Treatment with TNF-α induced a signicant increase NF-κB pro-
moter activity in primary human keratinocytes. Pre-treatment
with ACO oat extract resulted in a signicant inhibition of NF-κB
promoter levels over the TNF-α treated controls at 25 and 50 µg/
mL doses (P< 0.05) (Figure 2A).
UV-induced ROS Assay
Exposing human keratinocytes to UV light resulted in a signi-
cant increase in reactive oxygen species (ROS) generation by
the cells. There was a dose-dependent decrease in UV-induced
Population
Twenty-nine healthy female subjects between 18 and 60 years of
age completed the study. Mean age was 44.5. All subjects exhib-
ited bilateral mild to moderate itch with moderate to severe dry
skin on their lower legs. Exclusion criteria included those with
acute inammatory lesions on the lower leg area, individuals on
medications that could interfere with the outcome of the study
and those with known allergies/sensitivities to the test product.
Clinical Evaluations
Clinic evaluations were conducted at baseline, day 1, day 7, and
day 14. Participants also conducted self-evaluation questionnaires.
Clinical Grading
Subjects were clinically graded on the right or left lower legs
for the following objective irritation parameters: cracking, scal-
ing, dryness, erythema, and roughness. Subjects assessed the
following subjective irritation parameters on the lower leg area:
burning/stinging, tightness, and itching.
Corneometer Measurements
Triplicate Corneometer CM 825 (Courage + Khazaka Electric
GmbH) measurements were obtained on the lower leg (mid-
way between the major joints) to quantify moisture content in
the stratum corneum.
TEWL Measurements
Trans-epidermal water loss (TEWL) measurements were taken
on the lower leg area using the Dermalab (Cortex Technologies)
in conjunction with a computer.
Hi-Scope Images
A high-denition microscope [Hi-Scope KH-2400R videomi-
croscope (100x)] was used to capture images of the skin at
designated areas on the right and left lower lateral leg.
Biostatistics
Mean values for objective/subjective irritation parameters,
instrumental measurements (Corneometer and TEWL), and
self-assessment questionnaires at day 1, day 7, and day 14
were statistically compared to mean baseline values using a
paired t-test at the P≤ 0.05 signicance level. Average percent
TABLE 1.
Summary of Colloidal Oat Extracts
Extraction solvent Abbreviation Expected Phytochemistry IL-8 Production NF-κB Promoter ROS Production IL-8 Transcription
Hexanes HCO Oils and lipophilic compounds Decrease n/a n/a Decrease
80% Acetone ACO Mid-polar phenolics n/a Decrease Decrease Decrease
80% MeOH MCO Phenolics Decrease n/a n/a Decrease
Water WCO Proteins and carbohydrates Decrease n/a n/a n/a
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This document contains proprietary information, images and marks of Journal of Drugs in Dermatology (JDD).
No reproduction or use of any portion of the contents of these materials may be made without the express written consent of JDD.
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46
K. A. Reynertson, M. Garay, J. Nebus, et al.
ROS generation in cells treated with ACO oat extract (Figure
2B). At 10 and 100 µg/mL concentrations, there was a signicant
decrease (P = 0.0071 and P = 0.0004, respectively).
Clinical Results
Clinical evaluations of the subjects’ lower legs showed signi-
cant improvements (P< 0.05) in skin dryness, scaling, roughness,
itching and cracking after one day of use of the oat-containing
skin lotion when compared to baseline mean values (Figure 3).
This improvement increased over time (7 and 14 days). After 14
days, all subjects showed improvements in cracking, scaling, and
skin dryness. In addition, TEWL and corneometer measurements
improved signicantly (P< 0.05) at 7 and 14 days (Figure 4).
Study participants also completed surveys and assessments.
Subjects perceived a signicant (P< 0.05) mean reduction in itch
intensity after one day of using the colloidal oatmeal skin pro-
tectant lotion (Figure 5). Improvements in skin dryness, textural
parameters, and itch were maintained throughout the remain-
der of the study. Subjects also noticed signicant improvements
(P< 0.05) in roughness, dryness, itch and scaling of their skin as
early as the day 1 time point. And nally, high-resolution digital
imaging showed dramatic visible improvements in skin textural
properties including dryness and aking after two weeks of us-
ing the colloidal oatmeal skin protectant lotion (Figure 6).
DISCUSSION
Oats have been extensively used for dermatological treatment
of inammatory skin conditions, yet few studies have delin-
eated the anti-inammatory mechanism of action of colloidal
oatmeal. In the current study we report that extracts of colloi-
dal oatmeal show anti-inammatory and antioxidant activity,
as demonstrated by a decrease in NF-κB promoter activity,
decreased ROS generation, and reduced IL-8 production.
FIGURE 1. (A) At 50 µg/mL, HCO, MCO, and ACO decreased the mRNA transcript levels of interleukin-8 (IL-8) in primary human keratinocytes. (B) In
primary human keratinocytes induced with TNF-α, MCO and WCO led to inhibition of the production of the inammatory IL-8. (** P<0.01; ***P<0.001)
FIGURE 2. (A) At 25 and 50 µg/mL, ACO led to inhibition of TNF-α induced NF-κB promoter activity in primary human keratinocytes. Data expressed
as Relative Luminecence Units (RLU). (B) ACO led to a dose-dependent inhibition of UV-induced reactive oxygen species (ROS) in primary human
keratinocytes. Data expressed as Mean Fluorescence Intensity (MFI). (*P< 0.05; **P< 0.01; ***P< 0.001).
(A)
(A)
(B)
(B)
μ
(μg/mL)
α
μ
k
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47
K. A. Reynertson, M. Garay, J. Nebus, et al.
(pruritus) leads to scratching, which in turn causes damage
to skin barrier and integrity; this damage creates a feedback
loop and an increased release of pro-inammatory mediators
that will, in turn, increase the feeling of itch. Compositions
with colloidal oatmeal reduce skin inammation and there-
fore may help stop the itch-scratch cycle.
Our studies indicate that fractions rich in different oat phyto-
chemicals can also mediate biochemical pathways important in
assuaging clinical challenges. Oat has the highest oil content
(upwards of 10%) of any grain, and is rich in antioxidants and
other phytochemicals.7 Unlike other lipid-rich plants, whole oat
extracts contain components that not only protect oat lipids
from spoilage, they also form a synergistic mixture that en-
hances oat oil’s capacity to protect and nurture healthy skin.
HCO represents the oil fraction of colloidal oatmeal, and dem-
onstrated both a decrease in IL-8 transcription and cytokine
production. Phenolics (avonoids, avenanthramides) and
alcohol-soluble albumin proteins will be found in MCO and
ACO, which demonstrated decreases in ROS production, IL-8
FIGURE 3. At all evaluation timepoints, subjects using the colloidal
oatmeal skin protectant lotion showed signicant improvements in
cracking, scaling, dryness, and roughness. *P< 0.05
FIGURE 5. Signicant improvements with itchy skin were observed
over the course of the 14-day treatment (P< 0.05).
FIGURE 4. (A) Transepidermal water loss (TEWL) and (B) corneom-
eter measurements improved significantly (P< 0.05) over the course
of the 14 days study using the colloidal oat containing lotion.
(A)
(B)
NF-κB is a key nuclear receptor driving the expression of
many pro-inammatory and oxidative pathways during in-
ammation. Overexpression of the inammatory cytokine
IL-8 has been linked to itch and pruritic skin disease.15 Itch
FIGURE 6. High resolution digital images before and after use of the
colloidal oat skin protectant lotion.
Moderate to intense Itch
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48
K. A. Reynertson, M. Garay, J. Nebus, et al.
transcription and production, and NF-κB promoter activity. In
addition, WCO, rich in water-soluble oat proteins (globulins
and prolamines) and carbohydrates, demonstrated the stron-
gest decrease in IL-8 production of any extract (P= 0.0002). To
our knowledge this is the rst study to report that oat proteins
may directly contribute to the anti-inammatory skin benets
of oatmeal. Although these fractions were studied indepen-
dently in the current study, all of these extracts are contained
in whole colloidal oatmeal. It is expected that when formu-
lated into colloidal oatmeal-containing lotions, the proteins,
phenolic antioxidants, and lipids would contribute to the over-
all clinical efcacy of colloidal oatmeal.
The reduction of inammatory mediators in skin may assist in
the relief of a variety of skin conditions that result in dryness,
eczema, irritation, and inammation. Our clinical study demon-
strated that a colloidal oatmeal skin protectant lotion was able
to signicantly alleviate the itch (P< 0.05) and improve the con-
dition of moderate to severely dry skin, and was well tolerated
in subjects with compromised itchy, dry skin. Clinical evalua-
tions showed that the colloidal oatmeal skin protectant lotion
signicantly improved dryness, scaling and roughness as early
as 1 day after use, and these improvements were maintained
over the duration of the study with continued use of the lotion.
Subjects perceived a signicant mean reduction in itch intensi-
ty after only 1 day of use of the colloidal oatmeal skin protectant
lotion, and also reported that their itch signicantly improved
shortly after each application of the lotion as demonstrated
by comparison between pre-application and post-application
scores on days 0 through 3. Finally, transepidermal water loss
rates at days 7 and 14 showed a signicant improvement (P<
0.05) in skin barrier function when compared to baseline values.
Taken together, these in vitro results demonstrate that colloidal
oatmeal can reduce inammatory mediators associated with
skin inammation; the inhibition of inammatory mediator pro-
duction may in turn help enhance the skin benets of colloidal
oatmeal for dry, irritated, and eczematous skin.
ACKNOWLEDGMENTS
Portions of this manuscript were presented as posters at the
2006 and 2014 American Academy of Dermatology Annual
Meetings and the 2013 American Society of Pharmacognosy
Annual Meeting.
DISCLOSURES
All authors are employees of Johnson & Johnson Consumer
Companies, Inc., the manufacturer of Aveeno® products. The
preparation of this manuscript was sponsored in full by John-
son & Johnson Consumer Companies, Inc.
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AUTHOR CORRESPONDENCE
Kurt A. Reynertson PhD
E-mail:................……........................................ kreynert@its.jnj.com
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No reproduction or use of any portion of the contents of these materials may be made without the express written consent of JDD.
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