Assessment of dermal wound healing and in vitro antioxidant properties of

Avena sativa L.
E. Kpeli Akkol
a,
*
, I. Sntar
a
, I. Erdogan Orhan
a
, H. Keles
b
, A. Kan
c
, G. oksari
d
a
Department of Pharmacognosy, Faculty of Pharmacy, Gazi University, Etiler, 06330 Ankara, Turkey
b
Department of Pathology, Faculty of Veterinary Medicine, Afyon Kocatepe University, 03030 Afyonkarahisar, Turkey
c
Vocational School of Technical Sciences, Program for Food Technologies, Seluk University, 42070 Konya, Turkey
d
Department of Field Crops, Faculty of Agriculture, Seluk University, 42070 Konya, Turkey
a r t i c l e i n f o
Article history:
Received 9 April 2010
Received in revised form
15 December 2010
Accepted 24 January 2011
Keywords:
Avena sativa
Wound healing
Antioxidant
Total phenol and avonoid
a b s t r a c t
Avena sativa L. (Poaceae) has been reported to have traditional utilization against skin diseases and
inammation. Therefore, in this study, the n-hexane, ethyl acetate, ethanol, and water extracts of A. sativa
were investigated for their wound healing and antioxidant activities. Total phenol and avonoid contents
of the extracts were established spectrophotometrically. For the wound healing activity, linear incision
and circular excision models on rats and mice were evaluated with a standard ointment Madecassol

.
Antioxidant activity was evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, ferrous
ion-chelating, and ferric-reducing antioxidant power (FRAP) assays. Signicant wound healing activity
was observed with the ointment formulation of the ethanol extract at 1% concentration. The histo-
pathological examination results also supported the outcome of both linear incision and circular excision
wound models. All of the extracts exerted low antioxidant activity in the applied assays. The present
study provides a scientic evidence for the traditional usage of A. sativa in the management of wound
healing.
2011 Published by Elsevier Ltd.
1. Introduction
Avena sativa L. (Poaceae), known as common oat, is a cereal
grain species used also as food. It has been cultivated in Anatolian
peninsula sinceancient times whose cultivationwas recordedbythe
Greek scientist Galenus in the 17th century. The major cultivation
area of oat is the Marmara region and internal Anatolia inTurkey. In
some folk medicine systems belonging to different parts of the
world, it has been reported to be used traditionally against some
diseases. For instance; in the southern Appalachia region of the
United States, A. sativa was recorded to have utilization against
chickenpox, poison ivy, and other rashes externally (Cavender,
2006). In traditional medicine of Lebanon, its alcoholic macerate
was reported to be used as antirheumatic and antineuralgic (El
Beyrouthy et al., 2008). Ballabh et al. (2008) reported use of oat
against kidney and urinary disorders in traditional medicine of clod
desert Ladakhinthe remotest regionof the Indiansubcontinent. Oat
is an important crop plant containing high levels of proteins, lipids,
vitamins, dietary bers, and minerals as well as a food plant for both
humans and animals (Gajdosova et al., 2007). A. sativa was reported
to possess a variety of biological activities such as angiotensin-I
converting enzyme inhibitory (Cheung et al., 2009), anti-inam-
matory and anti-itchy (Sur et al., 2008), anti-HIV (Shun et al., 2008),
and cardioprotective activities (Ryan et al., 2007).
The aim of the present study was to investigate in vivo wound
healing activity of A. sativa to clarify its traditional use in a scientic
platform along with its in vitro antioxidant activity. The n-hexane,
ethyl acetate, ethanol, and water extracts of the plant were tested in
mice and rats for their wound healing activity using in vivo linear
incision and circular excision wound models, which have been used
together for conrmation of the wound healing activity. Addition-
ally, antioxidant activity was conducted in the extracts to exploit its
relation to wound healing activity. Antioxidant activity of the
extracts was determined using three different in vitro methods; 2,2-
diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, ferrous ion-
chelating, and ferric-reducing antioxidant power (FRAP) assays at
250, 500, and 1000 mg/ml concentrations. Total phenol and avo-
noid contents of the extracts were established spectrophotomet-
rically by FolineCiocalteau and AlCl
3
methods, respectively.
Abbreviations: ABTS, 2,2
0
azinobis-(3-ethylbenzthiazoline-6-sulphonic acid);
DPPH, 2,2-diphenyl-1-picrylhydrazyl; FRAP, Ferric-reducing antioxidant power;
S.E.M, Standard error mean.
* Corresponding author. Tel.: 90 312 2023185; fax: 90 312 2235018.
E-mail address: esrak@gazi.edu.tr (E.K. Akkol).
Contents lists available at ScienceDirect
Journal of Cereal Science
j ournal homepage: www. el sevi er. com/ l ocat e/ j cs
0733-5210/$ e see front matter 2011 Published by Elsevier Ltd.
doi:10.1016/j.jcs.2011.01.009
Journal of Cereal Science 53 (2011) 285e290
[280]
2. Experimental
2.1. Plant material
Cultivated sample of A. sativa was obtained from the experi-
mental farm of Seluk University, in Konya province, Turkey, at the
owering stage in June, 2009.
2.2. Preparation of plant extracts
After the plant material was driedinshade at roomtemperature, it
was weighedaccurately(200g) ina digital balancer (MettlereToledo)
and powdered in a mechanical grinder. Then, the powdered material
was subjected to successive extraction starting from n-hexane
(2 L 2), ethyl acetate (3 L 2), ethanol (3 L 2), and water (3 L 1).
After the corresponding organic phases in each extraction were
lteredthroughregular lter paper, they were evaporatedinvacuotill
dryness to give the crude extracts. The water extract was nally
lyophilized. The extract yields (w/w) were calculated as follows: The
n-hexaneextract; 2.09%, theethyl acetate extract; 25.98%, theethanol
extract; 15.61%, the water extract; 29.47%.
2.3. Wound healing activity tests
2.3.1. Animals
Male, SpragueeDawley rats (160e180 g) and Swiss albino mice
(20e25 g) were purchased fromthe animal breeding laboratories of
Experimental Animal Research Center of Gazi University (GDAM)
(Ankara, Turkey).
The animals were left for 3 days at room conditions for accli-
matization. They were maintained on standard pellet diet and
water ad libitum throughout the experiment. A minimum of six
animals were used in each group. Throughout the experiments,
animals were processed according to the suggested international
ethical guidelines for the care of laboratory animals under the audit
of Gazi University Commission of Animal Ethics.
2.3.2. Preparation of test samples for wound healing activity
Incision and excision wound models were employed to evaluate
the wound healing activity. For the in vivo wound models, test
samples were prepared in an ointment base (vehicle) consisting of
glycol stearate, 1,2-propylene glycol, liquid parafn (3:6:1) in 1%
concentration. 0.5 g of each test ointment was applied topically on
the wounded site immediately after the wound was created by
a surgical blade.
Animals of the vehicle group were treated with the ointment
base only, whereas animals of the reference drug group were
treated with 0.5 g of Madecassol

(Bayer, 00001199). Madecassol

contains a 1% extract of Centella asiatica.

2.3.3. Wound healing activity
2.3.3.1. Linear incision wound model. All the animals were anaes-
thetized with 0.15 cc Ketalar

and the back hair of the rats were

shaved by using a shaving machine. Five cm long, two linear-par-
avertebral incisions were made with a sterile surgical blade
through the full thickness of the skin at a distance of 1.5 cm from
the midline of each side of the vertebral column (Ehrlich and Hunt,
1968). The wounds were closed with three surgical interrupted
sutures 1 cmapart. The extracts, reference drug (Madecassol

), and
vehicle were topically applied once a day throughout 9 days.
Animals of the negative control group were not treated with any
material. All the sutures were removed on the 9th post wound day.
On the 10th day, all the animals were killed under anesthesia. One
linear-paravertebral incised skinwas measured using a tensiometer
(Zwick/Roell Z0.5, Germany) for its tensile strength, the other
incised skin was sent for histopathological examination (Lodhi
et al., 2006; Suguna et al., 2002).
Tensiometer measures the breaking strength in N (Newtons),
which is called tensile strength.
Tensile strengthTS of extract %
TS
extract
TS
vehicle
100
TS
vehicle
Tensile strength of reference %
TS
reference
TS
vehicle
100
TS
vehicle
Tensile strength of vechicle %
TS
vechicle
TS
negative
100
TS
negative
2.3.3.2. Circular excision wound model. A circular excision wound
model was used to monitor wound contraction and wound closure
time. Each group of animals (six animals in each) was anaesthetized
by 0.01 cc Ketalar

. The back hairs of the mice were depilated by

shaving. The circular wound was created on the dorsal inter-
scapular region of each animal by excising the skin with a 5 mm
biopsy punch; wounds were left open (Tramontina et al., 2002). The
extracts, the reference drug (Madecassol

, Bayer) and the vehicle

ointments were applied topically once a day till the wound was
completely healed. The progressive changes in wound area were
monitored by a camera (Fuji, S20 Pro, Japan) every other day. Later
on, wound area was evaluated by using AutoCAD program. Wound
contraction was calculated as percentage of the reduction in
wounded area. A specimen sample of tissue was isolated from the
healed skin of each group of mice for the histopathological exam-
ination (Sadaf et al., 2006).
2.3.4. Histopathological examination
The cross-sectional full-thickness skin specimens from each
group were collected at the end of the experiment to evaluate for
the histopathological alterations. Samples were xed in 10% buff-
ered formalin, processed and blocked with parafn and then
sectioned into 5 mm sections and stained with hematoxylin & eosin
(HE) and Van Gieson (VG) stains. The tissues were examined by
light microscope (Olympus CX41 attached Kameram

Digital Image
Analyze System) and graded as mild (), moderate () and severe
() for epidermal or dermal re-modeling. Re-epithelization or
ulcus in epidermis; broblast proliferation, mononuclear and/or
polymorphonuclear cells, neovascularization and collagen deposi-
tions in dermis were analyzed to score the epidermal or dermal re-
modeling. At the end of the examination, obtained results were
combined and staged for wound healing phases as inammation,
proliferation, and re-modeling in all groups.
Table 1
Effects of the extracts of A. sativa on linear incision wound model.
Materials Statistical
mean S.E.M.
a
(mm
2
)
(Tensile
strength %
b
)
Vehicle 14.86 1.81 12.1
Negative control 13.26 2.19 e
c
n-Hexane extract 14.31 2.03 e
Ethyl acetate extract 17.42 1.14 17.2
Ethanol extract 19.21 1.87 29.3
Water extract 17.91 1.21 20.5
Madecassol

23.45 1.75 57.8

a
Standard error meaning.
b
Percentage of the tensile strength values: The vehicle group was compared to
the negative control group; Extracts and the reference material were compared to
vehicle group.
c
No activity.
E.K. Akkol et al. / Journal of Cereal Science 53 (2011) 285e290 286
2.4. Antioxidant activity
2.4.1. DPPH radical scavenging assay
The hydrogen atom or electron donation capacity of the corre-
sponding extracts was computed from the bleaching property of
the purple-colored methanol solution of 2,2-diphenyl-1-picrylhy-
drazyl (DPPH). The stable DPPH radical scavenging activity was
determined by the method of Blois (1958). The samples and
reference dissolved in ethanol (75%) were mixed with DPPH solu-
tion (1.5 10
4
M). Remaining DPPH amount was measured at
520 nm using a Unico 4802 UVevisible double beam spectropho-
tometer (USA). The results were compared to that of gallic acid
employed as the reference. Inhibition of DPPH in percent (I%) was
calculated as given below:
I%
h
A
control
A
sample
.
A
control
i
100
where A
control
is the absorbance of the control reaction (containing
all reagents except the test sample), and A
sample
is the absorbance of
the extracts/reference. Experiments were run in triplicate and the
results were conveyed as average values with S.E.M. (Standard error
of mean).
2.4.2. Fe
2
eferrozine test system for iron chelating
The ferrous ion-chelating effect of the extracts by the
Fe
2
eferrozine test system was estimated by the method of Chua
et al. (2008). Accordingly, the samples were incubated with 2 mM
FeCl
2
solution. The reactionwas initiatedbythe additionof ferrozine
solution into the mixture and left standing at ambient temperature
for 10 min. The absorbance of the reaction mixture was measured at
562 nm. The ratio of inhibition of the ferrozineeFe
2
complex
formation was calculated as follows:
I%
h
A
control
A
sample
.
A
control
i
100
The control contained only FeCl
2
and ferrozine. Analyses were
run in triplicate and expressed as average values with S.E.M.
(Standard error of mean).
2.4.3. The ferric-reducing antioxidant power (FRAP)
The ferric-reducing antioxidant power (FRAP) of the extracts
and reference was tested using the assay of Oyaizu (1986). Different
concentrations of the extracts as well as chlorogenic acid as refer-
ence for comparative purposes was added to 2.5 ml of phosphate
buffer (pH 6.6) and 2.5 ml of potassium ferricyanide. Later, the
mixture was incubated at 50

C for 20 min and then trichloroacetic
acid (10%) was added. After the mixture was shaken vigorously, this
solution was mixed with distilled water and FeCl
3
(0.1%, w/v). After
30 min incubation, absorbance was read at 700 nm using a Unico
4802 UVevisible double beam spectrophotometer (USA). Analyses
were achieved in duplicate. Increase in absorbance of the reaction
indicated increase in reducing power of the extracts.
2.5. Determination of total phenol and total avonoid contents of
the extracts
Phenolic compounds were determined in accordance with
FolineCiocalteaus method (Singleton and Rossi, 1965). In brief, the
Table 2
Effects of the extracts of A. sativa on circular excision wound model.
Day Wound area (mm
2
) S.E.M.
a
(Contraction %
b
)
Vehicle Negative Control n-Hexane extract Ethyl acetate extract Ethanol extract Water extract Madecassol

0 19.14 1.89 19.60 2.09 19.86 1.22 20.05 1.03 20.28 1.68 19.25 1.26 19.21 1.44
2 17.22 1.40 17.30 1.56 17.01 2.24 16.29 1.69 16.05 1.10 17.04 2.37 15.29 2.09
(0.5) (1.2) (5.4) (6.8) (1.0) (11.2)
4 14.31 1.82 16.43 2.99 14.08 2.14 13.32 1.50 12.21 1.54 12.46 1.26 11.29 2.76
(12.9) (1.6) (6.9) (14.7) (12.9) (21.1)
6 12.53 1.07 13.42 1.62 12.27 1.38 10.62 1.63 9.96 1.32 11.10 1.17 8.43 1.64
(6.63) (2.1) (15.2) (20.51) (11.4) (32.7)*
8 8.77 1.37 10.78 1.54 8.80 1.43 7.53 0.36 6.46 0.11 8.51 0.94 3.90 1.71
(18.6) e
c
(14.1) (26.3) (2.9) (55.5)**
10 4.58 0.91 4.96 1.19 4.63 0.17 3.49 0.66 3.09 0.70 3.98 0.86 1.03 0.18
(7.7) e (23.8) (32.5)* (13.1) (77.5)***
12 2.50 0.06 2.76 0.17 2.59 0.19 1.76 0.26 1.48 0.03 2.24 0.16 0.00 0.00
(9.4) e (29.6) (40.8)* (10.4) (100.0)***
*p < 0.05.
***p < 0.001.
a
Standard error meaning.
b
Percentage of the contraction values: The vehicle group was compared to the negative control group; Extracts and the reference material were compared to vehicle group.
c
No activity.
Table 3
Wound healing processes and healing phases of the vehicle, negative control, A. sativa extracts and Madecassol

administered animals.
a
Groups Wound Healing Processes Healing Phases
S U RE FP CD MNC PMN NV I P R
Vehicle / / /
Negative Control / e / / e
n-Hexane extract / e e
Ethyl acetate extract / / / /
Ethanol extract e / / / / / / / /
Water extract / / / / / / /
Madecassol

e / / / / / /
a
HE and VG stained sections were scored as mild (), moderate () and severe () for epidermal and/or dermal re-modeling. S: Scab, U: Ulcus, RE: Re-epithelization,
FP: Fibroblast proliferation, CD: Collagen depositions, MNC: Mononuclear cells, PMN: Polymorphonuclear cells, NV: Neovascularization, I: Inammation phase, P: Proliferation
phase, R: Re-modeling phase.
E.K. Akkol et al. / Journal of Cereal Science 53 (2011) 285e290 287
samples were mixed with 750 mL of FolineCiocalteaus reagent and
sodium carbonate in test tubes. The tubes were then vortexed and
incubated at 40

C for 30 min. Afterward, absorption was measured
at 760 nm using a Unico 4802 UVevisible double beam spectro-
photometer (USA). Total avonoid content was calculated by the
aluminum chloride colorimetric method (Woisky and Salatino,
1998). To sum up, a number of dilutions of quercetin were
obtained to prepare a calibration curve. Then, the same dilutions
from the samples were also prepared and separately mixed with
95% ethanol, 10% aluminum chloride, 1 M sodium acetate and
distilled water. Following incubation for 30 min at room tempera-
ture, absorbance of the reaction mixture was measured at a wave-
length of 415 nm with a Unico 4802 UVevisible double beam
spectrophotometer (USA). The total phenol and avonoid contents
of the extracts were expressed as gallic acid and quercetin equiv-
alents (mg/g extract), respectively.
2.6. Statistical analysis of the data
The data on percentage wound healing was statistically
analyzed using one-way analysis of variance (ANOVA). The values
of p 0.05 were considered statistically signicant. Histopathologic
data were considered to be nonparametric; therefore, no statistical
tests were performed.
3. Results
3.1. Results of wound healing activity
The wound healing activity of the n-hexane, ethyl acetate,
ethanol, and water extracts of A. sativa was evaluated on rats and
mice by linear incision and circular excision wound models to
conrm the claimed folkloric usage of the plant on a scientic base.
The histopathological changes formed by these extracts were also
assessed. The results of the measurements of tensile strength were
shown in Table 1. Tensile strength of the animals treated with the
ethanol extract of the plant demonstrated the highest value (29.3%,
p < 0.05) on day 10. Topical application of the extract on the inci-
sion wound model expressed a signicant increase in wound
tensile strength as compared to other extract treated groups and
the control groups. On the other hand, reference drug Madecassol

demonstrated 57.8% (p < 0.001) tensile strength.

Measurements of the progress of wound healing induced by the
extracts, reference drug, vehicle groups, and negative control in the
circular excision wound model were shown in Table 2. Animals
treated with the ethanol extract revealed 32.5% (p<0.05) contrac-
tion on the 10th day, and the same extract demonstrated 40.8%
(p < 0.05) contraction on the 12th day. However, the rest of the
extracts demonstrated no signicant result on the given days.
Phases inwound healing processes (inammation, proliferation,
and re-modeling) were observed and recorded within the experi-
mental groups with different degree (Table 3). The negative control,
vehicle, and n-hexane groups demonstrated delayed wound heal-
ing processes. In comparison with these groups, faster re-modeling
Fig. 1. Histopathological view of wound healing and epidermal/dermal re-modeling in
the vehicle, negative control, A. sativa extracts and reference ointment Madecassol

administered animals. Skin sections show the hematoxylin & eosin (HE) stained
epidermis and dermis in A, and the dermis stained with Van Gieson (VG) in B. The
original magnication was 100 and the scale bars represent 120 mm for gures in A,
and the original magnication was 400 and the scale bars represent 40 mm for B. Data
is representative of 6 animals per group. 1) Vehicle group, 10 day old wound tissue
treated with only vehicle, 2) Negative Control group, 10 day old wound tissue,
untreated group, 3) n-Hexane extract group, 10 day old wound tissue treated with n-
Hexane extract, 4) Ethyl acetate extract group, 10 day old wound tissue treated with
ethyl acetate extract, 5) Ethanol extract group, 10 day old wound tissue treated with
Ethanol extract, 6) Water extract group, 10 day old wound tissue treated with water
extract, 7) Reference group, 10 day old wound tissue treated with Madecassol

.
Arrows pointing events during wound healing; s: scab, u: ulcus, re: re-epithelization, f:
broblast, c: collagen, mnc: mononuclear cells, pmn: polymorphonuclear cells, nv:
neovascularization.
E.K. Akkol et al. / Journal of Cereal Science 53 (2011) 285e290 288
was seen in the other groups. In experimental groups, best re-
modeling, particularly, re-epithelization was detected in the
Madecassol

treated group, and then, the same equal in both

ethanol and water extracts treated groups. Compared with these
groups, indecisive healing results and excessive neovascularization
were observed in the ethyl acetate-treated group. Histopathological
results are demonstrated in Fig. 1, which stained with HE and VG.
Accordingly, histopathological observations supported the out-
come of in vivo wound models of this study.
3.2. Antioxidant activity results and total phenol and avonoid
contents
Antioxidant activity of the extracts of A. sativa was tested in
three different in vitro assays; namely DPPH radical scavenging,
ferrous ion-chelating, and ferric-reducing antioxidant power
(FRAP) assays. As tabulated in Tables 4 and 5, the extracts showed
a low scavenging activity against DPPH radical and stumpy
chelating capacity on ferric ion. However, occurrence of moderate
FRAP was observed in the extracts.
For total phenol content of the extracts, the calibration equation
was calculated as y 0.9051x 0.0549 (r
2
0.9943) (gallic acid
equivalent), while it was y 5.5398x 0.1209 (r
2
0.9988)
(quercetin equivalent) for total avonoid content (Table 4). Our
ndings showed that the ethyl acetate extract was the richest in
terms of total phenol (75.79 4.02 mg/g extract) and avonoid
(77.59 6.71 mg/g extract) amounts as compared to the others.
4. Discussion
Wound healing is a multifarious procedure that includes
inammation, granulation tissue formation, re-epithelialization,
and matrix formation. Numerous studies have proved that oxida-
tion reactions, frequently initiated by various unwanted radicals in
biological systems, are associated with many pathophysiological
conditions including inammation and wound healing. In recent
years, oxidative stress and free radicals have been implicated in
impaired wound healing. Different mechanisms like free radical
scavenging, metal chelation as well as immune modulation may act
at different levels independently or in combination to bring about
the wound healing effects. Consequently, we also studied herein
antioxidant effects of A. sativa extracts besides their wound healing
activity. According to the literature survey; it was revealed that the
leaves, stems, and inorescences of A. sativa contained a range
of avonoid-type compounds including vitexin and isoswertisin
2
00
-rhamnosides, isovitexin and isoorientin 2
00
-arabinosides
(Chopin et al., 1977), which might be related to the wound healing
and antioxidant activities. Not only the avonoid type of constitu-
ents, but also vitamin E, phytic acid, and avenanthramides are the
most abundant constituents that exhibit antioxidant activity
(Peterson, 2001). Avenanthramides (substituted N-cinnamoylan-
thranilic acids), the alkaloid-class compounds specic to A. sativa,
have been reported to be strong antioxidants in both in vitro and in
vivo methods (Bratt et al., 2003; Chen et al., 2007; Dimberg et al.,
1993; Ji et al., 2003). Since avenanthramides were shown to
possess antioxidant activity, they might be responsible for antiox-
idant activity of the oat extracts as well. However, the amount of
avenanthramides in A. sativa samples may depend on several
factors such as genotype, soil factors, climate, irrigation, collection
period, etc. Therefore, their amounts might be supposed to be quite
low in our sample, which may have led to occurrence of low anti-
oxidant activity in our extracts. Nevertheless, the method distinc-
tion to assess antioxidant activity can also be speculated to
inuence level of activity. Wojdyo and Oszmainski (2007) analyzed
phenolic acids of the enzymatically hydrolyzed extract and the
methanol extract of oat and also studied their antioxidant activity
using scavenging tests against DPPH and 2,2
0
azinobis-(3- ethyl-
benzthiazoline-6-sulphonic acid) (ABTS) radicals. Although ferulic
Table 4
Total phenol and total avonoid contents and of DPPH radical scavenging activity of A. sativa extracts.
Materials Total phenol content S.E.M.
a
of the extracts
b
Total avonoid content S.E.M.
of the extracts
c
Inhibition % S.E.M. against DPPH radical
250 mg ml
1
500 mg ml
1
1000 mg ml
1
neHexane 26.10 2.31 40.72 4.81 3.72 1.24 4.69 0.12 7.04 0.42
Ethyl acetate 75.79 4.02 77.59 6.71 4.90 1.92 6.15 0.01 11.29 1.42
Ethanol 39.34 0.78 ND
d
6.51 0.50 7.03 0.08 11.35 0.17
Water 46.02 0.70 ND 4.02 0.67 5.97 1.42 6.50 1.17
Gallic acid (Reference for DPPH test) NT
e
91.61 0.06 92.57 0.10
a
Standard error meaning.
b
Data expressed in mg equivalent of gallic acid (GAE) to 1 g of extract.
c
Data expressed in mg equivalent of quercetin to 1 g of extract.
d
Not detected.
e
Not tested.
Table 5
Ferric ion-chelating capacity and ferric-reducing antioxidant power (FRAP) of A. sativa extracts.
Materials Ferric ionechelating capacity(Inhibition % S.E.M.
a
) Ferricereducing antioxidant power (FRAP)
(Absorbance at 700 nm S.E.M.)
250 mg ml
1
500 mg ml
1
1000 mg ml
1
250 mg ml
1
500 mg ml
1
1000 mg ml
1
neHexane 10.19 3.38 16.16 2.40 23.37 3.17 0.231 0.007 0.308 0.007 0.409 0.001
Ethyl acetate 11.97 1.89 11.77 1.04 11.29 0.03 0.268 0.001 0.324 0.007 0.475 0.001
Ethanol 3.96 2.19 7.95 0.06 8.13 0.54 0.234 0.011 0.261 0.023 0.332 0.038
Water 5.72 2.31 11.29 0.99 36.68 1.34 0.209 0.001 0.217 0.008 0.259 0.004
BHA
b
ND
d
21.71 1.10 26.94 1.48
Chlorogenic acid
c
ND 3.547 0.06 3.618 0.01
a
Standard error meaning.
b
Butylated hydroxyanisol; reference for ferric ion chelating capacity test.
c
Reference for FRAP.
d
Not determined.
E.K. Akkol et al. / Journal of Cereal Science 53 (2011) 285e290 289
acid was dominant among others, the rest of the phenolic acids
were in very small quantities. The authors stated that the hydro-
lyzed extract of oat had signicantly higher antioxidant activity
than the methanol extract.
On the other hand, there have been several studies on A. sativa,
which can be related to a wound healing effect. For instance, beta-
glucan obtained from A. sativa was revealed to enhance the resis-
tance to Staphylococcus aureus or Eimeria vermiformis infection in
mice (Yun et al., 2003). For wound healing activity, antimicrobial
agents are gaining importance, since they also provide a better and
rapid healing by forming a barrier against microbial contamination.
Boisnic et al. (2003) studied the anti-inammatory effect of
oatmeal extract oligomer on skin fragments stimulated by a neu-
romediator, vasoactive intestinal peptide and found out that after
treatment with oatmeal extract oligomer, the mean surface of
dilated vessels and edema were signicantly decreased. Thus, the
wound healing effect of A. sativa may be attributed to this mech-
anism. Moreover, a clinical study on 12 healthy individuals
demonstrated that, oatmeal extracts have preventive effects on
skin irritation in the sodium lauryl sulfate (SLS) irritation model
(Vie et al., 2002). In a randomized cross-over trial, effects of the
unprocessed oat bran having a total of 11 g dietary ber were
assessed on rectal epithelial cell proliferation of twenty subjects
with adenomas and it was concluded that total ber intake did not
display effects on proliferation (Macrae et al., 1997).
All these previous ndings can lead to an interpretation that the
signicant wound healing activity of the ethanol extract of A. sativa
may likely depend on some other mechanism rather than its
antioxidant effect. Besides, beta-glucans, complex polysaccharides
of D-glucose monomers linked by b-glycosidic bonds, have been
revealed to appear to enhance wound healing through several
mechanisms including the stimulation of collagen deposition,
activation of immune cells, increasing macrophage inltration into
the wound area, and by stimulating tissue granulation, collagen
deposition, re-epithelization, and tensile strength (Cerci et al.,
2008). Since beta-glucans have been found to exist commonly in
oat and other grains, wound healing activity of our oat ethanol
extract might be resulting from these components, at least partly.
5. Conclusion
Results of the present study have clearly demonstrated that the
aerial parts of A. sativa possess wound healing potential which
supports its traditional utilization in various folk medicines. In
a reference survey, no reports relating to the wound healing activity
of A. sativa extracts have been found so far. To the best of our
knowledge, we herein report for the rst time about wound healing
activity and antioxidant activity by ferrous ion-chelating and ferric-
reducing antioxidant power (FRAP) methods of the oat extracts.
The current results suggest that wound healing activity of the
ethanol extract of A. sativa may not be related to antioxidant
activity, but some other mechanisms appears to relate to its effect.
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