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Effects of Linum usitatissimum L. ingestion and oil topical application on hair growth in rabbit

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Flax (Linum usitatissimum) is an annual plant of the linaceae family with several biological properties such as an indirect effect on hair regrowth through the intermediary of γ-glutamyl transpeptidase. The aim of this study was to investigate its effect on hair growth process in rabbits. Two trials were carried out using two plant forms and administration routes: oil topical application and seed ingestion. For each trial animals were assigned into two groups, a control and a tested one. A 10cm/10cm area on the back of each rabbit was limited and every four weeks, 10 hairs were plucked to measure their lengths and widths, then the hair of the whole limited area was shaved and weighed. After 4 weeks of use, linseed oil topical application has a significant positive effect on hair width (39.00 µm against 27.2 µm for the control group). Like linseed oil application, the linseed supplementation showed a significant (P<0.10) beneficial effect on hair width, this effect appeared at the 12 th week of ingestion, and did not stopped 04 weeks after (P<0.5) supplementation withdraw. Both seed and oil had no significant positive effect on hair length and weight. These results suggest that flax-seed oil has some hair growth promoting potential. The mechanism of action, and the plant component(s) responsible of this activity, should be investigated.
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Int. J. Med. Arom. Plants, ISSN 2249 4340
RESEARCH ARTICLE
Vol. 3, No. 4, pp. 459-463, December 2013
*Corresponding author: (E-mail) s.halmi25 <@> yahoo.fr
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Effects of Linum usitatissimum L. ingestion and oil topical application
on hair growth in rabbit
BEROUAL K.1, MAAMERI Z.1, HALMI S.1*, BENLEKSIRA B.1, AGABOU A.3, HAMDI
PACHA Y.1, 2
1Laboratory of Toxicology and Pharmacology, Veterinary Science Institute, University of Constantine 1,
Algeria
2National Superior School of Veterinary, Algeria
3PDESCA Laboratory, Veterinary Science Institute, University of Constantine 1, Algeria
Article History: Received 27th October 2013, Revised 17th December 2013, Accepted 18th December 2013.
Abstract: Flax (Linum usitatissimum) is an annual plant of the linaceae family with several biological properties such as
an indirect effect on hair regrowth through the intermediary of γ-glutamyl transpeptidase. The aim of this study was to
investigate its effect on hair growth process in rabbits. Two trials were carried out using two plant forms and administra-
tion routes: oil topical application and seed ingestion. For each trial animals were assigned into two groups, a control and
a tested one. A 10cm/10cm area on the back of each rabbit was limited and every four weeks, 10 hairs were plucked to
measure their lengths and widths, then the hair of the whole limited area was shaved and weighed. After 4 weeks of use,
linseed oil topical application has a significant positive effect on hair width (39.00 µm against 27.2 µm for the control
group). Like linseed oil application, the linseed supplementation showed a significant (P<0.10) beneficial effect on hair
width, this effect appeared at the 12th week of ingestion, and did not stopped 04 weeks after (P<0.5) supplementation
withdraw. Both seed and oil had no significant positive effect on hair length and weight. These results suggest that flax-
seed oil has some hair growth promoting potential. The mechanism of action, and the plant component(s) responsible of
this activity, should be investigated.
Keywords: Linseed, ingestion; linseed oil; topical application; hair growth; rabbits.
Introduction
Phytotherapy is based on the use of herbal
remedies to treat and prevent diseases in hu-
mans and animals. Nowadays, the importance of
phytotherapy is increasing. Many patients prefer
herbal medicines and especially value their good
tolerability and low side effects profile. Fur-
thermore, herbal medicines are now approached
far more scientifically (Eichele 2010).
Alopecia, or hair loss, is a common and of-
ten distressing problem. Actually, no treatment
can completely cure alopecia except the hair-
transplants technique indicated in rebellious and
advanced alopecia cases (Clere 2010).
Various works have been undertaken to
document a variety of medicinal plants used to
improve hair growth in several animal species:
Hibiscus rosa-sinensis (Adhirajam ,et al., 2003),
Asiasari radix; (Rho ,et al., 2005), Amla (fruits
of Embelica officinalis), Brahmi (leaves of
Bacopa monnieri), Methi (seeds of Trigonella
foenumgraecum), Meetha Neem (Murraya
koenigii), Hibiscus rosa sinensis flowers,
(Purwal ,et al., 2008, Banerjee ,et al., 2009),
raspberry (Rubus idaeus) (Harda ,et al., 2008),
Eclipta alba (Roy ,et al., 2008),
Russelia equisetiformis (Awe and Makinde,
2009) and Abrus precatorius (Upadhyay ,et al.,
2012)
Flax (Linum usitatissimum) is an annual
plant of the family Linaceae. It is an oilseed
produced in more than 50 countries mainly in
the northern hemisphere. It contains about 40%
Lipids (most of them Omega-3 fatty acids), 30%
dietary fibers and 20 % protein (Rubilar et al.,
2010). After oil extraction from seeds, the lin-
seed meal is used as a supplement in animal
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Beroual et al.
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feeds (Cattles, rabbits) (Benatmane et al., 2010;
Bouchard 2010).
Flax showed several biological effects like
anti-inflammatory and antimicrobial properties
used successfully in mastitis and skin lesions
treatment (O’Neill et al., 2002). Its fixed oil has
interesting analgesic and antipyretic activities
(Kaithwas et al., 2011)
The aim of this study is to investigate the ef-
fect of linseed ingestion and oil topical applica-
tion on hair growth.
Materials and methods
Experiments procedures used in this study
were approved by the scientific council of the
Institute of Veterinary Sciences (University of
Constantine1. Algeria) and conform to the
guidelines of animal care and use in research
and teaching.
Vegetal material
Linseed and linseed oil were purchased from
a local herbalist. Specimens of the two products
are deposited at the laboratory of pharmacology-
toxicology-Institute of veterinary sciences, Uni-
versity of Constantine (Algeria).
Animals
The experiments have been carried out on
male New Zealand rabbits, weighing approxi-
mately 2.5 Kg and aged 06 months.
Animals were kept in individual standard
cages in the same room and under the same en-
vironmental conditions (temperature, relative
humidity and hygiene practices). Each morning
they received 180g of standard rabbits chow
during an acclimatization period of 07 days.
Experimental design
At the beginning, a limited skin zone (of
10cm/10cm) was shaved on the back of each
rabbit.
Two separate trials were conducted:
The first trial was undertaken (during 04
weeks) to evaluate the effect of linseed oil topi-
cal application (LSOA) on hair growth. Ani-
mals were divided into two groups. The first
group (04 rabbits) served as control group
(CRLo) without any applications while those of
the second group (04 rabbits) served as tested
group (LSOA group) with 01 ml of linseed oil
applied daily on their shaved areas (described
above).
The second trial is designed to evaluate the
effect of linseed Ingestion on hair growth during
a 16 weeks period. Animals were divided into
02 groups. The first group (08 rabbits) did not
receive any feed supplement and served as con-
trol (CRLigroup), the other one (09 rabbits)
served as tested group and daily received the
same feed as the previous group but supple-
mented with 3g of crashed linseed (LSI group)
during 12 weeks. At the end of this period meal
supplementation has been stopped and the kinet-
ic of linseed effects was studied till the 16th
week.
Hair samples
For all groups, hair was sampled every 04
weeks. 10 hairs were plucked with a pair of
tweezers to measure their lengths and widths
using respectively a ruler and a scaled microme-
ter, and then hair was shaved from the delimited
area and weighed with a high precision balance
(0.001g sensitivity. Kern PLS 510-3N. Germa-
ny).
Statistical analysis
Results are expressed as Mean. Statistical
significance was determined using student t-test.
Otherwise, the Wilcoxon T-test was used when
data are not normally distributed with unequal
variance. The software Matlab v7.7.0.2162 (Re-
lease 2008b) was used.
Results
Effect of linseed oil topical application on hair
growth
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Int. J. Med. Arom. Plants
Beroual et al.
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During 48 hours following linseed oil appli-
cation, animals were observed to detect any skin
irritation. Linseed oil application did not cause
any erythema or edema (as indicated by UNO
H, 1991); this attests that linseed oil is non irri-
tant to rabbit skin.
After 04 weeks of topical application, lin-
seed improved hair width but not hair length and
weight. A decrease in hair length was observed
in LSOA group, while its width has significantly
increased compared to the CTLogroup (α<9%)
(Table 1).
Table 1: Hair characteristics after 04 weeks top-
ical application.
Hair
Length (cm)
Width (µm)
Weight (gr)
CTLo
2.72
27.17
0.77
LSOA
2.38
39.00
1.17
P value
0.74
0.086
0.39
Effect of linseed ingestion on hair growth
Hair length
Compared to the control group, linseed in-
gestion had no beneficial effect on hair length
during the 12 weeks of ingestion and 04 weeks
after supplementation withdraw. A significant
transient positive effect was recorded in LSI
group at the 8th week compared the 4th one (Ta-
ble 2).
Table 2: Mean hair length (cm).
4 weeks
08 weeks
12 weeks
16 weeks
CTLi
1.73
2.74
2.32
2.72
LSI
1.66*
2.08*
2.04
2.15
P value
0.64
0.99
0.80
0.81
*P=0.006, α< 1%
Hair width (diameter)
During the first 08 weeks, hair mean width
has linearly decreased in the two groups. The
slight beneficial effect of supplementation was
significant (α<10%) after 12 weeks of intake.
However, this effect didn’t last; since it stopped
04 weeks after supplementation withdraw (cor-
responding to the 5th hair harvest) (α<5%) (Ta-
ble 3).
Table 3: Mean hair width (µm).
4 weeks
08 weeks
12 weeks
16 weeks
CTLi
49.90
45.32
32.60
27.20
LSI
50.09
37.06
40.30
38.00
P value
0.44
0.93
0.096
0.048
Hair weight
As for hair width, after 12 weeks ingestion,
hair weight, showed a non significant beneficial
effect of linseed which continued 04 weeks after
supplementation withdraw. At that time, hair
weight showed an acute decrease. A very signif-
icant positive effect (α<2%) was recorded at the
2nd hair harvest compared to the 1st one. This
effect vanished soon after (Table 4).
Results of the two administrations routes (ie
4 weeks of linseed oil topical application and 12
weeks of linseed ingestion) were compared. Oil
application is significantly (α<4%) more effec-
tive in promoting hair length than the ingestion
of the seeds (Table 5)
Table 4: Mean weight (g) of shaved hair.
4 weeks
08 weeks
12 weeks
16 weeks
CTLi
1.25
2.05
0.47
0.77
LSI
0.54*
1.47*
0.68
0.24
P value
0.95
0.77
0.30
0.75
* P=0.01
Table 5: Hair characteristics with tow use ap-
plication.
Length (cm)
Width (µm)
Weight (g)
LSOA
2.38
39.00
1.17
LSI
2.04
40.30
0.68
P value
0.035
0.55
0.40
Discussion
Several authors reported that fur quality de-
pends on some factors such as gender, environ-
mental conditions, season, photoperiodism and
sampling method (shaving or depilation)
(Charlet-Lery et al., 1985; Rochambeau and
Vrillon , 1985). This is why animals used in this
study were of the same sex and breed, and kept
at the same environmental conditions and during
the same season.
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Int. J. Med. Arom. Plants
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The important results of our study are main-
ly obtained with linseed oil topical application
and at a lesser degree with linseed ingestion,
which showed effect. Linseed ingestion im-
proves hair weight and insures a continuous ef-
fect once supplementation is withdrawn.
The exact mechanism of action or the com-
ponent(s) of flaxseed and flaxseed oil that pro-
mote the hair growth could not be established in
this study. However, it was shown that flaxseed
chutney diet doesn’t affect γ-glutamyl
transpeptidase load (Faseehuddin and Basavaraj
, 2007). This microsomal enzyme is an indicator
of hair growth (associated to alkaline phospha-
tase) (Kang Bong ,et al., 2011)
Furthermore, hair growth is inhibited by the
administration of paracrine growth factors such
as EGF (epithelial Growth factor) (Tsuboi
1997). This latter is inhibited by flaxseed use
(Tan ,et al., 2004).
In addition, the ALA (alpha linoleic acid) in
flaxseed oil can help in inhibiting the 5 alpha
reductase type 2 enzyme, responsible of con-
verting testosterone into dihydrotestosterone
(DHT). This male hormone shrinks hair follicles
and changes cyclic phase of hair growth cycle
(Galcera 2002; Brenner 2003); all these studies
may explain the beneficial effect observed in
our experiments.
Conclusion
Linseed ingestion has a slight beneficial ef-
fect on hair width, this result was more interest-
ing with linseed oil topical application, howev-
er, the right dosage should be determined. The
use of this plant may be a promising treatment
for alopecia and baldness but more studies are
necessary to assess its effects on skin histology,
hair structure and chemical composition. Fur-
thermore, the exact mechanism of action, and
the plant’s component(s) responsible of this ac-
tivity, should be investigated.
Acknowledgements: Authors are thankful to
Dr. Beghoul S. and Beroual L. for their
assistance during the implementation of the
experiments and also to Dr. Abdeldjelil M.C. for
his help in reviewing this article.
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... Therefore, an increase in the diameter of the fibre in the case of rabbit cover intended for use in the textile industry is considered an undesirable feature. Beroual et al. [41] conducted a study in which they analyzed the effect of adding flax grain into rabbit feed and rubbing linseed oil into the skin on hair growth and thickness. The authors observed that rubbing linseed oil had an effect on hair thickness, which in the experimental group increased after 4 weeks of the experiment by about 44% compared to the control group. ...
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