Citrullus colocynthis Schrad is a traditionally acclaimed hairtonic within Indian system associated ...

1. Effect of Citrullus coclocynthis Schrad. on hair growth
activity of albino rats.
Citrullus colocynthis Schrad is a traditionally acclaimed hair
tonic within Indian system associated with medicine. Scientific studies had been consequently
undertaken in
order to judge petroleum ether and also ethanolic extract involving C.
colocynthis regarding their particular impact on head of hair rise in albino rats. Your extracts
incorporated into oleaginous ointment base were applied topically on
shaved denuded skin of albino rats. The Particular occasion necessary pertaining to initiation of
hair growth too as conclusion involving locks growth cycle has been recorded.
Minoxidil 2% remedy had been applied topically and also served as positive
control. Locks growth initiation occasion ended up being significantly reduced to be able to half
on therapy with the extracts compared for you to manage animals. The Actual time
required with regard to total locks growth had been also considerably reduced. The
treatment has been productive throughout bringing a higher quantity of head of hair follicles
(>70%) throughout anagenic cycle than normal minoxidil (67%). the results
of remedy with 2% and 5% petroleum ether extracts had been comparable to
the positive control minoxidil.
Key words: alopecia, curly hair growth, Citrullus colocynthis, bitter
apple
Introduction
Hair loss is a dermatological disorder that has been recognised for
more than 2000 years. The idea is common throughout the world and contains been
estimated in order to impact nearly 2% of the world's population (Bertolino
2000, Olsen 1993). Apart coming from metabolic as well as hereditary causes alopecia

2. has been observed as a significant complication associated with anticancer,
immunosuppressant and several others drug treatments. Presently minoxidil
(useful in each male and female pattern baldness) (Goodman 1996) and
finasteride (useful inside male pattern baldness) (Libecco 2004) are generally two
USFDA approved synthetic drugs locating concomitant use regarding remedy of
androgenic alopecia although his or her unwanted effects get abbreviated their
usage.
Natural items are unequivocally advocated inside the cosmetic and
hair treatment business along with about 1000 different plant extracts get been
examined together with respect in order to head of hair growth activity; proanthocynidine from
grape seeds (Vitis Vinifera) along with [beta]-sitosterol within observed palmetto (Serenoa serrulata)
get proven remarkable effect (Takahashi 1998). There
are many items obtainable ready by combination of 1 or more
herbal drug that discover acceptability as hair tonics, curly hair growth
promoters, hair conditioners, curly hair cleansing agents, antidandruff agents
and for that therapy regarding alopecia and lice infection (Saraf 1991).
The traditional system of medication within India acclaims a new quantity of
herbal drugs for hair growth promotion even so insufficient audio scientific
evidence restrictions their use. Citrullus colocynthis (Cucurbitaceae) will be a
herb which usually may be recommended as being a locks growth promoter within traditional
literature. Your dried pulp of the unripe yet total grown fruit freed from
the rind, constitutes the particular drug colocynth along with has been included inside the
Pharmacopoeia involving India with regard to treating loss involving head of hair (The Prosperity of
India
1992). Your oil from your seeds involving the guarana plant offers reportedly been utilized by
ethnic tribes regarding putting examine to be able to premature falling as well as greying of

3. hairs (Kirtikar 1975, Hamdard Pharmacopoeia 1998).
Bhavpraksh, an Ayurvedic treatise, mentions the employment of your drug for
the therapy regarding indralupta i.e. a drug employed inside the treatment associated with hair
loss, most likely your Sanskrit name Indrayan signifying the locks growth
rejuvenating property with the drug (Chunekar 2002). Apart from its use as a
hair growth promoter C. colocynthis is used with regard to selection involving purposes
including the hypoglycaemic agent (Al-Ghaithi 2004), an antifertility
agent (Chaturvedi 2003) and an anti-inflammatory agent (Memon 2003).
In the actual existing research the effect of ethanolic and petroleum ether
extract regarding C. colocynthis in curly hair growth initiation along with marketing was
investigated.
Material and methods
Plant material
C. colocynthis fruits had been procured in the local market and
authenticated by simply researching the characteristics inside the Indian
Pharmacopoeia. Moderately coarse powder in the drug was used with regard to the
purpose involving extraction.
Preparation involving extracts
The dried coarse powder regarding C. colocynthis was fed inside a Soxhlet

4. extractor and also extracted initially together with petroleum ether (60-80oC) till
completely exhausted. The Particular solvent from your extract had been eliminated under
reduced pressure (yield 4.7% w/w). Your marc had been further extracted in
Soxhlet extractor together with ethanol (95%) till totally exhausted as well as was
freed regarding solvent below reduced pressure (yield 4.1% w/w).
Characterisation regarding extracts
Thin Layer Chromatography was performed upon precoated silica gel-G
plates (10X10) (Emerck, Germany) regarding characterisation of the extracts.
Chlorofrom : methanol : water (7:2.1:0.9 v/v) gave greatest resolution for
ethanolic extract. Petroleum ether extract had been best resolved in toluene : ethylacetate (84:16
v/v) as mobile stage (Wagner 2002). The Actual spots were
visualised making use of 50% ethanolic H2SO4 as derivatising agent.
Animals
Wistar strain albino rats regarding either sex weighing between 120-150 g
were fed on regular diet plan and also drinking water ad libitum. Your animals were housed
at room temperature (24 [+ or even -] two 0 C) on the normal day-night cycle
(06:00 hrs for you to 18:00 hrs).
Preparation regarding samples
The petroleum ether as well as ethanolic extracts were incorporated into
ointment base within concentration regarding 2% and also 5% (w/w) respectively.
Ointments had been created next your procedure offered in Pharmacopoeia of

5. India, 1996. Your extracts were then incorporated in the prepared
oleaginous base.
Treatment
Animals had been divided in sets of 6 rats each. Your following
treatment was handed to always be able to animals of numerous groups: group I Vehicle only
and served as control; group II topical application regarding 2% petroleum
ether extract within ointment base; Team III topical application regarding 5%
petroleum ether extract throughout ointment base; Team IV topical application
of 2% ethanolic extract throughout ointment base; group V topical application of
5% ethanolic extract within ointment base; group VI topical application 2%
alcoholic solution associated with minoxidil (Mintop(r)).
Hairs on the dorsal side with the rats were removed making use of hair
clippers and electrical shavers. Any marketed head of hair remover (Anne French)
was also accustomed to assure complete denual of six sq cm region (Roy 2006).
Toxicity studies
Toxicity studies had been carried out and the ethanolic as well as petroleum
extracts when applied in a concentration associated with approximately 10% did not present any
toxic negative effects or even erythema on skin surface. Thus the prepared
extracts were considered safe with regard to topical administration.
Permission from the institutional ethical committee has been taken

6. before animal experimentation.
Statistical treatment
Data is actually reported as imply [+ or even -] SEM. Statistical analysis associated with data
was carried out by one way ANOVA looking at most test teams vs control
followed through Dunnet's test making use of Instat v 2.1 computer software run on
Windows Xp residing in Pentium IV processor.
Qualitative hair growth study
Qualitative head of hair growth was evaluated simply by visual observation involving two
parameters a. curly hair growth initiation period i.e. minimum time to initiate
perceptible locks growth and b.) head of hair growth finalization time i.e. minimum
time taken to cover the particular denuded skin region together with new curly hair completely
(Adhirajan et al., 2001). Head Of Hair growth initiation as well as completion moment was
recorded with regard to each and also every group associated with animals.
Quantitative hair growth study
The method reported by Uno (1991) ended up being followed for that quantitative
evaluation regarding C. colocynthis extract. 1 rat via every group was
euthanicated following 10, twenty and also 30 days of treatment. Skin biopsies were
taken from your shaved area as well as specimens were preserved in 10% formalin.
The specimens were fixed in paraffin wax as well as blocks ready for
microtomy. Following fixation vertical sections regarding the skin were cut with
the aid of semiautomatic rotary microtome (Remi-1871, India).

7. The sections had been stained along with heamatoxylin along with eosin. The Particular number
of hair follicles per mm area involving skin and ratio of curly hair follicles in
different cyclic phases i.e. anagen (active growth phase) and also telogen (resting phase) were
established using your microscope. Head Of Hair folliculogram
was prepared by observing growth cycle of 100 hairs as well as duration associated with hair
follicle.
Global dispensary
Results
Qualititative studies about curly hair growth
Hair growth initiation along with conclusion time was considerably reduced
upon therapy together with petroleum ether extract regarding C. colocynthis. Within the
control group animals, head of hair growth ended up being initiated within denuded location
throughout the
second week, whereas it had been noted within the initial week within the petroleum
ether extract handled teams and also minoxidil treated groups. Curly Hair growth
was initiated about the 4th day along with 5% ointment of petroleum ether extract
of C. colocynthis. hair growth initiation ended up being recorded around the 5th day
with 2% ointment involving C. colocynthis whereas it absolutely was the actual 6th day within the
minoxidil handled manage group. Even though reduction inside hair growth
initiation time ended up being recorded inside the 2% along with 5% ethanolic extract treated
groups it hadn't been as pronounced as with the petroleum ether extract
treated groups (Table 1).

8. The period taken for complete hair growth around the shaved location ended up being also
affected with petroleum ether extract of C. colocynthis. Total hair
growth ended up being observed around the 18th day using 2% and the 16th day with 5%
petroleum ether extract treatment; about the 19th day together with minoxidil
treatment. Inside ethanolic extract as well as vehicle manage team animals
complete locks growth ended up being noted following 24 days. That had been further observed in
the petroleum ether extract treated team the texture regarding curly hair was
coarse, rough and also tough in comparison to the hairs in the minoxidil 2% treated
group which were brief and also silky (Fig I).
The results clearly show that the particular extract has been successful in
reducing time taken with regard to hair growth initiation as well as completion. The
results further declare that petroleum ether extract involving C. colocynthis
has better activity in comparison to become able to minoxidil as far as head of hair growth
initiation along with completion is actually concerned.
Quantitative studies upon hair growth
A considerable difference in cyclic cycle regarding head of hair growth was
observed inside teams handled with minoxidil as well as petroleum ether extract of
C. colocynthis. in vehicle handled manage group animals most of the
hair follicles have been in telogenic phase, only 1 or two are in catagenic
phase (PM I). Within the particular ethanolic extract handled group a new similar scenario
is noticeable along with nearly almost all of your follicles in telogenic stage along with merely a few
in anagenic stage without anagenic locks follicles (PM II).There is actually a
complete reversal involving the image inside petroleum ether extract and

9. minoxidil treated team where the vast majority involving locks follicles tend to be in
anagenic cycle using merely a few catagenic locks follicles. Telogenic
follicles are usually nearly negligible (PM III-IV).
An appreciable boost has been noted within the percentage anagenic
population (Table 2). Anagenic population of 47 [+ as well as -] 0.3% was found
in the car treated control group, whereas with 2% petroleum ether
extract it absolutely was 72 [+ or -] 0.2% and also along with 5% petroleum ether extract
treatment 75 [+ or -] 0.2% locks follicles were found in anagenic phase.
With minoxidil treatment, 67 [+ or -] 0.5% locks follicle had been observed
in anagenic phase whereas. Treatment Method along with ethanolic extract also
improved population involving anagenic head of hair follicles yet it wasn't as
appreciable as the petroleum ether extract treatment (Table 2).
Length involving head of hair follicles
The therapy using 2% along with 5% petroleum ether extract of C.
colocynthis stood a remarkable effect on the length regarding locks follicles. In
the control group merely 34 [+ or -] 0.4% had an average duration of 0.5mm,
whereas in the extract treated groups 46 [+ or -] 0.3% along with 48 [+ or -]
0.1% curly hair population with more than 0.5 mm was observed together with 2% along with 5%
extract remedy respectively. The Actual outcomes regarding remedy had been comparable
with your minoxidil team exactly where 49 [+ or perhaps -] 0.1% hair population had a
length involving 0.5 mm and also above (Fig. 3).
Discussion and conclusions

10. Topical application using the petroleum ether extract of C.
colocynthis fruit reduced the time needed regarding hair growth initiation
and has been superior for you to regular (minoxidil 2%) solution. The Actual quality of
hair inside the petroleum ether extract handled team had been coarse, rough and
hard whereas the minoxidil handled group resulted throughout soft as well as silky
hair.
Petroleum ether extract treatment method caused premature switching of
hair follicles through resting telogenic phase to end up being able to active anagenic phase. It
also assisted the handled group animals to retain anagenic hair follicles.
The percentage of head of hair follicle population throughout anagenic phase exhibited
marked improvement over the particular control group.
Petroleum ether extract has been your very greatest in inducing head of hair growth
initiation, with just about all the minoxidil treatment method next. Your research confirms that
the petroleum ether extract (5%) remedy reaches par with the 2%
minoxidil therapy within revitalising the expansion associated with locks within male rats. The
remarkable improvement long regarding locks follicles in addition supports the
hair growth advertising effects of the particular herb.
Androgenetic alopecia (AGA) is a dihydrotestosterone (DHT) mediated
process, characterised by constant miniaturisation involving androgen
reactive locks follicles as well as accompanied simply by perifollicular fibrosis of
follicular units throughout histological examination (Yoo 2006). Retention of
late anagenic follicles as well as rise in follicular size and
prevention of their own miniaturisation may for that reason become attributed to

11. 5[alpha]-reductase inhibitory activity. Even though comprehensive investigations
in this regard tend to be necessary to check out the possibility of this
mechanism.
The periodic transformation associated with locks follicles via telogen to
anagen stage inside the petroleum ether extract handled team is really a healthy
validation involving locks growth prospective regarding C. colocynthis. during the
treatment the secondary germ associated along with aggregated dermal papilla
cells within HOW TO MAKE YOUR HAIR GROW FAST telogen follicles was augmented as well as
their continuous growth and
differentiation may possess resulted inside construction of recent anagenic
follicles (Tanaka 1980).
The existing study validates the particular ethnomedical use of plants for hair
loss treatment. Further utilisation in the petroleum ether extract and
its incorporation in the formulation are warranted with regard to commercial
utilization associated with C. colocynthis.
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12. [FIGURE 3 OMITTED]
References
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herbal formulations for locks growth. Indian Drugs 38:11;559-63.
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vivo plus vitro evaluation of head of hair growth potential associated with Hibiscus rosa-sinensis
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effects involving Citrullus colocynthis within normal as well as diabetic rats. Mol Cell
Biochem 261:1-2;143-9.
Chaturvedi M, Mali PC, Ansari AS. 2003. Induction regarding reversible
antifertility with a crude ethanol extract involving Citrullus colocynthis
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alopecia. Expert Opin Pharmacother 5:4;933-40.
Memon U, Brohi AH, Ahmed SW, Azhar I, Bano H. 2003. Antibacterial
screening of Citrullus colocynthis. Pak J Pharm Sci 16:1;1-6.
Olsen EA. 1993. Androgenetic alopecia. In: E.A. Olsen (Ed.),
Disorders associated with curly hair growth: Diagnosis and treatment. New York: MacGraw
Hill Inc.
Pharmacopoeia regarding India. 1996. Vol II. New Delhi: Ministry of
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Saraf S, Pathak AK, Dixit VK. 1991. Locks growth promoting activity
of Tridax procumbens. Fitoterapia 62:495-8.
Takahashi T, Kamiya T, Yokoo Y. 1998. Proanthocyanidins through grape
seeds market proliferation regarding mouse locks follicle cells in vitro and
convert locks cycle inside vivo. Acta Derm Venereol 78:428-32.
Tanaka S, Saito M, Tabata M. 1980. Bioassay involving crude drugs pertaining to hair

14. growth marketing exercise in mice with a new straightforward method. Planta Med
S1:84-90.
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Uno H, Kurata S. 1993. Chemical agents and also peptides affect hair
growth. J Make Investments Dermatol 101;143-7S.
Uno H. 1991. Quantitative models for that study regarding head of hair growth in
vivo. In: Stenn KS, Messenger AY, Baden H P. Molecular and also Structural
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RK Roy, Mayank Thakur and also VK Dixit *
Department associated with Pharmaceutical Sciences
Dr HS Gour University
Sagar (MP) 470 003 India
* Corresponding Author

15. Prof VK Dixit, Department regarding Pharmaceutical Sciences Dr HS Gour
University, Sagar (MP) 470 003 India Telephone: +91 7582 264 582 +91
9826 497 030 Email: vkdixitl 11 @rediffmail.com
Table 1
Effect of Citrullus colocynthis Schrad extracts upon qualitative
hair growth
S. hair Growth (Days)
No. Remedy (topical) Initiation Period Finalization Time
1. Manage (vehicle 12 [+ or -] 0.82 24 [+ as well as -] 1.02
only)
2. Normal (% minoxi- 6 [+ as well as -] 0.41 *** twenty [+ or perhaps -] 1.08 ***
dil solution)
3. Petroleum ether 5 [+ or perhaps -] 0.28 *** 18 [+ or -] 0.56 ***
extract (2%
ointment)
4. Petroleum ether 4 [+ or -] 0.12 *** 18 [+ or -] 0.42 ***
extract (5%
ointment)
5. Ethanolic extract 11 [+ as well as -] 0.46 24 [+ or perhaps -] 1.34
(2% ointment)
6. Ethanolic extract ten [+ or perhaps -] 0.68 * 24 [+ or even -] 1.21

16. (5% ointment)
Value are imply [+ or perhaps -] SEM., n=6,
* p <0.05, ** p <0.01 along with *** p <0.001, significance vs control
Table 2
Hair growth population (anagen/telogen ratio) following treatment with
extracts of Citrullus colocynthis
Percentage of locks follicles
Following 10 Days
Treatment * Telogen Anagen
Control (Vehicle) 58 [+ or -] 0.4 42 [+ as well as -] 0.4
Standard (2% minoxidil) 54 [+ or -] 0.3 46 [+ or perhaps -] 0.3
2% pet. ether extract 52 [+ or -] 0.3 48 [+ or -] 0.3
5% pet. ether extract 46 [+ or perhaps -] 0.4 54 [+ or perhaps -] 0.4
2% ethanolic extract 55 [+ as well as -] 0.5 45 [+ or -] 0.5
5% ethanolic extract 54 [+ or -] 0.4 46 [+ or even -] 0.5
Percentage associated with locks follicles
Following 20 Days

17. Treatment * Telogen Anagen
Control (Vehicle) 55 [+ as well as -] 0.4 45 [+ or even -] 0.4
Standard (2% minoxidil) 37 [+ or perhaps -] 0.9 62 [+ or -] 0.9
2% pet. ether extract 37 [+ or even -] 0.1 63 [+ or -] 0.1
5% pet. ether extract 34 [+ as well as -] 0.2 66 [+ or even -] 0.2
2% ethanolic extract 50 [+ or even -] 0.6 50 [+ or -] 0.6
5% ethanolic extract 46 [+ or even -] 0.2 54 [+ or even -] 0.2
Percentage associated with hair follicles
Following 30 Days
T/A
Treatment * Telogen Anagen ratio
Control (Vehicle) 53 [+ or even -] 0.3 47 [+ or -] 0.3 1.21
Standard (2% minoxidil) 33 [+ or perhaps -] 0.5 67 [+ or -] 0.5 ** 0.49
2% pet. ether extract 28 [+ or -] 0.2 72 [+ as well as -] 0.2 ** 0.38
5% pet. ether extract 25 [+ as well as -] 0.2 75 [+ or perhaps -] 0.2 ** 0.33
2% ethanolic extract 46 [+ or perhaps -] 0.8 54 [+ as well as -] 0.8 0.85
5% ethanolic extract 43 [+ as well as -] 0.6 57 [+ or -] 0.6 0.75
T/A--Telogenic/Anagenic ratio after 30 days of treatment
* Almost All extracts were applied topically
Value are% mean [+ or -] SEM.

18. ** P< 0.05 (considered significant)