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Research
Paper

Protective effect of Aloe vera L. gel against sulphur mustard-

induced systemic toxicity and skin lesions
G. Anshoo, S. Singh, A.S. Kulkarni, S.C. Pant, R. Vijayaraghavan

ABSTRACT
Objective: Sulphur mustard (SM), chemically 2,2'-dichloro diethyl sulphide, is an incapacitating
and extremely toxic chemical warfare agent, and causes serious blisters on contact with human
skin. SM forms sulphonium ion in the body that alkylates DNA and several other macromolecules,
and induces oxidative stress. The aim of this study was to evaluate the protective effect of Aloe vera
Defence Research L. gel against SM-induced systemic toxicity and skin lesions.
and Development Materials and Methods: Aloe vera gel was given (250, 500 and 1000 mg/kg) orally to mice as three
Establishment, doses, one immediately after SM administration by percutaneous route, and the other two doses on
Gwalior 474002. the next two days. Protective index was calculated with and without Aloe vera gel treatment. Aloe
India
vera gel was also given orally as three doses with 3 LD50 SM and the animals were sacrificed for
Received: 8.7.2004 biochemical and histological evaluation, 7 days after SM administration. In another set of experi-
Revised: 20.9.2004 ment Aloe vera gel was liberally applied on the SM administered skin site and the animals were
Accepted: 29.9.2004 sacrificed after 14 days to detect its protective effect on the skin lesions induced by SM.
Results: The protection given by Aloe vera gel was marginal. 1000 mg/kg dose of Aloe vera gel
gave a protection index of 2.8. SM significantly decreased reduced glutathione (GSH), oxidised
Correspondence to:
R. Vijayaraghavan glutathione (GSSG) and WBC count, and significantly increased malondialdehyde (MDA) level,
E-mail: RBC count and Hb concentration. Aloe vera gel offered protection only in the increase of MDA
jai_vijay@hotmail.com level by SM. Severe damage was observed in the histology of liver, spleen and skin following SM
administration, and 1000 mg/kg of Aloe vera gel partially protected the lesions. However, topical
application of Aloe vera gel showed better protection of the skin lesions induced by SM.
Conclusion: The study shows that percutaneous administration of SM induces oxidative stress and
oral administration of Aloe vera gel could only partially protect it. Topical application of Aloe vera
gel may be beneficial for protecting the skin lesions induced by SM.

KEY WORDS: Aloe vera gel, glutathione, malondialdehyde, oxidative stress

Introduction
Bis (2-chloroethyl) sulphide, is commonly known as sul-
phur mustard (SM) or mustard gas, is an alkylating agent that
causes painful blisters on contact with human skin. It is a fre-
quently used chemical warfare agent and several reports of
its use are available.[1] SM forms sulphonium ion in the body
which leads to DNA strand break and cell death.[2] Due to the
high electrophilic property of the sulphonium ion, SM binds to
a variety of cellular macromolecules.[3] Eyes, skin and the res-
piratory tract are the principle targets of SM toxicity.[2,4-5] Be-
cause of its high lipophilic property, it rapidly penetrates into
the skin. Dermal contact of SM causes erythema, severe itch-
ing, blisters and ulceration. There are also several reports that
systemic toxicity of SM leads to oxidative stress.[6-7]
Aloe vera (Linn), also known as Aloe barbadensis of family
Liliaceae is widely distributed in Asia, Africa and other tropi-
cal areas. It has been used in folk medicine for the treatment
of skin ailments including wounds and burns, asthma and has
recently been investigated for its antioxidant effect.[8] Popular
interest and use of Aloe gel preparation has increased dra-
matically in the last decade. The mucilaginous gel from
parenchymatous cells in the leaf pulp of Aloe vera is incorpo-
rated in ointments, creams, lotions and other preparations
essentially for topical use. There are conflicting reports con-
cerning the efficacy of such preparations.[9-11]
In this study the dermal application of Aloe vera gel as a
protective natural product against SM-induced skin lesions,
and oral administration as an antioxidant for the systemic tox-
icity of SM has been evaluated.
Materials and Methods
Chemicals
Extreme care was taken during the synthesis of SM in the
declared facility of the Defence R & D Establishment. The syn-

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Anshoo G, et al.
thesized SM was found to be more than 99% pure by gas chro-
matographic analysis. The dilution of SM and its administra-
tion to animals was carried out in fume hoods with the per-
sonnel wearing butyl gloves. The animals were kept in a well-
ventilated area for 48 h,after SM administration and then trans-
ferred to the experimental animal room for further monitor-
ing. Polyethylene glycol (PEG-300) was purchased from Fluka
Biochemica (USA). The rest of the chemicals were purchased
from Merck India Ltd. Aloe vera gel was purchased from the
trade and certified by the Indian Aloe Science Council (Safed
Musli farms, Gwalior).
Animals
Randomly bred Swiss female mice (25 to 30 g, body weight)
from the Defence R & D Establishments animal facility were
used for the study. The animals were kept in polypropylene
cages on dust-free rice husk as the bedding material. Free
access to food (standard pellet diet, Amrut Ltd, India) and
water was allowed until two hours before the experiment. The
care and maintenance of the animals was as per the approved
guidelines of the Committee for the Purpose of Control and
Supervision of Experiments on Animals (CPCSEA), India. All
animal procedures were approved by the Animal Ethical Com-
mittee of the Establishment. A day before percutaneous expo-
sure of SM, the hair on the back of the animals was closely
clipped by a pair of scissors.
Protection studies against SM-induced lethality
SM was administered percutaneously (topical application)
in varying doses and Aloe vera gel was administered orally as
three doses. The first dose was given simultaneously with SM
and the remaining two doses were given daily on the next two
days. Three different doses of Aloe vera gel were used (250,
500, 1000 mg/kg, body weight) by dissolving the gel in dis-
tilled water. SM was diluted in polyethylene glycol (PEG 300)
and applied on the back of the mice. The dilutions were made
in such a manner that the quantity applied was not more than
100 l. Protection was expressed in terms of protection in-
dex, which is the ratio of the LD50 with treatment to that of
LD50 without treatment. For the determination of each LD50, 3
to 4 log doses were used, and for each dose 4 animals were
used. The animals were weighed daily and observed for mor-
tality for a 14-day period. LD50 was determined as per the
moving average method.[12]
Protection studies against SM-induced oxidative stress
To evaluate the protective effect of Aloe vera gel on oxidative
stress markers, the following groups of four mice each were
utilized.
I Distilled water, p.o. + PEG 300 dermal (control
group)
II Distilled water, p.o. + SM, 3 LD50 (19.4 mg/kg)
in PEG 300
III Aloe vera gel (250 mg/kg), p.o. + SM, 3 LD50 (19.4 mg/kg)
in PEG 300
IV Aloe vera gel (500 mg/kg), p.o. + SM, 3 LD50 (19.4 mg/kg)
in PEG 300
V Aloe vera gel (1000 mg/kg), p.o.+ SM, 3 LD50 (19.4 mg/kg)
in PEG 300
104 Indian J Pharmacol | April 2005 | Vol 37 | Issue 2 | 103-110
SM was diluted in polyethylene glycol (PEG 300) and ap-
plied on the back of the mice (percutaneous). The dilutions
were made in such a manner that the quantity applied was not
more than 100 l. Aloe vera gel dissolved in distilled water
was administered orally as three doses. The first dose was
given simultaneously with SM and the remaining two doses
were given daily on the next two days. Seven days post SM
application the animals were anaesthetized with ether for the
collection of blood from the orbital plexus and then killed by
cervical dislocation. The liver was dissected out and cleaned
for the biochemical estimations and for histology. Spleen and
skin samples were also taken out for histological examina-
tion.
Lipid peroxidation was studied by measuring the forma-
tion of thiobarbituric acid reactive substances according to
the method of Buege and Aust.[13] One hundred milligram of
liver tissue was directly homogenized in 5 ml thiobarbituric
reagent and boiled for 30 min. The content of the tube was
cooled and centrifuged. Optical density of clear supernatant
was measured at 535 nm in a spectrophotometer. The amount
of MDA formed was calculated, using a molar extinction coef-
ficient of 1.56 X 105 M-1cm-1 and expressed as nmol/gm of
tissue. Glutathione concentration of tissue was assayed by Hisin
and Hilf fluorometric method.[14] One hundred and fifty milli-
gram of tissue was homogenized in 0.1 M sodium phosphate
EDTA buffer (pH 8.0) and 25% meta phosphoric acid was added.
The content of the tube was centrifuged and supernatant was
used for GSH and GSSG estimation. Analysis of hematological
variables viz., WBC count, RBC count, Hb concentration were
carried out using a hematocytometer, Beckman Coulter Ana-
lyser. All the values are expressed in terms of percentage
change as compared to healthy control group.
Protection studies against SM-induced histopathology
Liver, spleen and skin samples were fixed in 10 % neutral
buffered formalin solution. After proper fixation, small pieces
were processed by dehydration and embedded in paraffin wax.
Sections of 5-6 m thickness were prepared and stained with
hematoxylin and eosin for light microscopy.[15] From each sam-
ple ten slides were prepared (Total 40 slides for 4 animals).
Out of 40 slides, six slides were selected randomly for lesion
evaluation. From each selected slide, 9 m2 area was identi-
fied randomly and lesions were marked and compared with
that of control. The severity of the lesions was characterized
using LEICA - Qwin - 500 Image Analyser and converted into
percentage.
Protection studies against SM-induced skin lesions
To evaluate the protective effect of Aloe vera gel on SM-
induced skin lesions a separate study was carried out. SM was
diluted in acetone and applied on the back of the mice (5 mice
per group). Two doses of SM were used, 5 mg/kg and 10 mg/
kg. After 24 h Aloe vera gel was applied liberally once daily for
14 days, the animals were sacrificed and skin samples were
dissected out for histological examination as explained above.
Statistical analysis
All the variables were analyzed by one-way ANOVA with
Dunnetts multiple comparison procedure. A probability of
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Aloe vera gel and sulphur mustard

<0.05 was considered as statistically significant. Table 1

Results Protective effect of Aloe vera gel against percutaneously
administered SM in mice
The LD50 of SM in PEG-300 through the percutaneous route
in mice was found to be 6.8 mg/kg when the animal mortality Group LD50 (mg/kg) PI+
was recorded till 14 days of SM administration. Oral Aloe vera (Confidence limits)
gel gave a dose-dependent protection against SM. A dose of Only SM 6.8 (2.1 - 22.7) -
1000 mg/kg of Aloe vera gel gave 2.8-fold protection (Table SM + Aloe vera gel, 250 mg/kg 9.7 (3.7 - 25.1) 1.4
1). The body weight of the animals administered with 3 LD50 SM + Aloe vera gel, 500 mg/kg 13.7 (7.8 - 23.8) 2.0
(19.4 mg/kg) of SM, decreased progressively. Body weight of SM + Aloe vera gel, 1000 mg/kg 19.3 (7.4 - 50.2) 2.8
the animals in the SM group was significantly decreased com-
pared to the control group from one day after administration Aloe vera gel was administered orally as three doses, with the first dose given
simultaneously with SM. The remaining two doses were given daily on the next
and was significantly decreased in all the groups on 7 days two days. Animals were observed for mortality for 14 days. PI += protective index
after administration. The body weight was 61.3+1.9% after 7
days in the SM group. Aloe vera gel failed to protect the de-
Table 2
crease in body weight even at 1000 mg/kg dose (Table 2).
Percutaneous administration of SM insignificantly de- Percent change in body weight following percutaneous
creased the WBC count and significantly increased the RBC administration of SM, 19.4 mg/kg (3 LD50) with and without Aloe
count and the hemoglobin content (Table 3). Oral administra- vera gel in mice
tion of Aloe vera gel failed to protect the changes induced by Group 1 day 4 days 7 days
SM even at 1000 mg/kg dose. SM also significantly decreased
Control 100.0+ 0.0 104.8+1.2 104.0+3.5
GSH and GSSG content. Seven days after SM administration
Only SM 95.8+ 0.8a 74.8+3.3a 61.3+1.9a
(3 LD50), the GSH and GSSG contents were 38.4+0.4 % and SM+Aloe vera gel, 250 mg/kg 97.6+ 1.1 93.0+6.4 72.1+2.6a
38.6 +2.4 % respectively. Oral administration of Aloe vera gel SM+Aloe vera gel, 500 mg/kg 93.5+ 0.7a 89.3+1.7 75.2+2.3a
failed to protect the depletion of GSH and GSSG caused by SM SM+Aloe vera gel, 1000 mg/kg 98.7+ 1.2 90.6+9.6 74.6+10.1a
even at 1000 mg/kg dose. The MDA content was significantly F 8.1 3.9 9.7
increased after SM administration. There was some improve- P < 0.01 < 0.05 < 0.001
ment in the MDA level after treatment with Aloe vera gel (Ta-
ble 4). Values are mean+SEM, n = 4 in each group. Aloe vera gel was administered
orally as three doses, with the first dose given simultaneously with SM. The
SM caused severe damage to the liver, spleen and the skin remaining two doses were given daily on the next two days. aSignificant from
(site of application). The liver of control mice showed normal control group.
hepatic cord arrangement, hepatic lobes and hepatocytes with
normal hepatic parenchyma (Figure 1a). Following SM appli- Table 3
cation, liver histopathology exhibited granulovacuolar degen-
eration of hepatocytes and perinuclear clumping of cytoplasm. Effect of percutaneous administration of SM, 19.4 mg/kg (3 LD50)
Varying degree of necrosis was observed in the hepatocytes. with and without Aloe vera gel on WBC, RBC and Hb of blood in
Hepatic parenchyma also revealed significant fatty changes mice
(Figure 1b). Treatment with 250 mg/kg of Aloe vera gel orally Group WBC RBC Hb
did not protect the hepatic lesions and 500 and 1000 mg/kg (%) (%) (%)
Aloe vera gel provided marginal protection to these lesions. Control 99.9+10.9 100.0+1.3 100.0+2.5
The hepatocytes were of normal shape, yet granulovacuolar Only SM 72.8+39.2 131.1+3.9a 129.9+7.4a
degeneration of cytoplasm still persisted (Figures 1c and 1d).
SM + Aloe vera gel, 250 mg/kg 91.9+23.3 137.6+2.9a 123.9+3.7a
Perinuclear clumping of cytoplasm and bile duct proliferation
SM + Aloe vera gel, 500 mg/kg 79.2+13.3 144.1+3.4a 134.4+5.0a
were still observed even after treatment with 1000 mg/kg of
SM + Aloe vera gel, 1000 mg/kg 132.5+72.3 131.9+0.7a 120.3+2.5a
Aloe vera gel. A comprehensive account of various histopatho-
logical lesions in the hepatic tissue and their protection by F 0.43 8.7 8.3
Aloe vera gel is given in Table 5. P NS < 0.001 < 0.001
Histological features of spleen from control mice showed Values are mean + SEM, n = 4 in each group; WBC = 13.4 + 0.7 x 103 cells/ml,
normal histoarchitecture with white pulp consisting of lym- RBC = 8.4 + 0.4 x 106 cells/ml, Hb= 13.2 + 0.4 g/dl. Aloe vera gel was administered
phatic follicles and red pulp consisting of sinusoids and other orally as three doses, with the first dose given simultaneously with SM. The
remaining two doses were given daily on the next two days. aSignificant from
elements of blood (Figure 2a). Spleen of SM-applied mice control group.
showed reduced density of white pulp follicles and congestion
in red pulp. The red pulp was also infiltrated with macrophages.
Obliteration of marginal zone and degeneration of peri arteri- ment gave protection as evidenced by reorganization of white
olar lymphatic sheath (PALS) was also observed following per- pulp follicle cells and the germinal center (Figure 2d). A com-
cutaneous administration of SM (Figure 2b). Treatment with prehensive account of various histopathological lesions in the
250 mg/kg of Aloe vera gel orally, did not give any protection splenic tissue and their protection by Aloe vera gel is given in
and 500 mg/kg gave minimal protection to SM-induced splenic Table 6.
lesions (Figure 2c). However, 1000 mg/kg Aloe vera gel treat- The histology of control mouse skin showed three distinct

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Anshoo G, et al.

Figure 1. Photomicrographs of control, SM-administered and Aloe vera gel-treated mouse liver; H & E 100x.
a. Control mouse liver showing normal hepatic parenchyma, hepatic lobules and hepatocytes,
b. SM-administered (3 LD50, p.c.) mouse liver showing granulovacuolar degeneration and perinuclear clumping of cytoplasm,
c. Aloe vera gel (500 mg/kg) treated mouse liver showing marginal protection of the lesions, and
d. Aloe vera gel (1000 mg/kg) treated mouse liver showing better protection that the hepatocytes are taking normal shape with less
granular degeneration.

Figure 2. Photomicrographs of control, SM-administered and Aloe vera gel treated mouse spleen, H & E 100x.
a. Control mouse spleen showing normal histology of germinal center, red and white pulp follicles,
b. SM-administered (3 LD50, p.c.) mouse spleen showing congestion, hypocellularity and splenic parenchymal degeneration,
c. Aloe vera gel (500 mg/kg) treated mouse spleen showing less protection,
d. Aloe vera gel (1000 mg/kg) treated mouse spleen showing protection as the hypocellularity of white pulp has been reduced and
reorganized.

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Aloe vera gel and sulphur mustard

Table 4 Table 6

Effect of percutaneous administration of SM, 19.4 mg/kg (3 LD50) SM (3 LD50)-induced splenic lesions and their protection by vari-
with and without Aloe vera gel on GSH, GSSG and MDA of liver ous doses of Aloe vera gel in mice
in mice Lesions SM only SM+Aloe SM+Aloe SM+Aloe
Group GSH GSSG MDA vera (250 vera (500 vera (1000
mg/kg) mg/kg) mg/kg)
(%) (%) (%)
White pulp hypocellularity +++ +++ ++ ++
Control 100.0+0.3 99.9+5.1 100.0+0.0
Atrophy of periarteriolar +++ +++ +++ ++
Only SM 38.4+0.4a 38.6+2.4a 136.0+15.3a
lymphatic sheath
SM + Aloe vera gel, 250 mg/kg 40.6+3.7a 42.5+2.4a 115.0+3.4
SM + Aloe vera gel, 500 mg/kg 27.5+3.6a 28.6+2.5a 126.0+1.3 Congestion and hemorrhage +++ +++ ++ +
SM + Aloe vera gel, 1000 mg/kg 35.0+6.4a 37.3+6.9a 109.1+5.4 Activation of Megakaryocytes ++ ++ + +
F 63.4 45.0 3.6 Condensation of chromatin ++++ ++++ ++ ++
P < 0.001 < 0.001 < 0.05 Necrosis/apoptosis ++++ ++++ +++ ++
Accumulation of fibrinoid +++ +++ +++ ++
Values are mean +SEM, n = 4 in each group. GSH =3.50+0.1 moles/gm of
tissue, GSSG = 1.86+0.3 moles/gm of tissue, MDA= 3.86+0.10 nanomoles/g material
of tissue. Aloe vera gel was administered orally as three doses, with the first
dose given simultaneously with SM. The remaining two doses were given daily Grading : - nil, + minimal (<12%), ++ mild (<22%), +++ moderate (<45%) and
on the next two days. aSignificant from control group. ++++ severe (>45%) n = 4 in each group. The animals were sacrificed 14 day
after SM administration. Aloe vera gel was given orally as three doses, with the
first dose given simultaneously and two more doses on the subsequent days.
Table 5

SM (3 LD50)-induced hepatic lesions and their protection by vari- lesions as skin histology showed reepithelization, reduced
ous doses of Aloe vera gel in mice number of inflammatory cells and dermoepidermal gaps (Fig-
ures 3c and 3d). There was a significant improvement in the
Lesions SM only SM+Aloe SM+Aloe SM+Aloe
vera (250 vera (500 vera (1000
lesions caused by SM by the topical application of Aloe vera
mg/kg) mg/kg) mg/kg) gel (Figure 4d). The reepithelization was better and the gaps
were also reduced.
Disturbed lobular pattern ++ ++ ++ +
Hemorrhage +++ +++ ++ + Discussion
Necrosis/apoptosis +++ +++ ++ +
The probability of accidental exposure to lethal concentra-
Obliteration of chromatin +++ +++ ++ ++
tions of SM exists during its decontamination program by the
material
declared State Parties, as part of the directive of the Organi-
Proliferation of bile duct +++ +++ ++ +
parenchyma
zation for the Prohibition of Chemical Weapons (OPCW, The
Perinuclear clumping of ++++ ++++ +++ ++
Netherlands). Keeping this point in view, mice were exposed
cytoplasm to SM by the percutaneous route which is closer to the actual
Hepatic vacuolation ++++ ++++ +++ ++ conditions, and the protection of Aloe vera gel against SM tox-
PMN. infiltration ++ ++ + + icity was evaluated. Following SM administration the animals
generally died within a period of 14 days.[16] The efficacy of
Grading : - nil, + minimal (<12%), ++ mild (<22%), +++ moderate (<45%) and Aloe vera gel was evaluated against SM toxicity and maximum
++++ severe (>45%), n = 4 in each group. The animals were sacrificed 14 days
after SM administration. Aloe vera gel was given orally as three doses, with the protection was observed with1000 mg/kg oral dose.
first dose given simultaneously and two more doses on the subsequent days. Oxidative stress is defined as a disturbance in the pro-
oxidant and antioxidant balance, in favor of the pro-oxidant
layers viz., epidermis, dermis and hypodermis. The epidermis state. Percutaneous (dermal) administration of SM can induce
consisted of three visible layers (i) the dark stained corneum oxidative stress in the experimental animals and antioxidants
layer having flattened dead cells, (ii) the stratum granulosum can protect it to some extent.[6, 7, 17] The alternative hypothesis
comprising dense basophilic granular cells and (iii) the stra- for SM toxicity is related to GSH which may act as an intracel-
tum spinosum consisting of large polyhedral cells. The dermis lular scavenger for SM.[6, 7, 18] A significant depletion of GSH
consisted of two layers of connective tissue, a superficial pap- was observed in the present study also. GSH has an impor-
illary and deeper reticular layer (Figure 3a). Percutaneous tant function in protecting the cell from oxyradicals and reac-
application of SM in mice caused severe epidermal atrophy as tive chemicals. It scavenges free radicals and its depletion is
evidenced by extensive epidermal necrosis in the stratum cor- related to cytotoxicity. It is also possible that in the absence of
neum layer. Spinous cells were swollen, enlarged and vacu- GSH the partially reduced oxygen species may be transformed
olated, and prominent dermoepidermal separation was also into highly toxic oxidants, which may further react with mem-
observed. A large number of inflammatory cells were observed brane phospholipids initiating lipid peroxidation. Since Aloe
in the dermal region. The degenerated dermoepidermal re- vera gel was giving marginal protection against the lethality of
gion was covered by exudates composed of fibrin, leucocytes SM, oxidative stress markers were evaluated. Oral adminis-
and RBCs (Figure 3b). Oral administration of Aloe vera gel 250 tration of Aloe vera gel did not show recovery of GSH and
mg/kg did not protect SM-induced skin injury. Oral adminis- GSSG, but the elevation of MDA level was protected, showing
tration of 500 and 1000 mg/kg Aloe vera gel reduced the skin marginal protection against SM-induced toxicity. Significant

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Anshoo G, et al.

Figure 3. Photomicrographs of control, SM-administered and Aloe vera gel-treated mouse skin, H & E 100x.
a. Control mouse skin showing normal epidermis and dermis with flattened fused cells, dense basophilic granular cells and large
polyhedral cells and well-defined cell boundaries.
b. SM-administered (3 LD50, p.c.) mouse skin showing invasion of the epidermis by acute inflammatory cells. Dermo-epidermal
separation and accumulation of necrotic debris were also seen,
c. Aloe vera gel (500 mg/kg) treated mouse skin showing reduction in the skin lesions,
d. Aloe vera gel (1000 mg/kg) treated mouse skin showing reduction in the accumulation of the inflammatory cells and reduction in
the dermo-epidermal gap.

Figure 4. Photomicrographs of control, SM-administered and Aloe vera gel-treated mouse skin, H & E 100x.
a. Control mouse skin showing normal epidermis and dermis with flattened fused cells, dense basophilic granular cells and large
polyhedral cells and well-defined cell boundaries.
b. Topical application of 100 l of acetone on the skin. The skin structure is similar to the normal skin.
c. SM-administered (10 mg/kg in acetone) mouse skin showing invasion of the epidermis by acute inflammatory cells. Dermo-
epidermal separation, complete erosion of dermis and accumulation of exudates composed of fibrinoid material and RBCs.
d. Aloe vera gel-applied mouse skin showing reepithelization of epidermal layers and reduction in the dermo-epidermal gap.

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protection on GSH, GSSG and MDA was observed following
SM toxicity only when the protection as measured in terms of
protection index was very high.[6,7] Since the maximum pro-
tection index obtained in the present study was 2.8 at 1000
mg/kg dose of Aloe vera gel, GSH and GSSG were not pro-
tected. The marginal protection offered by Aloe vera gel may
be due to the active components aloesin, aloeresin, and aloin.[8]
But Aloe vera gel has been found to be beneficial in protecting
the reduction of GSH and increase in MDA following gamma
radiation.[19] Following percutaneous administration of SM the
animals lost weight progressively and appeared extremely
weak and emaciated. Hemoconcentration was also observed.
It is generally expected that following SM administration there
should be a profound decrease in WBC count. But, due to
hemoconcentration the decrease in WBC count following SM
administration is not significant. Hemoconcentration occur-
ring due to the increased permeability of the endothelial cells
of the blood vessels may be the reason for the increased RBC
count and Hb content. Lipid peroxidation is attributed to the
increased permeability. Since the protection offered by Aloe
vera gel was only marginal there was no significant protection
in the hematological variables.
SM is a potent alkylating agent and causes serious blisters
on contact with human skin. Topical application of SM causes
erythema, followed by blisters and ulceration. In the animal
model percutaneously administered SM is extremely toxic.[20]
Though apparent blisters were not observed in animal mod-
els, ulceration appeared after 14 days of dermal application
of SM. Because of its lipophilic properties, SM rapidly pen-
etrates the skin and the best method of minimizing the toxic
effects on the skin as well the systemic toxicity is by decon-
tamination. The presently used decontaminants are Fullers
earth, a physical decontaminant and CC2 suspension, a per-
sonal-chemical decontaminant.[21,22] Even a shortest contact
period of SM on the skin is sufficient for the blister formation.
The presently used medical methods for the treatment of blis-
ters are the application of petroleum jelly, antimicrobial oint-
ments or povidone-iodine containing ointment which are symp-
tomatic measures.[23,24]
The mechanism of wound healing occurs in three stages;
inflammation, proliferation and remodelling. The wound heal-
ing process depends on the local blood circulation, as well as
the formation and deposition of collagen. Aloe vera gel con-
tains important constituents necessary for wound healing, such
as ascorbic acid, tocopherols, amino acids, and Zinc.[25] Ascor-
bic acid enhances the synthesis of collagen and counterbal-
ances collagen breakdown. Tocopherol is a fat-soluble vita-
min found in Aloe vera gel, that has proven antioxidant activ-
ity. It stabilizes lysosomal enzymes needed for the synthesis
of collagen and scavenges free radicals that may interfere in
the wound healing process. Zinc improves the tensile strength
and improves wound healing.[26] Aloe vera gel reduces dehy-
dration and necrosis, and also dilates the capillaries. Another
expected mechanism of Aloe vera gel for wound healing is the
mannose-containing product which increases macrophage
activity and fibroblast proliferation.[26]
When SM is diluted in PEG-300 and applied topically the
systemic toxicity is more than the local skin lesions due to the
combined lipophilic characteristics of SM and PEG-300. Hence
Aloe vera gel and sulphur mustard
to induce a prominent skin lesion SM was diluted in acetone
which is a less lipophilic solvent with a better spreading ef-
fect. When PEG-300 is used as a solvent no erythema is ob-
served and the lesions develop only after 14 days. While in
acetone, erythema develops in two days and the lesions also
appear in about 7 days. SM-administered mice skin showed
invasion of the epidermis by acute inflammatory cells,
dermoepidermal separation, erosion of dermis and accumu-
lation of exudates composed of fibrinoid material. An increased
number of polymorphonuclear cells were also observed in
guinea pig skin administered with liquid SM.[27] In the present
study topical application of Aloe vera gel protected the skin
lesions and they were less severe. As Aloe vera gel stimulates
and enhances vascularity around the wound area, the general
appearance of the wound was healthier than the untreated SM
wound. Aloe vera gel has also shown to be very effective in
burn injuries.[28-29] However, a clinical trial conducted to re-
lieve the side-effects on the skin site of breast cancer patients
following radiation therapy was not beneficial.[30-31]
The present study shows that percutaneously administered
SM can cause oxidative stress with a decrease in GSH and
GSSG concentrations and an increase in MDA content. Oral
administration of Aloe vera gel protected marginally. However,
dermal application of Aloe vera gel protected the skin lesions
induced by SM and improved the healing process.
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