Effects of aloe vera juice on histopathology of excision wound in
albino mice.
Abstract

This study is under taken to establish Comparasion between different conc.of
aloe vera juice(5%;25%,50% v/v topically) with respect to chemotherapeutic
agent like betadine solution on healing of dermal wound in albino mice.
Aloe vera juice used in the present work in the concentration of 5% , 25%,50% v/v have show
wounds healing effect on excision wound model in albino mice .The maximum effect was found with 5%
Aloe vera juice followed by , 25% and 50% respectively.
Histopathological studies revealed that wounds
treated with 25% and 50% Aloe vera juice, proliferation of fibroblast and mononuclear cell infiltration
was markedly higher than the Betadine (control) treated wound.But in case of 5% Aloe vera juice
treated wounds the fibroblast cells were found to be maximum as compared to all other
group .The collagen fibre in these treated group were thin ,fine and dispersed and markedly
prominent as compared to the Betadine treated (control) wound.The constitution of these
collagen fibre was found to be maximum with more thicker ,dense, and wavy fibres in the
healing wound tissue of 5% Aloe vera juice.
As per histochemical studies the collagen formation
appeared to be increasing gradually in wound of all group. However the collagen formation was
more marked and distinct in Aloe vera juice treated wound with maximum in 5% Aloe vera juice
as compared to 25% and 50% Aloe vera juice as well as Betadine (control) treated wound. The elastin
synthesis showed gradual increase in all the haeling wound tissue with maximum in 5% Aloe vera juice.
Aloe vera juice used in this study, 5% Aloe vera juice appeared to be the best in promoting the
wound healing.The stability profile and release characteristic of 5% Aloe vera juiceappear to be
desirable.
Key words: Aloe vera, Histopathology, Wound healing,
1.Introduction
Skin the “organ of beauty “ is the outermost covering of the body protecting it from the external
environment.A break in continuity of the skin result in wound and wound may have contamination
and finally produce different diseases in animal body .There are many preventive measures to
chcek the contamination of wound. There are two type of therapy ,firstly the PHYTOTHERAPY and
secondly the CHEMOTHERAPY .Due to unwanted effect of chemotherapy shifting to phytotherapy is
imperative. Due to less side effect ,the phytomedicine as atherapeutic tool is gaining importance in
treating diseases and improving health.Some herbal medicine like Pygeum is nuseful in prostatic
enlargement. Green tea extract in prevention of arteriosclerosis and cancer , kava for alleviating
anxiety and insomnia, cordyceps as an immunostimulant have been published in the recent past
b(Tyler et al., 1999) . Wound healing is widely discussed in the medical literature. Much research has
been carried out to develop more sophisticated dressing that expedite the healing process and diminish the
bacterial burden in wounds(Manafi et al 2009 )Burn wound healing is one of major indications of aloe
vera gel use in many countries. (Mohajeri et al.,2010 )
Wounds are sequels of injuries which result into opening or breaking of the skin (Meenakshi
et al., 2006). In other words wound is break in the epithelial integrity of the skin which may be
accompanied by disruption of the underlying structures and normal tissues resulting into
contusion, hematoma, laceration or an abrasion (Enoch and Jhon, 2005).
scar formation likely depends not only on the number of myofibroblasts but also on the
extracellular environment which regulates their function. Horsley et al. recently revealed
that the adipose tissue mediates fibroblast migration to promote wound healing [Schmidt et al.,
2013 ]. The delicate structu re and composition of the extracellular matrix (ECM) are also vital for
complete wound healing. African spiny mice are capable of skin autotomy and develop more porous
ECM during skin regeneration than non-skin regenerating mice [Seifert et al .,2012 ]. The slow ECM
deposition might allow different molecules to communicate and coordinate to restore the full skin
structure. Additionally, the skin regenerating ECM was dominated by collagen type III, while the
scar formation ECM was rich in collagen type I, which antagonizes skin appendage regeneration
[Satoh et al .,2011].
Aloe Vera (Aloe Barbadensis Miller) is a medicinal plant used traditionally due to its therapeutic
properties such as wound-healing properties, immunomodulatory, anti- inflammatory, antiviral,
antibacterial and antioxidant activities([Gupta et al (2012); Wozniak et al (2012). The most common
folk use of Aloe vera has been for the treatment of burn wounds and specifically to aid in the
healing process, reduce inflammation, and tissue scaring. The juice was described by Dioscorides
and used to treat wounds and mouth infections, soothe itching, and cure sores. ( Lans .,
Ethnomedicines and. Ethnobiol Ethnomed. 2006.) The majority of the abundant scientific
information regarding Aloe vera and its multiple biological activities have been largely attributed to
a complex carbohydrate or polysaccharide called Acemannan,which is the short name for poly –
beta-1,4 mono-acetyl mannose(Hamman .2008). It has been shown the activity of Aloe depends on
the Acemannan content.Accordingly the greater the Acemannan content, the greater bioactivity and
beneficial effect on skin care and wound healing. In fact,Acemannan is so important that the
International Aloe Science Council has determined that if a product does not contain Acemannan it
is not Aloe vera. wound healing involves the activity of an intricate network of blood cells, cytokines
and growth factors which ultimately leads to the restoration to the normal condition of the injured skin or
tissue( Gunde et al 2013 ., Bambal et al 2011) . Burn and wound healing effects of aloe vera are very
abundant with small quantity of solid material by providing essential micronutrients, antiinflammatory and antimicrobial effects (Chithra et al., 1998; Khorasani et al., 2009).
The historical application of aloe vera juice for the treatment
of wounds has been evaluated in surgical wounds and the randomized study concluded
that there was a significant delay in complete wound healing for the aloe vera juice
compared to standard treatment.(Schmidt et al ,1998; ) . The juice is clear, odorless, and
tasteless and should be free of leaf skin or yellow parts. No consistent standardization has
been established, but the International Aloe Science Council (IASC), a trade association of
internationally based aloe vera producers and marketers, requires adherence to certain
specifications for the product to be certified. Aloe vera may be effective in treatment of
wounds. Evidence on the effects of its sap on wound healing, however, is limited and
contradictory.Some studies, for example, show that aloe vera promotes the rates of
healing, while, in contrast, other studies show that wounds to which aloe vera juice was
applied were significantly slower to heal than those treated with conventional medical
preparations.( (Schmidt et al( 1991)) . Aloe vera based products showed remarkable
performance when used in burned skin, ranging in application from sun burn to radiation
induced dermatitsi (Reuter ,et.al 2008 ;Roberts et.al 1995 ). Aloe showed ability to
stimulate fibroblast formation and increased collagen, thus contributing to skin repair
(Yao ,et.al 2009 and Chithra , 1998 ).In fact Aloe showed significant acceleration of skin
repair following dermabration compared with no additional treatment.
Evidence supports the use of Aloe vera for the healing of first to second degree
burns ( Schmidt et al 1991). It has been shown that it does not have any toxic effects on
central nervous system cells in Albino mice models (KOSIF et. al., 2008). In the light of Av
use as a wound healing agent in folk and modern medicine, the present study was
undertaken to fully evaluate the dermal wound healing potential of this drug after topical
application of its aqueous extract on experimentally induced cutaneous wounds in rat
models. This study was undertaken to evaluate the wound healing properties of Aloe vera
on cutaneous wounds in albino mice.
Plane of work(Material and Method)
Materials:Test animal:- Swiss albino mice.
Test chemical:- Betadine
Plant material:- Aloe vera juice.
Method:The study was carried out in following part:-
(A)Extraction of Aloe vera juice and preparation of working formula.
(B) Creation of excision wound.
(C) Tretement of wounds.
(D) Estimation of wound healing.
(b) Histological and Histochemical study.
(E) Statistical analysis of the data Analysis .
Results:-
On 6th day of wound creation ,the microscopic examination of the healing tissue collected and
kept as control (T 1) showed that majority of wound area invaded by granulation tissue
comprising of macrophages , enlarged ,swollen , oval fusiform fibroblast cell as well as
proliferatng endothelial cell and newly formed blood vessels. Fine and scanty collagen fibre
were found to be interspersed between the fibroblast cell. Some area of fibrin rich clot and
coagulum invaded by neutrophils and macrophages were present on superficial surface and
tissue distant fromedges.The epidermis adjacent to wound edge showed hyperplasia.
The wound treated with Betadine (T2) solution showed similar type of histopathological picture
with almost same degree of fibroblast proliferation, neo-vascularization and reepithelialization.
25%(t 4) and 50%(T 5) Aloe vera juice treated cases showed more marked infiltration of fibroblast cells
and mononuclear cells as compared to the Betadine (T2) treated cases.The collagen fibres in these
groups were also thin , fine and dispersed and were comparatively more than the Betadine treated
wounds.But in the case of 5% Aloe vera treated wound (T3) ,the fibroblast cell were found to be
maximum as compared to all other groups.The collagen fibres were thick ,dense ,wavy ,and maximum in
number in this groups.
On 12th days of wound creation the microscopic examination of the wound tissue collected and
kept as control (T 1) showed marked increase in collagen accumulation and fibroblast cell
proliferation with the newly formed blood vessels. The fibroblast cellwerestill swollen, plumped
and oval. Collagen fibre were also thin ,fine and intermix with the matrix.
The the microscopic examination of Aloe vera juice treated wounds showed increase in the maturity
of grandulation tissue maximum in 5% (T3) treated wound. There is less prominent nuclei of fibroblast
cells ,more thick , dense and wavy collagen bundles ,comparatively less number of newly formed blood
vessels and lesser number of mononuclear cell infiltration.
The result were quantified visually by numbering them from 0 to 5 with the score 5 for the maximum
similarity and 0 for the least similarity from the normal tissue collected on the day of excision wound
formation in both the test and control groups. These are presented in Table -5 as well as illustrated in
figure 14 – 17.
On 6th day of wound creation ,infiltration was recorded higher in Aloe vera juice treated wound 5%
,2.041 ± 0.293 ; 25% ; 2.066 ± 0.318 , 50% 2.042 ± 0.310 ) as compared to Betadine ( 1.250 ±
0.446 ) treated wound . Infiltration on 12th days did not vary significantly but was higher in Aloe
vera treated wound.
On 6th day ,fibroblast proliferation was higher in Betadine treated (2.833 ± 0.562) wound than
the Aloe vera 5% (2.165 ± 0.204 ); 25% ( 1.961 ± 0.188 ) 50% (1.956 ± 0.246 ) treated wound .But
on 12th days ,the fibroblast proliferation was higher in Aloe vera juice treated 5% (3.166 ± 0.204 )
25% (3.042 ± 0.188); 50% (3.041 ± 0.188) as compared to the Betadine (2.042 ± 0.292 )
treated wound .
On 6th , the collagenization was higher in Aloe vera treated wound , maximum with 5% (3.169±
0.2.5) followed by 25% (2.208 ± 0.188 ) and 50% (2.208 ± 0.188 ) as compared to the Betadine
(1.165 ± 0.127 ) treated wound . On 12th day , the collagenization was much higher in 5% Aloe
vera ( 4.083 ± 0.204) followed by 25% Aloe vera ( 3.251 ± 0. 203) as compared to 50% Aloe vera
( 2.207 ± 0.245) ,Betadine ( 2.251 ± 0.158 ) treated wound .
On 6th day ,the neo-vascularization in the healing wound tissue was higher with 5% Aloe vera
(2.208 ± 0.188) than the other treatment group 25% (2.167 ± 0.129 ); 50% (2.083 ± 0.129 )
Betadine (1.082 ± 0.129 ). On 12th the neo-vascularization was higher with 5% Aloe vera
(3.207 ± 0.102 ) as compared to other group ( 25% , 2.208 ± 0.189 ; 50% ,2.167 ± 0.205
;Betadine ,1.26 ± 0.136 ).
The degree of epithelialization in the healed wound area was found in the range of 1.12 to 2.16
out of maximum 5 score in all the treatement group .
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