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2022;24(2):78-83doi:10.34172/jsums.2022.13
Effect of aqueous extract of fig fruit (Ficus Carica) on wound
healing in albino rabbits
Zahra Mokhtari Zarch1
ID
, Elham Salehi1*
ID
, Majid Morovati-Sharifabad1
ID
, Amin Paidar Ardakani2
ID
, Mohammad Saeed
Heydarnejad3
ID
1Department of Basic Sciences, Faculty of Veterinary Medicine, Ardakan University, Ardakan, Iran
2Department of Clinical Sciences, Faculty of Veterinary Medicine, Ardakan University, Ardakan, Iran
3Department of Animal Sciences, Faculty of Basic Sciences, Shahrekord University, Shahrekord, Iran
*Corresponding Author: Elham Salehi, Faculty of Veterinary Medicine, Ardakan University, Ardakan, Iran. Tel: 09132121299,
Email: esalehi@ardakan.ac.ir
http://j.skums.ac.ir
Original Article
Abstract
Background and aims: Skin ulcers are caused by various reasons such as physical, chemical, and biological damages. Wound healing has
long been considered one of the most important issues in surgery. Extensive research has been employed in this field and various chemical,
herbal, homeopathic, and physical methods such as laser therapy have been used for treatment. Nowadays, wound healing is one of the
most important aims of medical science. Furthermore, people prefer to use natural drugs because of fewer side effects. Therefore, the
present study was designed considering these issues.
Methods: For this study, 6 albino rabbits with an average weight of 3 kg were purchased. After preparing the rabbits, two pairs of full-
thickness wounds were created in a completely round shape with a diameter of almost 6.30 mm on both sides of the spine in two anterior
and posterior parts at a distance of 3 cm from the midline of the body. The animals were divided into 2 groups of 3 rabbits, including 1
experimental group and 1 control group. The wounds of the experimental group were treated with fig fruit extract while the wounds of the
control group received no treatment. The wounds were macroscopically examined at 4, 7, 12, 16, and 20 days after surgery.
Results: The results of t-test showed a significant difference (P < 0.05) among the groups on all days. Meanwhile, the average wound area
significantly decreased in the group which was treated with the extract of fig fruit.
Conclusion: This study showed that fig fruit extract contains phenolic compounds with antioxidant, antimicrobial, and anti-inflammatory
properties which can accelerate the wound healing process.
Keywords: Fig fruit extract, Skin wound, Albino rabbit, Surgery
Received: August 22 2021, Accepted: January 17 2022, ePublished: 2 June 2022
Introduction
A wound is a ruptured tissue that has lost its connectivity
due to various factors. Several reasons can cause skin
ulcers, including physical, chemical, and biological
damages (1). Wound healing is an essential process to
repair the structure and function of tissues that have been
injured by physical, chemical, bacterial, or viral damage.
It is also a complex process that causes restoration of
structural and functional integrity after injury (2).
Wound healing involves coordinated repair responses of
blood cells, extracellular matrix, and parenchymal cells
that occur after surgery or traumatic injury to the body
(3). Wound healing has long been considered one of the
most important issues in surgery. Extensive research
has been employed in this field and various chemical,
herbal, homeopathic, and physical methods such as laser
therapy have been used for treatment. The goal of these
methods is fast treatment, low complication rate as well
as low cost (4).
Today, standard topical antibiotics such as silver nitrate,
mafenide acetate, and silver sulfadiazine are commonly
used for treating the wounds. The use of mafenide acetate
can soothe pain or burning at the site of medication while
frequent consumption of silver nitrate due to the presence
of silver leads to permanent skin discoloration. Currently,
the use of sulfadiazine cream is one of the most common
methods of healing burn wounds; however, it has several
side effects, including non-penetration into the wound,
resistance to Gram-negative microorganisms, and delay in
the healing of small burn wounds. Reducing regimentation
and epithelialization is one of the goals of medical wound
healing in a shorter time, as well as reducing side effects (5).
Studies have shown that fig fruit as a laxative is useful
in the treatment of various diseases such as hemorrhoids,
gout, and epilepsy (6). Figs are also a good source of
potassium and thus are effective in lowering blood pressure.
Other benefits of figs include weight loss, prevention of
breast cancer, improvement in bone density, prevention
of muscle weakness, and improvement in nervous
system function (7). Although figs are known as edible
fruits, their leaves, roots, and sap are used as medicine,
the medicinal value of which has been proven through
Journal of Shahrekord University of Medical Sciences, Volume 24, Issue 2, 2022 79
Effect of g fruit extract on wound healing
scientific research (8). The biological activities of fig juice
are very diverse. In traditional medicine, fig juice is used
to treat stomach ulcers and skin infections such as viral
warts. Additionally, bioactive substances called 6-1-acetyl-
beta-glucose-betacitosterol and lupeol acetate have been
isolated from fig juice, which inhibit the proliferation of
cancer cells (9). Fig tree sap, which is a type of secretion
from the leaves and fruits of Ficus carica, has therapeutic
effects due to its antioxidant, antihyperlipidemic (10),
anticancer (11), and antimicrobial activities (12). It has
also been investigated by researchers due to the presence
of proteolytic enzymes, amino acids, minerals, sugars,
terpenes, and organic acids. Figs are free of fat, sodium,
and cholesterol. Among fruits, it has the highest amount
of minerals and especially contains a lot of calcium and
fiber (7). Figs can increase the metabolic activity of the
kidneys, which leads to an increase in the excretion of
harmful substances in the urine in patients who consume
figs. Uric acid combines with malic acid in figs to increase
bile secretion, which is effective in the treatment of liver
disease (13). Nowadays, wound healing is one of the most
important aims of medical science. Furthermore, people
prefer to use natural drugs because of fewer side effects.
Therefore, the present study was designed considering
these issues.
Materials and Methods
Preparation of fig fruit
Fresh fig fruit was collected from the gardens of Mehriz
city. It was identified and approved by the relevant experts
in the Medicinal Plants Research Centre.
Preparation of fig fruit extract
First, fig fruit was crushed with a mixer. Then, 100 g of
the crushed fruit was added to water (1 L) and kept in a
dark place for 72 hours. The resulting liquid was distilled
in a rotary evaporator by double vacuum distillation.
Afterwards, it was incubated at 38°C until finally a viscous
aqueous extract (10%) was obtained.
Animal preparation
In this experiment, 6 Albino rabbits with an average
weight of 3 kg were used. The animals were obtained from
the Animal Care Centre of Medical School and kept in
propylene cages. Animals were kept separately in special
cages under the same condition in terms of temperature
(20 ± 2°C) and light-dark cycle (12 hours). They were
fed with lettuce and carrots, and the drinking water was
provided ad libitum.
Making a wound
For surgery, the animals were anesthetized with xylazine
at a dose of 5 mg/kg and ketamine at a dose of 35 mg/kg by
intramuscular injection. After induction of anesthesia, the
animals were put in the prone position and the hairs on
the back on both sides of the thoracic and lumbar spines
were completely shaved. A skin punch was used to create
wounds of equal sizes (Figure 1 and Figure 2).
In the present study, two pairs of full-thickness wounds
were created on the skin on both sides of the spine.
Wounds on both sides (left and right) were created at a
distance of 3 cm from the midline of the body and the
distance between the wounds on both sides was 4 cm.
After scarring, the lesion areas were covered with
sterile dressings to prevent infection, and the rabbits were
examined until the end of full recovery. Then, each of
them was kept and treated in a separate cage.
Animal grouping
Rabbits were divided into two groups of three animals. In
the experimental group, fig ointment was applied to the
wounds while the control group received no treatment. A
Figure 1. Pre-operative preparation.
Figure 2. Sampling and wounding steps.
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Mokhtari Zarch et al
sufficient amount of fig ointment (1 mm thickness) was
applied daily to the wound.
Treatment duration
The wounds were treated for 21 days. The investigation
was initiated on day 0. Tissue sampling was performed on
histopathological sections on days 4, 9, and 21.
Macroscopic examination
Left anterior and posterior wounds were used for
macroscopic examination. Graphs were analyzed at days
0, 4, 8, 12,16, and 20 by a digital camera with a calibrated
ruler next to the wound. The images were then transferred
to a computer, and the area of the wounds was calculated
using TCapture software (Figure 3 and Figure 7).
Microscopic examination
Right anterior and posterior wounds were used for
microscopic evaluation of wounds. On days 4, 9, and, 21,
a part of the skin tissue with completely thick edges of the
wound, including healthy and damaged skin, was removed
by scalpel and after washing with physiological serum, it
was fixed in a container containing 10% formalin buffer.
Then, 5-μm sections of these samples were prepared by
microtome and stained with Fiber hematoxylin and eosin.
In the prepared slides, the histopathological evaluation of
the wound healing process was done based on evidence
such as inflammation, granulation tissue formation,
fibroblast cell maturation, collagen fiber arrangement,
vascularization, epithelial tissue rearrangement, and
wound healing rate.
Statistical analysis
In order to analyze the data obtained from macroscopic
evaluation, the average wound area in the treatment and
control groups was evaluated using student’s t test in SPSS
version 25.0. The significance level was considered to be
P < 0.05.
Results
On day 4, the damaged epidermal tissue (ep) was observed
in the control group. Furthermore, a large distance was
observed between the epidermis and the dermis. No trace
of granulation tissue was observed. Tissue hyperemia
was evident. Collagen fibers were disordered and the
inflammatory process was observed in this tissue. A slow
healing process was observed. Granulation tissue was
obvious. Inflammation was observed in the tissue. On day
4, the repair of the epidermis (ep) was not complete in the
treatment group. The fibroblasts matured. The number
of blood vessels (v) was small. Hair follicle formation
was observed. On day 4, granulation tissue was observed.
Inflammation was also observed in the tissue (Figure 3
and Figure 4).
On day of 9, the collagen fibers moved towards
parallelism and the maturation of fibroblasts was observed
in the control group. The repair of epidermal epithelial
tissue was still not complete. On day 9, increased blood
vessel formation, granulation tissue formation, and
maturation of fibroblasts were detected in the treatment
group (B). Collagen fibers had a more regular arrangement.
Angiogenesis (V) and rearrangement of epithelial tissue
were observed (Figure 5).
On day 21, granulation tissue was rarely seen in the
control group (A). The collagen fibers were more regular
and parallel. Limited evidence of angiogenesis (V) and
rearrangement of the epithelial tissue could be observed.
On day 21, angiogenesis was not observed in the treatment
group (B). The epithelial tissue was completely repaired
and limited traces of granulation tissue were observed (gt)
(Figure 6).
Figure 3. Results of Macroscopic Examination of the Control (a) and
Treatment Groups (b).
Figure 4. Results of microscopic examination of the control (A) and
treatment (B) groups (H&E, × 50) on day 4.
Figure 5. Results of Microscopic Examination of the Control (A) and
Treatment (B) Group (H&E, × 50) on Day 9.
Figure 6. Results of microscopic examination of control (A) and treatment
(B) group (H&E, × 50) on day 21.
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Effect of g fruit extract on wound healing
Statistical analysis
In the diagram, the average wound area was plotted
with standard error in terms of days in the control and
experimental groups. Then, the area under the curve was
calculated.
Tabl e 1 shows the statistical characteristics of the level
variable below the wound area chart in terms of days at the
experimental and control groups. In this table, the average
level below the chart is 48.355 for the control group and
40.605 for the target group. It can be said that the wound
area in terms of day in the target group is lower compared
to the control group. To determine the significance
and non-significance of the difference, t test with two
independent samples was used (Figure 7 and Tab le 1).
In this diagram, the average wound area was plotted
with standard error in terms of days in the control and
experimental groups. Then, the area under the curve was
calculated.
To determine the significance and non-significance of
the difference between the average surface variable below
the wound area diagram in comparison to day, t test with
two independent samples was used.
According to the t test, it was observed that at the error
level of less than 5%, the significance level of the Loon
test for the variable of the wound area was less than 5%.
(P = 0.023). Therefore, the condition of inequality of
variance was accepted. Due to the significant level of t-test
for this variable at an error level of less than 5%, the mean
wound area per day in the experiment and control groups
were significantly different from each other, which was
equal to 7.750. As a result, the mean difference between
the two groups was significant, so it could be said that
the wound area in the study period in the target group
compared to the control group had a significant decrease
(Tabl e 1 and Figure 7).
Discussion
In this study, the effect of aqueous extract of fig fruit on
the skin wound healing process in albino rabbits was
investigated. The results of macroscopic examinations
(measurement of wound area) at days 0 (day of wound
formation), 4, 8, 12, 16, and 20 in the control group and
the group treated with fig fruit showed that the wound
area in the whole period of time had a significant decrease
in the experiment group compared to the control (less
than 5%) and therefore this treatment was successful in
this group. Additionally, in the microscopic examination
performed in this study, the wound healing process during
the 4, 9, and 21 days was evaluated histologically and it
was shown that the speed and process of wound healing
in the group treated with fig fruit extract was higher than
in the control group. Wound healing is one of the most
fundamental issues that human beings have faced since
the beginning of creation. Numerous drugs and ointments
are used to repair open wounds, each of which has several
drawbacks, limitations, and side effects (3-5). Today,
the importance of healing is due to the high prevalence
of wounds in human and animal communities. Major
common wounds include traumatic, burn, chemical,
physical, and biological wounds. Research in the field of
wound healing is very important in terms of accelerating
the healing process of the wounds that heal slowly or
are under pressure and movement. The use of plant
extracts in the healing process of skin wounds has been
common from ancient times in traditional medicine. The
herbal medicine selected for wound healing based on its
physical and chemical properties should be able to ideally
improve one or more healing phases without causing side
effects. Numerous studies have shown positive effects of
herbal medicines on wound healing, some of which are
mentioned. The microscopic results of the present study
Table 1. The Results of t-test
Wound Area
Based on Day
Group Number Mean SD
Treatment 6 40.605 1.288
Control 6 48.355 1.188
Figure 7. Comparison of the average wound area between the control and treatment groups during different days.
Journal of Shahrekord University of Medical Sciences, Volume 24, Issue 2, 2022
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Mokhtari Zarch et al
showed that the number of blood vessels and granular
tissue of the group treated with fig fruit extract was
significantly different from the control, which is consistent
with a study by Yaghmaei et al. Yaghmaei et al showed that
the number of blood vessels in the group treated with
green tea extract was significantly different from control
group because green tea extract accelerated the healing
process, which is consistent with the present findings
(14). Additionally, the study by Rezvanipour et al on the
effect of mummy on some indicators in wound healing
of mice showed that the number of blood vessels in the
treated group was significantly different from the control
group and mummy could accelerate the healing process,
which is consistent with the present study (15). A study
by Azhdari-Zarmehri et al on the effect of hydroalcoholic
extract of Scrophularia striata on burn wounds in rats
showed that the granulation tissue and blood vessels of
the treated group were significantly different from the
control group and hydroalcoholic extract of this plant
accelerated the healing process; their results are consistent
with the present study (16). The obtained results of this
study showed the quality and speed of wound healing
and the amount of shrinkage of wound edges in the
treatment group were higher compared to the control
group. Avijgan et al investigated the effect of aloe vera
gel on chronic ulcers and showed that aloe vera gel was
much more effective and less expensive than conventional
treatments, which is similar to the present study (17).
Tramer et al in 2012, by studying medicinal plants, stated
that figs are plants whose different parts are medically
important and have been used extensively in traditional
Iranian medicine. They indicated that fig ointment has
an anti-inflammatory effect on tumors and can be used
to prevent cancer due to its antibacterial and antifungal
properties. Figs contain antioxidants that prevent damage
to cells and tissues. All antioxidant properties along with
antimicrobial properties are found in figs (18). A study
conducted by Hashemi and Abediankenari on mice
confirmed the anti-inflammatory effects of lavender.
They reported that the use of lavender was effective in
healing burns (19). Another study found that Ficus carica
stem extract contained large amounts of polyphenols and
carotenoids. On the other hand, the antioxidant activity
of aqueous extract and polysaccharide extracted from
figs was studied and researchers found that both aqueous
extract and polysaccharide obtained from figs had high
amounts of phenolic compounds and flavonoids with
remarkable inhibitory properties (2, 2 diphenyl-1-picryl
hydroxyl), which can be beneficial for human health
(20). In another study, the levels of phenolic compounds
including gallic acid, chlorogenic acid, epicatechin, and
catechins as well as the total antioxidant capacity of fig
juice were investigated, and it was found that fig juice can
be useful in preventing damage induced by free radicals
(21). Raj and Joseph stated that figs contain antioxidant
compounds including flavonoids and polyphenols and
some biologically active compounds such as arabinose,
β-amyrin, β-carotene, glycosides, etc (22). The results of
the phytochemical analysis of Ferula assa-foetida resin
show that this plant contains compounds such as ascorbic
acid, phenolic acid, oxalic acid, uric acid, maleic acid,
coumarins, alanine, tannins, alpha linoleic acid, and
glycoside. All these compounds have anti-inflammatory
effects and can accelerate the wound healing process,
which is consistent with the present study (23). Moreover,
Laitiff et al in 2010 showed that bitter gourd increases
the wound healing rate in diabetics due to the presence
of photochemical substances such as polyphenols,
flavonoids, and triterpenoids (24).
Conclusion
Wound healing is one of the most fundamental issues
that mankind has faced since the beginning of creation.
Numerous medications and ointments are used to heal
open wounds, each of which has several drawbacks,
limitations, and side effects. The use of herbal extracts
in the healing process of skin wounds has been common
in traditional medicine since ancient times. The herbal
medicine selected for wound healing based on its physical
and chemical properties should be able to ideally improve
one or more healing phases without causing side effects.
According to the present study, the aqueous extract of fig
fruit can improve the wound healing process.
Acknowledgments
The authors thank Mr. Mohsen Rashidi for providing histology
facilities.
Authors’ Contribution
MM, ES, and ZMZ contributed to the conception and design of
experimental work. APA contributed to surgical operations. All
authors read and approved the final manuscript.
Conflict of Interests
No conflict of interest exists.
Ethical Approval
This study was approved by the Ethics Committee of Ardakan
University (IR.YAZD.REC.1400.005).
Funding/Support
No funding was received for this study.
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