Download influence of honey-balm on the rate of scare formation during

Bull Vet Inst Pulawy 48, 311-316, 2004
INFLUENCE OF HONEY-BALM ON THE RATE
OF SCARE FORMATION DURING EXPERIMENTAL BURN
WOUND HEALING IN PIGS
AGATA KABAŁA-DZIK1, RAFAŁ STOJKO1, EWA SZAFLARSKA-STOJKO1,
IRENA WRÓBLEWSKA-ADAMEK1, ARTUR STOJKO2,
JERZY STOJKO2 AND BARBARA STAWIARSKA-PIĘTA1
1
2
Department of Pathology,
Department of Bioanalysis, Sanitary Sciences and Environmental Studies,
Faculty of Pharmacy in Sosnowiec, 41-200 Sosnowiec,
Silesian Medical University in Katowice, Poland
e-mail: [email protected]
Received for publication January 15, 2004.
Abstract
The pharmacological activity of honey balm, its
influence on wounds healing processes and the rate of scar
formation in comparison with control groups were assessed.
Clinical and histopathological studies showed that honeybalm not only shortened the period of wound healing, but
also had a positive impact on the general health condition of
the animals.
Key words: pig, burn wounds, honey-balm,
healing.
Wound healing is a complex process
influenced by a number of factors. An important factor
in the wound healing is correct blood supply. Tissue
ischaemia caused by blood vessels coagulation leads to
the development of a protein scab which is a perfect
ground for microorganisms development. Also,
impaired blood flow in wound tissues (especially in the
1st and 2nd degree wounds) may lead to further damage
and may cause the infection of a wound by bacteria
living in soft tissues (10, 12, 13).
Judging from the processes taking place
during wounds healing, it follows that the basic
conditions for damaged tissue restoration and healing
are:
wound
cleaning,
proper
macrophages
accumulation (they provide the growth factors),
proliferation of fibroblasts producing extracellular
matrix and the formation of new blood vessels
(neoangiogenesis). The activity of these processes may
be a marker of the reconstruction process occurring in
damaged tissues.
The current progress made in the analytical
studies and in the field of biotechnology has led to a
renewed interest in pharmacotherapy based on
biogenic components. More and more attention has
been given to apipharmacotherapy which utilizes the
healing properties of active fractions of bee products.
The great physician and philosopher Hippocrates, who
documented bee products’ therapeutical properties,
used strained honey or honeycombs with the crust in
the treatment of wounds difficult to cure (5). Clinical
and research centers – which are becoming better and
better equipped in modern analytical equipment – have
enriched the knowledge known for generations as far
as to the composition and properties of bee products.
Their studies have led to the isolation of active
substances in bee products and then to the
standardization of their pharmacological properties
(18).
The pharmacological activity of obtained
fractions in relation to human organisms arises from
the fact that bee products – to a greater or lesser extend
– include in their composition all basic substances
necessary for human body homeostasis (17, 18).
One of the newly discovered apitherapeutics
is the honey crust. Polish scientists have isolated in it
the active substance which has anti-bacterial and
regenerative properties, activating the immunological
system through the stimulation of properdine and the
complement activity.
312
Substances in the honey have a positive effect
on the correct functioning of an animal organism and
are also well-assimilated by it. They also determine a
number of honey's actions: antibacterial, regenerative,
detoxic, nutritious, immunomodulative, preservative,
anti-allergic as well as cardioprotective and soothing
(2, 3, 6 ,7, 9, 15, 16, 21, 23). Honey bioelements are
the activators of a number of immunological
mechanisms (7, 18, 22).
Having in mind the difficulties experienced
during the treatment of burn wounds as well as the
described properties of bee products, the aim of the
present study was to assess the healing rate and the
process of scar formation in burn wounds under the
influence of honey balm in pigs.
Material and Methods
The experimental part of the study was
conducted in the Central Experimental Animal
Quarters of the Silesian Medical University according
to the standards arising from the European Conference
and the standards of Good Laboratory Practice. The
experimental model consisted of identical after-burn
wounds of skin done according to the Hoekstr model.
This model is in accordance with the Dutch Act of
Animal Testing and the currently in force experimental
protocol of the Committee of Charity operating at the
University of Amsterdam (4, 11).
The tested pigs were of the "white lop-eared"
breed, aged 15-16 weeks, with weight 30-40 kg.
Throughout the experiment the animals lived in
identical zoohygienic conditions and were fed,
according to their age, the standard fodder. In order to
produce the burn wounds a hot brass block (size: 1.5 x
3 cm, temperature: 170°C) was placed for 10 s on the
skin of premedicated and then generally anaesthetized
animals. The wounds were performed on the sides,
along the spine of each animal (Fig.1).
Depending on the substance used, the burn
wounds were divided into the following groups:
Group I (control):
- subgroup IA: 9 wounds of pig no 1 - not treated
but only washed once a day with 0.9% NaCl
solution;
- subgroup IB: the remaining 9 wounds of pig no 1 treated with a thin layer of Dermazin (SSD-silver
sulphadiazine) applied once a day;
Group II (experimental):
- 9 wounds of pig no 2 treated with honey-balm
consisting honey crust. It was applied once a day
as a thin layer to each of the wounds.
Fig. 1. A pig after burn wounds execution. Burn wounds after therapeutics application and providing with sterile
dressing.
313
The clinical assessment was done throughout
the duration of the study and covered the general
health conditions as well as the reaction to
environment. The process of burn wound healing was
especially carefully assessed. The appearance and
processes taking place in the tissue surrounding the
wounds were evaluated as well as the granulation
tissue formation and the process of scar formation.
The histopathological assessment of the
healing process was done on the 3rd, 5th, 10th and 15th d
of the study. Before material samples were taken for
the histopathological tests, animals were subjected to
general anesthesia. The samples were collected from
the middle of the wounds, from their borders as well as
from the skin in the wound area. Paraffin sections were
stained with the standard haematoxylin – eosin
method.
The study was accepted by the Regional
Ethical Commission of Animal Experiments in
Katowice, Poland. The study was performed as part of
the NN-2-047/01. Project financed by the Scientific
Research Committee.
Results
Clinical observations:
Day
1
3
5
10
15
Honey-balm
NaCl
Dermazin
The clinical pictures of all the subgroups were similar. Along the necrotic changes, there were
inflammatory reactions with a large amount of exudate and a skin oedema around the wound.
The wounds were smaller with a No change in the clinical picture.
formed
scab.
Oedema
and
reddening at the wound were
visually smaller.
The wounds were clean with no Wounds were covered with thin, Wounds were covered with
non-fully formed scab.
exudate with a correctly developed poorly-formed scab.
Oedema and reddening on
scab.
skin around wounds.
Wounds were covered with smaller No changes when compared to Dry wounds. Formation of
scabs, and on the skin border on the day 5.
dry scabs, no changes in
surface of the wound there was a
skin around wounds.
pink scar, what suggests presence
of advanced epidermalization.
The wounds were covered with an Wounds covered with dry scabs Wound cavity filled with
fresh granulation tissue.
organized scar, and they could be of the same size.
Wound granulation process
treated as healed completely.
on the edges.
During palpation the animals did
not react defensively.
The clinical evaluation showed visible
differences in the process of wound healing after
applying the above mentioned medications.
In the case of the apitherapeutic treatment, the
differences in wound appearance and animal behaviour
were visible after 48 h. Wounds were more wet, skin at
the wounds was more elastic. Oedema and reddening
at the wound surroundings were smaller. On the 10th d
of the experiment the wound surface became visually
smaller. The scabs started to come off at the
boundaries of the wound surface, and a light-pink scar
was visible underneath. This suggested an advanced
epidermalization process. The wounds developed after
taking small segments to the histopathological tests
could be treated as healed completely. The applied
apitherapeutics not only shortened the period of wound
healing, but also has a positive impact on the general
health condition of the animals. During the time of the
experiment the animal did not show any signs of
excitation and did not react defensively.
After applying the honey balm, the signs of
granulation on the surface of the wound were observed
on the 5thday. The inflammatory infiltration with a
considerable amount of granulocytes was found. Their
presence in the wound surrounding indicated the
stimulation of cell migrating processes, intensified
wound cleaning and favourable conditions for the
epithelium migration. On the 10th d, the loss of
314
epithelium was observed in a part of the wound. In the
remaining part, the regenerating epithelium with the
mature granulation underneath was still visible. On the
15th
d the defect was completely covered with a great
amount of collagen fibers, which suggested the process
of scar formation (Fig.2).
Histopathological study:
Da
y
3
5
10
15
Honey balm
NaCl
Dermazin
Loss of stratified squamous epithelium, extensive coagulative necrosis and thick inflammatory infiltration.
First signs of granulation both in Loss of epithelium with
Loss of epithelium and
bottom and on rims of wound.
coagulative necrosis.
coagulative necrosis, under which
Inflammatory infiltration on the
there was inflammatory
deeper layers.
infiltration.
A regenerating epithelium was
Scab visible on the surface of
Inflammatory infiltration seen
seen on the edge of the wound.
epithelium. First phases of “fresh” under necrotic masses. Signs of
In the middle part of the wound
granulation seen on the bottom
granulation seen on the bottom
there was a granulating tissue
and edges of wound.
and edges of wound.
filing the whole defect.
The skin defect filled completely Wound cavity filled with fresh
Wound cavity filled with fresh
with granulation with a
granulation. Granulation
granulation. Granulation
considerable amount of collagen processes seen on the wound
processes seen on the wound
fiber (Fig.2).
edges (Fig. 3).
edges (Fig. 4).
Fig. 2. The 15th d after burn execution. Honey-balm. Granulation tissue with dominance of collagen fibers. (Area
100x, HE).
315
Fig. 3. The 15th d after burn execution. NaCl. A wound covered with crust. On border a epithelium visible. (Area
100x, HE).
Fig. 4. The 15th d after burn execution. Dermazin. Granulation filling a wastage. (Area 100x, HE).
316
Discussion
The histopathological study revealed that
pathological process in each of the sub-groups was
similar, the time from the moment the thermal wound
was made to the begining of repair processes in the
wound was different depending on the applied remedy.
In the case of honey-balm it was the 5th d, after
applying Dermazin (a standard medication used in
burn healing) the repairing process was observed at
day 15 after its application.
The therapeutic effectiveness of an
apitherapeutic, whose pharmacological activity results
from the physico-chemical characteristics of honey or
propolis, confirmed the previous report (14). The
effects of apitherapeutics on wound healing are well
known in the cases when the conventional methods of
healing are not effective (8).
The favourable effect of honey was also
substantiated in healing of the infected wounds
encontered after the cesarean section and histerectomy.
The application of honey not only shortens the healing
period. The hospitalization period in female patients,
who were treated with honey was 10 d shorter than in
those treated with the conventional methods (1). Honey
treatment influences scar formation and its cosmetic
appearance.
Subrahmanyam (19, 20) in his study showed
that healthy granulation tissue was observed earlier in
patients treated with honey. Of the wounds treated with
honey 87% healed within 15 d versus 10% in group
treated with silver sulfadiazine (SSD). Other
histological and clinical studies of wound healing
showed satisfactory epithelialization after 7 d in 84%
and after 21 d in 100% of wounds treated with honey.
In wound treated with SSD epithelialization occurred
by the 7th d in 72% of patients and by the 21st d in 84%
of patients.
Relief of pain, a lower incidence of
hypertrophic scar and postburn contracture, low cost
and easy availability make honey balm an ideal
dressing in the treatment of burn.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
21.
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