Received: 05/10/2024 Accepted: 25/12/2024 Published: 18/02/2025 1 of 8
https://doi.org/10.52973/rcfcv-e35540 RevistaCientíca,FCV-LUZ/Vol.XXXV
ABSTRACT
In cats, gingivitis treatment typically involves professional
dental cleaning under anesthesia, pain management, and
antibiotic administration to reduce inflammation. Considering
the disadvantages of antibiotic administration, the necessity
for alternative acute treatment protocols arises. It is believed
that enhancing the antibacterial, anti–inflammatory, and local
anesthetic effects of propolis with ozone could shorten the
treatment duration, help reduce the risks associated with gingivitis,
and also support the overall health of the cat. The study included
20 domestic cats diagnosed with the causative agent of gingivitis
presented to private clinics. The cats included in the study were
grouped based on bacterial agents (n=10; Female=4, Male=6)
and viral agents (n=10; Female=5, Male=5). Propolis extracted
with ozone–enriched oil was administered in spray form for 14
days (d). Before the application and on the 7
th
and 14
th
d of the
treatment, the gingival indices of the cats were recorded, scored,
saliva samples were collected, and photographs were taken. The
levels of VEGF and TNF–α in saliva were determined using a cat–
specic ELISA kit. When the data were evaluated, the application
of ozone–enriched propolis demonstrated a statistically signicant
reducing effect on the levels of TNF–α in saliva in both groups
(P<0.01). Salivary VEGF levels showed a signicant increase during
application, especially in gingivitis caused by bacterial agents
(P<0.05). In the viral group, application was found to be more
effective in increasing VEGF levels during the rst 7 d. In gingivitis
caused by bacterial agents, the gingival index (GI) and plaque index
(PI) decreased compared to pre–treatment values (P<0.05). In
gingivitis associated with viral agents, the decrease in the GI was
statistically signicant, while the decrease in the PI was found to
be non–signicant. In conclusion, this study demonstrated that
the application of ozone–enriched propolis might be an alternative
treatment option for cats with gingivitis.
Key words: Feline gingivitis; ozone–enriched propolis; saliva
VEGF; saliva TNF–α
RESUMEN
En los gatos, el tratamiento de la gingivitis suele consistir en una
limpieza dental profesional bajo anestesia, tratamiento del dolor y
administración de antibióticos para reducir la inflamación. Teniendo
en cuenta los inconvenientes de la administración de antibióticos,
surge la necesidad de protocolos alternativos de tratamiento agudo.
Se cree que potenciar los efectos antibacterianos, antiinflamatorios
y anestésicos locales del propóleo con ozono podría acortar la
duración del tratamiento, ayudar a reducir los riesgos asociados
a la gingivitis y también favorecer la salud general del gato. El
estudio incluyó 20 gatos domésticos diagnosticados con el agente
causante de la gingivitis presentados en clínicas privadas. Los gatos
incluidos en el estudio se agruparon por agentes bacterianos (n=10;
Hembra=4, Macho=6) y virales (n=10; Hembra=5, Macho=5). El
propóleo extraído con aceite enriquecido con ozono se administró
en forma de aerosol durante 14 días (d). Antes de la aplicación
y en los d 7 y 14 del tratamiento, se registraron y puntuaron los
índices gingivales de los gatos, se recogieron muestras de saliva y
se tomaron fotografías. Los niveles de VEGF y TNF–α en saliva se
determinaron mediante un kit ELISA especíco para gatos. Cuando
se evaluaron los datos, la aplicación de propóleos enriquecidos con
ozono demostró un efecto reductor estadísticamente signicativo
sobre los niveles de TNF–α en saliva en ambos grupos (P<0,01).
Los niveles de VEGF salival mostraron un aumento signicativo
durante la aplicación, especialmente en la gingivitis causada por
agentes bacterianos (P<0,05). En el grupo vírico, la aplicación
resultó más ecaz para aumentar los niveles de VEGF durante
los primeros 7 d. En la gingivitis causada por agentes bacterianos,
el índice gingival (IG) y el índice de placa (IP) disminuyeron en
comparación con los valores previos al tratamiento (P<0,05). En
la gingivitis asociada a agentes virales, la disminución del IG fue
estadísticamente signicativa, mientras que la disminución del
IP resultó no signicativa. En conclusión, este estudio demostró
que la aplicación de propóleo enriquecido con ozono podría ser
una opción de tratamiento alternativa para los gatos con gingivitis.
Palabras clave: Gingivitis felina; propóleo enriquecido con ozono;
VEGF salival; TNF–α salival
The therapeutic effect of ozone–enriched propolis oil extraction in cats
with gingivitis
Efecto terapéutico de la extracción de aceite de propóleo
enriquecido con ozono en gatos con gingivitis
Gamze Sevri Ekren–Aşıcı* , Umut Kal , Seda Berberoğlu , Funda Kıral
Aydın Adnan Menderes University, Faculty of Veterinary Medicine, Department of Biochemistry. Aydın, Türkiye.
*Corresponding author: gamze.ekren@adu.edu.tr
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INTRODUCTION
Problems with a pet’s oral and dental health can affect its overall
health and quality of life [1]. Although the prevalence of oral cavity
and periodontal diseases in cats (Felis catus) varies in the literature,
ranging from 70-85% [2] to 85-95% [3] depending on the age
groups of the studied population, all reports agree that these
conditions are frequently observed in cats older than two years,
with an incidence rate of up to 95% [4]. Gingivitis, the earliest stage
of periodontal disease, is an inflammation of the gums caused
by subgingival plaque bacteria or microbial by–products such
as cytotoxins and endotoxins in response to the host’s immune
response [5, 6, 7]. The presence of anaerobic species of gram–
negative bacteria is typically found in these bacterial plaques [8].
In cats with compromised immune systems, opportunistic
bacteria have been reported to cause gingivitis, with samples
from these cats primarily isolating gram–positive bacteria such
as Staphylococcus, Corynebacterium, and Streptococcus, as well
as gram–negative bacteria such as Pseudomonas spp., Proteus
mirabilis, and Klebsiella pneumoniae [9].
Gingivitis in cats can occur in all breeds and at any age with
varying severity; however, it is diagnosed more frequently in adult
or older cats. Its occurrence is not attributed to a single cause
[10, 11]. Gingivitis is more frequently observed in cats during
the periods of secondary teeth emergence (3-5 months), sexual
maturity (6-9 months), and later in life due to the accumulation of
tartar and dental calculus on the tooth surfaces [12]. Predisposing
factors include systemic diseases, particularly immune–
suppressing conditions such as feline immunodeciency virus
(FIV) and feline leukemia virus (FeLV), as well as diet. Additionally,
local dental diseases such as excessive tooth crowding (commonly
seen in many Persian cats), tooth morphology, feline odontoclastic
resorptive lesions (FORL), and fractured teeth are present [13].
The primary goal of treatment is to control gingivitis by cleaning
both supra – and subgingival dental plaque and tartar, as well as
addressing the predisposing factors [14, 15]. Broad–spectrum
antibiotics are utilized in the treatment of gingivitis. The most
commonly used antibiotics are amoxicillin–clavulanate acid,
cephadroxil, and clindamycin [16]. Although gingivitis is a reversible
inflammation, if treatment is not performed in a timely manner or if
resistant bacterial strains are involved, the ongoing infection can lead
to the loss of supporting dental tissues and progress to periodontitis.
In the advanced stages of periodontitis, it can lead to tooth loss, bone
infection, and allow pathogenic bacteria to enter the bloodstream,
potentially causing organ damage. Oral diseases can also be indicative
of other systemic conditions [7, 17]. It has even been suggested that
dental disease is a risk factor for the development of chronic kidney
disease in cats [18]. Therefore, easy–to–apply treatment options
that prevent the formation of antibiotic–resistant bacteria should
be developed for the treatment of gingivitis.
Propolis, a natural product whose use has become widespread
in various forms due to its pharmacological effects and benecial
properties in recent years, was chosen. Propolis has gained
popularity in recent years due to its therapeutic properties,
containing numerous natural compounds such as polyphenols,
phenolic aldehydes, sesquiterpene quinones, coumarins, amino
acids, and steroids [19]. Many studies have shown that propolis has
a wide range of biological and pharmacological effects, including
antimicrobial, antioxidant, anti–inflammatory, immunomodulatory,
antitumor, anticancer, anti–ulcer, hepatoprotective,
cardioprotective, and neuroprotective actions. These properties
have been studied for their potential use in veterinary medicine
and have shown promise in various formulations [20].
Most ozonated oils have been reported to be used in the
treatment of infections and skin diseases without causing side
effects [21, 22, 23]. It has been reported that ozone stimulates
the immune system by inducing leukocytosis and phagocytosis
at low doses [24], can promote tissue healing, and exhibits anti–
inflammatory effects [21]. Especially due to its biocompatibility
with epithelial and periodontal mucosal cells, positive results have
been achieved in the prevention, control, and treatment of oral
infections [22, 23]. Ozone therapy is used in veterinary medicine
for the local treatment of various lesions and neuromuscular
diseases, including mastitis, metritis, endometritis, fetal membrane
retention, vaginitis, urovagina, enteritis, and laminitis [24, 25, 26].
Saliva is an important fluid from a periodontal perspective. It
washes the inside of the mouth and protects the oral mucosa by
coating it against external factors. Pro–inflammatory cytokines
such as interleukin-1 beta (IL-1β) and tumor necrosis factor–alpha
(TNF–α) play a key role in the pathogenesis of periodontal diseases,
and the inhibition of these cytokines reduces bone loss associated
with periodontitis [27, 28]. TNF–α is a cytokine that mediates
some of the events that occur during periodontal disease. VEGF
stimulates the proliferation of vascular endothelial cells necessary
for angiogenesis, activates the release of proteolytic enzymes, and
intensies chemotaxis and migration [29]. Additionally, it plays a
signicant role in angiogenesis, bone formation, wound healing,
and the regeneration of oral epithelium.
As a result, the undesirable side effects of modern medications
have led to a preference for pharmaceuticals derived from natural
sources in recent years. Due to these negative aspects worldwide,
there has been a shift toward new explorations, and herbal therapy
research has gained importance. Literature studies have reported
that propolis is effective in the treatment of gingivitis in humans and
dogs; however, no such study has been found in cats. Therefore,
the aim of the study was to present an alternative treatment option
for gingivitis in cats.
MATERIALS AND METHODS
Research design
The domestic cats brought to private clinics located in the
Efeler district of Aydin province underwent careful oral and dental
examinations. Cats with naturally occurring varying degrees of
gingival inflammation, who had undergone a general examination
and had the causative factor of gingivitis identied, were included
in this study.
In the preliminary assessment for the oral and dental
examination, factors such as the number of teeth, presence of
dental caries, color and volume of the gingiva, halitosis, presence
of deciduous teeth, symmetry of the mouth, presence of lesions
in the oral mucosa, enamel defects, enamel hypoplasia, fractures,
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gingival hyperplasia and retraction, oral masses, oro–nasal stula,
and exposure of the pulp were taken into consideration.
The study material consisted of a total of 20 cats, with 10 having
a bacterial causative factor (female=4, male=6) and 10 having a
viral causative factor (female=5, male=5) in TABLE I. Although the
causative factors differed, all cats were diagnosed with chronic
gingivitis. Among the cats with viral–induced gingivitis, previuosly
2 were diagnosed with Feline Herpes Virus and Feline Calicivirus,
while 8 were diagnosed with Feline Immunodeciency Virus (FIV).
as well as on the 7
th
and 14
th
d. To collect the saliva sample, the
patients’ mouths were rinsed thoroughly with water. then cats
were placed on their side with their mouths closed and the saliva
accumulated in the oral cavity was collected approximately 0.5 mL
of saliva was obtained using a dropper into Eppendorf tubes. The
samples were then centrifuged (Nüve, Model:NF800R, Türkiye)
at 3500 g for 10 min to remove cellular components and plaque.
The samples were stored at -80
o
C (Nuaıre, Model: NU6617W35,
USA) until they were evaluated.
Measurement of plaque index (PI) and gingival index (GI)
Using a periodontal probe, the depth (periodontal pocket) was
measured by inserting it into at least three points on each tooth,
and the average value was recorded. To assess the severity of
gingivitis, the Plaque Index (PI) by Silness and Löe [30] and the
Gingival Index (GI) by Löe and Silness [31] were used.
Determination of TNF–α and VEGF levels based on ELISA method
In the study, the levels of TNF–α and VEGF in saliva samples
were measured using a cat–specic commercial VEGF ELISA kit
(BT LAB, E0139 Cat, China) and a TNF–α ELISA kit (BT LAB, E0031
Cat, China). The saliva samples were analyzed in accordance with
the procedures outlined in the kit, with two replicates performed
for each analysis. The results of the ELISA analyses were calculated
in ng/mL using CurveExpert Professional v. 2.7.3 software.
Exclusion criteria
Pregnant cats, cats that had recently undergone surgery, and
cats that had used antibiotics within the last 15 d were excluded
from the study, as their physiological conditions could lead to
different results. Additionally, cats that had received long–term
antibiotic and corticosteroid treatment were excluded from the
study due to the potential for differing levels of VEGF and TNF–α.
Statistical analyses
All statistical analyses were performed using the SPSS for
Windows Version 29.0 software package (SPSS Inc., Chicago, IL,
USA). A P–value of less than 0.05 was considered an indicator of
a statistically signicant difference in the decisions. The normal
distribution of the continuous variables was assessed using the
Shapiro–Wilks test. It was determined that all data, except for GI
and PI values, followed a normal distribution. To compare means
between the groups, all ELISA results were evaluated by using
repeated measures ANOVA. The Bonferroni correction was used
to adjust P–values in multiple comparison tests. The Friedman test
was applied to evaluate the PI and GI data. The Wilcoxon signed–
rank test was applied to assess differences between the groups.
RESULTS AND DISCUSSIONS
According to the data from a small survey conducted with
pet owners during our study, pet owners preferred the use of
oral spray over procedures that would distress and increase
the anxiety of their cats, such as administering medication and
taking blood samples. They were also more positively inclined
towards providing saliva samples instead of taking blood during
the treatment process. This study has presented a more easily
Ozonation and application of propolis
Ozone (O
2
/O
3
mixture) was generated from medical–grade oxygen
using ozonator equipment (Sorande; Model: SAIR-2,Türkiye).
The amount of ozone in the oily propolis was measured using a
UV spectrophotometer (Shımadzu,Model: UV-1601, Australia)
at 254nm. Propolis extracted in ozone–enriched (5%) olive oil
was placed in a spray bottle and applied once a day (d) for 14 d,
delivering 2-3 puffs (0.3 mL) each time.
Sampling
In this study, the effectiveness of the treatment was monitored
through photography on d 0, 1, 3, 5, 7, 10, and 14. Additionally,
plaque index (PI) and gingival index (GI) data were recorded.
Saliva samples were collected at the beginning of the treatment,
TABLE I
Distribution of cases according to race, age, gender and the causative agent
Case Rase Age (Year) Gender Diagnosis Causative Agent
1 Tabby 4 Male Gingivitis Bacterial
2 British 3 Female Gingivitis Bacterial
3 British 5 Male Calculus+Gingivitis Bacterial
4 Tabby 8 Female Calculus+Gingivitis Bacterial
5 British 2 Male Calculus+Gingivitis Bacterial
6 Tabby 6 Male Gingivitis Bacterial
7 Tabby 7 Female Calculus+Gingivitis Bacterial
8 Tabby 3 Male Calculus+Gingivitis Bacterial
9 Tabby 5 Female Gingivitis Bacterial
10 Tabby 4 Male Calculus+Gingivitis Bacterial
11 Tabby 1 Female Calculus+Gingivitis Viral
12 Tabby 6 Female Calculus+Gingivitis Viral
13 Tabby 7 Male Calculus+Gingivitis Viral
14 Tabby 1 Male Gingivitis Viral
15 Tabby 1 Male Calculus+Gingivitis Viral
16 Tabby 1 Female Calculus+Gingivitis Viral
17 British 2 Female Gingivitis Viral
18 Tabby 4 Male Calculus+Gingivitis Viral
19 Tabby 4 Male Calculus+Gingivitis Viral
20 Tabby 5 Male Calculus+Gingivitis Viral
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applicable alternative treatment method compared to other
treatment options. Prior to applying ozone–enriched oily propolis
The comparison of saliva TNF–α levels due to the application
of ozone–enriched propolis in gingivitis caused by both viral and
bacterial agents revealed a statistically signicant difference
between TNF–α levels on days 7 and 14 compared to the beginning
of the application process (P<0.01). (TABLE II, FIG 2).
Oral diseases in cats are an important issue in veterinary
medicine. Oral health is a critical factor that affects the overall
health of cats [7, 32]. Gingivitis is primarily caused by bacterial
plaques that colonize the gingival sulcus. These plaques produce
various toxins and by–products that lead to inflammation of the
gums. The immune system secretes cytokines in response to
these bacteria and by–products, including IL-1β, interleukin-8
(IL-8), prostaglandins, and TNF–α [7]. These cytokines lead to the
extraction to the cats meeting our criteria, PI and GI measurements
were taken and photographed (FIG 1).
Before treatment
process
14
th
day of
application
FIGURE 1. Images taken before and on the 14
th
day of application of ozone–enriched oily propolis extract
TABLE II
TNF–α levels (ng·L
-1
) in saliva samples collected at the beginning of
treatment, as well as on days 7 and 14, in gingivitis caused by viral and
bacterial agents following the application of ozone–enriched propolis
Application of ozone–
enriched propolis
TNF–α levels in gingivitis
caused by bacterial agents.
Mean ± SD (n=10)
TNF–α levels in gingivitis
caused by viral agents.
Mean ± SD (n=10)
Beforeapplication 126.26 ± 8.80 162.30 ± 18.65
7
th
dayofapplication 66.07 ± 9.72
***
128.25 ± 13.80
**
14
th
dayofapplication 37.09 ± 6.54
***
106.64 ± 11.37
**
**
: P<0.01,
***
: P<0.001indicatessignicancewhencomparingthebeginningofthe
treatmentwiththebeginningofthetreatmentaswellasondays7and14according
torepeatedmeasuresANOVA
FIGURE 2. The changes in salivary TNF–α levels in gingivitis caused by bacterial
agents (A) and viral agents (B) following the application of ozone–enriched olive
oil–extracted propolis (*P<0.05; **P<0.01; ***P<0.001)
A
B
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accumulation of inflammatory cells and initiate the inflammatory
process [1, 13]. There have been many studies in humans on the
relationship between some inflammatory mediators in saliva and
the pathogenesis of periodontitis. They reported that salivary
TNF–α levels were higher in individuals with periodontal disease
compared to healthy individuals [33, 34, 35]. Geng [36] and Rai
[37] have proposed that TNF–α levels in saliva may serve as a
potential biomarker for periodontal disease. In a study conducted
in dogs, TNF–α levels in saliva samples were found to be higher in
healthy dogs with gingivitis than in healthy dogs without gingivitis
[38]. In addition, TNF–α has been closely associated with tissue
destruction and immune response, and thus has been reported
to play an important role in periodontitis–induced bone loss [39].
When all these data are evaluated, it can be considered that the
decrease in TNF–α levels with ozone–enriched olive oil propolis
reflects the positive effects of the treatment.
When examining the VEGF levels in saliva, it was observed that
the levels increased during the application in gingivitis caused by
bacterial and viral agents, and this increase fluctuated throughout
the process. However, when the values obtained before and after
the nal application were examined, a signicant increase was
detected. Our results are summarized in FIG. 3 and Table III.
Current studies demonstrating the effects of propolis on oral
and dental health show that the flavonoids found in propolis
possess antimicrobial, anti–inflammatory, and immunomodulatory
properties, which are highly benecial in the treatment of aphthous
ulcers, candidiasis, gingivitis, and periodontitis [40, 41]. Thus, it
is believed that propolis will reduce oxidative stress and prevent
the formation of damage through its antioxidant effects.
In veterinary medicine, propolis is used as an ointment to control
mastitis in dairy cows. In pig herds, 5% propolis is added to the
milk as a prophylactic agent for respiratory and gastrointestinal
diseases. It is also added to the diets of rams, pigs, and calves
to stimulate growth. It is also used for wound healing and as a
local anesthetic for surgery [42]. The effects of propolis have
been studied in dogs for dermatophytosis, sarcoptic mange [43],
and fungal otitis [44]. Recently, it has also started to be used as a
treatment option for disinfection in regenerative endodontics [45].
No studies have been found regarding the use of propolis in cats.
In a study conducted in the early stages of gingivitis, it was
determined that VEGF levels decreased and returned to the level
before the onset of normal gingivitis with oral health practices [46].
Another study reported a decrease in salivary VEGF levels at the onset
of experimental gingivitis [47]. When our results are analyzed, it is
thought that VEGF levels increase especially in gingivitis caused by
bacterial agents, and VEGF levels, which decrease due to gingivitis,
increase due to the elimination of the agent during treatment.
Measurements taken with a periodontal probe during the study
were recorded according to the Löe and Silness gingival index
scoring system [31]. In cats with gingivitis caused by bacterial
agents, the GI was measured as 1.90 ± 0.82 at the beginning of
the application, 1.40 ± 0.69 on d 7, and 1.30 ± 0.48 on d 14. When
statistically evaluating the application, the difference between the
beginning and the end of the treatment was found to be statistically
signicant (P<0.05). When the pre–application and d 7 application
were statistically evaluated, the difference between them was
found to be statistically insignicant.
In cats with gingivitis caused by viral agents, the gingival
index values measured at the beginning of the application were
1.88 ± 0.33, remained at 1.88 ± 0.33 on d 7, and were recorded
as 1.44 ± 0.73 on d 14. When comparing the GI values between
the beginning of the application and d 7, no statistically signicant
difference was found between the groups (P>0.05). However,
the difference between the GI on d 7 and 14, was found to be
statistically signicant. When the GI values at the beginning and
the end of the study were evaluated, the difference between them
was found to be statistically signicant (P<0.05).
TABLE III
VEGF levels (ng·L
-1
) in saliva samples collected at the beginning of
treatment, as well as on days 7 and 14, following the application of
ozone–enriched propolis in relation to viral and bacterial agents
Application of ozone–
enriched propolis
VEGF levels in gingivitis
caused by bacterial agents.
Mean ± SD (n=10)
VEGF levels in gingivitis
caused by viral agents.
Mean ± SD (n=10)
Beforeapplication 267.54 ± 37.82 325.72 ± 39.87
7
th
dayofapplication 411.48 ± 53.14
*
415.98 ± 41.13
14
th
dayofapplication 511.56 ± 65.19
**
418.62 ± 29.71
*
*: P<0.05,**:P<0.01indicatessignicancewhencomparingthebeginningofthe
treatmentwiththebeginningofthetreatmentaswellasondays7and14according
torepeatedmeasuresANOVA
FIGURE 3. The changes in salivary VEGF levels in gingivitis caused by bacterial
agents (A) and viral agents (B) following the application of ozone–enriched olive
oil–extracted propolis (*
P<0.05; **P<0.01)
A
B
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Throughout the study, measurements taken with a periodontal
probe were statistically evaluated based on the Silness and Löe
plaque index (PI) [30]. In cats with gingivitis caused by bacterial
agents, the PI was measured as 1.70 ± 0.82 at the beginning of
the application, 1.30 ± 0.49 on d 7, and 1.30 ± 0.48 on d 14. When
statistically evaluating the application, the results from before
the application and on d 7 and 14 were found to be statistically
signicant (P<0.05). However, the difference between the results
on d 7 and d 14 was found to be statistically insignicant (P>0.05).
In cats with gingivitis caused by viral agents, the PI values were
measured as 1.89 ± 0.61 before the application, 1.88 ± 0.60 on d
7, and 1.78 ± 0.67 on d 14. When the PI was statistically evaluated
during the application period, the difference between the groups
was found to be statistically insignicant (P>0.05).
Since ozone therapy stimulates the wound healing process, it
has been reported to be effective in the treatment of various skin
disorders [48, 49]. It was reported to have benets such as faster
tolerance of hyperemia in the gums, quicker healing of gingival
bleeding, and rapid reduction of edema at the gingival margin.
Researchers applying ozone therapy concluded that it was highly
reliable and potentially effective due to the absence of observed
side effects and its natural origins. There are numerous studies
demonstrating the therapeutic effects of ozone application in the
treatment of gingivitis in humans. While there are studies [23]
proving that ozone application reduces plaque formation in dogs, no
studies have been found examining the effects of ozone application
on oral and dental health in cats. In light of this information, our
study is unique in investigating the therapeutic efcacy of both
propolis and ozone in cats.
The alcoholic extraction of propolis is not suitable for medical
use. The aqueous extraction has lower biological activity due to the
reduced content of phenolic and volatile oils. Studies have shown
that ozone itself does not penetrate cells and quickly reacts with
polyunsaturated fatty acids, leading to an increase in the formation
of oxidant products. However, when applied with oil, the entry of
ozone into the cells occurs in a slow and controlled manner. For
this purpose, the oily extract of propolis enriched with ozone was
chosen for application in our study.
CONCLUSION
This study will provide a new perspective on scientic research
related to herbal products used in veterinary medicine. The use
of herbal products is being conducted on animals; however, these
applications are not based on scientic data. Studies conducted in
this area are considerably insufcient compared to those in human
medicine. In this study, the data obtained from the oral application
of ozonated oily extract of propolis in cats diagnosed with gingivitis
will serve as an important resource for future research on the topic.
In our study, saliva samples were also collected to determine
the levels of TNF–α and VEGF present in the saliva. This results
also enhance its uniqueness as a study demonstrating that the
monitoring of periodontal diseases in animals can be conducted
through saliva samples. In summary, our study found that the
propolis extracted in olive oil enriched with ozone had a local
effect and reduced the inflammatory response.
Conflict of interest
There are no conflicts of interest, according to the authors,
regarding this article.
Credit author statement
G.S.E.A. was involved in the study design, animal handling,
laboratory experiments, and statistical analyses and interpretation
of the results. U.K. was involved in the literature review, the
application to animals and the evaluation of the results. S. B. was
involved in the literature review, the application to animals and
the evaluation of the results. F. K. was involved in the literature
review, interpretation of results and drafting of the manuscript.
Ethics approval and consent to participate
The study was approved by the Animal Experiments Local Ethics
Committee of Adnan Menderes University with the date January 19,
2023 and number 64583101/2023/20 and which were performed
in strict accordance with the guidelines of the Experimental Animal
Ethics Committee.
ACKNOWLEDGEMENT
The authors thank the Scientic and Technological Research
Council of Turkey (TUBITAK) for the nancial support.
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