Revista Cienﬁca, FCV-LUZ / Vol. XXXV Recibido: 22/08/2025 Aceptado: 09/10/2025 Publicado: 01/11/2025 UNIVERSIDAD DEL ZULIA Serbiluz Sistema de Servicios Bibliotecarios y de Información Biblioteca Digital Repositorio Académico 1 of 7 Revista Cienﬁca, FCV-LUZ / Vol. XXXV https://doi.org/10.52973/rcfcv-e35778 UNIVERSIDAD DEL ZULIA Serbiluz Sistema de Servicios Bibliotecarios y de Información Biblioteca Digital Repositorio Académico Eﬀect of Ankaferd blood stopper on Cadmium-induced heart damage in male rats Efecto de Ankaferd Blood Stopper sobre el daño cardíaco inducido por cadmio en ratas macho Mümin Polat Burdur Mehmet Akif Ersoy University, Faculty of Health Sciences, Burdur, Turkey. Correspondence author: mpolat@mehmetakif.edu.tr ABSTRACT Cadmium (Cd), a widespread environmental contaminant, is known to cause ssue injury through mechanisms involving oxidave stress, inﬂammaon, and apoptosis. The heart is parcularly vulnerable to such toxic eﬀects. Ankaferd Blood Stopper, a standardized herbal hemostac agent, also exhibits notable anoxidant, an-inﬂammatory, and an-apoptoc acvies. This study aimed to explore the potenal protecve eﬀects of Ankaferd Blood Stopper against cadmium-induced cardiac injury, evaluang whether Ankaferd Blood Stopper administraon migates histopathological and apoptoc damage in cardiac ssue caused by cadmium toxicity in rats. Thirtytwo male Wistar Albino rats (350–450 g) were divided into four groups (n=8 each): Control (saline, i.p.), Cd (2.5 mg/ kg CdCl₂, i.p.), Ankaferd Blood Stopper (0.5 mL ABS, i.p.), and Cd + Ankaferd Blood Stopper (CdCl₂ + Ankaferd Blood Stopper, both i.p.). Aﬅer treatment, cardiac ssues were collected and evaluated using histopathological staining and Caspase-3 immunohistochemistry. Blood samples were also collected for biochemical analysis. Cadmium exposure resulted in marked myocardial damage, including inﬂammatory inﬁltraon, vascular congeson, and disorganized myoﬁbers. Caspase-3 expression conﬁrmed increased apoptoc acvity. Ankaferd Blood Stopper administraon signiﬁcantly reduced histopathological lesions and Caspase-3 immunoreacvity in the Cd + Ankaferd Blood Stopper group, suggesng a protecve eﬀect. Ankaferd Blood Stopper eﬀecvely aenuated cadmium- induced cardiac injury, likely due to its anoxidant and an- inﬂammatory properes. These ﬁndings indicate that Ankaferd Blood Stopper may oﬀer therapeuc beneﬁts in prevenng or alleviang heavy metal-induced cardiovascular damage. Addional research is needed to validate these eﬀects in long- term and clinical models. Keywords: Cadmium; cardiac damage; ankaferd blood stopper; rat RESUMEN El cadmio (Cd), un contaminante ambiental ampliamente distribuido, es conocido por causar lesiones sulares a través de mecanismos que implican estrés oxidavo, inﬂamación y apoptosis. El corazón es parcularmente vulnerable a estos efectos tóxicos. Ankaferd Blood Stopper, un agente hemostáco herbal estandarizado, también presenta notables acvidades anoxidantes, aninﬂamatorias y anapoptócas. Este estudio tuvo como objevo explorar los posibles efectos protectores del Ankaferd Blood Stopper frente a la lesión cardíaca inducida por cadmio. Evaluar si la administración de Ankaferd Blood Stopper miga el daño histopatológico y apoptóco en el tejido cardíaco causado por la toxicidad por cadmio en ratas. Treinta y dos ratas macho Wistar Albino (350–450 g) se dividieron en cuatro grupos (n=8 cada uno): Control (solución salina, i.p.), Cd (2,5 mg/kg de CdCl₂, i.p.), Ankaferd Blood Stopper (0,5 mL de Ankaferd Blood Stopper, i.p.) y Cd + Ankaferd Blood Stopper (CdCl₂ + Ankaferd Blood Stopper, ambos i.p.). Tras el tratamiento, se recolectaron los tejidos cardíacos y se evaluaron mediante nción histopatológica e inmunohistoquímica de Caspasa-3. También se obtuvieron muestras de sangre para análisis bioquímico. La exposición al cadmio provocó un daño miocárdico marcado, que incluyó inﬁltración inﬂamatoria, congesón vascular y ﬁbras musculares desorganizadas. La expresión de Caspasa-3 conﬁrmó un aumento de la acvidad apoptóca. La administración de Ankaferd Blood Stopper redujo signiﬁcavamente las lesiones histopatológicas y la inmunorreacvidad de Caspasa-3 en el grupo Cd + Ankaferd Blood Stopper, lo que sugiere un efecto protector. El Ankaferd Blood Stopper atenuó de forma eﬁcaz la lesión cardíaca inducida por cadmio, probablemente gracias a sus propiedades anoxidantes y aninﬂamatorias. Estos hallazgos indican que el Ankaferd Blood Stopper podría ofrecer beneﬁcios terapéucos para prevenir o aliviar el daño cardiovascular inducido por metales pesados. Se recomienda realizar más invesgaciones para validar estos efectos en modelos clínicos y a largo plazo. Palabras clave: Cadmio, daño cardíaco, ankaferd blood stopper, rata

Eﬀect of Ankaferd Blood Stopper on Cadmium -Induced Heart Damage / Polat UNIVERSIDAD DEL ZULIA Serbiluz Sistema de Servicios Bibliotecarios y de Información Biblioteca Digital Repositorio Académico INTRODUCTION Ankaferd Blood Stopper® (ABS) is a therapeuc preparaon employed in the management of both minor and major bleeding owing to its hemostac acvity. The formulaon consists of extracts derived from the root of Urca dioica (nele), the dried leaves of Vis vinifera (grape) and Glycyrrhiza glabra (licorice), the dried rhizome of Alpinia oﬃcinarum (galangal), and the dried herb of Thymus vulgaris (thyme). In recent years, researchers have focused not only on the hemostac eﬀects of ABS but also on its anoxidant, anmutagenic, and an-inﬂammatory properes [1 , 2 ,[3]. The term heavy metal, which has gained increasing aenon in recent years and holds a signiﬁcant place in the literature, is widely used to describe metals or metalloids oﬅen associated with contaminaon and potenal toxicity or eco-toxicity. The deﬁnion of heavy metals typically refers to metals with a density greater than 5 g/cm³ [4 , 5]. Heavy metals can have toxic eﬀects, posing serious health risks, parcularly for fetuses, infants, and children, as these groups are more vulnerable to heavy metal exposure due to their higher consumpon relave to body weight. Prolonged exposure to high doses of heavy metals during pregnancy facilitates placental transfer, potenally causing severe and permanent brain damage in the fetus. As a result, later in life, aﬀected individuals may experience learning diﬃcules, memory impairment, and behavioral disorders such as aggression and hyperacvity [6 , 7]. Cadmium (Cd), a toxic heavy metal discovered by Friedrich Stromeyer in 1817, is naturally present in air, water, and soil as part of Group 2B on the periodic table. Cd is a pervasive environmental and occupaonal pollutant. Today, it is primarily used in the producon of nickel-cadmium baeries, pigments, and plasc stabilizers, while its applicaons in alloys, solders, and galvanizaon have declined. Addionally, Cd exposure has become inevitable due to its presence as a major compound in cigarees. Both natural and anthropogenic acvies contribute signiﬁcantly to environmental Cd levels. Occupaonal exposure to Cd primarily occurs in mining, baery producon, Cd- containing pigment manufacturing, Cd alloy processing, and electronic waste recycling. Due to its long biological half-life (15– 20 years) and persistence in the environment and body ssues, Cd remains a major public health concern, Moreover, previous experimental studies have shown that toxic exposures can result in cardiac necrosis and histopathological alteraons in rodents, highlighng the vulnerability of the heart to toxic agents [6 , 8]. Chronic Cd exposure, even at low doses, leads to its accumulaon in the liver, kidneys, and heart due to its long half- life (15–20 years). Regardless of the route of intake, Cd enters the bloodstream and binds to proteins and other blood components [9]. The Internaonal Agency for Research on Cancer (IARC) classiﬁes Cd as a human carcinogen. Although Cd is not a strong mitogen, it acts as a tumor promoter (iniator) due to its impact on gene expression. Cd contributes to carcinogenesis through several mechanisms, including abnormal gene expression, inhibion of DNA repair enzymes, oxidave stress inducon, and E-cadherin dysfuncon. Approximately 5% of ingested Cd is absorbed through the gastrointesnal tract, whereas about 90% of inhaled Cd is absorbed through the lungs. Once absorbed, Cd is rapidly distributed throughout the body, accumulang predominantly in the liver and kidneys, which together constute the majority of the total Cd body burden [10]. ABS (Ankaferd Health Products Ltd., Istanbul, Turkey) is a hemostac agent formulated by blending ﬁve herbal ingredients in speciﬁc proporons, which have been tradionally used in Turkish medicine for centuries. In addion to its hemostac properes, ABS modulates vascular endothelial funcon, blood cell acvity, angiogenesis, cell proliferaon, and vascular dynamics through the regulaon of several biological mediators. MATERIAL AND METHODS Animals and experimental protocols This study was approved by the Süleyman Demirel University Animal Experiments Local Ethics Commiee with the decision dated September 5, 2024 (No: 06/78). All experiments were conducted at the Süleyman Demirel University Animal Producon and Experimental Research Center in accordance with the ethical guidelines established by the commiee. A total of 32 male Wistar Albino rats (Raus norvegicus), weighing 350–450 g, were used in the study. The rats were obtained from the Süleyman Demirel University Animal Producon and Experimental Research Center (Isparta, Turkey). The animals were randomly divided into four experimental groups (n = 8 per group) as follows: • Group I (Control group): Received 0.5 mL of physiological saline intraperitoneally (i.p.). • Group II (Cadmium (CdCl₂) group): Received 2.5 mg/kg cadmium chloride (CdCl₂) dissolved in sterile saline at a volume of 1 mL/kg body weight, administered intraperitoneally (i.p.). • Group III (ABS group): Received 0.5 mL ABSi.p. • Group IV (Cd + ABS group): Received 2.5 mg/kg CdCl₂dissolved in sterile saline at a volume of 1 mL/kg body weight, followed by 0.5 mL of ABS administered intraperitoneally (i.p.). The ABS group received 0.5 mL/kg ABS (Ankaferd Health Products Ltd., Turkey) intraperitoneally from the right inguinal region on the day of the experiment. The Cd group received 2.5 mg/kg cadmium chloride (CdCl₂, Catalog No: 13667, Alfa Aesar, USA) dissolved in sterilsaline and administered intraperitoneally from the right inguinal region [11]. Forthe Cd + ABS group, treatment was administered as follows: CdCl₂ (2.5 mg/kg, i.p.) was injected ﬁrst,and one hourlater,0.5 mL/kg of ABS was administered intraperitoneally at the same site. One day (d) aﬅer Cd administraon, all rats were anesthezed using 90 mg/kg Ketamine HCl and 10 mg/kg Xylazine. Following euthanasia, blood and heart ssue samples were collected for biochemical and histopathological analyses. Histopathological examinaon The heart ssues collected from the animals were ﬁxed in 10% buﬀered formaldehyde soluon. Prior to ssue processing, the samples were washed overnight with running tap water, followed by dehydraon in graded Ethanol, clearing with Xylene, and embedding in paraﬃn. Secons of 4–5 μm thickness were obtained from the paraﬃn blocks using a rotary microtome 2 of 7

Revista Cienﬁca, FCV-LUZ / Vol. XXXV UNIVERSIDAD DEL ZULIA Serbiluz Sistema de Servicios Bibliotecarios y de Información Biblioteca Digital Repositorio Académico (Leica RM2235, Leica Biosystems, Wetzlar, Germany) and subsequently stained with hematoxylin and eosin (H&E). Histopathological examinaon and scoring were performed using a light microscope (Zeiss Primo Star, Carl Zeiss Microscopy GmbH, Jena, Germany). A modiﬁed semi-quantave scoring system was applied to evaluate the ﬁndings, categorized as follows: (0): No pathological ﬁndings (normal myocardial architecture). (+1): Mild ﬁndings, including slight myoﬁbrillar disorganizaon or focal nuclear pyknosis. (+2): Moderate ﬁndings, such as evident myoﬁbrillar loss, capillary congeson, or localized necrosis. (+3): Severe ﬁndings characterized by extensive myoﬁbrillar degeneraon, intersal edema, and widespread necrosis [12]. Immunohistochemical analyzes Apoptoc acvity was evaluated using immunohistochemical detecon of Caspase-3. Tissue secons (4–5 μm thick) were inially deparaﬃnized and dehydrated. Subsequently, the secons were sequenally incubated with 3% hydrogen peroxide, Ultra-V Block (Thermo Fisher Scienﬁc, Waltham, MA, USA), primary anbodies (Bioss Anbodies, Beijing, China), secondary anbody (Thermo Fisher Scienﬁc, Waltham, MA, USA), and streptavidin peroxidase (Thermo Fisher Scienﬁc, Waltham, MA, USA). The secons were then stained using 3,3’-diaminobenzidine (DAB) soluon (Vector Laboratories, Burlingame, CA, USA), followed by nuclear counterstaining with hematoxylin [13]. A previously reported semi-quantave scoring system was adapted to evaluate immunoreacvity. The intensity of immunostaining was scored as follows: 0: Negave 1: Weak 2: Moderate 3: Strong The percentage of posively stained cells was calculated and categorized as follows: 0%–4% = 1 5%–19% = 2 20%–39% = 3 40%–59% = 4 60%–79% = 5 80%–100% = 6 A composite “Quick” score was calculated by mulplying intensity and percentage scores to obtain a ﬁnal immunoreacvity value. Stascal analysis The data obtained were analyzed using GraphPad Prism 9 soﬅware. One-way ANOVA was performed to compare diﬀerences among the groups, and Tukey’s test was used to determine signiﬁcant diﬀerences between groups. A P-value < 0.05 was considered stascally signiﬁcant. RESULTS AND DISCUSSIONS Histopathology The histopathological ﬁndings are summarized in TABLE I. Longitudinal and cross-secons of the control and ABS groups exhibited normal myocardial histology, with regularly arranged muscle ﬁbers (FIGS. 1; A, E, B, F). No signiﬁcant diﬀerences were observed between the ABS group and the control group (all P > 0.05). In contrast, the Cd group demonstrated signiﬁcant histopathological alteraons, including pyknoc nuclei, myoﬁber damage (characterized by loss of striaons, eosinophilia, and disorganizaon), and capillary congeson, when compared to the control group (all P < 0.001) (FIGS. 1; I, J). However, a signiﬁcant reducon in myocardial degradaon, necrosis, and inﬂammatory inﬁltraon was observed in the Cd + ABS group compared to the Cd group (all P < 0.001) (FIGS. 1; M, N). TABLE I Histopathological ﬁndings in cardiac ssue of experimental rat groups Grup/parameter Control Abs Cd Cd+Abs Min. Max. Med. Min. Max. Med. Min. Max. Med. Min. Max. Med. Pyknoc nucleus 0 1 0 0 1 0 2 3 2 0 1 0 Myoﬁber damage 0 1 0 0 1 0 2 3 2.50 0 1 0.50 Capillary cogneson 0 0 0 0 0 0 2 3 3 0 2 1 ABS: Ankaferd Blood Stopper; Cd: Cadmium; Min.: Minimum; Max.: Maximum; Med.: Median. 3 of 7

Eﬀect of Ankaferd Blood Stopper on Cadmium -Induced Heart Damage / Polat UNIVERSIDAD DEL ZULIA Serbiluz Sistema de Servicios Bibliotecarios y de Información Biblioteca Digital Repositorio Académico The table shows the minimum, maximum, and median scores of histopathological alteraons observed in the cardiac ssue of rats from the Control, ABS, Cd, and Cd+ABS groups. Parameters evaluated included pyknoc nucleus, myoﬁber damage, and capillary congeson. Immunohistochemistry TABLE II presents the semi-quantave scoring results for Caspase-3 immunoreacvity across all groups. No immunoreacvity was observed in the control and ABS groups (FIGS. 1; C, D, G, H). Compared to the control group, Caspase-3 acvity was signiﬁcantly increased in the Cd group (P < 0.001) (FIGS. 1; K, L). However, a signiﬁcant reducon in Caspase-3 immunoreacvity was observed in the Cd + ABS group compared to the Cd group (P < 0.001) (FIGS. 1; O, P). TABLE II Mulplicave Quick score results of caspase-3 immunohistochemistry in cardiac ssue of control and experimental rat groups. Data are presented as median (Q25–Q75), minimum, and maximum values. Grup/parameter Control Abs Cd Cd+Abs Med. (Q25-Q75) Min. Max. Med. (Q25-Q75) Min. Max. Med. (Q25-Q75) Min. Max. Med. (Q25-Q75) Min. Max. Caspase-3 immunoreacvity 0 (0-0) 0 0 0 (0-0.5) 0 1 12(10-16.5) 10 18 3(2.5-4) 2 4 ABS: Ankaferd Blood Stopper; Cd: Cadmium; H&E: Hematoxylin and Eosin; Q25–Q75:Interquarle range (25th–75th percenles), represenng the middle 50% of the data distribuon. FIGURE 1. Photomicrographs of longitudinal and cross secons of cardiac muscles: In H&E staining, normal cardiac muscle histology with regularly arranged ﬁbrils was observed in the control (A, B) and ABS (E, F) groups. In contrast, the Cd group exhibited pyknoc nuclei (yellow arrow), diﬀuse vascular congeson (black arrow), and myoﬁbrillar damage charac- terized by loss of striaons, eosinophilia, and muscle disorganizaon (I, J). A reducon in these pathological ﬁndings was observed in the Cd + ABS group compared to the Cd group (M, N). In immunohistochemical analysis, no immunoreacvity was detected in the control (C, D) and ABS (G, H) groups. However, strong Caspase-3 immunoreacvity was observed in the Cd group (K, L). A signiﬁcant decrease in immunoreacvity was noted in the Cd + ABS group compared to the Cd group (O, P). (Scale bar = 50 µm). Tissue repair and wound healing are intricate processes involving inﬂammaon, ﬁbroplasia, neovascularizaon, wound contracon, and epithelial resurfacing of the wound defect [14]. The procoagulant pathway is acvated, inﬂammatory cells are recruited, and the process transions into phases of cellular proliferaon and ssue repair/resoluon of damage. One of the primary causes of ssue damage is oxidave reacon products and free radicals [15 ,[16]. The term “oxidave stress” refers to an imbalance in redox homeostasis, where the excessive generaon of free radicals exceeds the cellular defense mechanisms. Anoxidants are crucial in maintaining protecon against oxidave damage [17 ,[18]. The ﬁrst line of defense against oxidave damage consists of anoxidant enzymes such as glutathione peroxidase (GSH-Px), glutathione S-transferase (GST), superoxide dismutase (SOD), and catalase (CAT) [19]. These enzymes funcon synergiscally, and ﬂuctuaons in their acvity may push cells into a state of oxidave stress [20]. In this context, CAT and SOD acvies have been suggested as valuable biomarkers for assessing healing potenal [21 , 22 , 23]. While most previous experimental studies on hemorrhagic disorders have focused on the hemostac and wound repair eﬀects of therapeuc agents using clinical or histopathological approaches, the eﬀects of ABS on anoxidant enzyme acvies such as CAT and SOD have not been explored extensively [24 , 25]. To the best of current knowledge, no prior research has examined the eﬀects of ABS, a local hemostac agent used in surgery, on CAT and SOD acvies. Therefore, this study aimed to invesgate the potenal eﬀects of ABS on early-stage soﬅ ssue healing in a cadmium-induced injury model by evaluang its impact on CAT and SOD acvity. In this study, Cd was administered intraperitoneally (i.p.) as a single dose of 2.5 mg/kg. Non-biodegradable Cd and cadmium chloride (CdCl₂) enter the body and accumulate through bioaccumulaon in the food chain, adversely aﬀecng kidney funcon [26 , 27]. The kidneys, bones, and lungs are the primary target organs for Cd toxicity. Reports on CdCl₂ toxicity suggest that metal accumulaon in the renal cortex following both sub-chronic and chronic exposure signiﬁcantly impairs kidney funcon [28]. Loss of cardiac ssue structural integrity, characterized by complex matrix network formaon, focal necroc areas, ﬁbrosis, and irregular myoﬁbrillar branching, has been documented in previous studies [29 , 30 ,[31]. These studies suggest that Cdinduces oxidave stress, which mediates cardiovascular ssue damage. Furthermore, oxidave stress and inﬂammaon are closely associated with Cd exposure [32 , 33]. Similarly, in the present study, necrosis and ﬁbrosis were observed in cadmium- exposed cardiac ssue. 4 of 7

Revista Cienﬁca, FCV-LUZ / Vol. XXXV UNIVERSIDAD DEL ZULIA Serbiluz Sistema de Servicios Bibliotecarios y de Información Biblioteca Digital Repositorio Académico The ﬁndings of the present study suggest that i.p. administraon of ABS was more eﬀecve than oral gavage, in agreement with previous studies [11]. The 0.5 mL i.p. dose used. In the present study, the therapeuc eﬃcacy of ABS was veriﬁed. Thus, idenfying an eﬀecve dosage of ABS was essenal for reducing inﬂammaon. The ii.p administraon of ABS at 0.5 mg/kg was found to be eﬀecve and beneﬁcial in the present model, providing a suitable framework for future experimental studies requiring animal models. Moreover, the results of the present study indicated that ABS treatment signiﬁcantly reduced myoﬁbrillar damage in cadmium-exposed animals. Given the interrelaonship between inﬂammaon and oxidave stress, alteraons inanoxidant enzyme acvity observed in cadmium-treated rats may reﬂect an increased requirement for CAT and SOD acvies. These changes could serve as indirect indicators of systemic inﬂammaon in these animals [34]. ABS is a standardized extract derived from ﬁve medicinal plants (Thymus vulgaris, Glycyrrhiza glabra, Vis vinifera, Alpinia oﬃcinarum, and Urca dioica). Several studies have demonstrated that these plants exert beneﬁcial eﬀects on the anoxidant system. Cheel et al. [35] (demonstrated that Glycyrrhiza glabra exhibits potent anoxidant and free radical scavenging properes. Furthermore, a study by Hong et al. [36] reported that G. glabra treatment signiﬁcantly increased blood SOD, CAT, and GSH-Px levels, as well as total anoxidant acvity. In this study, ABS exhibited a greater wound-healing potenal than NHAA and demonstrated promising eﬀects on SOD acvity in the cadmium-treated group. CONCLUSION This study demonstrated that cadmium exposure induces signiﬁcant structural and funconal damage in cardiac ssue, primarily through mechanisms involving oxidave stress, inﬂammaon, and apoptosis. The observed histopathological and immunohistochemical changes, including myoﬁbrillar disorganizaon, vascular congeson, and elevated Caspase-3 expression, conﬁrm the cardiotoxic eﬀects of cadmium. Importantly, administraon of Ankaferd Blood Stopper markedly alleviated these pathological alteraons. Ankaferd Blood Stopper signiﬁcantly reduced myoﬁber degeneraon, inﬂammaon, and apoptoc acvity in the myocardium. These protecve eﬀects are likely due to the strong anoxidant and an-inﬂammatory properes of the phytochemicals contained in Ankaferd Blood Stopper. The ﬁndings of the present study conﬁrm that cadmium exposure induces signiﬁcant cardiac ssue damage, characterized by oxidave stress, inﬂammaon, and ﬁbrosis. However, Ankaferd Blood Stopper administraon signiﬁcantly reduced inﬂammatory responses and ﬁbroc changes in cadmium- exposed heart ssue. These results suggest that Ankaferd Blood Stopper may have therapeuc potenal in migang cadmium- induced cardiac damage by modulang oxidave stress and inﬂammaon. The results of the present study highlight the therapeuc potenal of Ankaferd Blood Stopper in counteracng heavy metal-induced cardiac injury. Future invesgaons are warranted to further elucidate the underlying mechanisms and explore clinical applicaons in occupaonal and environmental health contexts. ACKNOWLEDGMENTS The author gratefully acknowledge the Süleyman Demirel University Animal Producon and Experimental Research Center (Isparta, Turkey) for providing the experimental animals and for technical support during the study. Declaraons Ethics approval and consent to parcipate This study was approved by the Süleyman Demirel University Animal Experiments Local Ethics Commiee with the decision dated September 5, 2024 (No: 06/78). Consent for publicaon Not applicable. Availability of data and materials The datasets generated and analyzed during the current study are available from the corresponding author on reason- able request. Compeng interests The author declares that they have no compeng interests. Funding This research received no speciﬁc grant from any funding agency in the public, commercial, or not-for-proﬁt sectors. Conﬂict of interest The author menoned that they had no conﬂicts of interest. BIBLIOGRAPHIC REFERENCES [1] Gunes E. Anoxidant eﬀects of Ankaferd Blood Stopper- doped polyvinyl pyrrolidone in an experimental model created in insects. Food Chem. Toxicol. [Internet]. 2021; 148:111935. doi: hps://doi.org/p9hv [2] Ilhan I, Buyukbayram HI. Protecve role of Ankaferd Blood Stopper on cadmium-induced acute nephrotoxicity. Med. J. SDU. [Internet]. 2023; 30(1):111-118. doi: hps://doi. org/p9hw [3] Ugur A, Sarac N, Cankal DA, Ozle M. The anoxidant and anmutagenic acvies of Ankaferd Blood Stopper, a natural hemostac agent used in denstry. Turk. J. Med. Sci. [Internet]. 2016; 46(3):657-663. doi: hps://doi.org/ g9kbvr 5 of 7

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