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Öğe A comparative view to heavy metal pollution in soil and rainwater in Çanakkale, Turkey(ACG Publications, 2021) Menteşe, Sibel; Yayıntaş, Özlem Tonguç; Baş, Batuhan; İrkin, Latife Ceyda; Yılmaz, SelehattinOn Earth, atmosphere interact with crust and thus simultaneously monitoring of environmental pollution in both parts of the environment is important. Soil and rainwater samples were taken in different parts of Çanakkale, Turkey, in two seasons. Study sites laid along with the prevailing wind direction (from NE to SW) as wind has the potential to distribute pollutants emitted into the air throughout its path. The concentrations of selected elements were determined by Inductively Coupled Plasma-Mass Spectrometry (ICP-MS). Also, physical parameters such as pH, temperature, electrical conductivity of the rainwater and temperature and pH value of the soil samples were measured together with the meteorological parameters. Seasonal differences for the selected elements were insignificant in the soil samples (p>0.05), while some elements showed seasonal variations in the rainwater samples (p[removed] Pb > Mn> Cu> Cd > Ni> Cr in the rainwater samples and Mn > Zn > Cu > Pb > Cr > Ni > Co in the soil samples, respectively. The highest enrichments were found for Pb in the rainwater and As in the soil samples. Elevated As levels occurred in the samples can pose a great risk for public health and agriculture.Öğe Adverse Effects of Ruditapes decussatus (Linnaeus, 1758) Diet on Stomach Tissues in Rats(2021) İrkin, Latife Ceyda; Öztürk, ŞamilRuditapes decussatus (Linnaeus, 1758) belongs to the Veneridae family is given as nutrients to rats. R. decussatus was collected from three different locations from the Çanakkale Strait. Twenty-four male Wistar Albino rats (270-310 g) were used in the study. There were six rats for each experimental group. The first group fed with standard rat feed, the second group fed with 80% R. decussatus + 20% standard rat feed daily; the third group fed with 80% R. decussatus + 20% standard rat feed every other day, the fourth group fed with 80% R. decussatus + 80% standard rat feed every three days. After routine histopathological follow-up, gastric tissue samples of all subjects were stained and examined under the light microscope. There were no histopathological findings in the gastric tissues of rats in the control group of hematoxylin-eosin staining. In gastric tissues of rats fed with R. decussatus, chronic gastritis caused by mononuclear inflammation between lamina propria and occasional gastric glands was noted. It was observed that inflammation severity and distribution were high especially in tissues of rats fed with R. decussatus every day. It was observed that the most commonly consumed shellfish may cause pathological picture on the digestive system of rats. As a result, increasing environmental pollution threatens the life of water as well as land life, and the consumption of living organisms exposed to polluted environments continues to threaten and affect human life. It is important to pay attention to the conditions under which the consumed products are collected and how they are collected.Öğe Çanakkale Boğazı'nda yayılış gösteren bazı makroalglerin kimyasal kompoziyonunun aratırılması(Çanakkale Onsekiz Mart Üniversitesi, 2009) İrkin, Latife Ceyda; Erduğan, HüseyinArtan gıda ve hammadde gereksinimine karşılık olarak gelişen teknolojiler ile sadece karasal canlılardan yararlanma yöntemleri değil aynı zamanda denizel ürünlerden de yararlanma yöntemleri değişmiştir. Bu da alglerden farklı amaçlarla yararlanmayı beraberinde getirmiştir. İçerdikleri protein ile önemli bir gıda kaynağı olmaları, kimyasal içerikleriyle gübreden endüstriye her alanda yararlanılan etkili bir kaynak olmaları algler üzerindeki çalışmaları etkin kılmış ve bu amaçla bu çalışma planlanmıştır. Çalışmada, Çanakkale Boğazı'nın sekiz farklı noktasından alınan kahverengi, kırmızı ve yeşil alg gruplarına ait toplam 25 makro algde kimyasal kompozisyonun mevsimsel ve istasyonlara bağlı değişimleri araştırılmıştır. Analizler mevsimsel olarak (sonbahar, kış, ilkbahar ve yaz) ve iki tekrarlı gerçekleştirilmiştir. Çalışmada mevsimlere ve istasyonlara bağlı olarak alınan taksonlar için elde edilen bulgularda önemli farklılıklar saptanmıştır.Öğe Çanakkale Boğazı’nda yayılış gösteren Cystoseira barbata (Stackhouse) C. Agardh’nın kimyasal kompozisyonunun mevsimsel olarak incelenmesi(2014) İrkin, Latife Ceyda; Erduğan, HüseyinAlglerin kimyası üzerindeki çalışmalar 1900'lü yıllarda başlamış ve günümüzde artarak devam etmektedir. Yapılan çalışmalarda alglerden gıda kaynağı olarak yararlanmanın yolları araştırılmış ve önemli miktarda besleyici maddeler içerdikleri tespit edilmiştir (Lee ve diğ., 1977, Jeon ve diğ., 1980). Günümüz ve yapılan daha eski çalışmalar göz önünde bulundurularak, bu çalışmada Çanakkale Boğazı'nın dört farklı noktasından alınan Cystoseira barbata (Stackhouse) C. Agardh materyal olarak kullanılmıştır. Yıllık dağılış gösteren bu tür, Çanakkale Boğazı'nda bulunabilecek istasyon ve mevsimler için toplanmış, organik içerik bakımından en yaralı olabilecek lokasyon ve zaman bakımından değerlendirilmiştir. Çalışma sonunda yıllık yayılışı gözlenen türün kimyasal içeriğinin mevsimlere ve istasyonlara göre değişiklik gösterdiği saptanmıştır. Bu sonuçlara göre protein miktarı en yüksek ilkbahar Gelibolu'da (%17,42±0,92), en düşük ilkbahar Eceabat'ta (%6,77±0,42) saptanmıştır. Yağ miktarları incelendiğinde en yüksek miktar kış mevsiminde Havuzlar'da (%5,05±0,23), en düşük miktar yine Havuzlar'da ilkbahar mevsiminde kaydedilmiştir (%0,79±0,54). Kül miktarlarına bakıldığında en yüksek miktar Havuzlar mevkiinde sonbaharda (%38,3±0,52), en düşük miktar yaz mevsiminde Gelibolu'da (%15,43±0,77) saptanmıştırÖğe Effect of Mediterranean Mussels (Mytilus galloprovincialis) From Polluted Areas on Hepatotoicity in Rats by Immunuhistochemical Method(2021) İrkin, Latife Ceyda; Çakına, Suat; Özdemir, İlhan; Öztürk, ŞamilMussels (Mytilus galloprovincialis) are aquatic organisms that can accumulate all the factors that pollute the fresh andsaltwater environment. Although the Dardanelles has been exposed to environmental pollution for many years, it is also animportant region for crustaceans. We aimed to investigate the histopathological changes in the liver parenchyma which is animportant gland of the digestive system by feeding these mussels to rats. Twenty-four male Wistar albino were used in thestudy. The first group (control): fed with standard rat feed, the second group (experiment 1): 4/5 mussel + 1/5 standard ratfeed daily; third group (experiment 2): 4/5 mussel + 1/5 standard rat feed every other day, the fourth group (experiment 3):groups were formed with 4/5 mussel + 1/5 standard rat feed every three days. All liver tissue samples taken from theexperimental and control groups were stained with hematoxylin-eosin, and immuohitochemically staining inflammatorymarker TNF-? and NF-?B after routine histopathological follow-up and analyzed with a light microscope image analysissystem. It was observed that mononuclear cells caused inflammation of portal areas, increased sinusoidal dilatation andcongestion and degeneration due to vacuolization in hepatocytes in the liver parenchyma of mussel-fed rats. Besides,immunohistochemical staining, TNF-?, and NF-?B immunoreactivity were observed in the liver cells of especially in thesecond group of rats. As a result, it has been shown that the consumption of mussels obtained and marketed withoutconsidering environmental pollution may trigger important digestive system organs of liver diseases.Öğe Evaluation of Regeneration and Apoptosis in Experimental Hepatic Resection of Rats(2021) Öztürk, Şamil; İrkin, Latife CeydaObjective: Apoptosis and accompanying regeneration begins immediately after liver resection and transpalantation. It is known that DNA synthesis reaches its maximum level in the first 24-48 hours after resection. This study was carried out to examine the regeneration and apoptotic process in the liver of rats that underwent 70% liver resection on days 3 and 7. Materials and Methods: 21 adult male Wistar Albino rats were used. The laparotomy with an upper midline incision was performed on the subjects for liver resection. Three groups were formed as the control (Group I), the group that was sacrificed 3 days after resection (Group II), and the group that was sacrificed 7 days after resection (Group III). Results: Sinusoidal dilatation and vacuolization in hepatocytes were observed in the liver tissues of all experimental groups except the control group. However, these histopathological findings were observed more in group II than other groups. It was observed that the apoptotic index and proliferation index values reached the maximum level in the group II. In group III, it was observed that these index values gradually decreased. Values of these parameters were found to be quite low in control group. Conclusion: As a result, comparisons of the mechanisms accompanying the regeneration occurring after resection were determined by immunohistochemical and TUNEL methods, and findings that shed light on clinical applications were obtained day by day.Öğe Experimental 70% Hepatectomy Model: Apoptotic Index, Proliferative Index and Mitotic Index(2021) Öztürk, Şamil; İrkin, Latife CeydaLiver regeneration is known to begin after experimentally induced liver resection. At first 24- 48 hours, DNA synthesis reaches the maximum level after resection and significantly regeneration known to occur on the first 10 days. In this study, we were examined the regeneration of liver on the 1st and 14th days after the resection. In experiment 21 Wistar albino male rats were used. The rats were randomly divided into three groups. The first group was the control, the second group was sacrificed one day after hepatectomy, and the third group was sacrificed 14 days after hepatectomy. All experimentally groups realized midline incision with laparotomy for resection of liver. Pedicles of the left lateral and median lobes of the liver were applied 70% hepatectomy by 4/0 silk binding. At the end of 1st and 14th days, liver tissue removed for light misroscobic analysis. No histopathological findings were found in the control group. The tissue of all experimentally groups were showed some histopatological changes such as sinuzoidal dilatation, vacuolization in the hepatocytes. This histopatological findings were seem to increase at grup II more than group III and group I with routin H&E staining. Mitotic index, apoptotic index and proliferation index of values at group II was reached the maximum level. Group III dramatically reduced the value of these index and were seem to reach near to values of the control group. Relative liver weight that determined each of experimentally groups were statistically significant differences compared to the control group. As a result, in this study, liver regeneration was shown in liver transplants without any agent that would contribute to the clinic.Öğe Heavy Metal and Mineral Composition of Soil, Atmospheric Deposition, and Mosses with Regard to Integrated Pollution Assessment Approach(Springer, 2021) Menteşe, Sibel; Tonguç Yayıntaş, Özlem; Baş, Batuhan; İrkin, Latife Ceyda; Yılmaz, SelehattinThe fact that there are no real borders between the biosphere, atmosphere, lithosphere, and hydrosphere means that environmental pollution monitoring studies should not only include one of the environmental spheres. Thus, integrated environmental pollution assessment studies conducted in the biosphere, lithosphere, and atmosphere promote the “whole system” approach. In this study, the aim was to determine the pollution in the atmosphere, soil, and plants by taking advantage of the high pollution accumulation characteristics of the mosses. Prevailing wind has the potential to distribute pollutants emitted into the air throughout its path. With this regard, soil, mosses, and atmospheric deposition samples were collected in Çanakkale, Turkey, in two seasons. Concentrations of selected elements were measured by Inductively Coupled Plasma-Mass Spectrometry. The enrichment factor of the selected elements in the soil, moss, and deposition samples was calculated. The highest enrichments were found for Lead in atmospheric deposition, Arsenic in soil, and Mercury in moss samples. Cobalt and chromium accumulated more in mosses than in soil. Elevated arsenic levels found in the samples can pose a great risk for public health and agriculture. The study result showed that the elemental composition of the samples was influenced by the enhanced air plume dispersion of anthropogenic pollution sources along the Northeast-Southwest directions due to wind characteristics in the province. As expected, strong correlations were found among the moss, soil, and atmospheric deposition samples indicating the vital interactions between the environmental components.Öğe Histopathological Evaluation of Muscle Tissue of Horse Mackerel (Trachurus trachurus, Linnaeus, 1758) in Çanakkale Strait(2022) İrkin, Latife Ceyda; Öztürk, Şamil; Aldık, RuhayIt is supported by studies that heavy metals and other water pollutants can reach humans through the food chain because of accumulation in organs of digestion, respiration, and muscles of fish, and may cause toxic effects depending on the amount of pollution and accumulation. This study was carried out by obtaining Horse mackerel samples from the Çanakkale Strait, which are fish species with high economic and nutritional value, as well as frequently consumed, seasonally (spring, summer, autumn, and winter). In the study, the tissues dissected from the dorsolateral muscles of the fresh fish were taken into Bouin's fixative, and sections were taken after routine histological follow-up. The findings revealed degenerative and inflammatory findings in the muscle tissues of fish caught especially in the autumn season. In addition, a statistically significant difference was found in autumn samples in terms of immunoreactivity (p<0.05).Öğe Investigation of Apoptotic and Inflammatory Activity in Liver Tissue of Rats Fed with Clam (Pecten maximus, Linnaeus 1758)(Sciendo, 2021) İrkin, Latife CeydaOne of the most important threats for living things in aquatic ecosystems is environmental pollution. The changes in water quality caused by environmental pollution also reduce the quality of life for organisms in the environment. Among these, the crustaceans which are most affected by the negative changes of environment, fed by the filtra-tion method, are seen as pollution indicator. The consumption of these creatures reaches all steps of the pyramid, especially humans, through the food chain. People who frequently use seafood in their diets may be affected by these negative changes. Heavy metal contents of the clams obtained from Dardanelles were determined by the ICP-OES. Twenty-four female Wistar albino rats were fed for 30 days with the experimental diet using clams (Pecten maximus), which was dried and formed into pellets and added to the food in certain proportions. At the end of the study, the subjects were sacrificed under anesthesia, liver tissues were taken, and histochemical examination was performed. TUNEL method was performed to detect apoptotic activity, and immunohistochemical staining with TNF-α and NF-κB antibodies to determine inflammation. Concluding from the results, it was observed that the de-generation of vital digestive system tissues such as liver was inevitable in living creatures that frequently consume seafood obtained from unhealthy environment in their daily diets. The high analysis values of the heavy metal (P. maximus) in food additive can be considered as a reason for histopathological results.Öğe Investigation of Regeneration Capacity in Rat Liver After Hepatic Resection(2021) Öztürk, Şamil; İrkin, Latife Ceyda; Özdemir, İlhanThe liver is the largest gland of the body that secretes both endocrine and exocrine secretions and plays a major role in the regulation of metabolic functions. Numerous factors such as drugs, chemicals, accidents, alcohol, surgical procedures can cause damage to the liver tissue. In this study, we aimed to determine the regeneration capacity of liver tissue in order to recover the mass loss after hepatic resection. In our experiment 21 Wistar albino male rats were used. All experimental groups applied midline incision with laparotomy for resection of liver. At the end of 1 and 7th days, liver tissue removed for light microscopic analysis. The rats were divided three groups: Control, group 1: hepatectomy one day after liver resection, group 2: hepatectomy seven days after liver resection. The tissue of all experimental groups were showed some histopatological changes such as sinuzoidal dilatation, vacuolization in the hepatocytes. These histopathological differentiation was found to be severe in group II compared to other groups. At the end of the 7th day, it was observed that the regeneration increased significantly, and the mitotic index value reached almost the maximum level in the second group. It was observed that the mitotic index value gradually decreased in group I and reached values close to the control groupÖğe Investigation of spinal cord damage in rats fed with clam (tapesdecussatus) by tunel assay(Parlar Scientific Publications, 2021) İrkin, Latife CeydaToxicological and epidemiological evidence suggests that heavy metals cause neurodegenerative problems. Since crustacean are fed by filtering seawater, they can be exposed to factors such as heavy metals, pesticides, herbicides, toxic plankton, and household waste. It has been determined that aluminum and lead, which are metal groups with high neurotoxicity in humans, are particularly high in crustaceans. We found that experimental studies related to this subject were very limited and we did not find any studies on the histopathological effects of bivalve consumption on the medulla spinalis tissue. This study was conducted to determine cellular damage and apoptosis in the spinal cord tissue of rats fed with clams collected from determined locations in the Dardanelles Strait. The study was divided into four groups, the first group was the control group (n = 6) fed standard rat feed for three weeks, the second group (n - 6) 80% clams and 20% standard feed every day, the third group 80% clams every two days and 20% standard feed, the fourth group was determined as the experimental group to which 80% clams and 20% standard feed were applied every three days. At the end of the study, spinal cord tissue samples taken from rats were evaluated under a light microscope after routine histopathological procedures. While there is a decrease in the number of neurons in the experimental group, the increase in the number of astrocytes draws attention. TUNEL staining showed that apoptosis occurred intensely in glial cells, and mild apoptosis occurred in anterior and posterior horn motor and sensory neurons. As a result, it shows that long-term consumption of shellfish causes axonal damage of motor and sensory neurons and degeneration of glial cells. It is known that damage to glial cells will trigger neuronal damage. It is very important in terms of health that we feed ourselves with healthy and hygienically grown products.Öğe Investigation of the relationship between homalothecium sericeum(HedW.) schimp. And Cu/Zn sod enzyme in rat tissues with the immunohistochemical method(Parlar Scientific Publications, 2021) İrkin, Latife Ceyda; Yayıntaş, Özlem Tonguç; Çakina, SuatIn this study, the aim was to determine the production of Cu/Zn SOD (SOD1) enzyme in the heart, liver, and pancreas of rats administered Homalothecium sericeum extract. The effect of the use of moss extract has not been studied for Cu/Zn SOD enzyme production in different tissues of rats or for the cytotoxic effects on these tissues. In our study, 50 mg/kg (Group I, n: 6), 100 mg/kg (Group II, n: 6), 300 mg/kg (Group III, n: 6) and 500 mg/kg (Group IV, n: 6) doses of moss extract and 1 ml of distilled water (Group V, n: 6) were administered by gavage to 30 female rats for 30 days. Cu/Zn SOD enzyme immunoreactivity was investigated in liver, heart and pancreas tissues depending on the increase in moss extract dose. To determine Cu/Zn SOD immunoreactivity, after the application of Cu/Zn SOD primary antibodies using immunohistochemical staining methods, DAP was used as coloring and hematoxylin was used as inverse dye. The stained tissue samples were evaluated with an image analysis system under the research microscope and the study used the Kruskal-Wallis Test, one of the nonparametric tests, to detect the differences between the groups statistically.Öğe Levrek balığı (dicentrarchus labrax) yemlerinde sarımsak unu kullanımının büyüme, yem değerlendirme ve vücut kompozisyonuna etkileri(Çanakkale Onsekiz Mart Üniversitesi Eğitim Bilimleri Enstitüsü, 2014) İrkin, Latife Ceyda; Yiğit, MuratYemlerde başlıca protein kaynağı olan balık unu yerine alternatif protein kaynaklarının araştırılması oldukça yaygın bir çalışma alanıdır ve balık besleme ile ilgili araştırmaların birçoğu bu konu üzerinde yapılmaktadır. Yemin yüksek sindirilebilirliği ve değerlendirme oranı açısından yeme katılan immunostimulant (bağışıklık artırıcı) veya büyümeyi teşvik edici besin maddelerinin ilave edilmesi balıklardaki gelişmeyi ve büyüme hızını artırabilmektedir. Balıklarda iştahın ve stres faktörlerine karşı direncin artırılması ile alınan yemin yüksek oranda değerlendirilmesi sağlanırken aynı zamanda hastalıklara karşı da direnç sağlanıp balık sağlığı ve refahı da yükseltilmiş olacaktır. Yetiştiricilik ortamında strese maruz kalan balıklarda yemleme aktivitesi, büyüme performansı ve balık refahının olumsuz etkileneceği bildirilmektedir. Bu çalışmada da iştah artırıcı olarak ve aynı zamanda hazırlama tekniğine göre yüksek protein ihtiva edebilen sarımsak unu balık yemlerine çeşitli miktarlarda ilave edilerek, balıklarda yem tüketimi ve yem verimliliği, büyüme performansı ve balık vücudunun biyokimyasal kompozisyonu üzerine etkileri incelenmektedir.Öğe Seasonal evaluation of chemical composition of Codium fragile (Suringar) Hariot distributed in Canakkale Strait (Dardanelles), Turkey(Adem Yavuz SÖNMEZ, 2014) İrkin, Latife Ceyda; Erdugan, HüseyinThe significant increase of the World population in recent years has linked researchers to focus on the utilization of marine food resources additional to the terrestrial ones for human consumption. Being an important source of protein and an effective source used not only as fertilizer but also in the industry with its useful chemical content makes them a target topic for research. In the present study, temporal and spatial changes of the chemical compositions of Codium fragile were investigated. The analyses were carried out seasonally (fall, winter, spring and summer). Significant differences were recorded in the findings obtained for the species collected in relation to the seasons and stations.Öğe Seasonal variation in ash, lipid and protein contents of Scytosiphon lomentaria Lyngbye and Palisada perforata Bory de Saint-Vincent along Çanakkale Strait (Dardanelles), Turkey(Adem Yavuz SÖNMEZ, 2015) İrkin, Latife Ceyda; Erdugan, HüseyinAlgae are important components of the marine ecosystem community and have been a popular subject of research because of their biological and ecological roles in the ecosystem and several active algal-derived compounds. Nowadays, agricultural and industrial resources cannot meet the needs of rapidly increasing human population. In countries especially where agricultural production is insufficient, seafood utilization gains importance. Today, the focus of algal research is the utilization of algae as a rich protein source due to their high protein contents and for their nutritional values. Moreover, regular consumption of algae may prevent some diseases and may be important in the treatment of diseases. In the present study, seasonal and geographical changes in the lipid and protein content of two macro algae from Dardanelles were investigated. According to the results, for Scystosiphon simplicissimus, the highest protein level (23.56 ± 0.54%) was recorded in Gelibolu and the minimum protein level (9.87 ± 0.52%) was recorded in Eceabat. The minimum ash amount (20.40 ± 0.44%) was recorded in Eceabat, the highest ash amount (53.05 ± 0.13%) was recorded in Gelibolu. The highest lipid amount (3.53 ± 0.87%) was recorded in Eceabat, the minimum lipid amount (1.19 ± 0.28%) was recorded in İntepe. For Palisada perforata, the highest protein amount (14.81 ± 0.73%) was recorded in winter in Eceabat. The highest lipid level (5.59 ± 0.11%) was recorded in winter in Eceabat. The minimum lipid amount (0.33 ± 0.88%) was recorded in winter in Yapıldak. The minimum protein level (7.74 ± 0.34%) was recorded in summer in Yapıldak. The highest ash amount (33.52 ± 0.66%) was recorded in winter in Yapıldak and the minimum ash amount (22.47 ± 0.86%) was recorded in summer in Eceabat.Öğe Sıçanlarda Trikloroasetik Asit ile İndüklenen Deneysel İntrauterin Adezyonlarda Kök Hücre Tedavisi(2022) Öztürk, Şamil; Otcu, Serap Mutlu Özçelik; Özdemir, İlhan; İrkin, Latife CeydaKemik iliğinden elde edilen kök hücrelerin, sıçanlarda kimyasal kullanılarak oluşturulan intrauterin adezyon modellemesinde rahim içi yapışıklıkların giderilmesinde ve blastosistin endometriuma implantasyonunun yeniden sağlanmasında etkinliği ve rolü araştırıldı. Deney modeli, trikloroasetik asit kullanılarak tek uterin hornda meydana gelen hasara karşı, yalnızca kültür besiyeri (CM), kök hücre ve 48 saatlik Niş kullanılarak farklı grup ve iki alt grup oluşturulmuştur. Çalışmada toplam 30 dişi ve 3 erkek rat kullanıldı. Akut fazda model oluşumundan hemen sonra tedaviye başlandı ve 10 gün sonra denekler gebe kalmaları için erkek sıçanlarla aynı kafese yerleştirildi. Daha sonra gebelik durumu ve doğan yavru sayısı değerlendirildi. Histolojik değerlendirme hematoksilen-eozin boyaması ile yapıldı. Yenidoğan sayısına göre hem histolojik hem de morfolojik değerlendirmelerde kök hücre uygulanan gruplarda diğer gruplara göre istatistiksel olarak anlamlı fark (p [EN] The effectiveness and role of stem cells obtained from bone marrow in removing the adhesion formed in intrauterine adhesion modeling using chemicals in rats and restoring the implantation of the blastocyst to the endometrium were investigated. The experimental model was created in a single horn using trichloroacetic acid. Three different groups and two subgroups were formed as only culture medium (CM), stem cell and 48-hour medium (Niche). A total of 30 female and 3 male rats were used in the study. Treatment was started immediately after model formation in the acute phase, and 10 days later, the subjects were placed in the same cage with male rats for conception. Then, the pregnancy status and the number of puppies born were evaluated. Histological evaluation was performed with hematoxylin-eosin staining.A statistically significant difference (p