“Effect of Castor Oil on Esophageal Stricture in Rats; Expression of ST-2, IL-6, Neopterin Proteins in Corrosive Burn Model”

“Effect of Castor Oil on Esophageal Stricture in Rats; Expression of ST-2, IL-6, Neopterin Proteins in Corrosive Burn Model” | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article “Effect of Castor Oil on Esophageal Stricture in Rats; Expression of ST-2, IL-6, Neopterin Proteins in Corrosive Burn Model” Aybegum KALYONCU AYCENK, Onur YALCIN, Ahmet Burak GURPINAR, Ceren YAPAR GUMUS, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4630346/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 23 Oct, 2024 Read the published version in Scientific Reports → Version 1 posted 10 You are reading this latest preprint version Abstract Corrosive esophageal burn (CEB) is a disease with high mortality and morbidity rates. The aim of this study was to determine the efficacy of castor oil, in preventing stricture development at the experimental CEB model. In addition to studying standard histopathological damage data, neopterin, IL-33, and sSt-2 proteins were also studied for the first time. Fifty Wistar-Albino rats were divided into randomized 5 groups. Weight measurement, esophageal length, histopathological damage score (HDS) and stenosis score, tissue caspase-3 and VEGF staining, tissue hydroxyproline (HYP), blood TNF-Alpha, IL-6, IL-33, Neopterin, and sST-2 levels were measured. In the castor oil application groups, weight gain was observed, the acute phase reaction decreased, submucosal/tunica muscularis fibrosis and mucosal damage were reduced, and total stenosis scores and histopathological damage scores decreased. While no significant difference was detected in the ST-2 protein, which was used for the first time in this study model, a significant increase in neopterin protein was observed in the application groups. Results indicate the nutritional contribution of castor oil, as well as its tissue healing and esophageal stricture-preventing efficacy at histopathological and immune-histochemical levels. Biological sciences/Biochemistry Biological sciences/Chemical biology Biological sciences/Physiology Health sciences/Gastroenterology Health sciences/Medical research Corrosive esophageal burn Esophageal stricture Castor Oil ST-2 IL-6 Neopterin Figures Figure 1 Synopsis In this study, the effectiveness of castor oil in the treatment of corrosive esophageal burns was evaluated for the first time in the literature. Additionally, for the first time in this model, Neopterin, ST-2, and IL-6 proteins were studied in relation to inflammation, stricture, and tissue healing. Introduction Corrosive esophageal burn (CEB) is a disease with high mortality and morbidity rates, especially in children under the age of 5 years. According to World Health Organization data, 5–15 thousand pediatric corrosive burns require emergency admission in America ( 1 ). In studies conducted in Turkey, it has been found that 3.3–28.1% of child poisonings are due to corrosive substance ingestion ( 2 ). The average mortality rate following oral corrosive ingestion has been determined to be 4.1% in previous studies ( 3 ). After oral corrosive ingestion, tissue damage occurs in three phases. The acute necrotic phase, occurring in the first 1–3 days, involves hemorrhage and thrombosis, followed by local necrosis and angiogenesis. The subacute phase initiates the proliferative stage, characterized by ulceration, mucosal sloughing, fibroblast colonization, and granulation, and lasts 10–14 days. The third stage is the scarring phase, which typically commences approximately three weeks after ingestion and lasts approximately to 3–6 months ( 3 ). The primary goal of treatment is to prevent complications, which can have a high early mortality rate, such as aspiration pneumonia, upper gastrointestinal bleeding, esophageal perforation, mediastinitis, and the development of strictures and carcinoma in the later stages ( 4 ). The most common complication was stricture formation on CEB. Endoscopic dilation was performed as the initial treatment. Preventing stricture is the ideal goal in the treatment of CEB; however, there is still no accepted treatment protocol in current practice ( 3 ). Castor oil is a plant extract derived from Ricinuscommunis that has been widely used in traditional medicine since ancient times. In the contemporary medical literature, castor oil has been observed to exhibit antifungal, antiviral, and antimicrobial activities ( 5 , 6 , 7 ), as well as antioxidant, hepatoprotective, antidiabetic, wound healing, and bone regeneration effects ( 8 ). In current treatment approaches, castor oil is utilized for the treatment of blepharitis and dry eye problems ( 9 ), as a laxative for constipation treatment, and in various dermatological conditions owing to its wound-healing properties. It is also used to treat baldness, dermatitis, and regeneration in burn patients ( 10 ). This study aimed to determine the efficacy of castor oil in preventing stricture development in rats by using an experimental CEB model. In this study, tissue hydroxyproline levels, TNF-α and VEGF levels for tissue damage and inflammation, and Caspase-3 for apoptosis pathway-associated tissue damage were measured to evaluate wound healing. In addition to standard histopathological techniques, tissue damage, oxidative stress, and response to treatment were assessed for the first time by measuring neopterin, sST2, IL-6, and IL-33 proteins. Neopterin is a marker of oxidative stress induced by cellular immune response. Elevated levels of neopterin have been observed in viral infections, intracellular pathogens, autoimmune diseases, allograft rejection, trauma, burns, and malignant diseases ( 11 ). ST-2 (Suppression of Tumorigenicity 2 protein) is a protein belonging to the IL-1 family that exists in both soluble and transmembrane forms. The ligand for ST-2 is IL-33. ST-2 serves as an indicator of tissue damage in sepsis, is significant for mortality from trauma, serves as an important marker of heart failure, and is used in the prognosis of chronic kidney failure ( 12 ). Results 1.1. Laboratory Process: The laboratory process was completed by two researchers, who used the same laboratory clothes and equipment. At the end of the study, the number of subjects in each group was: G1 n = 8, G2 n = 8, G3 n = 8, G4 n = 6, and G5 n = 8, with a total of 38 subjects completing the process. The sham group appeared to be the healthiest, whereas the G4 group appeared to be the least healthy. Although no complications were observed during the gavage procedure in G5, castor oil administration was paused for one day due to the presence of loose stools in two subjects at two different times. 1.2. Results of Subjects' Weight Measurements and Surgery Duration The weights of the groups before and after decapitation (day 21) were not significantly different from the average weights of the groups before modeling (p = 0.060); a significant difference was found before decapitation (p < 0.029). In G5, the average weight before decapitation was significantly higher than that before modeling (P = 0.003). In all study groups, there was no significant difference between the esophageal lengths measured during and at the end of the study (p > 0.05). No statistically significant difference was found between the surgical durations of the groups (p = 0.060). 1.3. Biochemical evaluation: The changes in IL-6 (pg/mL), IL-33 (ng/L), ST2/IL-1 (ng/mL), TNF-α (ng/L), neopterin (nmol/L), and HYP (µg/g protein) levels measured in this study are presented in Table 1 . Before comparing the groups, we investigated whether there was a covariate effect on independent variables in the statistical analysis of the weights of the rats distributed to the groups based on randomization. There was no covariate effect of weight on any independent variable (p > 0.05). The mean IL-6 levels were not significantly different between the groups (P = 0.220). There was also no significant difference in IL-33 levels between the groups (p = 0.051). TNF-α and ST2/IL-1 levels were not significantly different between groups (p = 0.295 and p = 0.850, respectively). However, there was a significant difference in neopterin levels between groups (p 0.05); however, these groups had significantly higher neopterin levels than the other groups (p < 0.05). The Neopterin level in G1 was lower than that in all groups, except G3 (p < 0.05). There was also a significant difference in the HYP levels between the groups (p = 0.016). The HYP level in G5 was significantly higher than that in G1 (p < 0.05) (Fig. 1 ). Table 1 Variation in Biochemical Parameters According to Groups. n Mean. SS Median[IQR] RankMean. p IL-6 (pg/mL) Sham 8 37.18 16.34 0.220 * Control 8 44.37 9.83 EarlyStage 8 38.32 9.17 LateStage 6 31.39 13.57 Oral Application 8 28.65 18.21 IL-33 (ng/L) Sham 8 15.43 10.35 0.051 * Control 8 19.49 10.71 EarlyStage 8 18.85 11.19 LateStage 6 31.38 9.65 Oral Application 8 25.92 9.32 TNF-Alfa (ng/L) Sham 8 141.72 61.50 0.295 * Control 8 115.00 52.01 EarlyStage 8 87.14 50.48 LateStage 6 146.51 72.92 Oral Application 8 130.43 55.47 ST2/IL-1 (ng/mL) Sham 8 0.98 0.59 1.1[0.9] 18.75 0.850 ** Control 8 1.23 0.35 1.2[0.5] 23.00 EarlyStage 8 0.99 0.32 1.0[0.5] 16.63 LateStage 6 1.26 1.14 1.1[2.2] 19.50 Oral Application 8 1.12 0.38 1.1[0.7] 19.63 Neopterin (nmol/L) Sham 8 0.71 b 0.20 0.7[0.4] 9.00 d < 0.001 ** Control 8 1.33 a 0.42 1.2[0.7] 21.56 bc EarlyStage 8 1.59 ab 2.58 0.7[0.5] 11.88 cd LateStage 6 2.59 a 0.87 2.7[1.6] 31.67 a Oral Application 8 1.90 a 0.82 1.9[1.3] 26.44 ab HYP (ug/gr prot) Sham 8 581.45 102.08 584.9[145.2] 10.13 b 0.016 ** Control 8 1063.31 358.90 926.0[639.2] 25.63 a EarlyStage 8 864.12 505.95 723.4[938.1] 17.25 ab LateStage 6 862.01 672.15 694.4[649.0] 17.17 ab Oral Application 8 1288.87 579.14 1305.6[1178.1] 26.75 a *: One-way ANOVA; **: Kruskal-Wallis test There was a significant difference between groups with different letters (p < 0.05). 1.4. Histopathological Evaluation 1.4.1. Submucosal and Tunica Muscularis Fibrosis, Muscularis Mucosa Damage The presence of submucosal fibrosis (SF), muscularis mucosa damage (MMD), and tunica muscularis fibrosis (TMF) in the groups were compared and the results are presented in Table 2 (Figs. 2 ). The presence of SF varied significantly between the groups (p < 0.001). Although SF was absent in G1, it was present in both G2 and G3 groups. In G4, SF was observed in 66,7% of cases and in G5, it was observed in 50,0% of cases. Table 2 Comparison of the Presence of Submucosal Fibrosis, Muscularis Mucosa Damage, and Tunica Muscularis Fibrosis Among Groups *: Likelihood ratio chi-square value Sham Control Early Stage Late Stage Oral Application p * n % n % n % n % n % Submucosal Fibrosis No 8 100.0 0 0.0 0 0.0 2 33.3 4 50.0 < 0.001 Yes 0 0.0 8 100.0 8 100.0 4 66.7 4 50.0 Muscularis Mucosa Damage No 8 100.0 6 75.0 0 0.0 6 100.0 7 87.5 < 0.001 Yes 0 0.0 2 25.0 8 100.0 0 0.0 1 12.5 Tunica Muscularis Fibrosis No 8 100.0 3 37.5 5 62.5 5 83.3 4 50.0 0.025 Yes 0 0.0 5 62.5 3 37.5 1 16.7 4 50.0 The MMD showed significant differences among the groups (p < 0.001). MMD was absent in G1 and G4. However, it was present in all G3 cases. The incidence of MMD in G2 and G5 was 25.0% and 12.5%, respectively. The presence of TMF varied significantly among the groups (p < 0.025). TMF was not observed in the G1 phase. The group with the highest incidence of TMF was G2 (62.5%). Table 2 Comparison of the Presence of Submucosal Fibrosis, Muscularis Mucosa Damage, and Tunica Muscularis Fibrosis Among Groups Sham Control EarlyStage LateStage Oral Application p * n % n % n % n % n % Submucosal Fibrosis No 8 100.0 0 0.0 0 0.0 2 33.3 4 50.0 < 0.001 Yes 0 0.0 8 100.0 8 100.0 4 66.7 4 50.0 Muscularis Mucosa Damage No 8 100.0 6 75.0 0 0.0 6 100.0 7 87.5 < 0.001 Yes 0 0.0 2 25.0 8 100.0 0 0.0 1 12.5 Tunica Muscularis Fibrosis No 8 100.0 3 37.5 5 62.5 5 83.3 4 50.0 0.025 Yes 0 0.0 5 62.5 3 37.5 1 16.7 4 50.0 *: Likelihood ratio chi-square value 1.4.2 Total Stenosis Score, VEGF, Caspase-3 Ratios: The total stenosis score (TSS), VEGF, and caspase-3 ratios of the groups were compared (Fig. 3 , 4 ). TSS showed significant variation among the groups (p < 0.001). The lowest TSS was observed in G1 (median [IQR]: 0.0[0.0]) and the highest in G3 (median [IQR]:2.0[1.0]); this difference was statistically significant (p < 0.05). Both G4 and G5 had significantly lower TSS concentrations than G2 and G3 (P 0.05) but had a higher TSS than G1, G4, and G5 (p < 0.05). The VEGF ratio showed a significant variation among the groups (p = 0.001). The lowest VEGF ratio was observed in G1 (median [IQR]:1.0[1.0]) and the highest in G3 (median [IQR]:3.0[0.0]); this difference was statistically significant (p 0.05). There was also a significant difference in the caspase-3 ratio between groups (p = 0.005). The highest caspase-3 ratio was observed in G2 (median [IQR]:2.0[0.75]), which did not differ significantly from that in G4 (median [IQR]:1.5[1.0]) (p > 0.05). However, G2 had a significantly higher caspase-3 ratio than G1 (median [IQR]:1.0[0.0]), G3 (median [IQR]:1.0[0.0]), and G5 (median [IQR]:1.0[0.0]) (p < 0.05). 1.4.3. Histopathological Damage Score The histopathological damage scores (HDS) of the groups were compared, and a statistically significant difference was found between the groups (p < 0.001). The lowest HDS was observed in G1(median [IQR]:0.0[0.0]), and the highest in G2 (median [IQR]:4.0[1.0]); this difference was statistically significant (p 0.05), and both groups had significantly higher HDS than G1, G4, and G5 (p < 0.05). Discussion Corrosive esophageal burns (CEB) have high mortality and morbidity in the acute phase and become chronic in the long term if an esophageal stricture is present. CEB is one of the most common causes of upper gastrointestinal stenosis in children ( 14 ). In this study, we aimed to investigate the effectiveness of castor oil in preventing esophageal stricture formation in highly resistant and long-term treatment requiring in children, typically under 5 years of age. Esophageal strictures in the first three weeks can cause nutritional defects due to feeding difficulties, swallowing difficulties, and loss of appetite. In our study, we observed statistically significant weight gain in the group that received castor oil for 21 days (G5). We believe that this result can be attributed to the nutritional effects of castor oil. Additionally, we did not observe any weight gain in G4. We believe that this result could be due to the single-dose castor oil application and could be due to different reasons (weaker subjects in randomization) in G4. Akay et al. stated that St. John's wort oil had effects on nutrition and weight gain was observed in the subjects in the prevention of stricture. ( 15 ). In the CEB model, the esophageal tissue was damaged by NaOH. In the acute necrotic phase, hemorrhage and thrombosis develop within the first three days, followed by local necrosis and angiogenesis. In this phase, inflammation is expected to be triggered in the serum, oxidative stress markers to increase, necrosis to begin, and activation of apoptotic pathways ( 3 ). In our study, we used G3 to evaluate the early phases. In the G3 data, weight loss started, the esophageal length shortened, and IL-6, which only responds to the acute phase, increased differently; the tissue damage markers ST2/IL-1 and TNF-α decreased, while the oxidative stress and early inflammatory response marker neopterin started to rise. In the immunohistochemical data, an increase in the VEGF ratio and a decrease in caspase-3 protein related to apoptotic pathways were observed in G3. These data show that even a single dose of topically applied castor oil was effective. The ST2/IL-1 protein is effective in triggering tissue damage, inflammation, and remodeling. It plays a significant role in inflammation, heart disease, and tumor development ( 15 ). In our study, the proteins used for the first time in the CEB model did not show a statistically significant difference between the groups. We think this may be due to the fact that studies with ST2 so far have shown significant results in chronic diseases and our study had a limited number of subjects. IL-33 is a member of the IL-1 family and is found in endothelial cells, epithelial cells, and fibroblasts. IL-33 plays a role in neo-angiogenesis and in the response to cellular damage or stress. The IL-33-ST2 signaling pathway plays a pro-inflammatory role by activating the type 2 inflammatory response. Type 1 inflammatory responses occur during burn/trauma and/or bacterial infections ( 16 ). Considering this mechanism, we believe that no significant results were found for IL-33-ST2 values in our study. Trauma and burns cause T cell-macrophage interactions and increase neopterin levels. Increased neopterin activates the type 1 inflammatory response, and neopterin levels were measured for the first time in this study using the CEB model. Neopterin levels were significantly higher in G4. In our study, an increase in the inflammatory response was observed in the castor oil-treated groups, in contrast to the antioxidant and anti-inflammatory effects of castor oil. ( 17 , 18 ). When evaluating these results, it should be considered that neopterin can be triggered even in depression/stress situations ( 19 ). In many studies of esophageal stricture, tissue HYP levels have been evaluated in terms of stricture formation ( 20 ). Although low tissue HYP levels were considered positive in response to stricture treatment, elevated levels were also observed in the treatment groups as in the literature and same as our study ( 12 ). In this study, looking at the HYP levels, we observed that the oral application group (G5) had an elevated HYP level close to that of the control group. We thought that this increase could be secondary to esophageal mucosal damage caused by the daily gavage procedure, even though we tried to administer it carefully. In addition, when pathological evaluation was performed, the fibrosis rate in G5 was 50%, and the TSS was 1. Evaluation of esophageal tissue in pathological assessments is important. Effectiveness was assessed based on HHS, TSS, SF, MMD, and TMF. In this study, the esophagus was significantly better in G4 and G5 than in G2. The HHS was 4 in G2 and 1.5 in G5, and a statistically significant decrease in TSS was found in G4 and G5. These results support the anti-sclerotic and wound healing effects of castor oil. In pathological angiogenesis, VEGF facilitates the mobilization of inflammatory cells to damaged areas, inflammatory processes, and induction of pro-angiogenic factor synthesis ( 21 ). Hypoxia is the most important factor that triggers angiogenesis; however, other factors such as hypoglycemia, hypertension, low pH, mechanical stress, and chronic inflammation also trigger angiogenesis. Vascular response is the primary response mechanism in wound healing. At this stage, neo-angiogenesis begins and is completed within the first three days ( 22 ). In this study, VEGF and caspase-3 were measured on biopsy samples obtained from the CEB area. The VEGF ratio was the lowest in G1 and the highest in G3, and this difference was statistically significant. An increase in the VEGF ratio during the first three days of the vascular response phase was observed as an appropriate result in the wound healing response. However, no significant differences were observed among G2, G4, and G5. This result was thought to be secondary to the inflammatory process triggering VEGF synthesis, which is also observed in tissue healing ( 23 ). Caspase-3 is responsible for the fragmentation and death of the target molecules ( 22 ). In this study, the highest caspase-3 ratio was observed in G2, which was significantly higher than that in G5. This result demonstrates that oral application of castor oil supports remodeling. This is the first study to evaluate the efficacy of castor oil in an experimental CEB model and to use inflammatory proteins, such as ST-2/IL-1 and neopterin. Additionally, it includes a comprehensive project that incorporates immunohistochemical studies of VEGF and caspase-3 along with biochemical data. The results of this study showed that castor oil, consistent with literature data, is effective in tissue healing of esophageal corrosive burns and can be a preventive treatment alternative in the development of strictures. However, we believe that there are some limitations due to the experimental nature of the study and the limited number of subjects. Method 1.1. Animals, study design and 21-day follow-up period This study was conducted at our university's animal experimentation laboratory, with approval from the local ethics committee for animal experiments (2023/15). The researchers completed the study in accordance with the ARRIVE guidelines. Fifty Wistar-Albino rats were divided into 5 groups and an experimental CEB model were created by using the Gehanno and Guedon method ( 13 ). Group 1- Sham (n = 10): Instead of 1 ml corrosive 37,5% NaOH solution, 1 ml saline was used for modelling. Group 2– Control (n = 10): CEB modelling was performed. Group 3- Early Stage Topical Application (n = 10): After modelling and washing with saline, 1 ml. castor oil given for 90 s, without aspiration. Postoperative 3rd day decapitation was performed. Group 4- Late Stage Topical Application (n = 10): After modelling and washing with saline, 1 ml. castor oil given for 90 s, without aspiration. Postoperative 21th day decapitation was performed. Group 5- Oral Application (n = 10): On the first postoperative day, 1 ml castor oil was administered by oral gavage. The same dose was administered daily for 21 days. The procedures were paused in animals with diarrhea. Postoperative 21th day decapitation was performed. 1.2. Corrosive esophageal burn injury modelling After 12 h of fasting, rats were randomly divided into five groups. The rats were anesthetized with 100 mg/kg ketamine and 15 mg/kg 2% xylazine HCI. After anesthesia, the subjects’ weights were measured, and their abdomens were shaved. Fixed with drape on a 30-degree table with reverse Trendelenburg position, a 3 cm midline abdominal incision was made under sterile surgical conditions. The stomach was gently pulled, and the esophagus was reached. A 5 Fr orogastric catheter was advanced through the proximal esophagus and a 24 G cannula was placed from the stomach to the distal esophagus. A 2 cm esophageal segment was tied with 3/0 silk sutures over the proximal and distal catheters to prevent leakage. One milliliter of a 37.5% NaOH solution was added to the segment for 90 s. The fluid was then aspirated. After washing with saline, the silk sutures and cannulas were removed and the abdomen was sutured with a 4/0 atraumatic silk suture. In G3 and G4, 1 mL was washed with saline. castor oil for 90 s without aspiration, and the silk sutures and cannulas were removed. To prevent dehydration during the early stages, 10 ml of 0.09% saline was injected into the intraperitoneal cavity. All rats were fed a liquid diet on the first postoperative day and pellets were provided ad libitum on the second day. All rats were weighed on the day of modelling before decapitation. 1.3. Histopathological evaluation After decapitation, the distal esophagus of 50 rats was excised and fixed in a 10% neutral formalin solution for 24 h. Subsequently, the materials were embedded in paraffin blocks, and sections of four micrometers thickness were obtained and stained with Hematoxylin & Eosin and Masson's trichrome histochemical stains. A histopathological examination under light microscopy was conducted, and the stenosis index was assessed to evaluate the degree of inflammation and fibroblast proliferation. When determining the stenosis index, an increase in the amount of submucosal collagen was scored on a scale of 0 to 5 in three categories. These three categories include damage to the muscularis mucosa, tunica muscularis, and collagen deposition. Subsequently, to evaluate the changes in tissue damage before and after treatment, sections were stained with caspase 3 and VEGF immunohistochemically stains, and assessment was performed based on staining intensity in four categories within a high-magnification field. These categories were evaluated as follows: staining intensity 0–25%: 1; staining intensity 25–50%: 2; staining intensity 50–75%: 3; and staining intensity 75–100%: 4. The staining intensity and stenosis index in these categories were statistically evaluated. The histopathological damage score (HDS) for caustic esophageal burn injury was evaluated semi-quantitatively using esophageal collagen deposition and damage (Table 3 ). Table 3 The histopathological damage score for caustic esophageal burn criteria of injury score Criteria of injury Score Increase in submucosal collagen None 0 Mild (submucosal collagen at least twice the thickness of the muscularis mucosa) 1+ Marked (submucosal collagen more than twice the thickness of the muscularis mucosa) 2+ Muscularis mucosa damage None 0 Present 1+ Damage with collagen deposition in tunica muscularis None 0 Mild (collagen deposition around the smooth muscle fibers) 1+ Marked (same as mild, with collagen deposition replacing some muscle fibers) 2+ 1.4. Biochemical evaluation Blood samples were obtained from rats via intracardiac puncture and then sacrificed. The collected blood samples were placed in biochemical tubes without anticoagulants. After waiting for 30 min for blood clotting, the samples were centrifuged at 1800 × g for 10 min, and the obtained serum samples were stored at -80°C in a deep freezer until analysis. Following the collection of blood samples from the rats, esophageal tissue was excised, washed with isotonic sodium chloride (NaCl) solution, and then homogenized in a 0.1 M phosphate buffer solution containing 0.1mM EDTA at a ratio of 1/4 (weight/volume) and pH 7.0. The homogenized material was then centrifuged at 14000 rpm for 20 min at 4°C, and the supernatant was separated and stored at -80°C. Measurement of serum TNF-α, IL-6, ST-2, IL-33, neopterin, and tissue hydroxyproline levels: Measurements were performed using commercially available ELISA kits. In this method, measurements were performed using a sandwich ELISA. Each well of the microplate provided in the commercial kit was coated with specific antibodies. Standards and collected blood samples were added to plate wells and incubated at 37°C for 90 min. Biotin-labeled antibodies were added and incubated at 37°C for 1 h, followed by aspiration and washing. Biotinylated Avidin-Horseradish Peroxidase (HRP) conjugate was added to each well, incubated at 37°C for 30 minutes, aspirated again, and washed five times. A surfactant component was added to each well, and after incubation at 37°C for 15 min, the reaction was stopped by adding sulfuric acid solution, resulting in the formation of a yellow color in the wells. The absorbance was measured at an optical density of 450 nm, and a standard curve was drawn to calculate the concentrations of the unknown samples. Protein measurement of the esophageal tissue was conducted using a colorimetric method, and the levels of tissue hydroxyproline were reported as mg/L of tissue protein. 1.5. Statistical analysis A 5% statistical significance level was considered in the calculations and interpretations, and two-tailed p-values are presented. Before parametric tests, normal distribution control was performed with the Shapiro-Wilk test and homogeneity control of group variances was performed with the Levene's test. As a descriptive statistic; mean and standard deviation (SD) were calculated for continuous variables, median and interquartile range (IQR) for score variables, and frequencies (n, %) were calculated for categorical variables. Paired t-test or Wilcoxon signed-rank test was used to compare dependent groups. To compare independent groups, one-way ANOVA, Welch's ANOVA, or Kruskal-Wallis test was used, depending on the situation. As a multiple comparison test, the Tukey-Kramer test was used after ANOVA, the Games-Howell test was used after Welch ANOVA, and the Dunn test was used after the Kruskal-Wallis test. The multiple comparison tests were performed at α = 0.05, and the results were represented with letters. Chi-square test was used in the analysis of categorical variables. In the chi-square test, the Likelihood ratio test statistic was calculated instead of Pearson's since the assumptions were not met. SPSS v28 (IBM Corp.; Armonk, NY, USA) and MedCalc v22.005 (MedCalc Software Ltd, Ostend, Belgium) statistical software were used in the calculations. 1.6. Financial disclosure and conflict of interest This research was approved by the Local Ethics Committee at our university and adhered to the principles outlined in the Guide for the Care and Use of Laboratory Animals (U.S.). National Institute of Health Publication # 85 − 23, revised 1996). The authors affirmed that the study was conducted in accordance with the Declaration of Helsinki. The authors declare that they have no conflicts of interest or financial agreements with pharmaceutical or biomedical companies relevant to the subject matter discussed in this manuscript. Funding for this study was provided by the Scientific Research Fund of TUBITAK (The Scientific and Research Council of Türkiye), project number 123S428, ensuring no plagiarism or content duplication. M.D. Imge Benzes worked on this project as a scholarship student. Declarations Funding for this study was provided by the Scientific Research Fund of TUBITAK (The Scientific and Research Council of Türkiye), project number 123S428 and Ordu University scientific research projects support unit, project numberD-2307. Author Contribution A.K.A and T.N. conceived of the presented idea. A.K.A developed the theory. Y.K.A. verified the analytical methods and statistical analysis. A.K.A., O.Y., C.Y.G. carried out the experimental model.O.K.K. performed histopathological section of the study.A.B.G. and T.N. performed biochemical section of the study.A.K.A. wrote the manuscript with support from C.Y.G., O.K.K., A.B.G, Y.K.A. All authors discussed the results and contributed to the final manuscript. All authors discussed the results and commented on the manuscript. Acknowledgement This work supported by the Scientific Research Fund of TUBITAK (The Scientific and Research Council of Türkiye), project number 123S428 and Ordu University Scientific Research Fund, project number D-2307. M.D. Imge Benzes worked on this project as a scholarship student. Data Availability Detailed data of this study can be seen by searching the TUBITAK project database module with the project code numbered 123S28. Since the project is in the publication process, the data cannot yet be seen on the internet, but it will be visible on the TUBITAK platform after its publication (https://search.trdizin.gov.tr/tr/proje/ara?q=&searchName=&order=year-DESC&page=1&limit=20). References De Jong AL, Macdonald R, Ein S, Forte V, Turner A. 2001. “Corrosive Esophagitis in Children: A 30-Year Review”. Int J PediatrOtorhinolaryngol. 57(3):203–21111223452. Efe E., Altaş N., Dikmen Ş., Melikoğlu M. 2013. “TürkiyeKorozifMaddeAlımıNedeniyleYatırılan 139 OlgununRetrospektifDeğerlendirilmesi: EpidemiyolojikÇalışma”. GüncelPediatri; 11: 107 – 13 DOI: 10.4274/Jcp.11.58077 Sarma MS, Tripathi PR, Arora S. 2021. “Corrosive upper gastrointestinal strictures in children: Difficulties and dilemmas”. World J ClinPediatr. 2021;10(6):124–136. doi: 10.5409/wjcp.v10.i6.124 . eCollection 2021 Nov 9.PMID: 34868889. Chibishev A, Simonovska N, Shikole A. 2010. “Post-corrosive injuries of upper gastrointestinal tract”. Prilozi, 2010;31(1):297–316. Panghal, M., Kaushal, V., Yadav, J. 2011. “In Vitro Antimicrobial Activity of Ten Medicinal Plants Against Clinical İsolates of Oral Cancer Cases”. Ann. Clin. Microbiol. Antimicrob.10, 21. Gharibi R, Shaker A, Rezapour-Lactoee A, Agarwal S. 2021. “Antibacterial and Biocompatible Hydrogel Dressing Based on Gelatin- and Castor-Oil-Derived Biocidal Agent”. ACS BiomaterSci Eng. 2021;7(8):3633–3647. doi: 10.1021/acsbiomaterials.1c00706 . Epub 2021 Jul 1. PMID: 34196519. Elkousy RH, Said ZNA, Abd El-Baseer MA, Abu El Wafa SA. 2021. “Antiviral activity of castor oil plant (Ricinuscommunis) leaf extracts”. J Ethnopharmacol. 2021;271:113878. doi: 10.1016/j.jep.2021.113878 . Epub 2021 Jan 27.PMID: 33515683. De Morais JPP, Pacheco IKC, Filho ALMM, Ferreira DCL, Viana FJC, da Silva Reis F, de Matos JME, Dos Santos Rizzo M, Fialho ACV. 2021. “Polyurethane Derived From Castor Oil Monoacylglyceride (RicinusCommunis) For Bone Defects Reconstruction: Characterization and İn Vivo Testing”. J Mater Sci Mater Med. 2021;32(4):39. doi: 10.1007/s10856-021-06511-z.PMID : 33792773. Sandford EC, Muntz A, Craig JP. 2021. “Therapeutic Potential Of Castor Oil In Managing Blepharitis, Meibomian Gland Dysfunction And Dry Eye”. ClinExpOptom. 2021;104(3):315–322. doi: 10.1111/cxo.13148 . Epub 2021 Mar 10. PMID: 33037703 Review. Gholami H, Yeganeh H. 2020. “Vegetable Oil-Based Polyurethanes As Antimicrobial Wound Dressings: In Vitro And In Vivo Evaluation”. Biomed Mater. 2020;15(4):045001. doi: 10.1088/1748-605X/ab7387.PMID : 32031994. Pilanci O, Ergin S, Sirekbasan S, Ersin I, Habip Z, Yuksel P, Kuvat N, Aslan M, Dinc O, Saribas S, Kocazeybek B. 2016. “Serum Neopterin and Procalcitonin Levels in Relationship with Pediatric Burn Wound Infections”. ActaMicrobiolImmunol Hung. 2016;63(1):47–56. doi: 10.1556/030.63.2016.1.3. PMID: 27020868. Homsak E, Gruson D. 2020. “Soluble ST2: A complex and diverse role in several diseases”. ClinChimActa. 2020;507:75–87. doi: 10.1016/j.cca.2020.04.011 . Epub 2020 Apr 16. PMID: 32305537. Gehanno P, Guedon C. 1981. “Inhibition of experimental esophageal lye strictures by penicillamine”. Arch Otolaryngol. 1981;107:145–7. Vandenplas Y. 2017. “Management of Benign Esophageal Strictures in Children”. PediatrGastroenterolHepatolNutr 2017; 20: 211–215 [PMID: 29302501 DOI: 10.5223/pghn.2017.20.4.211] Akay MA, Akduman M, Tataroğlu AÇ, Eraldemir C, Kum T, Vural Ç, Yıldız GE. 2019. “Evaluation of the efficacy of Hypericumperforatum (St. John's wort) oil in the prevention of stricture due to esophageal corrosive burns”. Esophagus. 2019;16(4):352–361. doi: 10.1007/s10388-019-00671-2. Epub 2019 Apr 11. PMID: 30976959. Lucy W. Barrett, Vanessa S. Fear, Jason C. Waithman, Fiona M. Wood, Mark W. 2019. “Fear, Understanding acute burn injury as a chronic disease”. Burns & Trauma, Volume 7, 2019, s41038-019-0163–2, https://doi.org/10.1186/s41038-019-0163-2 . Ilavarasan R, Mallika M, Venkataraman S. Anti-inflammatory and free radical scavenging activity of Ricinuscommunis root extract. J Ethnopharmacol. 2006;103(3):478–80. doi: 10.1016/j.jep.2005.07.029 . Epub 2005 Nov 28. PMID: 16310994) Singh P. P., S. M. S. Chauhan. 2009. "Activity guided isolation of antioxidants from the leaves of Ricinuscommunis L.". Food chemistry.114.3 (2009): 1069–1072.) Cavaleri D, Bartoli F, Capogrosso CA, Guzzi P, Moretti F, Riboldi I, Misiak B, Kishi T, Rubin RT, Fuchs D, Crocamo C, Carrà G. 2022. “Blood concentrations of neopterin and biopterin in subjects with depression: A systematic review and meta-analysis”. ProgNeuropsychopharmacolBiol Psychiatry. 2023;120:110633. doi: 10.1016/j.pnpbp.2022.110633 . Epub 2022 Sep 9. PMID: 36089162. Türkyilmaz Z, Sönmez K, Karabulut R, Gülbahar O, Poyraz A, Sancak B, Başaklar AC.2009. “Mitomycin C decreases the rate of stricture formation in caustic esophageal burns in rats”. Surgery. 2009;145(2):219 – 25. doi: 10.1016/j.surg.2008.10.007 . PMID: 19167978. Melincovici CS, Boşca AB, Şuşman S, Mărginean M, Mihu C, Istrate M, Moldovan IM, Roman AL, Mihu CM. 2018. “Vascular endothelial growth factor (VEGF) - key factor in normal and pathological angiogenesis”. Rom J MorpholEmbryol. 2018;59(2):455–467. PMID: 30173249 Riwaldt S, Corydon TJ, Pantalone D, Sahana J, Wise P, Wehland M, Krüger M, Melnik D, Kopp S, Infanger M, Grimm D. 2021. “Role of Apoptosis in Wound Healing and Apoptosis Alterations in Microgravity”. Front BioengBiotechnol. 2021;9:679650. doi: 10.3389/fbioe.2021.679650 . PMID: 34222218; PMCID: PMC8248797. Garcia-Perez R, Munitiz V, Martinez-Caceres CM, Ruiz de Angulo D, Ortiz A, F Martinez de Haro L, Navas D, Parrilla P. 2017. “Histopathological and Immunohistochemical Analysis of the Use of Collagen Dressing as a Reinforcement of Esophagic Anastomosis in a Rat Experimental Model”. Cir Esp. 2017;95(10):588–593. English, Spanish. doi: 10.1016/j.ciresp.2017.08.008. Epub 2017 Nov 6. PMID: 29117903. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 23 Oct, 2024 Read the published version in Scientific Reports → Version 1 posted Editorial decision: Revision requested 29 Jul, 2024 Reviews received at journal 28 Jul, 2024 Reviewers agreed at journal 28 Jul, 2024 Reviews received at journal 23 Jul, 2024 Reviewers agreed at journal 12 Jul, 2024 Reviewers invited by journal 12 Jul, 2024 Editor assigned by journal 12 Jul, 2024 Editor invited by journal 02 Jul, 2024 Submission checks completed at journal 27 Jun, 2024 First submitted to journal 24 Jun, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4630346","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":329991079,"identity":"82260f81-ac27-4164-a8cd-32d394c77dbd","order_by":0,"name":"Aybegum KALYONCU AYCENK","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABDklEQVRIiWNgGAWjYFACHjBpwMAO4cqBiAMPiNLCDOEag7UkkKIlsQFE4tPCPyP34MOvbduM+Zt5DB8X1BxOnx92+CHQFjs53QbsWiRu5CUby7bdNpM4zGNsPOPY4dyNt9MMgFqSjc0O4LDmRo6ZtGTbbRuGwzxm0rwNQC2zE0BaDiRuw6FF/kaO+W+QFnmolnTD2ekf8GoxANrC+BHoMAOolgR56Rz8thieeZcszXDutrHhYbZiY55j6YYbpHMKDiQY4PaL3PHcgx9/lN02nHe8eeNjnhprefnZ6Zs/fKiwk8PpfYEEBmZI1HAYQJwKVmmAQzkI8B9gYPwBZrE/gARIAx7Vo2AUjIJRMCIBAJ4hY/ASsm7LAAAAAElFTkSuQmCC","orcid":"","institution":"Ordu University","correspondingAuthor":true,"prefix":"","firstName":"Aybegum","middleName":"KALYONCU","lastName":"AYCENK","suffix":""},{"id":329991082,"identity":"ab4637be-43f9-4ced-9801-6f2d75c8592f","order_by":1,"name":"Onur YALCIN","email":"","orcid":"","institution":"Ordu University","correspondingAuthor":false,"prefix":"","firstName":"Onur","middleName":"","lastName":"YALCIN","suffix":""},{"id":329991083,"identity":"7eb30bc4-b300-4dea-a654-c100f30eccd0","order_by":2,"name":"Ahmet Burak GURPINAR","email":"","orcid":"","institution":"Ordu University","correspondingAuthor":false,"prefix":"","firstName":"Ahmet","middleName":"Burak","lastName":"GURPINAR","suffix":""},{"id":329991086,"identity":"c5342ce5-c4d9-4c60-b5ae-564f17095d39","order_by":3,"name":"Ceren YAPAR GUMUS","email":"","orcid":"","institution":"Ordu University","correspondingAuthor":false,"prefix":"","firstName":"Ceren","middleName":"YAPAR","lastName":"GUMUS","suffix":""},{"id":329991088,"identity":"36e134b7-225f-4fdc-9a26-64f0d836c71a","order_by":4,"name":"Ozge KAYA KORKMAZ","email":"","orcid":"","institution":"","correspondingAuthor":false,"prefix":"","firstName":"Ozge","middleName":"KAYA","lastName":"KORKMAZ","suffix":""},{"id":329991093,"identity":"3da003e4-2923-4f4c-9445-5f869cb1ff15","order_by":5,"name":"Tevfik NOYAN","email":"","orcid":"","institution":"Ordu University","correspondingAuthor":false,"prefix":"","firstName":"Tevfik","middleName":"","lastName":"NOYAN","suffix":""},{"id":329991095,"identity":"6c9a2173-c1cb-40d0-b520-a264cda4c6a0","order_by":6,"name":"Yeliz KASKO ARICI","email":"","orcid":"","institution":"Ordu University","correspondingAuthor":false,"prefix":"","firstName":"Yeliz","middleName":"KASKO","lastName":"ARICI","suffix":""}],"badges":[],"createdAt":"2024-06-24 12:56:56","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4630346/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4630346/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41598-024-74914-3","type":"published","date":"2024-10-23T15:58:03+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":60902857,"identity":"6edcedb9-4612-47c8-9fdb-8791374afd86","added_by":"auto","created_at":"2024-07-23 11:17:55","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":43120,"visible":true,"origin":"","legend":"\u003cp\u003ea, violin plot illustrating the levels of Neopterin (nmol/L) in groups. The red dot represents the median, and the green bars represent the interquartile range (25th–75th quartiles). b, violin plot illustrating the levels of HYP (ug/gr prot) in groups. The red dot represents the median, and the green bars represent the interquartile range (25th–75th quartiles).\u003c/p\u003e","description":"","filename":"statisticsfigure1.png","url":"https://assets-eu.researchsquare.com/files/rs-4630346/v1/b82d04c38a3d752881cc6081.png"},{"id":67681977,"identity":"7b749cb4-793d-4b57-b6e2-325c522fef8f","added_by":"auto","created_at":"2024-10-28 16:11:59","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":616071,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4630346/v1/68d8a831-ba81-4e36-adc7-0ba8e5b5a8dc.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"“Effect of Castor Oil on Esophageal Stricture in Rats; Expression of ST-2, IL-6, Neopterin Proteins in Corrosive Burn Model”","fulltext":[{"header":"Synopsis","content":"\u003cp\u003eIn this study, the effectiveness of castor oil in the treatment of corrosive esophageal burns was evaluated for the first time in the literature. Additionally, for the first time in this model, Neopterin, ST-2, and IL-6 proteins were studied in relation to inflammation, stricture, and tissue healing.\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e"},{"header":"Introduction","content":"\u003cp\u003eCorrosive esophageal burn (CEB) is a disease with high mortality and morbidity rates, especially in children under the age of 5 years. According to World Health Organization data, 5\u0026ndash;15 thousand pediatric corrosive burns require emergency admission in America (\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e). In studies conducted in Turkey, it has been found that 3.3\u0026ndash;28.1% of child poisonings are due to corrosive substance ingestion (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e). The average mortality rate following oral corrosive ingestion has been determined to be 4.1% in previous studies (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAfter oral corrosive ingestion, tissue damage occurs in three phases. The acute necrotic phase, occurring in the first 1\u0026ndash;3 days, involves hemorrhage and thrombosis, followed by local necrosis and angiogenesis. The subacute phase initiates the proliferative stage, characterized by ulceration, mucosal sloughing, fibroblast colonization, and granulation, and lasts 10\u0026ndash;14 days. The third stage is the scarring phase, which typically commences approximately three weeks after ingestion and lasts approximately to 3\u0026ndash;6 months (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e). The primary goal of treatment is to prevent complications, which can have a high early mortality rate, such as aspiration pneumonia, upper gastrointestinal bleeding, esophageal perforation, mediastinitis, and the development of strictures and carcinoma in the later stages (\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe most common complication was stricture formation on CEB. Endoscopic dilation was performed as the initial treatment. Preventing stricture is the ideal goal in the treatment of CEB; however, there is still no accepted treatment protocol in current practice (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eCastor oil is a plant extract derived from Ricinuscommunis that has been widely used in traditional medicine since ancient times. In the contemporary medical literature, castor oil has been observed to exhibit antifungal, antiviral, and antimicrobial activities (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e), as well as antioxidant, hepatoprotective, antidiabetic, wound healing, and bone regeneration effects (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e). In current treatment approaches, castor oil is utilized for the treatment of blepharitis and dry eye problems (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e), as a laxative for constipation treatment, and in various dermatological conditions owing to its wound-healing properties. It is also used to treat baldness, dermatitis, and regeneration in burn patients (\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThis study aimed to determine the efficacy of castor oil in preventing stricture development in rats by using an experimental CEB model. In this study, tissue hydroxyproline levels, TNF-α and VEGF levels for tissue damage and inflammation, and Caspase-3 for apoptosis pathway-associated tissue damage were measured to evaluate wound healing. In addition to standard histopathological techniques, tissue damage, oxidative stress, and response to treatment were assessed for the first time by measuring neopterin, sST2, IL-6, and IL-33 proteins. Neopterin is a marker of oxidative stress induced by cellular immune response. Elevated levels of neopterin have been observed in viral infections, intracellular pathogens, autoimmune diseases, allograft rejection, trauma, burns, and malignant diseases (\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e). ST-2 (Suppression of Tumorigenicity 2 protein) is a protein belonging to the IL-1 family that exists in both soluble and transmembrane forms. The ligand for ST-2 is IL-33. ST-2 serves as an indicator of tissue damage in sepsis, is significant for mortality from trauma, serves as an important marker of heart failure, and is used in the prognosis of chronic kidney failure (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e).\u003c/p\u003e"},{"header":"Results","content":"\u003cp\u003e1.1. Laboratory Process:\u003c/p\u003e \u003cp\u003eThe laboratory process was completed by two researchers, who used the same laboratory clothes and equipment. At the end of the study, the number of subjects in each group was: G1 n\u0026thinsp;=\u0026thinsp;8, G2 n\u0026thinsp;=\u0026thinsp;8, G3 n\u0026thinsp;=\u0026thinsp;8, G4 n\u0026thinsp;=\u0026thinsp;6, and G5 n\u0026thinsp;=\u0026thinsp;8, with a total of 38 subjects completing the process. The sham group appeared to be the healthiest, whereas the G4 group appeared to be the least healthy. Although no complications were observed during the gavage procedure in G5, castor oil administration was paused for one day due to the presence of loose stools in two subjects at two different times.\u003c/p\u003e \u003cp\u003e1.2. Results of Subjects' Weight Measurements and Surgery Duration\u003c/p\u003e \u003cp\u003eThe weights of the groups before and after decapitation (day 21) were not significantly different from the average weights of the groups before modeling (p\u0026thinsp;=\u0026thinsp;0.060); a significant difference was found before decapitation (p\u0026thinsp;\u0026lt;\u0026thinsp;0.029). In G5, the average weight before decapitation was significantly higher than that before modeling (P\u0026thinsp;=\u0026thinsp;0.003).\u003c/p\u003e \u003cp\u003eIn all study groups, there was no significant difference between the esophageal lengths measured during and at the end of the study (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05). No statistically significant difference was found between the surgical durations of the groups (p\u0026thinsp;=\u0026thinsp;0.060).\u003c/p\u003e \u003cp\u003e1.3. Biochemical evaluation:\u003c/p\u003e \u003cp\u003eThe changes in IL-6 (pg/mL), IL-33 (ng/L), ST2/IL-1 (ng/mL), TNF-α (ng/L), neopterin (nmol/L), and HYP (\u0026micro;g/g protein) levels measured in this study are presented in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. Before comparing the groups, we investigated whether there was a covariate effect on independent variables in the statistical analysis of the weights of the rats distributed to the groups based on randomization. There was no covariate effect of weight on any independent variable (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003eThe mean IL-6 levels were not significantly different between the groups (P\u0026thinsp;=\u0026thinsp;0.220). There was also no significant difference in IL-33 levels between the groups (p\u0026thinsp;=\u0026thinsp;0.051). TNF-α and ST2/IL-1 levels were not significantly different between groups (p\u0026thinsp;=\u0026thinsp;0.295 and p\u0026thinsp;=\u0026thinsp;0.850, respectively).\u003c/p\u003e \u003cp\u003eHowever, there was a significant difference in neopterin levels between groups (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). There was no significant difference between G4 and G5 (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05); however, these groups had significantly higher neopterin levels than the other groups (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). The Neopterin level in G1 was lower than that in all groups, except G3 (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). There was also a significant difference in the HYP levels between the groups (p\u0026thinsp;=\u0026thinsp;0.016). The HYP level in G5 was significantly higher than that in G1 (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eVariation in Biochemical Parameters According to Groups.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eMean.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eSS\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eMedian[IQR]\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eRankMean.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003ep\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eIL-6 (pg/mL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSham\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e37.18\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e16.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0.220\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e44.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e9.83\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eEarlyStage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e38.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e9.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLateStage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e31.39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e13.57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOral Application\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e28.65\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e18.21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eIL-33 (ng/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSham\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e15.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e10.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0.051\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e19.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e10.71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eEarlyStage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e18.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e11.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLateStage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e31.38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e9.65\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOral Application\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e25.92\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e9.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eTNF-Alfa (ng/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSham\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e141.72\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e61.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0.295\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e115.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e52.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eEarlyStage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e87.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e50.48\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLateStage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e146.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e72.92\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOral Application\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e130.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e55.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eST2/IL-1 (ng/mL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSham\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.59\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.1[0.9]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e18.75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0.850\u003csup\u003e**\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.2[0.5]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e23.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eEarlyStage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.99\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.0[0.5]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e16.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLateStage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e1.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.1[2.2]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e19.50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOral Application\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.1[0.7]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e19.63\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eNeopterin (nmol/L)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSham\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.71\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.7[0.4]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e9.00\u003csup\u003ed\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0.001\u003c/b\u003e\u003csup\u003e**\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.33\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.2[0.7]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e21.56\u003csup\u003ebc\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eEarlyStage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.59\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e2.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.7[0.5]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e11.88\u003csup\u003ecd\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLateStage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.59\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2.7[1.6]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e31.67\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOral Application\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1.90\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1.9[1.3]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e26.44\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"4\" rowspan=\"5\"\u003e \u003cp\u003eHYP (ug/gr prot)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSham\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e581.45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e102.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e584.9[145.2]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e10.13\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u003cb\u003e0.016\u003c/b\u003e\u003csup\u003e**\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1063.31\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e358.90\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e926.0[639.2]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e25.63\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eEarlyStage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e864.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e505.95\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e723.4[938.1]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e17.25\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eLateStage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e862.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e672.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e694.4[649.0]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e17.17\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOral Application\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1288.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e579.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e1305.6[1178.1]\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e26.75\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"8\"\u003e*: One-way ANOVA; **: Kruskal-Wallis test\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThere was a significant difference between groups with different letters (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003col\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003e1.4. Histopathological Evaluation\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003e1.4.1. Submucosal and Tunica Muscularis Fibrosis, Muscularis Mucosa Damage\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003c/ol\u003e \u003c/p\u003e \u003cp\u003eThe presence of submucosal fibrosis (SF), muscularis mucosa damage (MMD), and tunica muscularis fibrosis (TMF) in the groups were compared and the results are presented in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e2\u003c/span\u003e (Figs.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The presence of SF varied significantly between the groups (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001). Although SF was absent in G1, it was present in both G2 and G3 groups. In G4, SF was observed in 66,7% of cases and in G5, it was observed in 50,0% of cases.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eComparison of the Presence of Submucosal Fibrosis, Muscularis Mucosa Damage, and Tunica Muscularis Fibrosis Among Groups *: Likelihood ratio chi-square value\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"15\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c13\" colnum=\"13\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c14\" colnum=\"14\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c15\" colnum=\"15\"\u003e\u003c/div\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" morerows=\"1\" nameend=\"c2\" namest=\"c1\" rowspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colspan=\"4\" nameend=\"c6\" namest=\"c3\"\u003e \u003cp\u003eSham\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c10\" namest=\"c9\"\u003e \u003cp\u003eEarly Stage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c12\" namest=\"c11\"\u003e \u003cp\u003eLate Stage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c14\" namest=\"c13\"\u003e \u003cp\u003eOral Application\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ep\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eSubmucosal Fibrosis\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e33.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e50.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0.001\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e66.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e50.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eMuscularis Mucosa Damage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e75.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e87.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0.001\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e25.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e12.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTunica Muscularis Fibrosis\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e37.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e83.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e50.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e\u003cb\u003e0.025\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e37.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e16.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e50.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe MMD showed significant differences among the groups (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001). MMD was absent in G1 and G4. However, it was present in all G3 cases. The incidence of MMD in G2 and G5 was 25.0% and 12.5%, respectively.\u003c/p\u003e \u003cp\u003eThe presence of TMF varied significantly among the groups (p\u0026thinsp;\u0026lt;\u0026thinsp;0.025). TMF was not observed in the G1 phase. The group with the highest incidence of TMF was G2 (62.5%).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eComparison of the Presence of Submucosal Fibrosis, Muscularis Mucosa Damage, and Tunica Muscularis Fibrosis Among Groups\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"15\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c13\" colnum=\"13\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c14\" colnum=\"14\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c15\" colnum=\"15\"\u003e\u003c/div\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" morerows=\"1\" nameend=\"c2\" namest=\"c1\" rowspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colspan=\"4\" nameend=\"c6\" namest=\"c3\"\u003e \u003cp\u003eSham\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c8\" namest=\"c7\"\u003e \u003cp\u003eControl\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c10\" namest=\"c9\"\u003e \u003cp\u003eEarlyStage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c12\" namest=\"c11\"\u003e \u003cp\u003eLateStage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c14\" namest=\"c13\"\u003e \u003cp\u003eOral Application\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ep\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eSubmucosal Fibrosis\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e33.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e50.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0.001\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e66.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e50.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eMuscularis Mucosa Damage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e75.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e87.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e\u003cb\u003e\u0026lt;\u0026thinsp;0.001\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e25.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e12.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTunica Muscularis Fibrosis\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e100.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e37.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e83.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e50.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e \u003cp\u003e\u003cb\u003e0.025\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e37.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e16.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c14\"\u003e \u003cp\u003e50.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c15\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"15\"\u003e*: Likelihood ratio chi-square value\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"15\"\u003e1.4.2 Total Stenosis Score, VEGF, Caspase-3 Ratios:\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThe total stenosis score (TSS), VEGF, and caspase-3 ratios of the groups were compared (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e,\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). TSS showed significant variation among the groups (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001). The lowest TSS was observed in G1 (median [IQR]: 0.0[0.0]) and the highest in G3 (median [IQR]:2.0[1.0]); this difference was statistically significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Both G4 and G5 had significantly lower TSS concentrations than G2 and G3 (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05). G2 (median [IQR]:2.0[1.5]) did not differ significantly from G3 (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05) but had a higher TSS than G1, G4, and G5 (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eThe VEGF ratio showed a significant variation among the groups (p\u0026thinsp;=\u0026thinsp;0.001). The lowest VEGF ratio was observed in G1 (median [IQR]:1.0[1.0]) and the highest in G3 (median [IQR]:3.0[0.0]); this difference was statistically significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). There was no significant difference in the VEGF ratio between the sham and control groups (median [IQR]:2.0[0.0]) or the Oral Application group (median [IQR]:2.0[0.8]) (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003eThere was also a significant difference in the caspase-3 ratio between groups (p\u0026thinsp;=\u0026thinsp;0.005). The highest caspase-3 ratio was observed in G2 (median [IQR]:2.0[0.75]), which did not differ significantly from that in G4 (median [IQR]:1.5[1.0]) (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05). However, G2 had a significantly higher caspase-3 ratio than G1 (median [IQR]:1.0[0.0]), G3 (median [IQR]:1.0[0.0]), and G5 (median [IQR]:1.0[0.0]) (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e1.4.3. Histopathological Damage Score\u003c/p\u003e \u003cp\u003eThe histopathological damage scores (HDS) of the groups were compared, and a statistically significant difference was found between the groups (p\u0026thinsp;\u0026lt;\u0026thinsp;0.001). The lowest HDS was observed in G1(median [IQR]:0.0[0.0]), and the highest in G2 (median [IQR]:4.0[1.0]); this difference was statistically significant (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). There was no significant difference between the G2 and G3 groups (median [IQR]: 3.0 [2.0] ) (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05), and both groups had significantly higher HDS than G1, G4, and G5 (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eCorrosive esophageal burns (CEB) have high mortality and morbidity in the acute phase and become chronic in the long term if an esophageal stricture is present. CEB is one of the most common causes of upper gastrointestinal stenosis in children (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn this study, we aimed to investigate the effectiveness of castor oil in preventing esophageal stricture formation in highly resistant and long-term treatment requiring in children, typically under 5 years of age.\u003c/p\u003e \u003cp\u003eEsophageal strictures in the first three weeks can cause nutritional defects due to feeding difficulties, swallowing difficulties, and loss of appetite. In our study, we observed statistically significant weight gain in the group that received castor oil for 21 days (G5). We believe that this result can be attributed to the nutritional effects of castor oil. Additionally, we did not observe any weight gain in G4. We believe that this result could be due to the single-dose castor oil application and could be due to different reasons (weaker subjects in randomization) in G4. Akay et al. stated that St. John's wort oil had effects on nutrition and weight gain was observed in the subjects in the prevention of stricture. (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn the CEB model, the esophageal tissue was damaged by NaOH. In the acute necrotic phase, hemorrhage and thrombosis develop within the first three days, followed by local necrosis and angiogenesis. In this phase, inflammation is expected to be triggered in the serum, oxidative stress markers to increase, necrosis to begin, and activation of apoptotic pathways (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e). In our study, we used G3 to evaluate the early phases. In the G3 data, weight loss started, the esophageal length shortened, and IL-6, which only responds to the acute phase, increased differently; the tissue damage markers ST2/IL-1 and TNF-α decreased, while the oxidative stress and early inflammatory response marker neopterin started to rise. In the immunohistochemical data, an increase in the VEGF ratio and a decrease in caspase-3 protein related to apoptotic pathways were observed in G3. These data show that even a single dose of topically applied castor oil was effective.\u003c/p\u003e \u003cp\u003eThe ST2/IL-1 protein is effective in triggering tissue damage, inflammation, and remodeling. It plays a significant role in inflammation, heart disease, and tumor development (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e). In our study, the proteins used for the first time in the CEB model did not show a statistically significant difference between the groups. We think this may be due to the fact that studies with ST2 so far have shown significant results in chronic diseases and our study had a limited number of subjects. IL-33 is a member of the IL-1 family and is found in endothelial cells, epithelial cells, and fibroblasts. IL-33 plays a role in neo-angiogenesis and in the response to cellular damage or stress. The IL-33-ST2 signaling pathway plays a pro-inflammatory role by activating the type 2 inflammatory response. Type 1 inflammatory responses occur during burn/trauma and/or bacterial infections (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e). Considering this mechanism, we believe that no significant results were found for IL-33-ST2 values in our study.\u003c/p\u003e \u003cp\u003eTrauma and burns cause T cell-macrophage interactions and increase neopterin levels. Increased neopterin activates the type 1 inflammatory response, and neopterin levels were measured for the first time in this study using the CEB model. Neopterin levels were significantly higher in G4. In our study, an increase in the inflammatory response was observed in the castor oil-treated groups, in contrast to the antioxidant and anti-inflammatory effects of castor oil. (\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e). When evaluating these results, it should be considered that neopterin can be triggered even in depression/stress situations (\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn many studies of esophageal stricture, tissue HYP levels have been evaluated in terms of stricture formation (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e). Although low tissue HYP levels were considered positive in response to stricture treatment, elevated levels were also observed in the treatment groups as in the literature and same as our study (\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e). In this study, looking at the HYP levels, we observed that the oral application group (G5) had an elevated HYP level close to that of the control group. We thought that this increase could be secondary to esophageal mucosal damage caused by the daily gavage procedure, even though we tried to administer it carefully. In addition, when pathological evaluation was performed, the fibrosis rate in G5 was 50%, and the TSS was 1.\u003c/p\u003e \u003cp\u003eEvaluation of esophageal tissue in pathological assessments is important. Effectiveness was assessed based on HHS, TSS, SF, MMD, and TMF. In this study, the esophagus was significantly better in G4 and G5 than in G2. The HHS was 4 in G2 and 1.5 in G5, and a statistically significant decrease in TSS was found in G4 and G5. These results support the anti-sclerotic and wound healing effects of castor oil.\u003c/p\u003e \u003cp\u003eIn pathological angiogenesis, VEGF facilitates the mobilization of inflammatory cells to damaged areas, inflammatory processes, and induction of pro-angiogenic factor synthesis (\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e). Hypoxia is the most important factor that triggers angiogenesis; however, other factors such as hypoglycemia, hypertension, low pH, mechanical stress, and chronic inflammation also trigger angiogenesis. Vascular response is the primary response mechanism in wound healing. At this stage, neo-angiogenesis begins and is completed within the first three days (\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e). In this study, VEGF and caspase-3 were measured on biopsy samples obtained from the CEB area. The VEGF ratio was the lowest in G1 and the highest in G3, and this difference was statistically significant. An increase in the VEGF ratio during the first three days of the vascular response phase was observed as an appropriate result in the wound healing response. However, no significant differences were observed among G2, G4, and G5. This result was thought to be secondary to the inflammatory process triggering VEGF synthesis, which is also observed in tissue healing (\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eCaspase-3 is responsible for the fragmentation and death of the target molecules (\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e). In this study, the highest caspase-3 ratio was observed in G2, which was significantly higher than that in G5. This result demonstrates that oral application of castor oil supports remodeling.\u003c/p\u003e \u003cp\u003eThis is the first study to evaluate the efficacy of castor oil in an experimental CEB model and to use inflammatory proteins, such as ST-2/IL-1 and neopterin. Additionally, it includes a comprehensive project that incorporates immunohistochemical studies of VEGF and caspase-3 along with biochemical data. The results of this study showed that castor oil, consistent with literature data, is effective in tissue healing of esophageal corrosive burns and can be a preventive treatment alternative in the development of strictures. However, we believe that there are some limitations due to the experimental nature of the study and the limited number of subjects.\u003c/p\u003e "},{"header":"Method","content":"\u003cp\u003e1.1. Animals, study design and 21-day follow-up period\u003c/p\u003e \u003cp\u003e This study was conducted at our university's animal experimentation laboratory, with approval from the local ethics committee for animal experiments (2023/15). The researchers completed the study in accordance with the ARRIVE guidelines.\u003c/p\u003e \u003cp\u003eFifty Wistar-Albino rats were divided into 5 groups and an experimental CEB model were created by using the Gehanno and Guedon method (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eGroup 1- Sham (n\u0026thinsp;=\u0026thinsp;10): Instead of 1 ml corrosive 37,5% NaOH solution, 1 ml saline was used for modelling.\u003c/p\u003e \u003cp\u003eGroup 2\u0026ndash; Control (n\u0026thinsp;=\u0026thinsp;10): CEB modelling was performed.\u003c/p\u003e \u003cp\u003eGroup 3- Early Stage Topical Application (n\u0026thinsp;=\u0026thinsp;10): After modelling and washing with saline, 1 ml. castor oil given for 90 s, without aspiration. Postoperative 3rd day decapitation was performed.\u003c/p\u003e \u003cp\u003eGroup 4- Late Stage Topical Application (n\u0026thinsp;=\u0026thinsp;10): After modelling and washing with saline, 1 ml. castor oil given for 90 s, without aspiration. Postoperative 21th day decapitation was performed.\u003c/p\u003e \u003cp\u003eGroup 5- Oral Application (n\u0026thinsp;=\u0026thinsp;10): On the first postoperative day, 1 ml castor oil was administered by oral gavage. The same dose was administered daily for 21 days. The procedures were paused in animals with diarrhea. Postoperative 21th day decapitation was performed.\u003c/p\u003e \u003cp\u003e1.2. Corrosive esophageal burn injury modelling\u003c/p\u003e \u003cp\u003eAfter 12 h of fasting, rats were randomly divided into five groups. The rats were anesthetized with 100 mg/kg ketamine and 15 mg/kg 2% xylazine HCI. After anesthesia, the subjects\u0026rsquo; weights were measured, and their abdomens were shaved. Fixed with drape on a 30-degree table with reverse Trendelenburg position, a 3 cm midline abdominal incision was made under sterile surgical conditions. The stomach was gently pulled, and the esophagus was reached. A 5 Fr orogastric catheter was advanced through the proximal esophagus and a 24 G cannula was placed from the stomach to the distal esophagus. A 2 cm esophageal segment was tied with 3/0 silk sutures over the proximal and distal catheters to prevent leakage. One milliliter of a 37.5% NaOH solution was added to the segment for 90 s. The fluid was then aspirated. After washing with saline, the silk sutures and cannulas were removed and the abdomen was sutured with a 4/0 atraumatic silk suture. In G3 and G4, 1 mL was washed with saline. castor oil for 90 s without aspiration, and the silk sutures and cannulas were removed. To prevent dehydration during the early stages, 10 ml of 0.09% saline was injected into the intraperitoneal cavity.\u003c/p\u003e \u003cp\u003eAll rats were fed a liquid diet on the first postoperative day and pellets were provided ad libitum on the second day. All rats were weighed on the day of modelling before decapitation.\u003c/p\u003e \u003cp\u003e1.3. Histopathological evaluation\u003c/p\u003e \u003cp\u003eAfter decapitation, the distal esophagus of 50 rats was excised and fixed in a 10% neutral formalin solution for 24 h. Subsequently, the materials were embedded in paraffin blocks, and sections of four micrometers thickness were obtained and stained with Hematoxylin \u0026amp; Eosin and Masson's trichrome histochemical stains. A histopathological examination under light microscopy was conducted, and the stenosis index was assessed to evaluate the degree of inflammation and fibroblast proliferation. When determining the stenosis index, an increase in the amount of submucosal collagen was scored on a scale of 0 to 5 in three categories. These three categories include damage to the muscularis mucosa, tunica muscularis, and collagen deposition.\u003c/p\u003e \u003cp\u003eSubsequently, to evaluate the changes in tissue damage before and after treatment, sections were stained with caspase 3 and VEGF immunohistochemically stains, and assessment was performed based on staining intensity in four categories within a high-magnification field. These categories were evaluated as follows: staining intensity 0\u0026ndash;25%: 1; staining intensity 25\u0026ndash;50%: 2; staining intensity 50\u0026ndash;75%: 3; and staining intensity 75\u0026ndash;100%: 4. The staining intensity and stenosis index in these categories were statistically evaluated.\u003c/p\u003e \u003cp\u003eThe histopathological damage score (HDS) for caustic esophageal burn injury was evaluated semi-quantitatively using esophageal collagen deposition and damage (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe histopathological damage score for caustic esophageal burn criteria of injury score\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCriteria of injury\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eScore\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eIncrease in submucosal collagen\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMild (submucosal collagen at least twice the thickness of the muscularis mucosa)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMarked (submucosal collagen more than twice the thickness of the muscularis mucosa)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eMuscularis mucosa damage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePresent\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eDamage with collagen deposition in tunica muscularis\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMild (collagen deposition around the smooth muscle fibers)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMarked (same as mild, with collagen deposition replacing some muscle fibers)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e1.4. Biochemical evaluation\u003c/p\u003e \u003cp\u003eBlood samples were obtained from rats via intracardiac puncture and then sacrificed. The collected blood samples were placed in biochemical tubes without anticoagulants. After waiting for 30 min for blood clotting, the samples were centrifuged at 1800 \u0026times; g for 10 min, and the obtained serum samples were stored at -80\u0026deg;C in a deep freezer until analysis. Following the collection of blood samples from the rats, esophageal tissue was excised, washed with isotonic sodium chloride (NaCl) solution, and then homogenized in a 0.1 M phosphate buffer solution containing 0.1mM EDTA at a ratio of 1/4 (weight/volume) and pH 7.0. The homogenized material was then centrifuged at 14000 rpm for 20 min at 4\u0026deg;C, and the supernatant was separated and stored at -80\u0026deg;C.\u003c/p\u003e \u003cp\u003eMeasurement of serum TNF-α, IL-6, ST-2, IL-33, neopterin, and tissue hydroxyproline levels: Measurements were performed using commercially available ELISA kits. In this method, measurements were performed using a sandwich ELISA. Each well of the microplate provided in the commercial kit was coated with specific antibodies. Standards and collected blood samples were added to plate wells and incubated at 37\u0026deg;C for 90 min. Biotin-labeled antibodies were added and incubated at 37\u0026deg;C for 1 h, followed by aspiration and washing. Biotinylated Avidin-Horseradish Peroxidase (HRP) conjugate was added to each well, incubated at 37\u0026deg;C for 30 minutes, aspirated again, and washed five times. A surfactant component was added to each well, and after incubation at 37\u0026deg;C for 15 min, the reaction was stopped by adding sulfuric acid solution, resulting in the formation of a yellow color in the wells. The absorbance was measured at an optical density of 450 nm, and a standard curve was drawn to calculate the concentrations of the unknown samples.\u003c/p\u003e \u003cp\u003eProtein measurement of the esophageal tissue was conducted using a colorimetric method, and the levels of tissue hydroxyproline were reported as mg/L of tissue protein.\u003c/p\u003e \u003cp\u003e1.5. Statistical analysis\u003c/p\u003e \u003cp\u003eA 5% statistical significance level was considered in the calculations and interpretations, and two-tailed p-values are presented. Before parametric tests, normal distribution control was performed with the Shapiro-Wilk test and homogeneity control of group variances was performed with the Levene's test. As a descriptive statistic; mean and standard deviation (SD) were calculated for continuous variables, median and interquartile range (IQR) for score variables, and frequencies (n, %) were calculated for categorical variables. Paired t-test or Wilcoxon signed-rank test was used to compare dependent groups. To compare independent groups, one-way ANOVA, Welch's ANOVA, or Kruskal-Wallis test was used, depending on the situation. As a multiple comparison test, the Tukey-Kramer test was used after ANOVA, the Games-Howell test was used after Welch ANOVA, and the Dunn test was used after the Kruskal-Wallis test. The multiple comparison tests were performed at α\u0026thinsp;=\u0026thinsp;0.05, and the results were represented with letters. Chi-square test was used in the analysis of categorical variables. In the chi-square test, the Likelihood ratio test statistic was calculated instead of Pearson's since the assumptions were not met. SPSS v28 (IBM Corp.; Armonk, NY, USA) and MedCalc v22.005 (MedCalc Software Ltd, Ostend, Belgium) statistical software were used in the calculations.\u003c/p\u003e \u003cp\u003e1.6. Financial disclosure and conflict of interest\u003c/p\u003e \u003cp\u003e This research was approved by the Local Ethics Committee at our university and adhered to the principles outlined in the Guide for the Care and Use of Laboratory Animals (U.S.). National Institute of Health Publication # 85\u0026thinsp;\u0026minus;\u0026thinsp;23, revised 1996). The authors affirmed that the study was conducted in accordance with the Declaration of Helsinki. The authors declare that they have no conflicts of interest or financial agreements with pharmaceutical or biomedical companies relevant to the subject matter discussed in this manuscript. Funding for this study was provided by the Scientific Research Fund of TUBITAK (The Scientific and Research Council of T\u0026uuml;rkiye), project number 123S428, ensuring no plagiarism or content duplication. M.D. Imge Benzes worked on this project as a scholarship student.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eFunding\u003c/h2\u003e \u003cp\u003efor this study was provided by the Scientific Research Fund of TUBITAK (The Scientific and Research Council of T\u0026uuml;rkiye), project number 123S428 and Ordu University scientific research projects support unit, project numberD-2307.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eA.K.A and T.N. conceived of the presented idea. A.K.A developed the theory. Y.K.A. verified the analytical methods and statistical analysis. A.K.A., O.Y., C.Y.G. carried out the experimental model.O.K.K. performed histopathological section of the study.A.B.G. and T.N. performed biochemical section of the study.A.K.A. wrote the manuscript with support from C.Y.G., O.K.K., A.B.G, Y.K.A. All authors discussed the results and contributed to the final manuscript. All authors discussed the results and commented on the manuscript.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eThis work supported by the Scientific Research Fund of TUBITAK (The Scientific and Research Council of T\u0026uuml;rkiye), project number 123S428 and Ordu University Scientific Research Fund, project number D-2307. M.D. Imge Benzes worked on this project as a scholarship student.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eDetailed data of this study can be seen by searching the TUBITAK project database module with the project code numbered 123S28. Since the project is in the publication process, the data cannot yet be seen on the internet, but it will be visible on the TUBITAK platform after its publication (https://search.trdizin.gov.tr/tr/proje/ara?q=\u0026amp;searchName=\u0026amp;order=year-DESC\u0026amp;page=1\u0026amp;limit=20).\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eDe Jong AL, Macdonald R, Ein S, Forte V, Turner A. 2001. \u0026ldquo;Corrosive Esophagitis in Children: A 30-Year Review\u0026rdquo;. Int J PediatrOtorhinolaryngol. 57(3):203\u0026ndash;21111223452.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEfe E., Altaş N., Dikmen Ş., Melikoğlu M. 2013. \u0026ldquo;T\u0026uuml;rkiyeKorozifMaddeAlımıNedeniyleYatırılan 139 OlgununRetrospektifDeğerlendirilmesi: Epidemiyolojik\u0026Ccedil;alışma\u0026rdquo;. G\u0026uuml;ncelPediatri; 11: 107\u0026thinsp;\u0026ndash;\u0026thinsp;13 DOI:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.4274/Jcp.11.58077\u003c/span\u003e\u003cspan address=\"10.4274/Jcp.11.58077\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSarma MS, Tripathi PR, Arora S. 2021. \u0026ldquo;Corrosive upper gastrointestinal strictures in children: Difficulties and dilemmas\u0026rdquo;. World J ClinPediatr. 2021;10(6):124\u0026ndash;136. doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.5409/wjcp.v10.i6.124\u003c/span\u003e\u003cspan address=\"10.5409/wjcp.v10.i6.124\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. eCollection 2021 Nov 9.PMID: 34868889.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChibishev A, Simonovska N, Shikole A. 2010. \u0026ldquo;Post-corrosive injuries of upper gastrointestinal tract\u0026rdquo;. Prilozi, 2010;31(1):297\u0026ndash;316.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePanghal, M., Kaushal, V., Yadav, J. 2011. \u0026ldquo;In Vitro Antimicrobial Activity of Ten Medicinal Plants Against Clinical İsolates of Oral Cancer Cases\u0026rdquo;. Ann. Clin. Microbiol. Antimicrob.10, 21.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGharibi R, Shaker A, Rezapour-Lactoee A, Agarwal S. 2021. \u0026ldquo;Antibacterial and Biocompatible Hydrogel Dressing Based on Gelatin- and Castor-Oil-Derived Biocidal Agent\u0026rdquo;. ACS BiomaterSci Eng. 2021;7(8):3633\u0026ndash;3647. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1021/acsbiomaterials.1c00706\u003c/span\u003e\u003cspan address=\"10.1021/acsbiomaterials.1c00706\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. Epub 2021 Jul 1. PMID: 34196519.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eElkousy RH, Said ZNA, Abd El-Baseer MA, Abu El Wafa SA. 2021. \u0026ldquo;Antiviral activity of castor oil plant (Ricinuscommunis) leaf extracts\u0026rdquo;. J Ethnopharmacol. 2021;271:113878. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.jep.2021.113878\u003c/span\u003e\u003cspan address=\"10.1016/j.jep.2021.113878\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. Epub 2021 Jan 27.PMID: 33515683.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDe Morais JPP, Pacheco IKC, Filho ALMM, Ferreira DCL, Viana FJC, da Silva Reis F, de Matos JME, Dos Santos Rizzo M, Fialho ACV. 2021. \u0026ldquo;Polyurethane Derived From Castor Oil Monoacylglyceride (RicinusCommunis) For Bone Defects Reconstruction: Characterization and İn Vivo Testing\u0026rdquo;. J Mater Sci Mater Med. 2021;32(4):39. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s10856-021-06511-z.PMID\u003c/span\u003e\u003cspan address=\"10.1007/s10856-021-06511-z.PMID\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e: 33792773.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSandford EC, Muntz A, Craig JP. 2021. \u0026ldquo;Therapeutic Potential Of Castor Oil In Managing Blepharitis, Meibomian Gland Dysfunction And Dry Eye\u0026rdquo;. ClinExpOptom. 2021;104(3):315\u0026ndash;322. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/cxo.13148\u003c/span\u003e\u003cspan address=\"10.1111/cxo.13148\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. Epub 2021 Mar 10. PMID: 33037703 Review.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGholami H, Yeganeh H. 2020. \u0026ldquo;Vegetable Oil-Based Polyurethanes As Antimicrobial Wound Dressings: In Vitro And In Vivo Evaluation\u0026rdquo;. Biomed Mater. 2020;15(4):045001. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1088/1748-605X/ab7387.PMID\u003c/span\u003e\u003cspan address=\"10.1088/1748-605X/ab7387.PMID\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e: 32031994.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePilanci O, Ergin S, Sirekbasan S, Ersin I, Habip Z, Yuksel P, Kuvat N, Aslan M, Dinc O, Saribas S, Kocazeybek B. 2016. \u0026ldquo;Serum Neopterin and Procalcitonin Levels in Relationship with Pediatric Burn Wound Infections\u0026rdquo;. ActaMicrobiolImmunol Hung. 2016;63(1):47\u0026ndash;56. doi: 10.1556/030.63.2016.1.3. PMID: 27020868.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHomsak E, Gruson D. 2020. \u0026ldquo;Soluble ST2: A complex and diverse role in several diseases\u0026rdquo;. ClinChimActa. 2020;507:75\u0026ndash;87. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.cca.2020.04.011\u003c/span\u003e\u003cspan address=\"10.1016/j.cca.2020.04.011\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. Epub 2020 Apr 16. PMID: 32305537.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGehanno P, Guedon C. 1981. \u0026ldquo;Inhibition of experimental esophageal lye strictures by penicillamine\u0026rdquo;. Arch Otolaryngol. 1981;107:145\u0026ndash;7.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVandenplas Y. 2017. \u0026ldquo;Management of Benign Esophageal Strictures in Children\u0026rdquo;. PediatrGastroenterolHepatolNutr 2017; 20: 211\u0026ndash;215 [PMID: 29302501 DOI: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.5223/pghn.2017.20.4.211]\u003c/span\u003e\u003cspan address=\"10.5223/pghn.2017.20.4.211]\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAkay MA, Akduman M, Tataroğlu A\u0026Ccedil;, Eraldemir C, Kum T, Vural \u0026Ccedil;, Yıldız GE. 2019. \u0026ldquo;Evaluation of the efficacy of Hypericumperforatum (St. John's wort) oil in the prevention of stricture due to esophageal corrosive burns\u0026rdquo;. Esophagus. 2019;16(4):352\u0026ndash;361. doi: 10.1007/s10388-019-00671-2. Epub 2019 Apr 11. PMID: 30976959.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLucy W. Barrett, Vanessa S. Fear, Jason C. Waithman, Fiona M. Wood, Mark W. 2019. \u0026ldquo;Fear, Understanding acute burn injury as a chronic disease\u0026rdquo;. Burns \u0026amp; Trauma, Volume 7, 2019, s41038-019-0163\u0026ndash;2, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1186/s41038-019-0163-2\u003c/span\u003e\u003cspan address=\"10.1186/s41038-019-0163-2\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIlavarasan R, Mallika M, Venkataraman S. Anti-inflammatory and free radical scavenging activity of Ricinuscommunis root extract. J Ethnopharmacol. 2006;103(3):478\u0026ndash;80. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.jep.2005.07.029\u003c/span\u003e\u003cspan address=\"10.1016/j.jep.2005.07.029\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. Epub 2005 Nov 28. PMID: 16310994)\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eSingh P. P., S. M. S. Chauhan. 2009. \"Activity guided isolation of antioxidants from the leaves of Ricinuscommunis L.\". Food chemistry.114.3 (2009): 1069\u0026ndash;1072.)\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCavaleri D, Bartoli F, Capogrosso CA, Guzzi P, Moretti F, Riboldi I, Misiak B, Kishi T, Rubin RT, Fuchs D, Crocamo C, Carr\u0026agrave; G. 2022. \u0026ldquo;Blood concentrations of neopterin and biopterin in subjects with depression: A systematic review and meta-analysis\u0026rdquo;. ProgNeuropsychopharmacolBiol Psychiatry. 2023;120:110633. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.pnpbp.2022.110633\u003c/span\u003e\u003cspan address=\"10.1016/j.pnpbp.2022.110633\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. Epub 2022 Sep 9. PMID: 36089162.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eT\u0026uuml;rkyilmaz Z, S\u0026ouml;nmez K, Karabulut R, G\u0026uuml;lbahar O, Poyraz A, Sancak B, Başaklar AC.2009. \u0026ldquo;Mitomycin C decreases the rate of stricture formation in caustic esophageal burns in rats\u0026rdquo;. Surgery. 2009;145(2):219\u0026thinsp;\u0026ndash;\u0026thinsp;25. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.surg.2008.10.007\u003c/span\u003e\u003cspan address=\"10.1016/j.surg.2008.10.007\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. PMID: 19167978.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMelincovici CS, Boşca AB, Şuşman S, Mărginean M, Mihu C, Istrate M, Moldovan IM, Roman AL, Mihu CM. 2018. \u0026ldquo;Vascular endothelial growth factor (VEGF) - key factor in normal and pathological angiogenesis\u0026rdquo;. Rom J MorpholEmbryol. 2018;59(2):455\u0026ndash;467. PMID: 30173249\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eRiwaldt S, Corydon TJ, Pantalone D, Sahana J, Wise P, Wehland M, Kr\u0026uuml;ger M, Melnik D, Kopp S, Infanger M, Grimm D. 2021. \u0026ldquo;Role of Apoptosis in Wound Healing and Apoptosis Alterations in Microgravity\u0026rdquo;. Front BioengBiotechnol. 2021;9:679650. doi: \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3389/fbioe.2021.679650\u003c/span\u003e\u003cspan address=\"10.3389/fbioe.2021.679650\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e. PMID: 34222218; PMCID: PMC8248797.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGarcia-Perez R, Munitiz V, Martinez-Caceres CM, Ruiz de Angulo D, Ortiz A, F Martinez de Haro L, Navas D, Parrilla P. 2017. \u0026ldquo;Histopathological and Immunohistochemical Analysis of the Use of Collagen Dressing as a Reinforcement of Esophagic Anastomosis in a Rat Experimental Model\u0026rdquo;. Cir Esp. 2017;95(10):588\u0026ndash;593. English, Spanish. doi: 10.1016/j.ciresp.2017.08.008. Epub 2017 Nov 6. PMID: 29117903.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Corrosive esophageal burn, Esophageal stricture, Castor Oil, ST-2, IL-6, Neopterin","lastPublishedDoi":"10.21203/rs.3.rs-4630346/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4630346/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eCorrosive esophageal burn (CEB) is a disease with high mortality and morbidity rates. The aim of this study was to determine the efficacy of castor oil, in preventing stricture development at the experimental CEB model. In addition to studying standard histopathological damage data, neopterin, IL-33, and sSt-2 proteins were also studied for the first time. Fifty Wistar-Albino rats were divided into randomized 5 groups. Weight measurement, esophageal length, histopathological damage score (HDS) and stenosis score, tissue caspase-3 and VEGF staining, tissue hydroxyproline (HYP), blood TNF-Alpha, IL-6, IL-33, Neopterin, and sST-2 levels were measured. In the castor oil application groups, weight gain was observed, the acute phase reaction decreased, submucosal/tunica muscularis fibrosis and mucosal damage were reduced, and total stenosis scores and histopathological damage scores decreased. While no significant difference was detected in the ST-2 protein, which was used for the first time in this study model, a significant increase in neopterin protein was observed in the application groups. Results indicate the nutritional contribution of castor oil, as well as its tissue healing and esophageal stricture-preventing efficacy at histopathological and immune-histochemical levels.\u003c/p\u003e","manuscriptTitle":"“Effect of Castor Oil on Esophageal Stricture in Rats; Expression of ST-2, IL-6, Neopterin Proteins in Corrosive Burn Model”","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-07-23 11:17:51","doi":"10.21203/rs.3.rs-4630346/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-07-29T21:24:24+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-07-28T12:06:26+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"320906011617992357977122255445144439847","date":"2024-07-28T07:31:01+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-07-23T06:15:07+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"198910032115408437026727216874142423011","date":"2024-07-12T07:20:23+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-07-12T06:19:35+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-07-12T06:13:14+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2024-07-02T18:15:39+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-06-27T11:15:43+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2024-06-24T12:55:34+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"4d89b3f8-98fa-414f-902b-7a96b49431bb","owner":[],"postedDate":"July 23rd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":34928089,"name":"Biological sciences/Biochemistry"},{"id":34928090,"name":"Biological sciences/Chemical biology"},{"id":34928091,"name":"Biological sciences/Physiology"},{"id":34928092,"name":"Health sciences/Gastroenterology"},{"id":34928094,"name":"Health sciences/Medical research"}],"tags":[],"updatedAt":"2024-10-28T16:04:35+00:00","versionOfRecord":{"articleIdentity":"rs-4630346","link":"https://doi.org/10.1038/s41598-024-74914-3","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2024-10-23 15:58:03","publishedOnDateReadable":"October 23rd, 2024"},"versionCreatedAt":"2024-07-23 11:17:51","video":"","vorDoi":"10.1038/s41598-024-74914-3","vorDoiUrl":"https://doi.org/10.1038/s41598-024-74914-3","workflowStages":[]},"version":"v1","identity":"rs-4630346","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4630346","identity":"rs-4630346","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}