Effect of Propolis on Salivary Glands After Radiotherapy.

OBJECTIVES: To investigate the effects of increasing propolis doses on salivary glands exposed to radiotherapy (RT). METHODS: Thirty-seven rats were divided into 4 groups: The control group (G0, n: 7), G1 group (n: 10), G2 group (n: 10), and G3 group (n: 10). The rats in the G1 group received 15 Gray (Gy) RT only to the head and neck area. The rats in the G2 and G3 groups received 15 Gy RT for the head and neck area along with 100 mg/kg/ml and 200 mg/kg/ml of propolis. The parotid, submandibular, and sublingual glands of rats were immunohistochemically stained with aquaporin-1 (AQP-1) and aquaporin-5 (AQP-5). They were also evaluated for malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPO), total antioxidant (TAS), and total oxidant status (TOS). RESULTS: AQP-1 and AQP-5 values were highest in G0 group followed by G3, G2, and G1 groups in decreasing order. The MDA and TOS values were highest in G1 group, which was followed by G2, G3, and G0 groups. The highest GPO, SOD, and TAS values were observed in G0 group followed by G3, G2, and G1 groups in decreasing order. CONCLUSION: It was found that propolis increased antioxidant products and decreased oxidative products in the salivary glands receiving RT in parallel with the dose increase. Similarly, in the groups receiving propolis, an increase in the immune expression of aquaporin molecules was detected in a dose-dependent manner. Based on these data, it could be stated that propolis has a healing effect on the salivary glands exposed to RT.

Effects of ketamine on penile tissues in an experimental priapism model in rats.

BACKGROUND: This study aimed to evaluate the histopathological and biochemical effects of ketamine on penile tissues following ischemia-reperfusion injury induced by priapism. METHODS: Twenty-four male rats were randomized into three groups. Group 1 served as the control group. Group 2 underwent the priapism model to induce ischemia-reperfusion injury. Group 3, the treatment group, experienced a similar ischemia-reperfusion model as Group 2; additionally, 50 mg/kg of ketamine was administered intraperitoneally just before reperfusion. Blood biochemical analyses and penile histopathological evaluations were performed. RESULTS: In Group 3, significant improvements were observed in all histopathological scores, including desquamation, edema, inflammation, and vasocongestion compared to Group 2 (p<0.001). Blood biochemical analyses showed that the malondialdehyde (MDA) levels were recorded as 10 in Group 2, with a significant decrease in Group 3 (p=0.013). Similarly, proinflammatory cytokine levels, including interleukin-1 beta (IL-1ß), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α), were found to be suppressed in Group 3 compared to Group 2 (p=0.003, p=0.022, and p=0.028, respectively). Antioxidant enzyme activities, such as glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD), were higher in Group 3 compared to Group 2 (p=0.016 and p=0.024, respec-tively). CONCLUSION: Ketamine is an effective anesthetic agent in alleviating the effects of penile ischemia-reperfusion injury.

Effects of systemic Anatolian propolis administration on a rat-irradiated osteoradionecrosis model.

OBJECTIVE: Radiotherapy after head and neck cancer is associated with the risk of osteonecrosis development. This study aims to investigate the effectiveness of systemic propolis application to prevent the disease as it has no definite treatment protocol despite the proposed treatment methods and significantly decreases individuals' quality of life. METHODOLOGY: In total, 29 male Wistar-Albino rats were divided into control, 35 Gy irradiation (Group 1), 35 Gy irradiation+100 mg/kg/ml propolis administration (Group 2), and 35 Gy irradiation+200 mg/kg/ml propolis administration groups (Group 3). Propolis was first applied on the day after radiotherapy, except for the control group. Right first and second molars were extracted from all rats three weeks following radiotherapy. Samples were collected seven weeks after radiotherapy. Osteoblast and osteoclast counts were calculated by histomorphometric analysis. Immunohistochemical analysis determined bone morphogenic protein-2 (BMP-2) and transforming growth factor beta-3 (TGFß-3). RESULTS: Group comparison found non-significant differences regarding osteoblast (p=0.130) and osteoclast (p=0.063) counts. However, Group 1 showed the lowest mean osteoblast (OBL: 82.63 [±13.10]) and highest mean osteoclast counts (OCL: 12.63 [±5.55]). OBL/OCL ratio showed significant differences between groups (p=0.011). Despite the significant difference between the Control and Groups 1 (p=0.006) and 2 (p=0.029), Group 3 showed a non-significant difference (p=0.091). For BMP-2 and TGFB3, the control group showed significant differences with the other two groups (p<0.001), except for Group 3. CONCLUSION: Anatolian propolis showed beneficial effects in a radiotherapy-mediated osteonecrosis model, highlighting its potential as a promising intervention.

Effects of thymoquinone and memantine alone and in combination on memory and hippocampal morphology in rats with streptozotocin-induced Alzheimer's disease.

BACKGROUND: Alzheimer's disease (AD) is a progressive neurodegenerative disease. Thymoquinone (TQ) has broad biological functions, including antiinflammatory, antioxidant, neuroprotective properties. Memantine (MEM) is indicated for the symptomatic treatment of moderate to severe AD. We aimed to evaluate the effect of TQ alone or in combination with MEM on memory and hippocampal morphology in an STZ-induced rat AD model. METHODS: Thirty male rats were included in this study. The AD model was created by giving ICV STZ. The rats were divided into 5 groups (n = 6 each). Group 1 (control group): The rats received only ICV-STZ 3 mg/kg for 2 weeks. Group 2 (sham group): In addition to ICV STZ, 9% NaCl, 1 mL/day i.p. for 2 weeks of injection, was applied. Group 3 (TQ group): In addition to ICV STZ, rats received TQ 10 mg/kg i.p. for 2 weeks. Group 4 (MEM group): In addition to ICV STZ, rats were given MEM at a dose of 5 mg/kg for two weeks. Group 5 (TQ+MEM group): In addition to ICV STZ, this group was given TQ (10 mg/kg/day, i.p.) and MEM (5 mg/kg/day, i.p.) for 2 weeks. On the 15th day, passive avoidance learning (PAL) was applied to all groups. Then, rats were sacrificed, neurons in the hippocampal CA1, CA2, CA3 regions were evaluated. RESULTS: Groups 3, 4, 5 had longer latency periods than groups 1 and 2. The neuron density in the CA1, CA2, CA3 regions had decreased in groups 1 and 2 compared to groups 3, 4, 5. There were significantly more neurons in groups 3, 4, 5 than in groups 1 and 2. DISCUSSION: We found that TQ alone and in combination with MEM showed ameliorative effects on memory and hippocampal morphology. TQ may offer a promising treatment strategy for AD.

The effects of the pulsed magnetic field on sepsis-induced liver tissue injury in rats.

Sepsis is the host's response to infection and can lead to severe and life-threatening cases. We aimed to investigate the effects of pulsed magnetic field (PMF) on septic liver tissue injury. A total of 28 adult Wistar albino rats were divided equally into four study groups: Sham, PMF-1, PMF-2, and Sepsis, with seven rats in each. Sepsis was performed using the CLP method. PMF-1 and PMF-2 were exposed to 7.5 Hz and 15 Hz PMF, respectively, for 24 hours. After having their livers removed, liver tissues were analysed using histological techniques. We observed remarkable healing in PMF groups. Apoptotic cells decreased in the PMF-treated groups compared with the Sepsis group (p < 0.05). Immune expressions of Acas-3, Bax, and HIF-1 increased in the Sepsis group, while Bcl-2 expression decreased (p < 0.05). The results imply that PMF application has anti-apoptotic, antiinflammatory, and therapeutic effects on septic liver tissue injury.

Protective role of vitamin B12 on acetic acid induced colitis in rats.

Objectives: Inflammatory bowel disease (IBD) is a chronic, relapsing, and remittent inflammatory disease of the gastrointestinal tract. Nutritional deficiency may be instrumental in and attributable to this disease. We examined the effect of VitB12 supplementation on acetic acid (AA)-induced colitis in rats. Material and Methods: Five minutes after the application of acetic acid to the rats to create a colitis model, VitB12 was administered 1 mg/kg, i.p concentration, then the application continued for three consecutive days. Control groups were included for colitis and VitB12. After 4d, the rats were sacrificed, and colonic tissues were harvested for macroscopic and microscopic examination of colonic damage. TNF-α, IL-1ß, IL-6, MDA, GSH and SOD values were measured biochemically. Results: There was statistically significant macroscopic improvement in damage to the colon tissues (p <0.05). The severity of inflammation reduced in the VitB12 treated rat group compared with the control group, but was not significantly. The levels of TNF-α, IL-1ß, MDA, and SOD did not differ between AA control and VitB12 treated AA colitis group. However, the levels of IL-6 and GSH were statistically significant different in rats with AA-induced colitis after VitB12 injection (p <0.05). Conclusion: Nutritional deficiencies might contribute to the pathogenesis of IBD, and the efficacy of VitB12 supplementation has controversial effects on the intestinal mucosa.

Microanatomic analyses of extratemporal facial nerve and its branches, hypoglossal nerve, sural nerve, and great auricular nerve.

OBJECTIVE: To investigate microanatomic organizations of the extratemporal facial nerve and its branches, hypoglossal nerve, sural nerve, and great auricular nerve. METHODS: Nerve samples were dissected in 12 postmortem autopsies, and histomorphometric analyses were conducted. RESULTS: There was no significant difference between the right and left sides of the nerve samples for the nerve area, fascicle area, number of fascicles and average number of axons. The lowest mean fascicle number was found in the hypoglossal nerve (4.9 ±â€¯1.4) while the highest was in great auricular nerve (11.4 ±â€¯6.8). The highest nerve area (3,182,788 ±â€¯838,430 µm2), fascicle area (1,573,181 ±â€¯457,331 µm2) and axon number (14,772 ±â€¯4402) were in hypoglossal nerve (p < 0.05). The number of axons per unit nerve area was higher in the facial nerve, truncus temporofacialis, truncus cervicofacialis and hypoglossal nerve, which are motor nerves, compared to the sural nerve and great auricular nerve, which are sensory nerves (p < 0.05). The number of axons per unit fascicle area was also higher in motor nerves than in sensory nerves (p < 0.05). CONCLUSION: In the present study, it was observed that each nerve contained a different number of fascicles and these fascicles were different both in size and in the number of axons they contained. All these variables could be the reason why the desired outcomes cannot always be achieved in nerve reconstruction.

Microanatomic analyses of extratemporal facial nerve and its branches, hypoglossal nerve, sural nerve, and great auricular nerve

Abstract Objective: To investigate microanatomic organizations of the extratemporal facial nerve and its branches, hypoglossal nerve, sural nerve, and great auricular nerve. Methods: Nerve samples were dissected in 12 postmortem autopsies, and histomorphometric analyses were conducted. Results: There was no significant difference between the right and left sides of the nerve samples for the nerve area, fascicle area, number of fascicles and average number of axons. The lowest mean fascicle number was found in the hypoglossal nerve (4.9 ± 1.4) while the highest was in great auricular nerve (11.4 ± 6.8). The highest nerve area (3,182,788 ± 838,430 μm2), fascicle area (1,573,181 ± 457,331 μm2) and axon number (14,772 ± 4402) were in hypoglossal nerve (p < 0.05). The number of axons per unit nerve area was higher in the facial nerve, truncus temporofacialis, truncus cervicofacialis and hypoglossal nerve, which are motor nerves, compared to the sural nerve and great auricular nerve, which are sensory nerves (p < 0.05). The number of axons per unit fascicle area was also higher in motor nerves than in sensory nerves (p < 0.05). Conclusion: In the present study, it was observed that each nerve contained a different number of fascicles and these fascicles were different both in size and in the number of axons they contained. All these variables could be the reason why the desired outcomes cannot always be achieved in nerve reconstruction.

Effects of systemic Anatolian propolis administration on a rat-irradiated osteoradionecrosis model

Abstract Objective Radiotherapy after head and neck cancer is associated with the risk of osteonecrosis development. This study aims to investigate the effectiveness of systemic propolis application to prevent the disease as it has no definite treatment protocol despite the proposed treatment methods and significantly decreases individuals' quality of life. Methodology In total, 29 male Wistar-Albino rats were divided into control, 35 Gy irradiation (Group 1), 35 Gy irradiation+100 mg/kg/ml propolis administration (Group 2), and 35 Gy irradiation+200 mg/kg/ml propolis administration groups (Group 3). Propolis was first applied on the day after radiotherapy, except for the control group. Right first and second molars were extracted from all rats three weeks following radiotherapy. Samples were collected seven weeks after radiotherapy. Osteoblast and osteoclast counts were calculated by histomorphometric analysis. Immunohistochemical analysis determined bone morphogenic protein-2 (BMP-2) and transforming growth factor beta-3 (TGFβ-3). Results Group comparison found non-significant differences regarding osteoblast (p=0.130) and osteoclast (p=0.063) counts. However, Group 1 showed the lowest mean osteoblast (OBL: 82.63 [±13.10]) and highest mean osteoclast counts (OCL: 12.63 [±5.55]). OBL/OCL ratio showed significant differences between groups (p=0.011). Despite the significant difference between the Control and Groups 1 (p=0.006) and 2 (p=0.029), Group 3 showed a non-significant difference (p=0.091). For BMP-2 and TGFB3, the control group showed significant differences with the other two groups (p<0.001), except for Group 3. Conclusion Anatolian propolis showed beneficial effects in a radiotherapy-mediated osteonecrosis model, highlighting its potential as a promising intervention.

The Effects of Probiotics and Omega-3 Fatty Acids in Liver Steatosis Induced in Rats by High-Fructose Corn Syrup.

Aims: This study was designed to reveal the effect of probiotics and omega-3 fatty acids in a fatty liver model in rats induced by high-fructose corn syrup (HFCS). Methods: In the study, 40 male Wistar Albino rats were used, and these rats were divided into five groups. HFCS was added to the drinking water (30% solution) of four groups (Groups 2, 3, 4, and 5) for three weeks, and the animals were fed ad libitum. At the end of three weeks, the rats in Groups 3, 4, and 5 were administered omega-3 fatty acids (400 mg/kg) and probiotics (1.5 × 109 cfu/mL/day) with the gavage method for four weeks. The body weights of rats were weighed and recorded before starting the experiment, at the end of the third week, and before the animals were sacrificed at the last week, all at the same hour. By subtracting the remaining amount of food and water from the daily food and water amount, the amount of food and water consumed was calculated. These values were recorded for seven weeks. At the end of the seven weeks, the rats were sacrificed after blood specimens and tissues were taken. Results: Analyzing the changes in the food intake of each group within itself throughout the experiment, it was observed that there was an increase in the food intake in the control group; from the starting week to the last week, the food intake amount of the HFCS group began to decrease particularly after the second week; and it began to decrease after the third week in the groups that were administered probiotics and omega-3 fatty acids. The changes in the sacrifice weights in the HFCS + omega-3 fatty acid, HFCS + probiotic, and HFCS + probiotic + omega-3 fatty acid groups were found to be lower than that in the HFCS group. The maximum levels of glucose, ALT, ALP, serum cholesterol, triglyceride and AST were found to be in the HFCS group. It was determined that the minimum mean steatosis level was in the control group, while the maximum steatosis level was in the HFCS group. Conclusions: As a result, there was a protective effect of probiotic and omega-3 fatty acid.

The effect of systemic application of propolis on tongue damage and oral mucositis in rats exposed to radiation.

PURPOSE: In this experimental study, the effect of dose-dense systemic application of propolis on oral mucosity, histological changes in papilla, and inflammatory and hypoxic markers in rats exposed to radiation was investigated. METHODS: Seven rats were in the control and 30 rats in the experimental group. Three experimental groups were formed. In Group 1 RT (15 Gy) was delivered only to the head and neck region. In Group 2, RT (15 Gy) and systemic administration of 100 mg/kg/ml propolis, in Group 3, RT (15 Gy) and systemic administration of 200 mg/kg/ml propolis were applied. Oral mucositis index (OMI) was scored in control and experimental groups. Proinflammatory markers [interleukin-6 (IL-6), myeloperoxidase (MPO), tumor-necrosis factor-α (TNF-α)] hypoxia markers [glucose transporter-1 (GLUT-1), hypoxia-inducible factor 1α (HIF-1α)] were studied histomorphologically. RESULTS: The significantly highest OMI score was observed in the G1. OMI score was statistically significantly decreased in experimental groups receiving systemic propolis, especially in G3. Proinflammatory markers increased significantly only in the experimental RT group, G1. Serum levels of MPO and TNF-α significantly decreased in the dose-dense systemic propolis arm. The highest levels of hypoxia markers (HIF-1α and GLUT-1) were detected in the RT group, then in G2, G3, and control groups in order of decreasing frequency. However, the difference between the groups did not reach the level of statistical significance. CONCLUSION: Systemic propolis can be reduced acute mucositis with its anti-inflammatory effect without developing resistance to RT (tumor protection). However, greater number of clinical studies should be designed to arrive at definitive conclusions.

Sinapic acid reduces ischemia/reperfusion injury due to testicular torsion/detorsion in rats.

This study aimed to investigate the protective effect of sinapic acid (SA) on biochemical and histopathological changes in an experimental testicular torsion-detorsion rat model. Twenty-four rats were randomised into four groups: sham group, ischemia/reperfusion (IR) group subjected to testicular torsion for 2 hr and then detorsion for 4 hr, and two groups treated with SA1 and SA2 (10 mg/kg and 20 mg/kg, by single intraperitoneal injection, 30 min before reperfusion). Serum testosterone, follicle-stimulating hormone (FSH), and luteinizing hormone (LH) were measured by an autoanalyzer, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), protein carbonyl (PC), and nitric oxide (NO) oxidative stress parameters by spectrophotometric methods, and tumour necrosis factor (TNF-α), interleukin-1 beta (IL-1ß), and interleukin 6 (IL-6) parameters by the Elisa method. In addition, immunohistochemical and histopathological examinations were performed on testicular tissues. There was no significant difference between the groups in terms of serum testosterone, FSH and LH levels (p > .05). SA significantly reduced increased testicular damage, oxidative stress, inflammation, cell death and also restored decreased antioxidant enzyme activities (p < .05). Pre-treatment of rats with SA reduced testicular dysfunction and morphological changes IRI. SA's antioxidant, anti-inflammatory, and antiapoptotic properties were found to be protective against testicular IR.

3,4,5-Trihydroxybenzoic Acid Attenuates Ligature-Induced Periodontal Disease in Wistar Rats.

BACKGROUND: 3,4,5-Trihydroxybenzoic acid, which is also known as gallic acid, is an antiinflammatory agent that could provide beneficial effects in preventing periodontal inflammation. The present study aimed to evaluate the anti-inflammatory effects of gallic acid on experimental periodontitis in Wistar rats. Alveolar bone loss, osteoclastic activity, osteoblastic activity, and collagenase activity were also determined. METHODS: Thirty-two Wistar rats were used in the present study. Study groups were created as following: Healthy control (C,n=8) group; periodontitis (P,n=8) group; periodontitis and 30 mg/kg gallic acid administered group (G30,n=8); periodontitis and 60 mg/kg gallic acid administered group (G60,n=8). Experimental periodontitis was created by placing 4-0 silk sutures around the mandibular right first molar tooth. Morphological changes in alveolar bone were determined by stereomicroscopic evaluation. Mandibles were undergone histological evaluation. Matrix metalloproteinase (MMP)-8, tissue inhibitor of MMPs (TIMP)-1, bone morphogenetic protein (BMP)-2 expressions, tartrateresistant acid phosphatase (TRAP) positive osteoclast cells, osteoblast, and inflammatory cell counts were determined. RESULTS: The highest alveolar bone loss was observed in the periodontitis group. Both doses of gallic acid decreased alveolar bone loss as compared to the P group. TRAP-positive osteoclast cell counts were higher in the P group, and gallic acid successfully lowered these counts. Osteoblast cells also increased in gallic acid administered groups. Inflammation in the P group was also higher than those of C, G30, and G60 groups supporting the role of gallic acid in preventing inflammation. 30 and 60 mg/kg doses of gallic acid decreased MMP-8 levels and increased TIMP-1 levels. BMP levels increased in gallic acid administered groups, similar to several osteoblasts. CONCLUSION: Present results revealed an anti-inflammatory effect of gallic acid, which was indicated by decreased alveolar bone loss and collagenase activity and increased osteoblastic activity.

Protective effect of famotidine on ischemia-reperfusion injury following testicular torsion in rats.

INTRODUCTION: In testicular torsion, testicular blood flow is impaired, resulting in ischemic changes. Torsion must be corrected urgently with surgical treatment. Detorsioning and restoration of blood supply to the testis cause reperfusion injury. OBJECTIVE: In this experimental study, we aimed to investigate the effect of famotidine on ischemia-reperfusion injury in a rat model of testicular torsion. STUDY DESIGN: The rats were randomly divided into three groups; Group I (control, no torsion) (n = 8), Group II (torsion + detorsion) (n = 8), Group III (torsion + detorsion + famotidine) (n = 8). Levels of oxidative stress markers, such as malondialdehyde (MDA) and nitric oxide (NO), and antioxidants, such as superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), were measured for biochemical analysis. Testicular tissues were assessed by Johnsen Scoring for spermatogenic evaluation. Tissues were also examined with TUNEL staining to determine the extent of apoptosis. RESULTS: Average MDA level was higher in Group II than Groups I and III. The difference was only significant between Group I and II (p = 0.03). Average NO level was significantly higher in Group II than Groups I and III (p = 0.03; p = 0.04; respectively). Conversely, average SOD level was lower in Group II than Groups I and III. The difference was only significant between Group II and III (p < 0.001). Average GSH-Px level was lower in Group II than Groups I and III, but the differences were not significant (p = 0.37; p = 0.35; respectively). The average Johnsen score in Group II was significantly lower than the scores in Groups I and III (p < 0.001; p < 0.001; respectively). The apoptotic index of Group II was significantly higher than those of Groups I and III (p < 0.001; p < 0.001; respectively). DISCUSSION: Famotidine prevented increases in oxidative stress markers and reductions of antioxidants during ischemia-reperfusion injury in our study. Spermatogenesis was less affected and DNA injury was reduced in rats treated with famotidine. The antioxidant characteristics of famotidine and its protective effects have been shown in our study. CONCLUSION: Famotidine may prevent oxidative tissue injury during ischemia-reperfusion.

Hyaluronic acid with antioxidants improve wound healing in rats.

Hyaluronic acid (HA) is found in connective tissue and participates in wound healing. We investigated the efficacy of a HA gel (2% hyaluronic acid; 1% antioxidants, coenzyme Q10 and vitamin E; and 5% benzocaine) on healing of palatal wounds in rats. We established two groups of rats: a control group treated with vehicle and an HA group treated with HA gel. The control group was divided into five subgroups and the HA group was divided into four subgroups according to the day on which animals were sacrificed. Wounds were created by elevating 5 mm diameter full thickness flaps. Healed and unhealed wound areas were measured using photographs. Transforming growth factor (TGF)-ß, insulin-like growth factor (IGF), and collagen I and III expressions were determined using immunohistochemistry. The number of fibroblasts increased and inflammatory cells decreased from day 0 to 21 in both groups. The HA group exhibited more fibroblasts by day 7 compared to controls; (TGF)-ß and IGF levels were similar between HA and control groups. HA groups exhibited fewer inflammatory cells than controls on days 3 and 7. We found significant differences in TGF-ß and IGF levels among HA groups between days 3 and 21, and among control groups between days 0 and 21. Collagen I and III levels were greater for the day 3 HA group compared to controls. We observed improved wound healing in HA treated rats within 7 days.

Cinnamic acid decreases periodontal inflammation and alveolar bone loss in experimental periodontitis.

BACKGROUND AND OBJECTIVE: Periodontitis is the chronic destructive disease of the periodontium, which causes severe inflammation in the tissues. Cinnamic acid as an unsaturated carboxylic acid might prevent inflammation and periodontal destruction. The present study aimed to evaluate the effects of cinnamic acid in two different forms as free cinnamic acid and cinnamic acid liposome on experimental periodontitis in Wistar rats. METHODS: Thirty-two female rats were used in the present study. Four main groups were created as follows: C: control group; P: periodontitis group; C-P: free cinnamic acid-administered periodontitis group; and CL-P: cinnamic acid liposome applied group. Periodontitis was induced via ligating 4-0 silk sutures around lower first molar teeth on both right and left mandibles. The study duration was 30 days, and the ligatures were removed from half of the rats in the periodontitis-induced groups. The other half carried the ligatures throughout 30 days, and all rats were euthanized at 30th day. Mandibles were removed and evaluated via stereomicroscope and underwent histological procedures. Inflammatory cell counts, osteoblast, and osteoclast cell counts were determined in hematoxylin-eosin-stained slides, and peroxisome proliferator-activated receptor (PPAR)-γ, cyclooxygenase (COX)-2, receptor activator of nuclear factor κ-B (RANKL), and osteoprotegerin (OPG) expressions were evaluated by immunohistochemistry. RESULTS: Control group had the lowest bone loss, and periodontitis group which kept ligatures had the highest bone loss compared to the other groups. Ligature removal provided significant improvement in bone measurements. Cinnamic acid groups also showed lower bone loss compared to the periodontitis group. The inflammatory cell and osteoclast counts were also higher in the periodontitis group, and both applications of cinnamic acid decreased these values. Osteoblast cells were the lowest in the periodontitis group, and cinnamic acid increased these counts. PPAR-γ and COX-2 levels were higher in the periodontitis group, and cinnamic acid decreased these levels but not to a significant level except for the cinnamic acid liposome ligature removal group, which had significantly lower values in the PPAR-γ and COX-2. OPG levels were lower in the periodontitis group compared to the other groups. Cinnamic acid significantly decreased RANKL and increased OPG levels. CONCLUSION: Periodontitis caused increased inflammation and bone destruction accompanied by increased PPAR-γ, COX-2, and RANKL levels and osteoclast counts. Cinnamic acid decreased osteoclast counts and inflammation and increased osteoblast counts and OPG expression in the present animal model of periodontitis.

Quercetin Decreased Alveolar Bone Loss and Apoptosis in Experimentally Induced Periodontitis Model in Wistar Rats.

BACKGROUND: Quercetin is a flavonoid which has potent anti-inflammatory, antibacterial, and antioxidant effect. Purpose of this study was to evaluate effects of quercetin on alveolar bone loss and histopathological changes in ligature-induced periodontitis in rats. METHODS: Wistar rats were divided into four experimental groups: non-ligated control (C, n=8) group; periodontitis (P, n=8) group; ligature and low dose quercetin group (75 mg/kg/day quercetin, Q75 group, n=8); ligature and high dose quercetin group (150 mg/kg/day quercetin, Q150 group, n=8). Silk ligatures were placed at gingival margin of lower first molars of mandibular right quadrant. Study duration was 15 days, and animals were sacrificed end of this period. Changes in alveolar bone levels were clinically measured and tissues were immunohistochemically examined, matrix metalloproteinase 8 (MMP 8), inducible nitric oxide synthase (iNOS), tissue inhibitor of metalloproteinase 1 (TIMP 1), Cysteine-aspartic proteases 3 (Caspase 3), and tartrate-resistant acid phosphatase (TRAP) positive osteoclast cells, osteoblast, and neutrophil counts were also determined. RESULTS AND DISCUSSION: Alveolar bone loss was highest in P group, and differences among P, Q75, and Q150 groups were significant. Both doses of quercetin decreased TRAP+ osteoclast cells and increased osteoblast cells. Inflammation in P group was also higher than those of C, Q75, and Q150 groups indicating anti-inflammatory effect of quercetin. iNOS, MMP-8, and caspase-3 levels were highest, and TIMP-1 expression was lowest in P group; differences were statistically significant. CONCLUSION: Within limits of this study, it can be suggested that quercetin administration may reduce alveolar bone loss by increasing osteoblastic activity, decreasing osteoclastic activity, apoptosis, and inflammation in an experimental model of periodontitis.

Histological evaluation of peri-implant mucosal and gingival tissues in peri-implantitis, peri-implant mucositis and periodontitis patients: a cross-sectional clinical study.

Objective: Aim of present study was to evaluate gingival tissue samples obtained from healthy and diseased sites of teeth and dental implants in terms of hypoxia and collagenase activity.Methods: Four study groups were created as Group-1; healthy individuals (H), Group-2; periodontitis patients with stage 3 grade B (P), Group-3; patients with peri-implant mucositis. Group-4; patients with peri-implantitis (P-IMP). Plaque index (PI), gingival index (GI) and probing pocket depth (PPD) were recorded. Gingival and peri-implant mucosal biopsies were obtained. Fibroblast and inflammatory cells were counted. Hypoxia-inducible factor (HIF)-1α, prolyl hydroxylase (PH), matrix metalloproteinase (MMP)-8, tissue inhibitor of MMPs (TIMP)-1, cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) levels were determined via immunohistochemistry.Results: Healthy controls had highest fibroblast cell counts and lowest inflammatory cell counts compared to other groups. Peri-implantitis and periodontitis samples had similar fibroblast and inflammatory cell counts, while peri-implant mucositis had higher fibroblast cells and lowered inflammatory cells compared to periodontitis and peri-implantitis samples. HIF-1α, COX-2 and iNOS levels were lowest in healthy controls and increased in other groups. Peri-implant mucositis samples had significantly lower expressions of HIF-1α, COX-2 and iNOS compared to peri-implantitis and periodontitis groups. PH expressions were lower in periodontitis and peri-implantitis groups compared to healthy controls and peri-implant mucositis groups. MMP-8 levels were lower in healthy group compared to other groups while levels were similar in periodontitis, peri-implant mucositis and peri-implantitis groups. TIMP levels were similar in groups.Conclusion: Periodontitis, peri-implantitis, and peri-implant mucositis samples exhibited higher inflammation and lower fibroblast cell counts and tend to have increased tissue collagenase activity, hypoxia and inflammation compared to healthy samples.

Evaluation of apoptosis and hypoxia-related factors in gingival tissues of smoker and non-smoker periodontitis patients.

OBJECTIVE: Smoking causes pathological changes in all tissues, including gingiva and alveolar bone. The aim of present study was to evaluate apoptotic tissue alterations and tissue destruction in smoker and non-smoker periodontitis patients and healthy individuals. METHODS: Gingival biopsy samples from 15 systemically and orally healthy individuals (Group 1), 15 systemically healthy periodontitis patients (Group 2), 15 systemically and orally healthy smokers (Group 3), and 15 systemically healthy smoker periodontitis patients (Group 4) were enrolled in the present study. Clinical periodontal measurements as plaque index (PI), gingival index (GI), and clinical attachment levels (CAL) were recorded, and gingival biopsies were obtained. Biopsy samples were fixed in formalin solution and embedded in paraffin. Fibroblast and inflammatory cell counts were determined via histomorphometrically. Hypoxia-inducible factor alpha (HIF-1α), vascular endothelial growth factor(VEGF), tissue inhibitor of matrix metalloproteinase-1(TIMP-1), matrix metalloproteinases-8(MMP-8) expressions, Bax, Bcl-2, and caspase-3 expressions were evaluated via immunohistochemistry. RESULTS: Demographic data of the study groups were similar. Smoking levels of the smokers were also similar. The highest fibroblast cell counts were observed in healthy controls and the counts were similar in other groups. The highest inflammatory cell counts were found in smoker periodontitis group, and the lowest counts were found in healthy control groups. The differences were statistically significant. HIF-1α and Bax expressions were elevated and Bcl-2 decreased in smoker periodontitis patients compared with healthy individuals. However, there were no differences in VEGF, MMP-8, and TIMP-1 expressions. CONCLUSION: Within limits of present study, it can be suggested that both smoking and periodontitis caused similar decrease in fibroblast counts while causing a dramatic increase in inflammatory cell counts. Increased apoptosis and hypoxia also accompanied to the increased inflammation.

Free and liposome form of gallic acid improves calvarial bone wound healing in Wistar rats

Objective: To evaluate the effect of free and liposome form of gallic acid on bone regeneration in critical defects in Wistar rats. Methods: Thirty-two female Wistar rats were divided into four study groups: group 1, negative control; group 2, positive control; group 3, gallic acid powder; group 4, gallic acid liposome. A critical-sized defect was created in all rats. Groups 2 to 4 had xenograft, autograft and membrane placement while negative control rats did not receive any treatment. The defect area was sutured and rats were kept alive for 30 d. At the end of the study, a bone specimen including the defect area was removed from calvaria. All specimens were evaluated under the stereomicroscope, then underwent histological analysis. Inflammatory cell counts, osteoblast, osteoclast counts, receptor activator of nuclear factor κ-B (RANKL), osteoprotegerin (OPG), runt-related transcription factor 2 (Runx2), bone morphogenetic protein-2 (BMP-2), and alkaline phosphatase were determined. Results: The biggest unhealed defect area was observed in the negative control group and the smallest was observed in the gallic acid liposome group. There were no differences between the positive control group vs. the gallic acid powder group and the gallic acid powder group vs. the gallic acid liposome group. The severity of inflammation was the highest in the negative control group and the lowest in the gallic acid liposome group with significant differences between the groups. All groups had similar osteoblast counts while osteoclast counts were the highest in the positive control group. Gallic acid groups had a lower number of osteoclasts compared with the positive control group. Runx2 and alkaline phosphatase levels were similar in the groups while OPG and BMP-2 levels exhibited a significant increase compared with the negative control group and the positive control group. RANKL was similar in the negative control group, the positive control group, and the gallic acid powder groups but decreased in the gallic acid liposome group. Conclusions: Gallic acid powder and liposome significantly improve bone regeneration in Wistar rats with calvarial defects. The improvement in healing is evident with decreased inflammation and RANKL expressions and increased OPG and BMP-2 expressions.