Protective effects of passiflora incarnata on ovarian ischemia/reperfusion damage in rats with ovarian torsion.

BACKGROUND: This study aimed to investigate whether Passiflora Incarnata (PI) has a protective effect against ischemia-reperfu-sion (IR)-induced oxidative and inflammatory ovarian damage. METHODS: The effects of PI on ovarian ischemia-reperfusion injury were investigated in female Wistar albino rats. The animals were randomly divided into three groups: Group 1 (sham), Group 2 (IR), and Group 3 (IR+PI). RESULTS: The mean levels of Malondialdehyde (MDA), Myeloperoxidase (MPO), and Total Oxidant Status (TOS) were higher in the IR group (p=0.025, p<0.001, and p=0.016, respectively). The Total Antioxidant Status (TAS) levels were lower in the IR group (p=0.005). Immunostaining revealed significant differences across the groups for Tumor necrosis factor-alpha (TNF-α): 13.84%, 49.51%, and 22.51% for Groups 1, 2, and 3, respectively (p<0.01). Bax: 10.53%, 46.74%, and 26.46% for Groups 1, 2, and 3, respectively (p<0.01). Annexin V: 12.24%, 44.86%, and 23.28% for Groups 1, 2, and 3, respectively (p<0.01). The mean scores for hemorrhage, inflammation, follicular degeneration, and congestion showed significant variations among the groups, all registering p<0.001. CONCLUSION: Passiflora Incarnata exhibited antioxidant, anti-inflammatory, and anti-apoptotic properties, promoting cell survival, histologically protecting ovarian tissue, and ameliorating IR injury by reducing oxidative stress.

Wound healing activity of Salvia huberi ethanolic extract in streptozocin-induced diabetic rats.

Objective: The aim of this study was to examine the in vivo wound healing potential of Salvia huberi Hedge (endemic to Turkey) on excision and incision wound models in diabetic rats. Method: Male Wistar albino rats, 3-4 months old and weighing 180-240g were used. The animals were randomly divided into five groups including Control, Vehicle and Fito reference, and two different concentrations (0.5% and 1% weight/weight (w/w)) of ethanol extract of Salvia huberi were investigated in both wound models on streptozocin-induced diabetic rats using macroscopic, biomechanical, biochemical, histopathological, genotoxic and gene expression methods over both seven and 14 days. Fito cream (Tripharma Drug Industry and Trade Inc., Turkey) was used as the reference drug. Results: A total of 60 rats were used in this study. Salvia huberi ointments at 0.5% and 1% (w/w) concentrations and Fito cream showed 99.3%, 99.4% and 99.1% contraction for excision wounds, and 99.9%, 97.0% and 99% contraction for incision wounds, respectively. In Salvia huberi ointments and Fito cream groups, re-epithelialisation increased dramatically by both day 7 and day 14 (p<0.05). By day 14, low hydroxyproline and malondialdehyde (MDA) levels, and high glutathione (GSH) levels were observed in the Salvia huberi ointment groups. After two application periods, damaged cell percent and genetic damage index values and micronucleus frequency of Salvia huberi ointment treatment groups were lower than Control and Vehicle groups (p<0.001). A growth factor expression reached a high level by day 7 in the Control group; in Salvia huberi-treated groups it was decreased. Conclusion: The study showed that application of Salvia huberi ointments ameliorated the healing process in diabetic rats with excisional and incisional wounds and may serve as a potent healing agent.

Effects of Carvacrol in an Experimentally Induced Esophageal Burn Model: Expression of VEGF and Caspase-3 Proteins.

INTRODUCTION: We investigated the therapeutic effects of carvacrol in an experimental esophageal burn rat model with immunohistochemical techniques. Materials and Methods: Three groups were included in this study, composed of eight Wistar albino rats each. The control group was given 1 mL 0.9% (wt/vol) NaCl; esophageal burns were induced in groups 2 and 3 by administration of 1 mL 40% NaOH in the distal 2 cm of the esophagus. The treatment group was administered 75 mg/kg carvacrol in 2 mL 0.9% NaCl for 10 days. After a routine histological examination of the tissues, sections were stained with vascular endothelial growth factor (VEGF) and caspase-3 for immunohistochemical analysis and were examined under a light microscope. Results: In the control group, there were regular cells in the cornified epithelial tissue and cylindrical cells in the basal layer, which faced toward the apical surface in the mitotic phase. The burn group displayed wide degeneration, necrosis, and abundant apoptotic cells in the epithelial tissue as well as intense inflammatory cell infiltration. In the treatment group, there was an increase in mitotic activity in the basal cells of the epithelial layer and degenerative changes, but a preserved epithelial layer and significant cornified structures. The treatment group showed positive caspase-3 expression in some apoptotic cells within the epithelial layer and in connective tissue, and there were only a small number of degenerated cells in the muscle layer. Additionally, in the treatment group, VEGF expression was evident in small numbers of inflammatory cells in the papillary region of the epithelium, and in dilated vascular endothelial cells. Conclusions: Carvacrol may contribute to a reduction in fibrosis by decreasing inflammation and preventing cell apoptosis.

The effect of graft application and simvastatin treatment on tibial bone defect in rats. A histological and immunohistochemical study.

PURPOSE: To evaluate histologically and immunohistochemically the bone regeneration after application of simvastatin on tibial bone defects in rats. METHODS: Sixty Wistar albino rats were divided into 3 groups as control (6 mm tibial bone defect), defect + graft (allograft treatment), and defect + graft + simvastatin (10 mg/kg/day) for 28 days. RESULTS: Histopathological examination revealed inflammation in control group (defect group), congestion in blood vessels, and an increase in osteoclast cells. In defect + graft group, osteoclastic activity was observed and osteocyte cells were continued to develop. In defect + graft + simvastatin group, osteocytes and matrix formation were increased in the new bone trabeculae. Osteopontin and osteonectin expression were positive in the osteclast cells in the control group. Osteoblasts and some osteocytes showed a positive reaction of osteopontin and osteopontin. In defect + graft + simvastatin group, osteonectin and osteopontin expression were positive in osteoblast and osteocyte cells, and a positive expression in osteon formation was also seen in new bone trabeculae. CONCLUSION: The simvastatin application was thought to increase bone turnover by increasing the osteoinductive effect with graft and significantly affect the formation of new bone.

The effect of graft application and allopurinol treatment on calvarial bone defect in rats1.

PURPOSE: To investigate the effects of allopurinol administration on osteoinductive reaction and bone development with graft material. METHODS: Thirty-six Wistar albino rats were divided into 3 groups. In the control group, calvarial bone defect was only created without any treatment. In the Defect + Graft group, allograft treatment was performed by forming 8 mm calvarial bone defect. In the Defect + Graft + Allopurinol group, alloplastic bone graft was placed in the calvarial bone defect and then, allopurinol (50 mg/kg/day) treatment was intraperitoneally applied for 28 days. RESULTS: Histopathological examination revealed inflammation, congestion in the vessels, and an increase in osteoclast cells in the defect area. We also observed that new osteocyte cells, increase in connective tissue fibers, and new bone trabeculae. Osteopontin expression was positive in osteoblast cells and lacunated osteocyte cells were located in the periphery of the new bone trabeculae. Osteopontin expression was also positive in osteoblasts and osteocytes cells of new bone trabeculae in the graft site. CONCLUSION: It has been shown that allopurinol treatment in rat calvaria defects may induce osteoblastic activity, matrix development, mature bone cell formation and new bone formation when used with autogenous grafts.

Wound healing properties, antimicrobial and antioxidant activities of Salvia kronenburgii Rech. f. and Salvia euphratica Montbret, Aucher & Rech. f. var. euphratica on excision and incision wound models in diabetic rats.

Diabetic patients suffer from persistent and non-healing wounds. Salvia species are traditionally used for the treatment of wounds and colds. The aims of the present study were to evaluate the in vivo wound healing potential, in vitro antimicrobial and antioxidant activities, and total phenolic and flavonoid contents of the aerial parts of two endemic taxa, Salvia kronenburgii Rech. f. (SK) and Salvia euphratica Montbret, Aucher & Rech. f. var. euphratica (SE). Two different concentrations (0.5% and 1% (w/w)) of ethanol extracts were investigated in incision and excision wound models on Streptozotocin-induced diabetic rats using biomechanical, biochemical, histopathological, macroscopic, and genotoxic methods for 7 and 14 days. Antimicrobial activity was evaluated against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Acinetobacter baumannii, Aeromonas hydrophila, Mycobacterium tuberculosis, Candida glabrata, Candida parapsilosis, and Candida tropicalis using the broth microdilution and the resazurin microtiter assay plate methods. Fito®, Ampicillin, Ethambutol, Isoniazid, and Fluconazole were used as reference drugs. Antioxidant capacities and total phenolic and flavonoid contents of both extracts were detected using DPPH free radical scavenging assay, Folin-Ciocalteu, and Al(NO3)3 methods, respectively. SK ointment at 0.5% and 1% (w/w) concentrations and SE ointment at 1% (w/w) concentration showed 99.9%, 99.5%, and 99.7% contraction, respectively for excision wounds, and SK and SE ointments at 1% (w/w) concentration showed 99.4% and 99.2% contraction for incision wounds while Fito® showed 98.9% and 98.5% contraction, respectively. Increased re-epithelialization (P < 0.01 and P < 0.001), angiogenesis, and decreased dermal inflammation (P < 0.001) were determined for SK and SE ointments at both 7 and 14 days. SE ointment on day 7 and SK ointment on day 14 reduced oxidative damage to DNA when compared to control (P < 0.01 and P < 0.001). Both tested plants had greater antibacterial activity against A. baumannii (62.5 µg/mL MIC value) and SE had greater antimycobacterial activity against M. tuberculosis (0.24 µg/mL MIC value) when compared to reference drugs Ampicillin, Isoniazid, and Ethambutol (125, 0.97, and 1.95 µg/mL MIC values, respectively). Antioxidant capacities, total phenolic and flavonoid contents of SE and SK were 87.08%, 76.21 µg GAE/mg, 43.43 µg QE/mg and 72.17%, 41.81 µg GAE/mg, 33.62 µg QE/mg, respectively. SK and SE had strong wound healing effects while SK found to be more effective than SE at both 7 and 14 days.

The effect of graft application and simvastatin treatment on tibial bone defect in rats. A histological and immunohistochemical study

Abstract Purpose: To evaluate histologically and immunohistochemically the bone regeneration after application of simvastatin on tibial bone defects in rats. Methods: Sixty Wistar albino rats were divided into 3 groups as control (6 mm tibial bone defect), defect + graft (allograft treatment), and defect + graft + simvastatin (10 mg/kg/day) for 28 days. Results: Histopathological examination revealed inflammation in control group (defect group), congestion in blood vessels, and an increase in osteoclast cells. In defect + graft group, osteoclastic activity was observed and osteocyte cells were continued to develop. In defect + graft + simvastatin group, osteocytes and matrix formation were increased in the new bone trabeculae. Osteopontin and osteonectin expression were positive in the osteclast cells in the control group. Osteoblasts and some osteocytes showed a positive reaction of osteopontin and osteopontin. In defect + graft + simvastatin group, osteonectin and osteopontin expression were positive in osteoblast and osteocyte cells, and a positive expression in osteon formation was also seen in new bone trabeculae. Conclusion: The simvastatin application was thought to increase bone turnover by increasing the osteoinductive effect with graft and significantly affect the formation of new bone.

The effect of graft application and allopurinol treatment on calvarial bone defect in rats

Abstract Purpose: To investigate the effects of allopurinol administration on osteoinductive reaction and bone development with graft material. Methods: Thirty-six Wistar albino rats were divided into 3 groups. In the control group, calvarial bone defect was only created without any treatment. In the Defect + Graft group, allograft treatment was performed by forming 8 mm calvarial bone defect. In the Defect + Graft + Allopurinol group, alloplastic bone graft was placed in the calvarial bone defect and then, allopurinol (50 mg/kg/day) treatment was intraperitoneally applied for 28 days. Results: Histopathological examination revealed inflammation, congestion in the vessels, and an increase in osteoclast cells in the defect area. We also observed that new osteocyte cells, increase in connective tissue fibers, and new bone trabeculae. Osteopontin expression was positive in osteoblast cells and lacunated osteocyte cells were located in the periphery of the new bone trabeculae. Osteopontin expression was also positive in osteoblasts and osteocytes cells of new bone trabeculae in the graft site. Conclusion: It has been shown that allopurinol treatment in rat calvaria defects may induce osteoblastic activity, matrix development, mature bone cell formation and new bone formation when used with autogenous grafts.

Ultrastructural Alterations of Liver Tissue Cells in Methotrexate-Treated Balb/c Mice.

AIM: The current study investigated the efficacy of methotrexate (MTX) on liver tissue cells of Balb/c mice at the ultrastructural level using transmission electron microscopy. BACKGROUND: This agent is well known and used as a chemotherapeutic agent for a long time and not selective for cancer cells so, healthy cells beside cancer cells are also affected by MTX. MATERIALS AND METHODS: Experimental animals were divided into two groups; the first group was kept without treatment and served as the control, the second group was treated with 115 mg/kg MTX i.p. once weekly for 4 weeks and sacrificed under anesthesia after the 4th week. The liver tissues were osmium fixed and embedded in araldite, sectioned and observed under transmission electron microscope. RESULTS: Normal cell ultrastructure was determined in the control group whereas the liver cells of the MTX-treated group revealed ultrastructural alterations, such as the increase in lipid droplets, discontinuity of rough endoplasmic reticulum cisternae and vacuole formation. In addition, the loss of cytoplasmic material in hepatocytes was also evident. Condensation of nuclear chromatin and fusion of nucleic membranes were observed in the liver cells of the treated group. CONCLUSIONS: Results of the study indicated that MTX, used for different types of medical treatment, disturbed liver cell ultrastructure.

An ultrastructural study, effects of Proteus vulgaris OX19 on the rabbit spleen cells.

Effects of Proteus vulgaris OX19 on the spleen cells of rabbits were investigated. Control group (n=5) and Proteus treated group (n=5) of New Zealand male rabbits were used in this study. Bacteria were injected to the rabbits in five days periods with increasing dosages for one month. Thin sections were examined by transmission electron microscope (Jeol 100CXII). Ultrastructural changes were defined in spleen tissue cells due to the antigenic stimulation of bacteria. Spleen cells observed in control group were in normal structure and cells were in close contact with each other. However, spleen cells of Proteus treated group displayed structural changes with regard to the control group in electron microscopic examinations. Chemotaxis of macrophages, forming of pseudopodia and presence of phagocytic vacuoles were observed. Lymphocytes, the major cells of spleen revealed mitotic activity. In addition, chromatin condensation in nucleus and dilatations in perinuclear space were significant. Interactions of lymphocytes and macrophages were noteworthy.

Microscopic investigations in a diabetic rat urinary bladder infected with Trichosomoides crassicauda.

Rats are widely used laboratory animals and have several parasites. One of these are helminths, known not only to cause serious effects on the experimental results in healthy subjects, but also in subjects with heavy infections. One of the relatively pathologic helminth is Trichosomoides crassicauda, which lives in the nodules of the urinary bladder. It is known that diabetics are more prone to infections with several microorganisms. Observations in a diabetic rat bladder showed T. crassicauda eggs inside the transitional epithelium, and structural changes in the bladder epithelium were evident. Urinary-bladder tissues taken from streptozotocin-injected diabetic subjects and citrate buffer-injected control subjects were fixed, embedded in araldite and investigated under a light microscope. Distinct changes in the histological structure of a diabetic urinary bladder transitional epithelium were observed after T. crassicauda infection. Many papillomas were formed and the epithelial tissues were completely degenerated. In addition, electron microscopic examinations also revealed degeneration of the subepithelial tissues.