Protective effects of Silymarin on testicular torsion/detorsion in rats.

OBJECTIVE: The present research aimed to study the possible protective effects of Silymarin on testicular I/R injury in a rat model evaluated through histopathology and biochemical parameters. MATERIALS AND METHODS: This research investigated the impact of Silymarin on IR damage in male Wistar albino rats. Animals were divided into three groups: group 1 (sham), group 2 (IR), and group 3 (IR+Silymarin). RESULTS: There were no notable differences in the levels of malondialdehyde (MDA), myeloperoxidase (MPO), and glutathione (GSH) across the three groups (p=0.260, p=0.486 and p=0.803, respectively). Contrarily, the total antioxidant status (TAS) levels exhibited significant variations between groups (p=0.001). The total oxidant status (TOS) levels also differed significantly between groups (p=0.004). The tissue evaluations uncovered substantial differences in the Johnson score, which is used to gauge testicular damage. A distinct contrast was seen between Group 1 and Group 2, and also between Group 2 and Group 3, with an all-encompassing p-value lower than 0.01. The same significant disparities were found for the percentages of Bax and Annexin V immunostaining (p<0.01 for each), reflecting the inflammation and apoptosis brought about by ischemia-reperfusion and the protective effects of the treatment. CONCLUSIONS: The outcomes of the current investigation showed that Silymarin could be a valuable agent for reducing testicular tissue damage following I/R injury.

Effects of alloplastic graft material combined with a topical ozone application on calvarial bone defects in rats.

BACKGROUND: This study presents the evaluation of the damage in the bone tissue resulting from a calvarial defect in rats and the efficiency of exposure to an ozone application with an alloplastic bone graft on the calvarial bone damage. MATERIALS AND METHODS: Wistar male rats (n = 56) were divided into four groups: a control group (n = 14), defect and ozone group (n = 14), defect and graft group (n = 14), and defect, graft, and ozone group (n = 14). Under anaesthesia, a circular full-thickness bone defect was created in all groups, and the experimental groups were further divided into two sub-groups, with 7 rats in each group sacrificed at the end of the 4th and 8th weeks. Bone samples were dissected, fixed in 10% formalin solution, and decalcified with 5% ethylene-diamine-tetraacetic acid (EDTA). After the routine follow-up on tissues, immunostaining of osteopontin and osteonectin antibodies was applied to sections and observed under a light microscope. RESULTS: The control group exhibited osteopontin and osteonectin expression in fibroblasts and inflammatory cells at the end of the 4th week with an acceleration at the 8th week. Ozone administration elucidated new trabecular bone formation by increasing osteoblastic activity. Lastly, our observations underscore that a combination of allograft and ozone application increased the osteoblast, osteocyte, and bone matrix development at the 4th and 8th weeks. CONCLUSIONS: Exposure to an ozone application with an alloplastic bone graft on calvarial bone damage may induce osteoblastic activity, matrix development, mature bone cell formation, and new bone formation in rats.

Biochemical and immunohistochemical investigations on bone formation and remodelling in ovariectomised rats with tamoxifen citrate administration.

BACKGROUND: Osteoporosis results with the imbalance between osteoblastic formation and osteoclastic resorption, resulting in susceptibility to bone fractures. Ovariectomy leads to osteoporosis by triggering alterations in bone formation and structure. Tamoxifen as an anti-oestrogen is used for adjuvant therapy especially in metastatic diseases and known to have a bone mass protective effect after ovariectomy. MATERIALS AND METHODS: An animal model of ovariectomy induced osteoporosis after tamoxifen citrate administration was studied via biochemical and immunohistochemical methods. Female Wistar albino rats (n = 45), selected according to their oestrous cycle, were divided into three groups; I - control, II - ovariectomy, III - ovariectomy + tamoxifen. Following ovariectomy, tamoxifen citrate (10 mg/kg) was given intraperitoneally daily for 8 weeks. At the end of the period, animals were sacrificed under anaesthesia, blood samples were taken to measure oestrogen, calcium, and alkaline phosphate. Tibia bone samples were fixed in formalin solution and decalcified with 5% ethylene-diamine tetra acetic acid. After the routine histological follow up, samples were embedded in paraffin and cut with a microtome for semi-thin sections. Primary antibodies osteonectin and osteopontin were applied to sections and examined under light microscope. RESULTS: As a consequence, when oestrogen and calcium data were compared there was a decrease in ovariectomy group with an increase in alkaline phosphatase. In ovariectomy + tamoxifen group, these values were close to the control group. Osteonectin was observed to promote bone formation by influencing collagen fibre formation, extracellular matrix development, osteoblast differentiation and the capacity to affect osteoclast activity. CONCLUSIONS: It has been suggested that osteopontin, the cytokine and cell binding protein, stimulates cellular signalling pathways, induces bone remodelling and acts in osteoporosis.

Neuroprotective effects of allopurinol on spinal cord injury in rats: a biochemical and immunohistochemical study.

BACKGROUND: Lesion in spinal cord causes a cascade of events such as the apoptosis of neurons and eventually, neurological dysfunction. Neurologic damage developing after acute spinal cord injury is also related with necrosis and free radical formation. Allopurinol, a xanthine oxidase inhibitor, was shown to have protective effects in several studies. B-cell lymphoma 2 (Bcl-2) family proteins regulate apoptosis. Apoptosis causes the death of neuronal cells, particularly neurons and oligodendrocytes in the spinal cord after lesion. Glial fibrillary acidic protein (GFAP) takes part in astrocyte and neuronal interconnection and synaptic transmission. MATERIALS AND METHODS: Male Sprague Dawley rats (n = 30) were divided as control, trauma, and trauma + allopurinol (i.p., 50 mg/kg of body weight) groups. Animals were applied a surgical procedure causing spinal cord injury and treated for 7 days then sacrificed under anaesthesia. The spinal cords were dissected, measurements of myeloperoxidase, malondialdehyde and glutathione were performed, remaining parts were fixed in 10% formaldehyde solution for histological and immunohistochemical evaluations. RESULTS: Biochemical results exhibited an increase in myeloperoxidase levels in trauma group but a decrease in the allopurinol treatment group similar to malondialdehyde levels. Degenerative changes in multipolar and bipolar neurons together with apoptotic changes in some glial cells were observed in the trauma group whereas, mild degenerative changes were observed after allopurinol treatment. In the trauma group, negative GFAP expression in multipolar versus bipolar neuronal processes with a reduction in glial processes around blood vessels and positive GFAP expression were observed but, a regular and parallel positive GFAP expression of glial processes around blood vessels in the allopurinol treated group was apparent. Trauma group depicted a positive Bcl-2 expression in glial cells and in motor and bipolar neurons. On the contrary, negative Bcl-2 expression was noticed in the trauma + allopurinol group. CONCLUSIONS: This study is of importance to understand the effects of allopurinol in preventing degenerative changes in nerve and glial cells related to spinal cord injuries.

Effects of formaldehyde on vascular endothelial growth factor, matrix metallopeptidase 2 and osteonectin levels in periodontal membrane and alveolar bone in rats.

BACKGROUND: The objective of this study was to investigate whether long term formaldehyde inhalation may affect periodontal membrane and alveolar bone loss leading to periodontitis. The negative effects of formaldehyde were described using vascular endothelial growth factor (VEGF), matrix metallopeptidase 2 (MMP-2) and osteonectin antibodies involved in the extracellular matrix and angiogenetic development. MATERIALS AND METHODS: Thirty adult Wistar albino rats were used in this study. Rats were divided into two groups: a control group (n = 15) and formaldehyde administered group (n = 15). Formaldehyde group was exposed to inhalation of 10 ppm formaldehyde 8 hours a day, 5 days a week for 5 weeks. Maxillary bone regions were dissected under anaesthesia. After fixation in 10% formaldehyde solution, tissues were passed through graded ethanol series to obtain paraffin blocks. Five-micrometre histological sections were cut with RM2265 rotary microtome stained with Masson trichrome and VEGF, MMP-2 and osteonectin antibodies for examination under Olympus BH-2 light microscopy. RESULTS: The present study revealed that congestion in blood vessels, degeneration of collagen fibres and alveolar matrix around alveolar bone were observed to be more significant in formaldehyde group than the control group (p ≤ 0.001). Interestingly, VEGF expression in the formaldehyde group was the most significant finding between the two groups (p < 0.001). When compared inflammation, MMP-2 and osteonectin expressions were significant (p < 0.01) in the formaldehyde group. CONCLUSIONS: It was suggested that formaldehyde toxicity decreased the expression of MMP-2 and in osteoblasts as well as affecting the retention of MMP levels in tooth cavity, which is very low in collagen fibres. But, vice versa for the expression of VEGF in dilated vascular endothelial cells and osteocytes in alveolar bone. As a conclusion, formaldehyde disrupts the periodontal membrane and may cause collagen fibres degeneration by affecting the alveolar bone matrix.