Biotechnic & Histochemistry最新文献

We aimed to investigate TLR-4 receptor activity in the development of endometriosis and the effect of trastuzumab in experimentally induced endometriotic tissue via TLR-4 in this study. Twenty-eight female Wistar-Albino rats were divided into four groups: Group 1 (Control Group), Group 2 (Endometriosis Group), Group 3 (Endometriosis + Trastuzumab Group), and Group 4 (Trastuzumab Group). All animal tissue samples were collected. Histopathological, immunohistochemical, and biochemical analyses were performed. In histopathological analysis, there was a significant difference between Group 2 and other groups in terms of connective tissue edema, inflammation, hemorrhage, epithelial damage, and mast cell density. In immunohistochemical analysis with TLR-4, Group 2 exhibited strong staining. In biochemical analysis, it was found that there was a highly significant difference between Group 2 and Group 1 considering the Malondialdehyde (MDA) levels in plasma samples. There was no significant difference in terms of the MDA levels among other groups. Considering the glutathione levels in the plasma samples, it was found that there was a highly significant difference between Group 2 and Groups 3 and 4. Trastuzumab may play a role in the treatment of histopathological damage and fibrosis in experimentally induced endometriotic implants by showing anti-inflammatory and antiproliferative activity.

Paracetamol (PAR) is a drug that is widely used throughout the world and has limited treatment options in case of use-related hepatotoxicity. Apilarnil (AP), a bee product has high levels of antioxidant properties, which result from the rich polyphenols found in its structure. Despite it being shown that AP treatment might have a protective effect on liver damage induced by carbon tetrachloride and lipopolysaccharide, there is no study investigating the possible role of this agent in PAR-induced hepatotoxicity using an experimental in vivo model. Therefore, we aimed to investigate the therapeutic effects of AP on paracetamol-induced hepatotoxicity and its relationship with apoptosis and oxidative stress. Our results indicated that PAR administration caused irregularities in hepatocyte cords, bleeding and dilatation of sinusoids, and inflammatory cell infiltration in the portal area and liver parenchyma. PAR caused an increase in p53 and caspase3 expressions and malondialdehyde (MDA) levels, while it caused a decrease in catalase (CAT) and glutathione peroxidase (GSHpx) levels. AP treatment significantly improved histopathological changes in liver tissues and decreased p53 and caspase3 expressions. Our data suggest that AP alleviates paracetamolinduced hepatotoxicity by regulating p53 and caspase-3 expressions and modulating oxidative stress mechanisms.

Tendon injuries remains a challenge to treat owing to its poor intrinsic reparative ability. It is hypothesised that hypoxic conditioning of mesenchymal stem cells (MSC) through the activation of hypoxia-inducible factor-1 alpha (HIF-1α), may enhance tendon repair process by promoting cellular proliferation and tenogenic differentiation. To demonstrate this, a study using roxadustat, a specific hypoxia mimetic mediator and HIF-1α inducer was conducted on adipose-derived mesenchymal stromal cells (AD-MSCs). Cellular morphology, proliferation rates, tenogenic protein and gene expression levels in untreated AD-MSCs (Group 1), roxadustat pre-conditioned AD-MSCs (Group 2), AD-MSCs subjected to CAY10585 (Group 3), roxadustat pre-conditioned AD-MSCs with CAY10585 (Group 4) and untreated primary tenocytes (Group 5) were evaluated. MSCs pre-conditioned with 12.5µM roxadustat for 24 hours showed the highest expression of HIF-1α without affecting the proliferation rates of AD-MSCs. However, significant reduction of HIF-1α levels was observed when the cells were treated with 3.5µM CAY10585. Roxadustat significantly up-regulated collagen I and III expressions by 6.6 and 6.3-fold respectively. HIF-1α promoted Scleraxis, Tenascin-C and Collagen III expressions, resulting in an increase of 6, 7, and 3 folds respectively. Conversely, using CAY10585 reduced these expressions to 3, 2 and 1 folds respectively. These trends were observed in the gene expression levels across Groups 1 to 4. However, the expression of these genes in Group 2 was significantly lower as compared to Group 5. Conclusion: HIF-1α accumulation promotes superior cell proliferation and tenogenic differentiation of AD-MSCs, indicating that roxadustat may be a potential therapeutic mediator in tendon repair strategies.

The anti-inflammatory effect of the ethanol extract of Momordica charantia in two different chemically induced inflammatory bowel disease models, which are frequently used in experimental studies, was investigated. For this purpose, IBD models were created with acetic acid (AA) and 2,4,6 trinitrobenzene sulphonic acid (TNBS) in rats and 300 mg/kg M.charantia extract was given by oral gavage for 10 days. In the animal experiment phase, a total of 42 animals in six groups were arranged so that two different experimental models could be studied simultaneously. Colon tissues were examined at light and electron microscopy levels. In the microscopic examination, areas of inflammation extending to the muscularis externa were observed in the macroscopically severely damaged areas in both IBD model groups, and epithelial damage, mucosal inflammation, and crypt abscess were observed in the macroscopically less damaged areas. Microscopic large intestine damage was significantly reduced in M.charantia administered groups compared to disease models. TNF-α and IL-1β expression, which was determined to be increased in the AA and TNBS groups immunohistochemically, was observed to decrease in the treatment groups. The surface epithelium was evaluated by electron microscopic observations. This study demonstrates the positive effect of M.charantia ethanol extract on colon histopathology in two different IBD models and highlights the importance of considering inflammation-related cell populations in the treatment of this disease.

Corneal injuries are common in human and veterinary ophthalmology. There are many studies which have investigated the treatment of corneal epithelial defects. This study aimed to investigate the effects of Neutral Protamine Hagedorn (NPH) insulin as an ointment for wound healing in experimental corneal defects. First, a superficial keratectomy was performed on 12 rabbits using a corneal trephine. The animals were divided into two groups, Group I (treated, n = 6) and Group II (vehicle control, n = 6). Insulin ointment was applied topically once daily in the treated group, and saline ointment was applied in the same manner in the vehicle control group. Corneal defects were observed and photographed, and changes in wound surface were recorded on days 7, 14, 21, 30, and 60. In both groups, a significant reduction in the wound surface area was noticeable on the 30th day after defect creation. Between the 30th and 60th days, while the changes in the wound surface area in Group II remained limited, the decrease continued rapidly in Group I. At the end of the study, the corneal opacity score observed was lower in Group I than in Group II. In conclusion, we determined that topical NPH insulin may accelerate corneal wound healing after superficial lamellar keratectomy. A new alternative treatment may be developed for treating corneal wounds through continuing studies on this subject.

The main symptoms of depression, a chronic mental illness, include sadness, low self-esteem, and a diminished sense of enjoyment in life. Many factors have been suggested to be associated with depression, one of which is low testosterone in men. The serotonin reuptake inhibitor fluoxetine (FLU), used to treat depression, has been reported to potentially have detrimental effects on spermatogenesis in rats after long-term use. The multimodal antidepressant vortioxetine (VTX) offers new promise for the treatment of depression. The chronic unpredictable mild stress (CUMS) model is widely known as an experimental paradigm used to study depression-like behaviors in rodents. Stress leads to various neurochemical and immune changes, affecting multiple organs. Our study aims to examine the histopathological findings in testicular tissue induced by CUMS and the immunohistochemical expression of Cas-8, IL-6, and RANKL using a depression model in rats. Rats were split into 4 groups of 7 animals each at random. Group 1 (control) did not experience any stress. Group 2 (CUMS) was exposed to chronic, unpredictable mild stress using a specific procedure. Group 3 (CUMS+VTX) and Group 4 (CUMS+FLU) underwent CUMS and received intraperitoneal drug treatment at a dose of 10 mg/kg during the final three weeks of the study. The rat testicles collected during necropsy were evaluated histopathologically and immunohistochemically for Cas-8, IL-6, and RANKL expressions using a light microscope. In Group 1, histological analysis showed normal tissue architecture in the testicles and epididymis. In Group 2, there was significant depletion of spermatozoa in the seminiferous tubules and empty tubules in the epididymis. In Groups 3 and 4, FLU and VTX treatment led to improvements in the testicles. Cas-8, RANKL, and IL-6 immunohistochemistry revealed increased expression in Group 2, primarily in interstitial cells. In Groups 1, 3, and 4, no or very slight expression of these markers was observed. The results of this study showed that sperm production in the testes is negatively affected in CUMS-induced depression and that Cas-8, IL-6, and RANKL expression is increased, particularly in interstitial cells. VTX and FLU, used in the treatment of depression, suggest potential for mitigating the adverse effects of CUMS on the testes.

Dexmedetomidine has been shown to exert protective and curative effects on various tissues and organs in different pathological processes. This study aimed to investigate the effect of dexmedetomidine on the regeneration process after making holes in the parietal bones of rabbits. Twenty-four male Oryctolagus cuniculus rabbits were allocated to three groups, and an 8-mm circular parietal critical-sized bone defect was induced in each animal. Group_C (control) received saline; Group_LD (low dose) was given dexmedetomidine 2.75 µg/kg; Group_HD (high dose), dexmedetomidine 5.5 µg/kg; all were administered intraperitoneally for 7 days. After 8 weeks the bones were examined by micro-computed tomography (micro-CT) and histomorphometry. The results indicated that regeneration was improved in both the dexmedetomidine-treated groups. The lower dose increased the bone volume ratio (BV/TV) more than the higher dose. Trabecular thickness, connectivity value, and connectivity density were also higher in Group_LD than in Group_HD. Significant intramembranous ossification was observed in the dexmedetomidine-treated groups, and active osteoblasts were seen at the margins of new bone trabeculae. We conclude that dexmedetomidine, especially at the lower dosage, increases osteoblastic activity and regeneration quality.

Numerous studies reported about potential effects of L-carnosine in regulation of tumor growth and metabolism. We evaluated the effects of different concentrations of L-carnosine from Karnozin EXTRA® supplement on mitochondrial respiratory chain complexes of human embryo lung fibroblasts (MRC-5) and human breast cancer cells (MCF-7), with different energy pathways. Also, we analyzed the proliferation index and expression of various markers of oxidative stress. Treatment with Karnozin EXTRA^® (concentration of L-carnosine were 2, 5 and 10 mM) for 24 hours gradually decreased the number of cells and changed their morphological features. In both cell lines, a dose-dependent reduction of cell viability was recorded compared to the control group. Also, experimental groups showed a concentration-dependent decrease in fluorescence intensity of SOD2 expressions in MCF-7, while in MRC-5 we noticed higher fluorescence intensity in Carnosine 2 mM group. Treated cells, in both cell lines, showed different intensity of iNOS cytoplasmic immunopositivity in a concentration-dependent manner. In all experimental groups, we noticed an increased expression of marker of oxidative stress-cytochrome P450 2E1 (CYP2E1). The effects of Karnozin EXTRA® capsule on mitochondrial respiration, assessed with the Clark-type electrode, were manifested as a reduction of: basal cell respiration, maximum capacity of electron transport chain and mitochondrial ATP-linked respiration. Also, significant decrease in the activity of complex I (NADH-ubiquinone oxidoreductase), complex II (succinate dehydrogenase) and complex IV (cytochrome c oxidase) was observed in both cell lines. Bearing in mind that Karnozin EXTRA® is a potential regulator of energy metabolism of MCF-7 and MRC-5, these results provide a good basis for further preclinical and clinical research.

Anterior cruciate ligament (ACL) injury and subsequent reconstruction induce marrow adipose tissue (MAT) accumulation accompanied by bone loss. Short-term immobilization or non-weightbearing after ACL reconstruction further promotes MAT accumulation. However, it is unclear if combining immobilization and non-weightbearing synergistically promotes MAT accumulation. Additionally, it is unknown whether MAT increase induced by immobilization or non-weightbearing can be reversed through remobilization or reloading. We aimed to address these questions. ACL-reconstructed rats were divided into four groups: no intervention, immobilization, non-weightbearing, or immobilization plus non-weightbearing. Immobilization and non-weightbearing were applied for 2 weeks, after which all rats were allowed to move unrestricted. Intact rats were used as controls. The marrow adiposity and trabecular bone in the proximal tibia were histologically assessed at 2-, 4-, and 12-weeks post-surgery. ACL reconstruction induced MAT accumulation and trabecular bone loss accompanied by increased osteoclastogenesis. Two weeks of immobilization and non-weightbearing after ACL reconstruction individually promoted MAT accumulation, but the combined use of these interventions had a similar impact on MAT accumulation as either of each intervention. Importantly, the increased MAT induced by immobilization or non-weightbearing did not reverse even after remobilization or reloading. Neither immobilization, non-weightbearing, nor both conditions combined after ACL reconstruction further decreased trabecular bone compared to no intervention. These findings suggest no synergistic effect of immobilization and non-weightbearing on MAT accumulation, and MAT accumulation induced by 2 weeks of both immobilization or non-weightbearing did not decrease even after at least 10 weeks of remobilization or reloading. MAT accumulation due to both immobilization and non-weightbearing did not have negative effects on trabecular bone.

Cervicovaginal (CV) microbiota is critical for the well-being of host. We investigated the relationship between the ratio of Lactobacilli (LB) and cocci/coccobacilli (C/CB)-type microbial cells with biofilm formation of CV mixed cultures of women with no inflammation/infection or any epithelial abnormalities in Pap-stained smears Group 1 (G1) corresponds to the flora with LB-type cells alone, whereas G2 corresponds to the LB-dominated flora. G3 contains balanced LB and C/CB cells and G4 is dominated with C/CB. G5 corresponds to a flora with C/CB-type cells alone. Biofilm formation of CV mixed cultures was assessed by crystal violet binding assay and optical density (OD)≥0.8 were defined as biofilm producers. G1 and G3 exist in higher frequencies compared to the other smear groups. However, although the frequency of G5 dominated with C/CB-type cells were the lowest (4%); biofilm formation in that group was observed in the highest frequency (42.9%). The least biofilm formation frequency was observed in G3 smears with balanced flora (1%). Biofilm formation in healthy CV flora increases when there becomes an imbalance between LB and C/CB-type cells and an increase in C/CB-type cells. Our approach may enable early detection of vaginal dysbiosis in healthy flora prone to biofilm-associated CV infections such as bacterial vaginosis (BV).