Effects of alpha‑tocopherol on gingival expression of inducible nitric oxide synthase in the rats with experimental periodontitis and diabetes

Background: The aim of this study was to investigate effects of α‑tocopherol and/or insulin on the number of gingival inducible nitric oxide synthase (iNOS) positive cells in rats with experimental periodontitis with or without streptozotocin (STZ)‑induced diabetes.Materials and Methods: A total of 60 Sprague‑Dawley rats were divided into three groups: Group I: The group without diabetes; Group II: The group with STZ‑induced diabetes; Group III: The group with STZ‑induced diabetes receiving insulin therapy. All animals received anesthesia, and 3/0 silk suture was inserted around the mandibular molar teeth. All groups were divided into subgroups receiving saline (Groups IA, IIA, IIIA) and α‑tocopherol injection (Groups IB, IIB, IIIB). After a period of 3 weeks, all rats were sacrificed, and the number of gingival iNOS positive cells was analyzed using image analysis software.Results: Applying α‑tocopherol suppressed the number of gingival iNOS positive cells in Groups IB, IIB, and IIIB compared to application of saline (Groups IA, IIA, and IIIA) (P < 0.05). Numbers of gingival iNOS positive cells were found to be similar in the Groups I and III (P > 0.05).Conclusions: Within limitations of the current study, this is the first study one may suggest that α‑tocopherol may reduce oxidative damage in the gingiva of the rats with periodontitis with or without STZ‑induced diabetes and increase effects of insulin.Keywords: Diabetes, experimental periodontitis, inducible nitric oxide synthase, α‑tocophero

Etanercept treatment in the endotoxin-induced uveitis of rats

This study was conducted to investigate therapeutic value of a soluble tumor necrosis factor-alpha (TNF-alpha) receptor, etanercept, in a rat model of endotoxin-induced uveitis (EIU). Forty-two inbred male Lewis rats were divided into seven equal groups. 200 mug of Escherichia coli 055:1355 lipopolysaccharide, (LPS) was injected in one hind footpad of the Groups 2, 3, 4, 5, 6, and 7 rats. Group 5, 6, and 7 rats also received subcutaneous etanercept 24 hr prior to LPS injection at a dose of 0(.)4 mg kg(-1). Group 1 rats were used as controls. Eight, 24, and 48 hr after treatment clinical uvetis scores (miosis, iris hyperemia, and hypopyon) were assessed by a masked observer and the rats were euthanized. Neutrophil leukocytes, CD8 +, CD4 +, and CD45RO + cells in the anterior uveal tissue were counted either after hematoxylin-eosin or monoclonal antibody staining. TNF-alpha. levels were also measured in the aqueous humor samples by an ELISA method. Etanercept treatment significantly improved clinical uveitis scores at all examination points compared to the LPS injected animals. The improvement was almost complete expect for the miosis score, since no significant difference was detected between the controls and LPS + Etanercept treated animals at all examination points. Cell counts were also at significantly lower levels in LPS + Etanercept treated animals at all examination points, except for CD8 + and CD45RO + cell counts at 24 hr examination point. There was no significant difference between the controls and LPS + Etanercept treated animals at all examination points as with CD4 + and CD45RO + cell counts at 48 hr. Our data showed that etanercept had a definite effect on the treatment of EIU. Further studies should clarify its efficacy on clinical uveitis conditions. (C) 2004 Elsevier Ltd. All rights reserved

Immunohistochemical analysis of orbital connective tissue specimens of patients with active graves ophthalmopathy

Purpose: To explore the immune mechanism of Graves ophthalmopathy (GO) by analyzing infiltrating cells in orbital connective tissue (OCT) specimens of patients with active GO using immunohistochemical methods. Methods: Five OCT specimens obtained from patients with active GO and five control specimens obtained from forensic cadavers who died from nonmedical reasons were stained with anti-CD3, CD4, CD8, CD45RO, HLA-Dr, CD25, and TNF-alpha monoclonal antibodies. Positively stained cells were counted and results were interpreted as cell counts/mm(2). Four of five GO patients had never been treated with any immunomodulating therapy. Only one had received oral prednisolone prior to tissue sampling, but this treatment had ceased 5 months before surgery. Results: The retro-orbital tissue specimens obtained from forensic cadavers did not show any significant positive staining for any monoclonal antibody tested. However, the specimens from GO patients showed positively stained means of 36.66 +/- 4.61 HLA- Dr(+), 12.8 +/- 3.42 CD8(+), 11.8 +/- 1.78 CD4(+), 16.6 +/- 1.81 CD3(+), 21.2 +/- 3.12 CD45RO(+), 10.4 +/- 2.07 TNF-alpha(+), 7.2 +/- 1.48 CD25(+), 3.2 +/- 1.09 CD4(+) CD8(+), 4.6 +/- 1.67 CD4(+) CD45RO(+), 2.8 +/- 0.83 CD8(+) CD45RO(+), 1.6 +/- 0.89 CD4(+) CD25(+), and 1.8 +/- 1 0.83 CD8(+) CD25(+) cells/ mm(2). Conclusions: Our study supports that most of the infiltrating lymphocytic cells in the active stage of GO are T cells, and a significant proportion of them are CD45RO+ cells. Infiltration of OCT by HLA- Dr+, CD25+, and TNF-alpha cells suggests that Th1-type immune reaction with the interference of proinflammatory cytokine(s) (TNF-alpha) may be important in the pathogenesis of disease. Further studies are needed to understand the disease pathogenesis and may provide a scientific basis for future treatment alternatives for the disease (e. g., anticytokine treatment)

immunohistochemical evaluation

Objective: Although it has been shown that rifamycin is an effective agent for bone graft decontamination, no information exists on the effects of rifamycin decontamination on bone graft incorporation. The aim of this study was to evaluate the influence of rifamycin decontamination on the incorporation of autologous onlay bone grafts quantitatively.Design: In 30 rats, a standardized 5.0-mm-diameter bone graft was harvested from the right mandibular angle, contaminated with saliva, decontaminated with rifamycin solution, and augmented to the left as an onlay graft. Ten animals were sacrificed at 7, 14, and 21 days after surgery. In the control group (10 rats), the onlay grafts were neither contaminated nor decontaminated, and the rats were sacrificed at 21 days after surgery. Histological slides were prepared from each grafted site for both immunohistochemistry analysis (bone morphogenetic protein-2 (BMP-2) and vascular endothelial growth factor (VEGF) antibodies) and histometric analysis. Images obtained from the graft incorporation area with the light microscope were transferred to a PC, and theywere evaluated using Clemex PE 3.5 image analysis software.Results: The grafts were fully incorporated in all specimens. The results showed that rifamycin decontamination has no detrimental effect on graft incorporation and the findings revealed a tendency for earlier revascularization and osteogenesis in the decontamination group. Data were analyzed using variance analysis and Tukey's test.Conclusions: Rifamycin decontamination has no detrimental effect on autogenous graft incorporation, and it can be used for graft decontamination with confidence. (C) 2014 Elsevier Ltd. All rights reserved