L-glutathione 1% promotes neuroprotection of nitrergic neurons and reduces the oxidative stress in the jejunum of rats with Walker-256-bearing tumor.

AIMS: Our main goals were to investigate the effects of L-glutathione (1%) treatment in Walker-256 tumor-bearing rats by analyzing immunoreactive neurons (IR), responsive to the nNOS enzyme and 3-Nitrotyrosine, in their jejunum myenteric plexus. Moreover, the oxidative state and inflammatory process in these animals were investigated. METHODS: Four experimental groups were utilized: control (C), control treated with L-glutathione (CGT), Walker-256 tumor-bearing rats (TW), and Walker-256 tumor-bearing rats treated with L-glutathione (TWGT). After 14 days of tumor inoculation, the jejunum was collected for immunohistochemical techniques and assessment of oxidative status. Plasma was collected to evaluate oxidative status and measure cytokines. RESULTS: The TW group exhibited a decrease of reduced glutathione in their jejunum, which was prevented in the L-glutathione treated TWGT group. TW animals presented pronounced oxidative stress by increasing levels of lipoperoxidation in their jejunum and malondialdehyde in their plasma; however, the L-glutathione treatment in TWGT group was not able to avoid it. The total antioxidant capacity was altered in groups TW and TWGT, yet the last one had a better index in their plasma. The IL-10, and TNF-α levels increased in TWGT animals. The nNOS-IR neuron density decreased in the jejunum myenteric plexus of the TW group, which was avoided in the TWGT group. The nNOS +3-Nitrotyrosine neurons quantification did not show significative alterations. CONCLUSION: The treatment with L-glutathione (1%) imposed an important defense to some parameters of oxidative stress induced by TW-256, leading to neuroprotection to the loss in the nNOS-IR neuron density.

Oxidative stress markers in the saliva of patients with oral lichen planus.

OBJECTIVE: To evaluate the levels of oxidative stress markers in the saliva of patients with oral lichen planus (OLP). METHODS: A cross-sectional study was conducted with 22 patients diagnosed both clinically and histologically with OLP (reticular or erosive) and 12 individuals without OLP. Non-stimulated sialometry was performed and oxidative stress (myeloperoxidase - MPO and malondialdehyde - MDA) and antioxidant (superoxide dismutase - SOD and glutathione - GSH) markers were determined in the saliva. RESULTS: Among the patients with OLP, most were women (n = 19; 86.4%) and reported to have experienced menopause (63.2%). Patients with OLP were mostly in the active stage of the disease (n = 17; 77.3%) and the reticular form was predominant (n = 15; 68.2%). No statistically significant differences were observed when comparing SOD, GSH, MPO and MDA values between individuals with and without OLP, as well as between erosive and reticular forms of OLP (p > 0.05). Patients with inactive OLP presented higher SOD when compared to those with active disease (p = 0.031). CONCLUSION: Oxidative stress markers in the saliva of patients with OLP were similar to those found in people without OLP, which can be related to the high exposure of the oral cavity environment to several physical, chemical and microbiological stimuli, important generators of the oxidative stress.

Effect of p-cymene and rosmarinic acid on gastric ulcer healing - Involvement of multiple endogenous curative mechanisms.

BACKGROUND: p-Cymene and rosmarinic acid are secondary metabolites found in several medicinal plants and spices. Previous studies have demonstrated their anti-inflammatory, antioxidant, and cytoprotective effects. PURPOSE: To evaluate their gastroduodenal antiulcer activity, gastric healing and toxicity in experimental models. METHODS: Preventive antiulcer effects were assessed using oral pre-treatment on HCl/ethanol-induced gastric lesions and cysteamine-induced duodenal lesions models. Gastric healing, the underlining mechanisms and toxicity after repeated doses were carried out using the acetic acid-induced gastric ulcer rat model and oral treatment for 14 days. RESULTS: In the HCl/ethanol-induced gastric ulcer and cysteamine-induced duodenal injury, p-cymene and rosmarinic acid (50-200 mg/kg) decreased significantly the ulcer area, and so prevented lesions formation. In the acetic acid-induced ulcer model, both compounds (200 mg/kg) markedly reduced the ulcerative injury. These effects were related to an increase in the levels of reduced glutathione (GSH) and interleukin (IL)-10, and due to a decrease in malondialdehyde (MDA), IL-1ß, tumor necrosis factor (TNF)-α, total and mitochondrial reactive oxygen species (ROS) levels. Downregulation of factor nuclear kappa B (NFκB) and enhanced expression of suppressor of cytokine signaling (SOCS)3 were also demonstrated. Furthermore, positive vascular endothelial growth factor (VEGF), metalloproteinase (MMP)-2, and cyclooxygenase (COX-2)-stained cells were increased in treated groups. Treatment also upregulated the platelet-derived growth factor (PDGF), basic fibroblast growth factor (bFGF), transforming growth factor (TGF)-ß and epidermal growth factor receptor (EGFR) in gastric tissues. In isolated gastric epithelial cells this healing effect seems to be linked to a modulation of apoptosis, proliferation, survival and protein phosphorylation, such as the extracellular signal-regulated kinases (ERK)1/2 and p38 mitogen-activated protein kinase (MAPK). Oral toxicity investigation for 14 days revealed no alterations in heart, liver, spleen, and kidneys weight nor the biochemical and hematological assessed parameters. p-Cymene and rosmarinic acid also protected animals from body weight loss maintaining feed and water intake. CONCLUSIONS: Data altogether suggest low toxicity, antiulcer and gastric healing activities of p-cymene and rosmarinic acid. Antioxidant and immunomodulatory properties seem to be involved in the curative effect as well as the induction of different factors linked to tissue repair.

Effect of Dexamethasone-Loaded PLGA Nanoparticles on Oral Mucositis Induced by 5-Fluorouracil.

Oral mucositis (OM) is characterized by the presence of severe ulcers in the oral region that affects patients treated with chemotherapy. It occurs in almost all patients who receive radiotherapy of the head and neck, as well as patients who undergo hematopoietic cell transplantation. The pathophysiology of OM is complex, and there is no effective therapy. The aim of this study was to evaluate the effect of dexamethasone-loaded poly(d,l-Lactic-co-glycolic) nanoparticles (PLGA-DEX NPs) on an OM model induced in hamsters. The NPs were synthesized using the emulsification-solvent evaporation method and were characterized by the size, zeta potential, encapsulation efficiency, atomic force microscopy, physicochemical stability, and the in vitro release. The OM was induced by the administration of 5-FU on the first and second days and mechanical trauma on the 4th day of the experiment. PLGA-DEX NPs were administered to treat OM. The animals were euthanized on the 10th day. Macroscopic and histopathological analyses were performed, measurement of malonaldehyde (MDA) and ELISA was used to determine the levels of IL-1ß and TNF-α. Immunoexpressions of NF-κB, COX-2, and TGF-ß were determined by immunohistochemistry, and qRT-PCR was used to quantify the gene expression of the GILZ, MKP1, and NF-κB p65. The PLGA-DEX NPs (0.1 mg/kg) significantly reduced macroscopic and histopathological scores, decreased MDA, TNF-α and IL-1ß levels, immunostaining for NF-κB, COX-2, TGF-ß, and suppressed NF-κB p65 mRNA expression, but increased GILZ and MKP1 expression.

p-Cymene and Rosmarinic Acid Ameliorate TNBS-Induced Intestinal Inflammation Upkeeping ZO-1 and MUC-2: Role of Antioxidant System and Immunomodulation.

p-Cymene (p-C) and rosmarinic acid (RA) are secondary metabolites that are present in medicinal herbs and Mediterranean spices that have promising anti-inflammatory properties. This study aimed to evaluate their intestinal anti-inflammatory activity in the trinitrobenzene sulphonic acid (TNBS)-induced colitis model in rats. p-C and RA (25-200 mg/kg) oral administration reduced the macroscopic lesion score, ulcerative area, intestinal weight/length ratio, and diarrheal index in TNBS-treated animals. Both compounds (200 mg/kg) decreased malondialdehyde (MDA) and myeloperoxidase (MPO), restored glutathione (GSH) levels, and enhanced fluorescence intensity of superoxide dismutase (SOD). They also decreased interleukin (IL)-1ß and tumor necrosis factor (TNF)-α, and maintained IL-10 basal levels. Furthermore, they modulated T cell populations (cluster of differentiation (CD)4+, CD8+, or CD3+CD4+CD25+) analyzed from the spleen, mesenteric lymph nodes, and colon samples, and also decreased cyclooxigenase 2 (COX-2), interferon (IFN)-γ, inducible nitric oxide synthase (iNOS), and nuclear transcription factor kappa B subunit p65 (NFκB-p65) mRNA transcription, but only p-C interfered in the suppressor of cytokine signaling 3 (SOCS3) expression in inflamed colons. An increase in gene expression and positive cells immunostained for mucin type 2 (MUC-2) and zonula occludens 1 (ZO-1) was observed. Altogether, these results indicate intestinal anti-inflammatory activity of p-C and RA involving the cytoprotection of the intestinal barrier, maintaining the mucus layer, and preserving communicating junctions, as well as through modulation of the antioxidant and immunomodulatory systems.

Estragole prevents gastric ulcers via cytoprotective, antioxidant and immunoregulatory mechanisms in animal models.

BACKGROUND: Estragole is an aromatic organic compound belonging to the class of phenylpropanoids derived from cinnamic aldehydes and present in essential oils of plant species, such asRavensara anisata (madeira), Ocimum basilicum (manjericão/alfavaca) and Croton zehntneri (canelinha). Pharmacological studies report its anti-inflammatory, antioxidant and vasorelaxant activity. HYPOTHESIS/PURPOSE: This study aimed to evaluate the acute non-clinical toxicity, gastroprotective activity and the related mechanisms of action. METHODS: Acute toxicity was assessed according to OECD guide 423 in mice. Ethanol, stress, piroxicam and pylorus ligation-induced gastric ulcer models were used to investigate antiulcer properties. The related mechanisms of action were using the ethanol-gastric lesions protocol. RESULTS: In the acute oral toxicity assay, doses of 300 or 2000 mg/kg of estragole administered orally in Swiss mice did not induce any behavioral changes. However, the dose of 2000 mg/kg showed a decrease in water and feed intake. Lethal dose 50 % (LD50) was set to be equal to or greater than 2500 mg/kg, according to OECD. In all evaluated protocols, estragole (31.25, 62.5, 125 and 250 mg/kg) significantly reduced the area of ​​ulcerative lesion when compared to control groups. To investigate the mechanisms involved in the gastroprotective activity, the antisecretory or neutralizing of gastric secretion, cytoprotectant, antioxidant and immunoregulatory effects were evaluated. Results showed that treatment with estragole (250 mg/kg) reduced (p < 0.05) the volume of the gastric juice. Besides, sulfhydryl groups, nitric oxide, mucus and prostaglandins seems to be involved in the gastroprotective property. Treatment also increased (p < 0.001) levels of reduced glutathione (GSH), interleukin-10 (IL-10) and positive cells marked for glutathione peroxidase (GPx) and cyclooxygenase 2 (COX-2). It also reduced (p < 0.001) malondialdehyde (MDA), myeloperoxidase (MPO), interleukin-1 beta (IL-1ß), tumor necrosis factor-alpha (TNF-α) and inducible nitric oxide synthase (iNOS) (p < 0.05) levels. CONCLUSION: Thus, it is possible to infer that estragole presents gastroprotective activity related to antisecretory, cytoprotective, antioxidant and immunomodulatory mechanisms.

Efficacy of antimicrobial photodynamic therapy with chloro-aluminum phthalocyanine on periodontal clinical parameters and salivary GSH and MDA levels in patients with periodontitis.

The purpose of this study was to evaluate the efficacy of antimicrobial photodynamic therapy (aPDT) with chloro-aluminum phthalocyanine (AlClPc) on several periodontal parameters includingsalivary glutathione (GSH) and MDA (malondialdehyde) levels in periodontal sites presenting with periodontitis. Randomized clinical trial, comprising 40 test group (TG) sites and 23 control group (CG) sites. The TG was treated with scaling and root planning (SRP) and aPDT, and CG, only with SRP. Visible plaque index [VPI], gingival bleeding index [GBI], bleeding on probing [BOP], probing depth [PD] and clinical attachment level [CAL] were calculated and saliva samples were taken at baseline (T0), three (T3) and six months (T6). Data was analyzed by the Wilcoxon and Mann-Whitney tests. An intragroup analysis indicated significant differences at T3 and T6 for GBI, CAL and GSH only in the CG (p < 0.05). For BOP, a significant decrease was observed only in the TG between T0 and T6 (p = 0.008). No significant differences were observed for VPI, BOP and MDA. In the intergroup analysis, significant differences were observed for GBI at T6 (p = 0.041), and for GSH at T3 (p = 0.031), being higher in the TG. Although aPDT with AlClPc did not present statistically proven benefits, but the employed periodontal treatment resulted in decreased BOP, PD, CAL and MDA after 3 and 6 months of treatment. In addition, the lower need for glutathione production may initially suggest an additional benefit of AlClPc aPDT in the early reestablishment of the balance between oxidative and non-oxidative agents related to oxidative stress.

Gliclazide Prevents 5-FU-Induced Oral Mucositis by Reducing Oxidative Stress, Inflammation, and P-Selectin Adhesion Molecules.

Oral mucositis (OM) is one of the main side effects of the head and neck cancer treatment, particularly radiotherapy and/or chemotherapy. OM is characterized by ulcers, erythema, dysphagia, xerostomia, and increased susceptibility to opportunistic infections. In the perspective of finding pharmacological therapies to prevent inflammation and ulceration of OM, the investigation of the pleiotropic effect of commercial drugs is needed, among them gliclazide, an antidiabetic drug. This study aimed to evaluate the effect of gliclazide in an experimental OM model induced by 5-fluorouracil. Male hamsters were pre-treated with oral gliclazide (1, 5, or 10 mg/kg) for 10 days. Cheek pouch samples were subjected to histopathological and immunohistochemical analysis (COX2, iNOS, MMP-2, NFκB P65, GPx) and imunofluorescence (P-selectin). IL-1ß and TNF-α levels, Myeloperoxidase activity (MPO) and malondialdehyde (MDA) levels were investigated by ultraviolet-visible spectroscopy analysis. NFκB NLS P50 protein levels were analyzed by western blotting. The group treated with gliclazide at a dose of 10 mg/kg showed presence of erythema, no evidence of erosion, and absence of mucosal ulceration with a score of 1 (1-2) (p < 0.01). Histopathological data for the group treated with gliclazide 10 mg/kg showed re-epithelialization, discrete mononuclear inflammatory infiltrate and absence of hemorrhage, edema, ulcers and abscesses with a score of 1 (1-1) (p < 0.01). Treatment with gliclazide 10 mg/kg reduced MPO activity (p < 0.001), MDA levels (p < 0.001) and NFκB NLS P50 (p < 0.05) protein levels, resulting in low immunostaining to Cox-2, iNOS (p < 0.05), NFκB P65 (p < 0.05), and negative immunoreaction to MMP-2 (p < 0.001). However, it appeared that for Gpx1, the staining was restored in the GLI 10-FUT group compared with 5FUT/saline (p < 0.05). Immunofluorescence revealed decreased levels of P-selectin (p < 0.001) after treatment with gliclazide 10 mg/kg (p < 0.05). In summary, gliclazide accelerated mucosal recovery and reduced oxidative stress and inflammation in the 5-FU-induced OM in hamsters.

Crude extract from Libidibia ferrea (Mart. ex. Tul.) L.P. Queiroz leaves decreased intra articular inflammation induced by zymosan in rats.

BACKGROUND: Libidibia ferrea (L. ferrea) has been used in folk medicine to treat several conditions and to prevent cancer. This study performed a chromatographic analysis of the crude aqueous extract of Libidibia ferrea (Mart. ex. Tul.) L.P. Queiroz (LfAE) leaves and evaluated its in vivo antioxidant and anti-inflammatory potential. METHODS: Polyphenols present in LfAE were characterized by high performance liquid chromatography (HPLC). Anti-inflammatory activity was studied in an experimental model of zymosan-induced intra-articular inflammation, conducted in Wistar rats treated with LfAE at the doses of 100, 200 and 300 mg/kg by gavage. Synovial fluid was collected for global leukocyte count, for spectrocopical UV/VIS analysis of myeloperoxidase (MPO) activity, total glutathione and malondialdehyde (MDA), and for quantification of inflammatory cytokines IL1-ß and TNF-α by enzyme-linked immunosorbent assay. Synovial membrane was collected for histological analysis. The level of statistical significance was p < 0.05. RESULTS: HPLC detected concentrations of 1.56 (0.77) %m/m for ellagic acid and 1.20 (1.38) %m/m for gallic acid in LfAE leaves. Treatment with LfAE at all doses significantly decreased the leukocyte influx into the synovial fluid (p < 0.001) and myeloperoxidase activity (p < 0.001), an important marker of neutrophils. LfAE at doses of 100 (p < 0.05), 200 and 300 mg/kg (p < 0.001) also reduced the levels of MDA. LfAE at doses of 200 and 300 mg/kg significantly decreased the levels of IL-1ß (p < 0.05) and TNF-α (p < 0.001). All doses of LfAE resulted in increased levels of total glutathione (p < 0.001). Histopathological findings confirmed a reduction of the inflammatory infiltrate in the rats treated with LfAE at a dose of 200 mg/kg (p < 0.05). CONCLUSION: LfAE has an important anti-oxidant and anti-inflammatory effect on intra-articular inflammation.

Intestinal Anti-Inflammatory Activity of the Aqueous Extract from Ipomoea asarifolia in DNBS-Induced Colitis in Rats.

Inflammatory bowel disease is triggered by an uncontrolled immune response associated with genetic, environmental, and intestinal microbiota imbalance. Ipomoea asarifolia (IA), popularly known as "salsa" or "brave salsa", belongs to the Convolvulaceae family. The aim of this approach was to study the preventive effect of IA aqueous extract in 2,4-dinitrobenzene sulfonic acid (DNBS)-induced colitis in rats. Rats pretreated with IA extract or sulfasalazine (SSZ) received intracolonic instillation of DNBS in 50% ethanol (v/v). IA extract presented a protective effect against intestinal inflammation, with improvement in the disease activity index and macroscopic damage. IA or SSZ significantly reduced myeloperoxidase activity, and also down-regulation of the gene expression of JNK1, NF-κß-p65, STAT3, and decreased levels of TNFα, IL-1ß, and increased IL-10, associated with a significant improvement of oxidative stress, in addition to a reduction in MDA and an increase of glutathione in colonic tissue. The protective effect of the extract was also confirmed in histological evaluation, showing preservation of the colonic cytoarchitecture. Immunohistochemical analysis revealed down-regulation of NF-κß-p65, iNOS, IL-17, and up-regulation of SOCs-1 and MUC-2. IA extract presents antioxidant and anti-inflammatory intestinal properties, and proved to be a potential application for preventing damage induced by DNBS.

Photodynamic therapy using chloro-aluminum phthalocyanine decreases inflammatory response in an experimental rat periodontal disease model.

BACKGROUND AND OBJECTIVE: Emerging evidence suggests that photodynamic therapy (PDT) can exhibit immunomodulatory activity. The purpose of the present study was to analyse cytokine profiles after application of PDT in gingival tissues of rats with ligature-induced periodontal disease (PD). STUDY DESIGN/MATERIAL AND METHODS: Periodontal disease was induced through the introduction of a cotton thread around the first left mandibular molar, while the right side molars did not receive ligatures. After 7days of PD evolution, ligatures were removed from the left side, and the animals were randomically divided into the following treatment groups: I, rats without treatment; II, rats received chloro-aluminum phthalocyanine (AlClPc); III, rats received low-level laser alone; and IV, rats received AlClPc associated with low-level laser (PDT). The animals were killed 7days after the treatments, and the mandibles were histologically processed to assess morphological and immunohistochemical profile, while gingival tissues were removed for quantification of tumor necrosis factor (TNF)-α, interleukin (IL-)1ß and IL-10 expression (by ELISA). RESULTS: Histomorphological analysis of periodontal tissues demonstrated that PDT-treated animals show tissue necrosis, as well as lower TNF- α expression, compared to ligatured animals treated with AlClPc alone. CONCLUSIONS: It was concluded that PDT using AlClPc entrapped in a lipid nanoemulsion may be useful in therapies, because of immunomodulatory effects that decreased the inflammatory response and cause tissue destruction.

Efeitos da terapia fotodinâmica com aluminio - cloro ftalocianina sobre a peroxidação lipídica e produtos antioxidantes em tecidos inflamados animais / Effects of Photodynamic therapy aluminum - chloro phthalocyanine on lipid peroxidation products and antioxidant in animal tissues inflamed

A terapia fotodinâmica (TFD) envolve a administração de um agente fotossensibilizador (FS) seguida pela aplicação do laser, com comprimento de onda adequado, resultando em uma sequência de processos fotoquímicos e fotobiológicos, que geram espécies reativas de oxigênio (EROs). Este estudo avaliou, os efeitos da TFD com nanoemulsão de alumínio-cloro ftalocianina (AlClFc) sobre os níveis de malondildeído (MDA), glutationa (GSH), superóxido dismutase (SOD) e glutationa peroxidase (GPx), que representam indicadores envolvidos no estresse oxidativo e defesas antioxidantes. Para tanto, o estudo utilizou 120 ratas da espécie Rattus norvegicus, raça Wistar, distribuídas em 5 grupos experimentais: Saudáveis (S), com doença periodontal (DP), com doença periodontal e tratamento com o FS (F), com doença periodontal e tratamento com o laser (L) e com doença periodontal e tratamento com TFD (FL). Foi utilizado um modelo experimental de doença periodontal (DP) induzida por ligadura. Após sete dias, da indução da DP, foram instituídos os tratamentos, conforme os grupos. No grupo tratado, com TFD, foi aplicado 40μl do FS (5μM), seguida da irradiação do laser diodo fosfeto de índio-gálio-alumínio (InGaAlP - 660nm, 100J/cm2). As ratas sofreram eutanásia no 7º e no 28º dia, após o tratamento. Os espécimes teciduais foram removidos para análises histológicas, ensaios bioquímicos e imuno-histoquímica. Os resultados histológicos exibiram, alterações inflamatórias, desorganização do tecido conjuntivo e perda óssea alveolar, nos grupos com DP induzida. Os ensaios demonstraram, que os níveis de MDA estavam mais elevados, nos grupos com DP induzida, que no grupo saudável. Não existindo diferenças estatísticas significantes (p>0,05). Níveis elevados de GSH foram encontrados nas ratas saudáveis, com diferenças estatísticas significativas, nos grupos L (p=0,028) e FL (p=0,028), quando comparados com o grupo DP. Imuno-histoquimicamente, a SOD apresentou maior imunomarcação nos grupos L e FL, comparados ao grupo saudável. Sem diferenças estatísticas significativas (p>0,05). GPx mostrou imunomarcação significativamente menor no grupo DP, comparado ao saudável (p=0,05) e no grupo F, comparado ao DP (p<0,05). Os resultados apresentados sugerem discreta participação da TFD mediada por nanoemulsão contendo AlClFc no estresse oxidativo. O protocolo utilizado, neste experimento, demonstrou maior influência da TFD sobre os mecanismos antioxidantes.

Photodynamic therapy (PDT) consists of a non-toxic photosensitizing agent (FS) administration followed by a laser source resulting in a sequence of photochemical and photobiological processes that generate reactive oxygen species (ROS) that damaging cells. The present work evaluated the effects of PDT nanoemulsion-aluminum chloride phthalocyanine (AlClFc) mediated on malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels, which represent indicators involved in oxidative stress and antioxidant defenses. For this purpose, this study used 120 female rats of the Rattus norvegicus species, Wistar race, divided into 5 groups: Healthy (H), with periodontal disease (PD), with periodontal disease and treatment with FS (F), with periodontal disease and treatment with the laser (L); and periodontal disease and treatment with PDT (FL). An experimental model for represent periodontal disease (PD) was induced by ligature (split-mouth). Seven days later the induction of PD, the treatments were instituted according to the groups. In the group treated with PDT was applied 40μl FS (5μM) followed by laser irradiation diode InGaAlP (660nm, 100J / cm2). The rats were sacrificed on the 7th and 28th day after treatment and tissue specimens were removed and subjected to histological, immunohistochemical methods and enzymatic colorimetric measurements with detection by UV / VIS spectroscopy. Inflammatory changes, connective tissue disorganization and alveolar bone loss were displaying in groups with PD induced. The enzyme dosages showed that MDA levels were higher in PD induced groups, with no statistically significant differences (p> 0.05). High levels of GSH were found in groups L (p = 0.028) and FL (p = 0.028) compared with PD group, with statistically significant differences. Immunohistochemistry for SOD showed higher immunostaining in L and FL groups, compared to the PD group without statistically significant differences (p> 0.05). GPx showed lower immunoreactivity in the DP group when compared to the other groups and statistically significant differences were observed between the DPxL groups (p <0.05). TFD administered in this experiment did not induce elevation of MDA levels significantly increased the GSH levels and showed intense immunostaining pada SOD and GPx, showing that this therapy does not accentuated lipid peroxidation, however, it was able to induce effects on the antioxidant defenses processes. The LBI therapy appeared to show photomodulatory promoting effects reduction of the MDA levels, increasing GSH levels and with intense immunostaining for SOD and GPx, demonstrating that laser therapy induced antioxidant effects.

Efeitos da terapia fotodinâmica com aluminio – cloro ftalocianina sobre a peroxidação lipídica e produtos antioxidantes em tecidos inflamados animais / Effects of Photodynamic therapy aluminum - chloro phthalocyanine on lipid peroxidation products and antioxidant in animal tissues inflamed

A terapia fotodinâmica (TFD) envolve a administração de um agente fotossensibilizador (FS) seguida pela aplicação do laser, com comprimento de onda adequado, resultando em uma sequência de processos fotoquímicos e fotobiológicos, que geram espécies reativas de oxigênio (EROs). Este estudo avaliou, os efeitos da TFD com nanoemulsão de alumínio-cloro ftalocianina (AlClFc) sobre os níveis de malondildeído (MDA), glutationa (GSH), superóxido dismutase (SOD) e glutationa peroxidase (GPx), que representam indicadores envolvidos no estresse oxidativo e defesas antioxidantes. Para tanto, o estudo utilizou 120 ratas da espécie Rattus norvegicus, raça Wistar, distribuídas em 5 grupos experimentais: Saudáveis (S), com doença periodontal (DP), com doença periodontal e tratamento com o FS (F), com doença periodontal e tratamento com o laser (L) e com doença periodontal e tratamento com TFD (FL). Foi utilizado um modelo experimental de doença periodontal (DP) induzida por ligadura. Após sete dias, da indução da DP, foram instituídos os tratamentos, conforme os grupos. No grupo tratado, com TFD, foi aplicado 40μl do FS (5μM), seguida da irradiação do laser diodo fosfeto de índio-gálio-alumínio (InGaAlP - 660nm, 100J/cm2). As ratas sofreram eutanásia no 7º e no 28º dia, após o tratamento. Os espécimes teciduais foram removidos para análises histológicas, ensaios bioquímicos e imuno-histoquímica. Os resultados histológicos exibiram, alterações inflamatórias, desorganização do tecido conjuntivo e perda óssea alveolar, nos grupos com DP induzida. Os ensaios demonstraram, que os níveis de MDA estavam mais elevados, nos grupos com DP induzida, que no grupo saudável.

Não existindo diferenças estatísticas significantes (p>0,05). Níveis elevados de GSH foram encontrados nas ratas saudáveis, com diferenças estatísticas significativas, nos grupos L (p=0,028) e FL (p=0,028), quando comparados com o grupo DP. Imuno-histoquimicamente, a SOD apresentou maior imunomarcação nos grupos L e FL, comparados ao grupo saudável. Sem diferenças estatísticas significativas (p>0,05). GPx mostrou imunomarcação significativamente menor no grupo DP, comparado ao saudável (p=0,05) e no grupo F, comparado ao DP (p<0,05). Os resultados apresentados sugerem discreta participação da TFD mediada por nanoemulsão contendo AlClFc no estresse oxidativo. O protocolo utilizado, neste experimento, demonstrou maior influência da TFD sobre os mecanismos antioxidantes. (AU)

Photodynamic therapy (PDT) consists of a non-toxic photosensitizing agent (FS) administration followed by a laser source resulting in a sequence of photochemical and photobiological processes that generate reactive oxygen species (ROS) that damaging cells. The present work evaluated the effects of PDT nanoemulsion-aluminum chloride phthalocyanine (AlClFc) mediated on malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD) and glutathione peroxidase (GPx) levels, which represent indicators involved in oxidative stress and antioxidant defenses. For this purpose, this study used 120 female rats of the Rattus norvegicus species, Wistar race, divided into 5 groups: Healthy (H), with periodontal disease (PD), with periodontal disease and treatment with FS (F), with periodontal disease and treatment with the laser (L); and periodontal disease and treatment with PDT (FL). An experimental model for represent periodontal disease (PD) was induced by ligature (split-mouth). Seven days later the induction of PD, the treatments were instituted according to the groups. In the group treated with PDT was applied 40μl FS (5μM) followed by laser irradiation diode InGaAlP (660nm, 100J / cm2). The rats were sacrificed on the 7th and 28th day after treatment and tissue specimens were removed and subjected to histological, immunohistochemical methods and enzymatic colorimetric measurements with detection by UV / VIS spectroscopy. Inflammatory changes, connective tissue disorganization and alveolar bone loss were displaying in groups with PD induced. The enzyme dosages showed that MDA levels were higher in PD induced groups, with no statistically significant differences (p> 0.05). High levels of GSH were found in groups L (p = 0.028) and FL (p = 0.028) compared with PD group, with statistically significant differences. Immunohistochemistry for SOD showed higher immunostaining in L and FL groups, compared to the PD group without statistically significant differences (p> 0.05).

GPx showed lower immunoreactivity in the DP group when compared to the other groups and statistically significant differences were observed between the DPxL groups (p <0.05). TFD administered in this experiment did not induce elevation of MDA levels significantly increased the GSH levels and showed intense immunostaining pada SOD and GPx, showing that this therapy does not accentuated lipid peroxidation, however, it was able to induce effects on the antioxidant defenses processes. The LBI therapy appeared to show photomodulatory promoting effects reduction of the MDA levels, increasing GSH levels and with intense immunostaining for SOD and GPx, demonstrating that laser therapy induced antioxidant effects (AU)

Immunoexpression of HIF-1α and VEGF in Periodontal Disease and Healthy Gingival Tissues.

Hypoxia-inducible factor 1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF) are proteins that stimulate the proliferation and migration of endothelial cells. These proteins have been described in many pathologic and inflammatory conditions, but their involvement in the development of periodontitis has not been thoroughly investigated. This study compared the immunohistochemical expression of these proteins, involved in angiogenesis and hypoxia, by immunostained inflammatory and endothelial cells in periodontal disease and healthy gingival tissues. Gingival tissue samples were divided as follows: 30 samples with chronic periodontitis, 30 with chronic gingivitis, and 30 of healthy gingiva. Results were analyzed statistically by the Kruskal-Wallis, Mann-Whitney and Spearman correlation tests (p=0.01). Inflammatory and endothelial cells were found to express these proteins. Periodontitis showed median percentage of HIF-1α-positive cells of 39.6%, 22.0% in cases of gingivitis and 0.9% in the healthy gingiva group (p=0.001). For VEGF, median percentage of immunopositive cells was 68.7% for periodontitis, 66.1% in cases for gingivitis, and 19.2% for healthy gingival specimens (p<0.001). Significant correlation between VEGF and HIF-1α was also observed in healthy gingiva (p<0.001).The increased expression of HIF-1α and VEGF in periodontitis, compared to gingivitis and healthy gingiva, suggests possible activation of the HIF-1α pathway in advanced periodontal disease. The correlation between HIF-1α and VEGF expression in healthy gingiva suggests a physiological function for these proteins in conditions of homeostasis. In periodontal disease, HIF-1α and VEGF expression may be regulated by other factors, in addition to hypoxia, such as bacterial endotoxins and inflammatory cytokines.

Immunoexpression of HIF-1α and VEGF in Periodontal Disease and Healthy Gingival Tissues

Abstract Hypoxia-inducible factor 1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF) are proteins that stimulate the proliferation and migration of endothelial cells. These proteins have been described in many pathologic and inflammatory conditions, but their involvement in the development of periodontitis has not been thoroughly investigated. This study compared the immunohistochemical expression of these proteins, involved in angiogenesis and hypoxia, by imunnostained inflammatory and endothelial cells in periodontal disease and healthy gingival tissues. Gingival tissue samples were divided as follows: 30 samples with chronic periodontitis, 30 with chronic gingivitis, and 30 of healthy gingiva. Results were analyzed statistically by the Kruskal-Wallis, Mann-Whitney and Spearman correlation tests (p=0.01). Inflammatory and endothelial cells were found to express these proteins. Periodontitis showed median percentage of HIF-1α-positive cells of 39.6%, 22.0% in cases of gingivitis and 0.9% in the healthy gingiva group (p=0.001). For VEGF, median percentage of immunopositive cells was 68.7% for periodontitis, 66.1% in cases for gingivitis, and 19.2% for healthy gingival specimens (p<0.001). Significant correlation between VEGF and HIF-1α was also observed in healthy gingiva (p<0.001).The increased expression of HIF-1αα and VEGF in periodontitis, compared to gingivitis and healthy gingiva, suggests possible activation of the HIF-1α pathway in advanced periodontal disease. The correlation between HIF-1α and VEGF expression in healthy gingiva suggests a physiological function for these proteins in conditions of homeostasis. In periodontal disease, HIF-1 and VEGF expression may be regulated by other factors, in addition to hypoxia, such as bacterial endotoxins and inflammatory cytokines.

Resumo O fator induzível por hipóxia 1 alfa (HIF-1α) e o fator de crescimento endotelial vascular (VEGF) são proteínas que estimulam a proliferação e a migração de células endoteliais. Estas proteínas têm sido descritas em muitas condições patológicas e inflamatórias, mas o envolvimento dessas no desenvolvimento de periodontite não foi completamente investigado. Este estudo comparou a expressão imunohistoquímica destas proteínas, envolvidas na angiogênese e hipóxia, na doença periodontal e em tecidos gengivais saudáveis por meio de contagem de células inflamatórias e endoteliais imunomarcadas. As amostras de tecido gengival foram divididas da seguinte forma: 30 amostras com periodontite crônica, 30 com gengivite crônica e 30 de gengiva saudável. Os resultados foram analisados estatisticamente pelos testes de Kruskal-Wallis e Mann-Whitney (p=0.01). As células inflamatórias e endoteliais foram positivas para estas proteínas. A porcentagem média de células positivas para HIF-1α foi de 39,6% nos casos de periodontite, 22,0% nos casos de gengivite e de 0,9% no grupo de gengiva saudável (p = 0,001). A porcentagem média de células imunopositivas para o VEGF foi de 68,7% nos casos de periodontite, 66,1% nos casos de gengivite, e 19,2% em espécimes gengivais saudáveis (p<0,001). Correlação significativa entre o VEGF e HIF-1α foi observada em gengival. A expressão elevada do HIF-1α e VEGF em periodontite, comparada a gengivite e gengiva saudável, sugere ativação da via do HIF-1α, na doença periodontal avançada. A correlação entre o HIF-1α e expressão de VEGF na gengiva saudável, sugere uma função fisiológica para estas proteínas em condições de homeostase. Na doença periodontal, a expressão de HIF-1α e VEGF pode ser regulada por outros fatores, além da hipóxia, tais como endotoxinas bacterianas e citocinas inflamatórias.

Immunohistochemical expression of GLUT-1, GLUT-3, and carbonic anhydrase IX in benign odontogenic lesions.

BACKGROUND: Some benign odontogenic lesions have a distinct biological behavior with high recurrence rates and local aggressive behavior. To determine whether glucose transporters proteins (GLUT-1 and GLUT-3) and carbonic anhydrase IX (CA IX) are associated with the development of as dentigerous cyst (DC), odontogenic keratocyst (OK), and ameloblastoma (AM), we evaluated the immunohistochemical expression of these proteins in these lesions. MATERIALS AND METHODS: Immunoexpression of GLUT-1, GLUT-3, and CA IX was evaluated semiquantitative fields in each of the 20 cases of OK, AM, and DC. The cases were classified according to the scores: 0 (0% positive cells), 1 (<10% of positive cells), 2 (10-50% of positive cells), and 3 (>50% of positive cells). The statistical analysis was performed using Pearson's chi-square, Kruskal-Wallis and Mann-Whitney tests. RESULTS: All cases were positive for GLUT-1 and 65% of OK showed scored 3. Staining was diffuse in 90% of OK and 85% of DC cases (P < 0.001). In 50% of OK and AM, staining was only observed in the membrane (P = 0.01). Most of the samples (66.7%) were negative for GLUT-3. Staining intensity for anhydrase was higher in the epithelium of DC when compared to OK (P = 0.01). Strong staining was observed in 55% of DC and 20% of OK samples (P = 0.01). CONCLUSIONS: These results suggest that GLUT-1 may be involved in the metabolic regulation of glucose in odontogenic lesions studied. In addition, CA IX appears to influence the development of AM, OK, and DC which can explain the differences their biological behavior.

Effects of photodynamic therapy mediated by nanoemulsion containing chloro-aluminum phthalocyanine: a histologic and immunohistochemical study in human gingiva.

BACKGROUND: Photodynamic therapy (PDT) uses photosensitizing agents, which are delivered in target cells, followed by local application of visible light in specific wavelengths. This reaction produce reactive oxygen species able to induce cell death by apoptosis or necrosis, injured to the local vasculature, and exert important effects on the immune system. OBJECTIVE: The present work evaluated the clinical findings, histomorphological alterations and immunodetection of VEGF after PDT using chloro-aluminum phthalocyanine (AlClPc) entrapped in a lipid nanoemulsion in a split-mouth clinical trial. MATERIAL AND METHODS: Eight healthy volunteers with clinical indication for extraction were included in the study. Seven days before the extraction 40 ul of nanoemulsion AlClPc 5µM was injected into gingival tissue followed by irradiation with diode laser, the contralateral side was used as control. Tissue specimens were removed seven days after the PDT and divided into two groups (test and control groups) for histological and immunohistochemical analysis. Patients were monitored at days, 0, 7, 14 and 30 to assess adverse effects of the therapy. RESULTS: The therapy was well tolerated by all patients. Adverse effects were short-time and completely reversible. Areas of edema, vascular congestion, and intense vascularization were viewed in gingival samples that received PDT. Additionally, dystrophic calcification was observed in subepithelial region. VEGF showed moderate to strong immunostaining in specimens subjected to PDT. CONCLUSIONS: Taken together, the results showed that the protocol used in this study mediated by nanoemulsion containing AlClPc is safe for clinical application in gingival tissue and suggests that VEGF is increased after PDT.