Effect of nitro-conjugated linoleic acid on the inflammatory response of murine macrophages activated with lipopolysaccharide derived from Prevotella intermedia.

Nitro-conjugated linoleic acid (NO2-CLA) has been observed to manifest salutary signaling responses, including anti-inflammatory and antioxidant properties. Here, the authors have explored the influence and underlying mechanisms of NO2-CLA on the proinflammatory reaction of murine macrophages that were challenged with lipopolysaccharide (LPS) derived from Prevotella intermedia, a putative periodontopathic bacterium. Treatment of LPS-activated RAW264.7 cells with NO2-CLA notably dampened the secretion of iNOS-derived NO, IL-1ß and IL-6 as well as their gene expressions and significantly enhanced the markers for M2 macrophage polarization. NO2-CLA promoted the HO-1 expression in cells challenged with LPS, and tin protoporphyrin IX, an HO-1 inhibitor, significantly reversed the NO2-CLA-mediated attenuation of NO secretion, but not IL-1ß or IL-6. We found that cells treated with NO2-CLA significantly increased mRNA expression of PPAR-γ compared to control cells, and NO2-CLA significantly reverted the decrease in PPAR-γ mRNA caused by LPS. Nonetheless, antagonists to PPAR-γ were unable to reverse the NO2-CLA-mediated suppression of inflammatory mediators. In addition, NO2-CLA did not alter the p38 and JNK activation elicited by LPS. Both NF-κB reporter activity and IκB-α degradation caused by LPS were notably diminished by NO2-CLA. NO2-CLA was observed to interrupt the nuclear translocation and DNA binding of p50 subunits caused by LPS with no obvious alterations in p65 subunits. Further, NO2-CLA attenuated the phosphorylation of STAT1/3 elicited in response to LPS. We propose that NO2-CLA could be considered as a possible strategy for the therapy of periodontal disease, although additional researches are certainly required to confirm this.

Effect of telmisartan on an experimental model of periodontitis in mice.

OBJECTIVE: This study was performed to explore the impact of telmisartan on experimental periodontitis in mice, in terms of alveolar bone destruction, by using micro-computed tomography analysis. MATERIALS AND METHODS: Male BALB/c mice were divided into four groups of 7 to 9 mice each: control (C) group; experimental periodontitis (E) group; experimental periodontitis-plus-telmisartan 5 mg/kg (ET5) group; and experimental periodontitis-plus-telmisartan 10 mg/kg (ET10) group. The mice in Group C were not subjected to experimental periodontitis. The other mice from Groups E, ET5 and ET10 were exposed to periodontitis. Periodontitis was induced by inoculation with Porphyromonas gingivalis. RESULTS: Telmisartan significantly suppressed both the reduction in alveolar bone height and increase of root exposure caused by P. gingivalis infection. When mice were treated with telmisartan, the decrease in the bone volume fraction induced by the infection was notably recovered. In addition, telmisartan reversed P. gingivalis-induced alterations in the microstructural parameters of trabecular bone, except trabecular thickness.No significant difference was evident between Groups ET5 and ET10 in both the extent of alveolar bone loss and microstructural parameters assessed, except bone volume fraction and trabecular number. CONCLUSION: Telmisartan may have potential benefits as a host modulation agent for the therapy of periodontitis.

Carbon monoxide-releasing molecule-401, a water-soluble manganese-based metal carbonyl, suppresses Prevotella intermedia lipopolysaccharide-induced production of nitric oxide in murine macrophages.

CONTEXT: Many reports in the literature have suggested the therapeutic value of carbon monoxide-releasing molecules (CORMs) against various diseases. However, to date, little is known about their possible influence on periodontal disease. OBJECTIVE: This study was performed to investigate the influence of CORM-401 on the generation of nitric oxide (NO) in murine macrophage cells activated with lipopolysaccharide (LPS) derived from Prevotella intermedia, a pathogen associated with periodontal disease. MATERIALS AND METHODS: LPS was isolated by the hot phenol-water method. Culture supernatants were analyzed for NO. Real-time PCR and immunoblotting were conducted to quantify mRNA and protein expression, respectively. NF-κB-dependent SEAP levels were estimated by reporter assay. DNA-binding of NF-κB was also analyzed. RESULTS: CORM-401 caused an apparent suppression of NO production through inhibition of iNOS at both the mRNA and protein levels in RAW264.7 cells stimulated with P. intermedia LPS. CORM-401 upregulated the expression of both the HO-1 gene and its protein in LPS-activated cells, and treatment with the HO-1 inhibitor significantly reversed the attenuating influence of CORM-401 against LPS-induced generation of NO. CORM-401 caused an apparent attenuation of NF-κB-dependent SEAP release induced by LPS. IκB-α degradation and nuclear translocation of NF-κB p50 subunit induced by LPS were significantly reduced by CORM-401. Additionally, CORM-401 significantly attenuated DNA-binding of p65 and p50 induced by LPS. CORM-401 attenuated NO generation induced by P. intermedia LPS independently of PPAR-γ, JNK, p38 and STAT1/3. CONCLUSION: The modulation of host inflammatory response by CORM-401 might be of help in the therapy of periodontal disease.

Effects of Rubus coreanus byproducts on intestinal microbiota and the immune modulation.

OBJECTIVE: Although the efficacy of Rubus coreanus (RC) byproducts as a feed additive has been recognized, its effects on intestinal microorganisms and the immune system are still unknown. METHODS: Six-week-old male rats were treated with 0.5% RC (T1), 1.0% RC (T2), and 1.5% RC (T3) for 4 weeks. RESULTS: We found that treatment with RC byproducts significantly increased the daily gain of body weight and feed intake. Treg-cell differentiation was enhanced in the mesenteric lymph nodes and spleen from the rats fed with RC byproducts. Illumina sequencing showed that bacteria in the phylum Firmicutes decreased and while those in the phylum Bacteroidetes increased in RC-treated groups. Particularly, the pathogenic microorganisms in the family Peptococcaceae decreased, and the non-pathogenic families Lachnospiraceae and S24-7 increased. Quantitative polymerase chain reaction analysis showed that the RC byproducts increased the lactic acid bacteria Bifidobacterium spp., Oscillospira spp., Leuconostoc citreum, and Weissella cibaria in a concentration-dependent manner. CONCLUSION: RC byproducts may be effective in immunomodulation by affecting intestinal microorganisms.

Effect of nitric oxide-releasing derivative of indomethacin on Prevotella intermedia lipopolysaccharide-induced production of proinflammatory mediators in murine macrophages.

The purpose of this study was to investigate the influences of NCX 2121, a nitric oxide (NO)-releasing derivative of indomethacin, upon the generation of proinflammatory mediators using murine macrophages activated by lipopolysaccharide (LPS) isolated from Prevotella intermedia, which is one of the pathogens implicated in periodontal diseases. Inducible NO synthase (iNOS)-derived NO, IL-1ß and IL-6 as well as their relevant mRNA were significantly attenuated by NCX 2121 in RAW264.7 cells activated by P. intermedia LPS. NCX 2121 was much more effective than the parental compound indomethacin in reducing these proinflammatory mediators. NCX 2121 triggered induction of heme oxygenase-1 (HO-1) in cells exposed to P. intermedia LPS, and its inhibitory influence upon P. intermedia LPS-elicited NO generation was notably blocked by SnPP treatment. NCX 2121 attenuated NF-κB-dependent SEAP release induced by P. intermedia LPS. NCX 2121 did not display inhibitory action towards IκB-α degradation triggered by LPS. Instead, it significantly diminished nuclear translocation as well as DNA-binding action of NF-κB p50 subunit elicited by P. intermedia LPS. Further, NCX 2121 significantly up-regulated SOCS1 mRNA expression in cells challenged with P. intermedia LPS. In summary, NCX 2121 down-regulates P. intermedia LPS-elicited generation of NO, IL-1ß and IL-6 in murine macrophages in a mechanism that involves anti-inflammatory HO-1 induction as well as decrement of NF-κB activation, which may be associated with SOCS1 expression. NCX 2121 may have potential benefits as a host immunomodulatory agent for the therapy of periodontal disease.

The Edible Red Alga Porphyra yezoensis Promotes Neuronal Survival and Cytoarchitecture in Primary Hippocampal Neurons.

The edible red alga Porphyra yezoensis is among the most popular marine algae and is of economic and medicinal importance. In the present study, the neurotrophic and neuroprotective activities of the ethanol extract of P. yezoensis (PYE) were investigated in primary cultures of hippocampal neurons. Results revealed that PYE significantly increased neurite outgrowth at an optimal concentration of 15 µg/mL. PYE dose-dependently increased viable cells, significantly accelerated the rate of neuronal differentiation in cultures, promoted axodendritic arborization, and eventually induced synaptogenesis. In addition to morphological development, PYE also promoted functional maturation as indicated by the staining of live cultures with FM 1-43. Moreover, PYE increased neuronal survivability, which was attributed to reduced apoptosis and its ROS scavenging activity. Taurine, a major organic acid in PYE (2.584/100 mg of dry PYE) promoted neurite outgrowth in a dose-dependent manner, and this promotion was suppressed by the taurine antagonist isethionic acid. The study indicates that PYE and its active component, taurine, facilitate neuronal development and maturation and have a neuroprotective effect.

Cloning and Characterization of an Endoglucanase Gene from Actinomyces sp. Korean Native Goat 40.

A gene from Actinomyces sp. Korean native goat (KNG) 40 that encodes an endo-ß-1,4-glucanase, EG1, was cloned and expressed in Escherichia coli (E. coli) DH5α. Recombinant plasmid DNA from a positive clone with a 3.2 kb insert hydrolyzing carboxyl methyl-cellulose (CMC) was designated as pDS3. The entire nucleotide sequence was determined, and an open-reading frame (ORF) was deduced. The ORF encodes a polypeptide of 684 amino acids. The recombinant EG1 produced in E. coli DH5α harboring pDS3 was purified in one step using affinity chromatography on crystalline cellulose and characterized. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis/zymogram analysis of the purified enzyme revealed two protein bands of 57.1 and 54.1 kDa. The amino terminal sequences of these two bands matched those of the deduced ones, starting from residue 166 and 208, respectively. Putative signal sequences, a Shine-Dalgarno-type ribosomal binding site, and promoter sequences related to the consensus sequences were deduced. EG1 has a typical tripartite structure of cellulase, a catalytic domain, a serine-rich linker region, and a cellulose-binding domain. The optimal temperature for the activity of the purified enzyme was 55°C, but it retained over 90% of maximum activity in a broad temperature range (40°C to 60°C). The optimal pH for the enzyme activity was 6.0. Kinetic parameters, Km and Vmax of rEG1 were 0.39% CMC and 143 U/mg, respectively.

Inhibitory effect of marine green algal extracts on germination of Lactuca sativa seeds.

The allelopathic potential of nine green seaweed species was examined based on germination and seedling growth of lettuce (Lactuca sativa L.). Out of nine methanol extracts, Capsosiphon fulvescens and Monostroma nitidum extracts completely inhibited germination of L. sativa at 4 mg/filter paper after 24 hr of treatment. Water extracts of these seaweeds generally showed low anti-germination activities than methanol extracts. Of the nine water extracts, Enteromorpha linza extract completely inhibited L. sativa germination at 16 mg/filter paper after 24 hrs. To identify the primary active compounds, C. fulvescens. powder was successively fractionated according to polarity, and the main active agents against L. sativa were determined to be lipids (0.0% germination at 0.5 mg of lipids/paper disc). According to these results, extracts of C. fulvescens can be used to develop natural herbicidal agents and manage terrestrial weeds.

Physiological properties of Scomber japonicus meat hydrolysate prepared by subcritical water hydrolysis.

The health-beneficial biological activities, including antioxidant and tyrosinase inhibitory activities, of Scomber japonicus muscle protein hydrolysates prepared by subcritical water hydrolysis were investigated. After 5 min of subcritical hydrolysis at 140 degrees C, 59.76% of S. japonicus muscle protein was hydrolyzed, the highest degree of hydrolysis in all the groups were tested. According to the response surface methodology results, as the reaction temperature and reaction time became lower and shorter, the yield became higher. The highest 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity (90.63%) occurred in hydrolysates treated at 140 degrees C for 5 min, and the highest tyrosinase inhibitory activity (65.54%) was identified in hydrolysates treated at 200 degreesC for 15 min. Changes in the molecular weight distribution of S. japonicus muscle proteins after subcritical water hydrolysis were observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Subcritical water hydrolysis is a suitable technique for obtaining S.japonicus muscle protein hydrolysates with useful biological activities, within a short time (5-15 min).

DHA suppresses Prevotella intermedia lipopolysaccharide-induced production of proinflammatory mediators in murine macrophages.

Several reports have indicated that dietary intake of DHA is associated with lower prevalence of periodontitis. In the present study, we investigated the effect of DHA on the production of proinflammatory mediators in murine macrophage-like RAW264.7 cells stimulated with lipopolysaccharide (LPS) isolated from Prevotella intermedia, a pathogen implicated in inflammatory periodontal disease, and its mechanisms of action. LPS was isolated from lyophilised P. intermedia ATCC 25,611 cells using the standard hot-phenol-water protocol. Culture supernatants were collected and assayed for NO, IL-1ß and IL-6. Real-time PCR analysis was carried out to detect the expression of inducible NO synthase (iNOS), IL-1ß, IL-6 and haeme oxygenase-1 (HO-1) mRNA. Immunoblot analysis was carried out to quantify the expression of iNOS and HO-1 protein and concentrations of signalling proteins. DNA-binding activities of NF-κB subunits were determined using an ELISA-based assay kit. DHA significantly attenuated the production of NO, IL-1ß and IL-6 at both gene transcription and translation levels in P. intermedia LPS-activated RAW264.7 cells. DHA induced the expression of HO-1 in cells treated with P. intermedia LPS. Selective inhibition of HO-1 activity by tin protoporphyrin IX significantly mitigated the inhibitory effects of DHA on LPS-induced NO production. DHA significantly attenuated the phosphorylation of c-Jun N-terminal kinase induced by LPS. In addition, DHA suppressed the transcriptional activity of NF-κB by regulating the nuclear translocation and DNA-binding activity of NF-κB p50 subunit and inhibited the phosphorylation of signal transducer and activator of transcription 1. Further in vivo studies are needed to better evaluate the potential of DHA in humans as a therapeutic agent to treat periodontal disease.

Inhibitory effects of seaweed extracts on the growth of the vaginal bacterium Gardnerella vaginalis.

Of 44 species of seaweed screened for potential anti-Gardnerella vaginalis activity, 27 (61.4%) showed antimicrobial activity by the agar disk-diffusion method. Among them, the strongest activities against the pathogen were exhibited by Chlorophyta, with Ulva pertusa producing an 11.3-mm zone of inhibition at 5 mg disk⁻¹. The MIC values of U. pertusa extracts against both G. vaginalis KCTC 5096 and KCTC 5097, the main cause of vaginosis, were 312 µg ml⁻¹, while the MIC values against both Candida albicans KCTC 7270 and KCTC 7965, the main cause of candidiasis, were 2.5 mg ml⁻¹. Against Lactobacillus gasseri KCTC 3173 and Lactobacillus jensenii KCTC 5194, members of the normal vaginal microflora, no inhibitory effect was seen even at 10 mg ml⁻¹. To identify the primary active compounds, a U. pertusa powder was successively fractionated according to polarity, and the main active agents against G. vaginalis KCTC 5096 were determined to be nitrogenous compounds (156 µg ml⁻¹ of the MIC value). According to these results, it was suggested that extracts of the seaweed U. pertusa are valuable for the development of natural therapeutic agents for treating women with bacterial vaginosis.

Seasonal variation of antibacterial activities in the green alga Ulva pertusa Kjellman.

The present study was performed to screen out the extracts of algae and assess the seasonal variation in antimicrobial activity of Ulva pertusa against Gardnerella vaginalis. Seasonal variation in antibacterial activity was observed, with the extracts showing no activity during summer and autumn, and showing antibacterial activity from early winter (December) to middle spring (April). The maximum value of antimicrobial activity (6.5 mm inhibition zone at 5 mg disk(-1)) of U. pertusa against G. vaginalis was observed in April. Otherwise, for both chlorophyll a and b, the highest content (2.87 mg g(-1) and 1.37 mg g(-1)) was observed in March 2009. These results may reflect variation in cellular chemical compositions such as secondary metabolite(s) rather than chlorophyll and biological activities according to season.

The marine alga Gelidium amansii promotes the development and complexity of neuronal cytoarchitecture.

Neurotrophic factors are vital not only to support neuronal development but also to protect mature neurons from atrophy in neurodegenerative diseases. As an effort to explore natural sources that possess neurotrophic activity, we screened common marine algae for their neuritogenic activity in the developing rat hippocampal neurons in culture. Of the 22 seaweed species examined, ethanol extracts of Gelidium amansii (GAE) exhibited potent neuritogenic activity, followed by Undaria pinnatifida and Sargassum fulvellum extracts. The effects of GAE were dose dependent with an optimum concentration of 15 µg/mL. The GAE significantly promoted the initial neuronal differentiation from the stage I into the stage II and increased the indices of axonal and dendritic development such as the length, the numbers of primary processes, and branching frequencies by a minimum of twofold compared with the vehicle control. These results show that marine algae are promising candidates for neurotrophic potentials.

Effects of seaweed Laminaria japonica extracts on skin moisturizing activity in vivo.

Twelve species of edible seaweed from the coast of Korea were screened for skin moisturizing activity. We placed the lead of a Corneometer on an approximately 6-cm2 test area of the forearm and measured both untreated skin (control) and skin treated with test moisturizing creams either containing or not containing 5% water:propylene glycol (50:50) extracts of seaweeds. Over the 8-h observation period, the strongest activity of the Laminaria japonica extracts occurred at the 2-h period. For the 10% extract, hydration with the L. japonica extract increased by 14.44% compared with a placebo. Transepidermal water loss (TEWL) was also measured using a test cream with 10% L. japonica extract. For up to 8 h after applying the creams, TEWL was decreased to 4.01 g/cm2, which was approximately 20% of that seen with the control. We suggest that the L. japonica extract hydrates skin via the humectants and hydrocolloids that it contains. To confirm the safety of L. japonica extracts, we performed a patch test on human skin. The results suggested that at moderate doses humans can safely use the extracts. For commercial applications, we evaluated the physicochemical characteristics of the test cream products, including Hunter L, a, and b values; pH; refractive index; and coefficient of viscosity. L. japonica extract did not affect overall formulations of the test cream product in any of the tested aspects. These results suggest that L. japonica extract is a promising ingredient in moisturizing formulations.

Effects of Lycopersicon esculentum extract on hair growth and alopecia prevention.

To evaluate the potential hair growth-promoting activity and the expression of cell growth factors of Lycopersicon esculentum extracts, each 3% (w/w) of ethyl acetate extract (EAE), and supercritical CO2 extract (SCE) of L. esculentum and isolated lycopene Tween 80 solution (LTS) and test hair tonic (THT) containing LTS were applied on the dorsal skin of C57BL/6 mice, once a day for 4 weeks. At week 4, LTS and THT exhibited hair growth-promoting potential similar to that of 3% minoxidil as a positive control (PC). Further, in the LTS group, a significant increase of mRNA expression of vascular endothelial growth factor (VEGF), keratinocyte growth factor, and insulin-like growth factor-1 (IGF-1) was observed than PC, as well as the negative control (NC). In the THT group, increases in IGF-1 and decrease in VEGF and transforming growth factor-ß expression were significant over the NC. In a histological examination in the THT group, the induction of anagen stage of hair follicles was faster than that of NC. In the Draize skin irritation study for THT, no observable edema or erythema was observed on all four sectors in the back skin after exposure for 24 or 72 h for any rabbit. Therefore, this study provides reasonable evidence that L. esculentum extracts promote hair growth and suggests that applications could be found in hair loss treatments without skin irritation at moderate doses.

The antibacterial activity of various saturated and unsaturated fatty acids against several oral pathogens.

The antibacterial activity of various saturated fatty acids (SFA) and unsaturated fatty acids (USFA) against different oral pathogens which are implicated in the cause of dental caries, stomatitis, gingivitis, and periodontitis was examined. The saturated fatty acids Pa, StA and ArA, and the unsaturated omega-7 fatty acids PLA and omega-9 fatty acids OA showed either none to low antimicrobial activity against all of the 12 oral pathogenic strains used in this study. In contrast, the omega-3 PUFAs, ALA, SDA, EPA and DHA, and the omega-6 PUFAs, LA, GLA, and AA showed considerable antimicrobial activity against 8, 7, 6 and 5 strains, and 6, 10 and 5 strains, respectively. In particular, the omega-3 and omega-6 PUFAs showed strong antimicrobial activity against Porphyromonas gingivalis KCTC 381, the cause of periodontitis, and against Aggregatibacter segnis KCTC 5968, Fusobacterium nucleatum subsp. Polymorphum KCTC 5172 and Prevotella intermedia KCTC 25611, all organisms implicated in the cause of gingivitis. To date, no bacterial resistance to free fatty acids has been encountered and no resistance phenotype has emerged. Therefore, these results suggest that PUFAs may be useful in the development of therapeutic agents for oral diseases, and in particular, in the development of agents that have minimal side effects and against which there is no bacterial resistance.

Single oral dose toxicity test of polycalcium, a mixed composition of polycan and calcium lactate-gluconate 1:9 (G/G) in SD rat.

The object of this study was to obtain acute oral toxicity information of Polycalcium, a mixed composition of Polycan and Calcium lactate-gluconate 1:9 (g/g), in Sprague-Dawely (SD) rats. In order to investigate the toxicity and identify target organs, Polycalcium were once orally administered to female and male SD rats at dose levels of 2000, 1000, 500 and 0 (control) mg/kg body weights. The mortality, changes on body weight and clinical signs were monitored during 14 days after treatment with gross observation, changes on the organ weights and histopathology of principle organs and treatment sites based on the recommendation of KFDA Guidelines [2009-116, 2009]. As the results of single oral treatment of Polycalcium, no treatment related mortalities were observed within 14 days after end of treatment up to 2000 mg/kg, the limited dosage of rodents in the both genders. In addition, no Polycalcium treatment related changes on the body and organ weights, clinical signs, necropsy and histopathological findings were detected. The results obtained in this study suggest that the Polycalcium is non-toxic in rats. The LD50 and approximate LD in rats after single oral dose of Polycalcium were considered over 2000 mg/kg in both female and male, respectively.

Nifedipine attenuates alveolar bone destruction and improves trabecular microarchitectures in mice with experimental periodontitis.

Studies have shown that nifedipine exerts anti-inflammatory and immunosuppressive actions in addition to being a calcium channel blocker. The present study was performed to explore the influence of nifedipine on alveolar bone destruction in mice with experimental periodontitis by evaluating morphological information acquired from micro-computed tomography analysis. BALB/c mice were randomly assigned into four groups: control (C) group; experimental periodontitis (E) group; experimental periodontitis + 10 mg/kg dose of nifedipine (EN10) group; and experimental periodontitis + 50 mg/kg dose of nifedipine (EN50) group. Periodontitis was induced by oral inoculation with Porphyromonas gingivalis over a 3-week time period. Nifedipine significantly mitigated the loss of alveolar bone height as well as increase of root surface exposure induced by experimental periodontitis. Additionally, the reduction in the bone volume fraction associated with P. gingivalis infection was significantly recovered upon nifedipine treatment. Further, nifedipine attenuated P. gingivalis-induced deteriorations in the trabeculae-associated parameters. Significant difference was evident between Groups EN10 and EN50 in both the extent of alveolar bone loss and microstructural parameters assessed, except trabecular separation and trabecular number. Nifedipine appeared to have good performance in ameliorating bone loss in mice with induced periodontitis. Nifedipine may be utilized in the clinical management of periodontitis, though further research is indicated to verify the therapeutic effect.

Inhibition of oral pathogens and collagenase activity by seaweed extracts.

Fifty-seven species of common seaweed from the Coast of Korea were screened for antimicrobial (i.e. inhibition of Prevotella intermedia and Porphyromonas gingivalis growth) activity. As a source of bioactive compounds, seaweeds can produce many secondary metabolites with a variety of activities. Using the agar diffusion method, only 17 species (29.8%) showed inhibitory activity. Of these, methanol extracts of Enteromorpha linza, Sargassum sagamianum, and Ulva pertusa showed strong inhibitory effects against both P. intermedia and P. gingivalis. The MIC values of E. linza, S. sagamianum, and U. pertusa extracts against P. intermedia were 625, 78 and 625 microg ml(-1) and those against P. gingivalis were 312, 156 and 625 microg ml(-1), respectively. When these three species' extracts were separated into five fractions according to their polarity, the main active agents were determined to be phenolic compounds. We then compared the antimicrobial activities of these phenolic compounds against various periodontal pathogens using a MIC test. Phenolic compound containing extracts at concentrations of 10 to 100 microg ml(-1) showed a moderate to significant inhibitory effect on collagenase 1,2 and 3 activity.

Nitrooleic acid inhibits macrophage activation induced by lipopolysaccharide from Prevotella intermedia.

The hypothesis of the present study was that nitro-fatty acids (NO2-FAs) would suppress inflammation associated with periodontal disease. To test this hypothesis, we investigated the influence of nitrooleic acid, a prototypical NO2-FA, on the inflammatory response of murine macrophages activated with lipopolysaccharide (LPS) from Prevotella intermedia, a pathogen associated the etiology of different types of periodontal diseases. LPS was prepared from P. intermedia cells by using phenol-water protocol. Culture supernatants were assayed for nitric oxide (NO), interleukin-1ß (IL-1ß), and IL-6. Real-time polymerase chain reaction and immunoblotting analyses were performed to quantify messenger RNA and protein expression, respectively. The secreted embryonic alkaline phosphatase reporter assay was performed to measure NF-κB activation. The transcription factor assay kit was used to measure DNA-binding of NF-κB subunits. Findings obtained from the present study revealed that nitrooleic acid suppresses the generation and messenger RNA expression of inducible NO synthase-derived NO, IL-1ß, and IL-6 in RAW264.7 cells activated with P. intermedia LPS and promotes macrophage polarization toward anti-inflammatory M2 phenotype. We also found that nitrooleic acid exerts its effect via heme oxygenase-1 induction and suppression of NF-κB signaling. The inhibition of NO and proinflammatory cytokine production by nitrooleic acid was independent from PPAR-γ, JNK, p38, and STAT1/3. Nitrooleic acid may represent a novel class of agent as a host modulator which has therapeutic benefit in periodontal disease, though more work is needed to confirm this.