Bioactive fraction of geopropolis from Melipona scutellaris decreases neutrophils migration in the inflammatory process: involvement of nitric oxide pathway

FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOThe aim of this study was to evaluate the activity of the ethanolic extract of geopropolis (EEGP) from Melipona scutellaris and its fractions on the modulation of neutrophil migration in the inflammatory process, and the participation of nitric oxide (NO) pathway, as well as to check the chemical profile of the bioactive fraction. EEGP and its aqueous fraction decreased neutrophil migration in the peritoneal cavity and also the interaction of leukocytes (rolling and adhesion) with endothelial cells. The levels of chemokines CXCL1/KC and CXCL2/MIP-2 were not altered after treatment with EEGP and the aqueous fraction. It was found that the injection of NO pathway antagonists abolished the EEGP and the aqueous fraction inhibitory activity on the neutrophil migration. The expression of intercellular adhesion molecule type 1 (ICAM-1) was reduced, and nitrite levels increased after treatment with EEGP and aqueous fraction. In the carrageenan-induced paw edema model, EEGP and the aqueous fraction showed antiedema activity. No pattern of flavonoid and phenolic acid commonly found in propolis samples of Apis mellifera could be detected in the aqueous fraction samples. These data indicate that the aqueous fraction found has promising bioactive substances with anti-inflammatory activity2013FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPESP [2009/12352-3, 2010/20214-7]2009/12352-3; 2010/20214-

Salivary IL-21 and IgA responses to a competitive match in elite basketball players

Athletes engaged in strenuous training might experience transient immune suppression that could lead to greater incidence of upper respiratory tract infections (URTI). Since interleukin 21 (IL-21) stimulates immunoglobulin A (IgA) secreting cells and a low level of this immunoglobulin is associated with increased incidence of URTI, the aim of the present study was to investigate the effect of a basketball match on salivary cortisol (sC), salivary IL-21 (sIL-21) and salivary IgA (sIgA) levels. Twenty male basketball players participated in an official game in two teams (10 players in each team). The saliva samples were collected before the warm-up and approximately 10-15 min after the end of the match and were analysed by ELISA methods. sC concentration increased significantly after the match while sIL-21 level was reduced (p < 0.05). In opposition to the study’s hypothesis, sIgA level did not change in response to the match. The present findings suggest that a basketball match is sufficiently stressful to elevate sC concentration and attenuates the sIL-21 output without compromising the sIgA level. It is reasonable to speculate that the stability of sIgA acute responses to the match, despite the decrement in sIL-21, indicates that other mechanisms rather than IL-21 stimulating B cell proliferation/differentiation might modulate IgA concentration and secretion rate.FAPESP 2008/10404-

Effect of silver nanoparticles on the physicochemical and antimicrobial properties of an orthodontic adhesive

ABSTRACT Orthodontic treatment with fixed brackets plays a major role on the formation of white spot lesions. Objective This study aimed to incorporate silver nanoparticle solutions (AgNP) in an orthodontic adhesive and evaluate its physicochemical and antimicrobial properties. Material and Methods Silver nanoparticle solutions were added to a commercial adhesive in different concentrations (w/w): 0%, 0.11%, 0.18%, and 0.33%. Shear bond strength (SBS) test was performed after bonding metal brackets to enamel. Raman spectroscopy was used to analyze in situ the degree of conversion (DC) of the adhesive layer. The surface free energy (SFE) was evaluated after the measurement of contact angles. Growth inhibition of Streptococcus mutans in liquid and solid media was determined by colony-forming unit count and inhibition halo, respectively. One-way ANOVA was performed for SBS, DC, SFE, and growth inhibition. Results The incorporation of AgNP solution decreased the SBS (p<0.001) and DC in situ (p<0.001) values. SFE decreased after addition of 0.18% and 0.33% AgNP. Growth inhibition of S. mutans in liquid media was obtained after silver addition (p<0.05). Conclusions The addition of AgNP solutions to Transbond&#8482; XT adhesive primer inhibited S. mutans growth. SBS, DC, and SFE values decreased after incorporation up to 0.33% AgNP solution without compromising the chemical and physical properties of the adhesive

The role of endogenous opioid peptides in the antinociceptive effect of 15-deoxy (delta 12,14)-prostaglandin J(2) in the temporomandibular joint

CNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOCAPES - COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL E NÍVEL SUPERIORFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOWe have previously demonstrated that peripheral administration of 15d-PGJ(2) in the Temporomandibular joint (TMJ) of rats can prevent nociceptor sensitization, mediated by peroxisome proliferator activated receptor-gamma (PPAR-gamma), and kappa- and delta- opioid receptors. However, the mechanism that underlies the signaling of PPAR-gamma (upon activation by 15d-PGJ(2)) to induce antinociception, and how the opioid receptors are activated via 15d-PGJ(2) are not fully understood. This study demonstrates that peripheral antinociceptive effect of 15d-PGJ(2) is mediated by PPAR-gamma expressed in the inflammatory cells of TMJ tissues. Once activated by 15d-PGJ(2), PPAR-gamma induces the release of beta-endorphin and dynorphin, which activates kappa- and delta-opioid receptors in primary sensory neurons to induce the antinociceptive effect. (C) 2016 Elsevier Ltd. All rights reserved.We have previously demonstrated that peripheral administration of 15d-PGJ(2) in the Temporomandibular joint (TMJ) of rats can prevent nociceptor sensitization, mediated by peroxisome proliferator activated receptor-gamma (PPAR-gamma), and kappa- and delta- opioid receptors. However, the mechanism that underlies the signaling of PPAR-gamma (upon activation by 15d-PGJ(2)) to induce antinociception, and how the opioid receptors are activated via 15d-PGJ(2) are not fully understood. This study demonstrates that peripheral antinociceptive effect of 15d-PGJ(2) is mediated by PPAR-gamma expressed in the inflammatory cells of TMJ tissues. Once activated by 15d-PGJ(2), PPAR-gamma induces the release of beta-endorphin and dynorphin, which activates kappa- and delta-opioid receptors in primary sensory neurons to induce the antinociceptive effect1102734CNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOCAPES - COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL E NÍVEL SUPERIORFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOCAPES - COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL E NÍVEL SUPERIORFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO483988/20121830/20102011/00683-

The indirect antinociceptive mechanism of 15d-PGJ(2) on rheumatoid arthritis-induced TMJ inflammatory pain in rats

FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPEMIG - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE MINAS GERAISInflammation of the temporomandibular joint (TMJ) induced by rheumatoid arthritis (RA) have resulted in persistent pain and caused distress to many patients. Considering that not all patients respond to traditional drugs therapy to RA and it has demonstrated that 15-deoxy-(Delta 12,14)-prostaglandin J(2) (15d-PGJ(2)) into TMJ has a potential peripheral antinociceptive effect, the aim of this study was to evaluate the peripheral effect of 15d-PGJ2 in RA-induced TMJ inflammatory hypernociception. Antigen-induced arthritis (AIA) was generated in rats with methylated bovine serum albumin (mBSA). RA-induced TMJ hypernociception was assessed by measuring the behavioural nociceptive responses. After behavioural experiments, the animals were terminally anaesthetized and periarticular tissues were removed and homogenized. The supernatants were used to evaluate the levels of tumour necrosis factor (TNF)-alpha, interleukin (IL)-1 beta and keratinocyte-derived chemokine (KC) by enzyme-linked immunosorbent assay as well the expression of PKC epsilon and PKA by western blotting analysis. The intra-articular injection of mBSA, but not phosphate buffered saline (control), in immunized rats induced dose-and time-dependent behavioural nociceptive responses in which the peak of nociceptive responses were obtained by using 10 mu g/TMJ of mBSA after 24 h. Pretreatment with 15d-PGJ(2) (30, 100 and 300 ng/TMJ) inhibited the RA-induced TMJ inflammatory hypernociception. In addition, 15d-PGJ(2) reduced the RA-induced release of TNF-alpha, IL-1 beta and KC (p < 0.05) as well the expression of PKA and PKC epsilon (p < 0.05). In the present study, we demonstrated that 15d-PGJ(2) was able to reduce the RA-induced TMJ inflammatory hypernociception by an indirect mechanism. This antinociceptive effect is in part due to decrease of TNF-alpha, IL-1 beta and KC levels and PKA/PKC epsilon expression in the TMJ16811061115FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPEMIG - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE MINAS GERAISFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPEMIG - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE MINAS GERAISFAPESP [2011/00683-5]Fundacao de Amparo a Pesquisa do Estado de Minas Gerais, Brazil [FAPEMIG PPM 097/09]2011/00683-5PPM097/0

Gonadal hormones decrease temporomandibular joint kappa-mediated antinociception through a down-regulation in the expression of kappa opioid receptors in the trigeminal ganglia

FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOWe have previously demonstrated that activation of kappa-opioid receptor located in the temporomandibular joint (TMJ) of rats induces a significantly greater TMJ antinociception in diestrus females than in proestrus females (higher estradiol serum levels than diestrus) and males. These findings indicate that gonadal hormones decrease TMJ kappa-mediated antinociception. The aim of this study was to investigate some of the mechanisms by which gonadal hormones decrease TMJ kappa-mediated anti nociception. Western blot analysis demonstrated a significantly lower kappa-opioid receptor expression in the trigeminal ganglia of intact males than in intact and ovariechtomized (OVX) females and orchidectomized (ORX) males. In females, kappa-opioid receptor expression in the trigeminal ganglia was significantly lower in proestrus than in diestrus and OVX females. Taken together these findings suggest that gonadal hormones, especially male gonadal hormones, down-regulate kappa-opioid receptor expression. Co-application of the NOS inhibitor L-NMMA or the NO-sensitive guanylyl cyclase inhibitor ODQ with the kappa-opioid receptor agonist U50,488 blocked TMJ kappa-mediated antinociception in males and females. These findings suggest that antinociception induced by activation of kappa opioid receptors in the TMJ region is mediated by the L-arginine/NO/cGMP pathway in both sexes. Despite the involvement of the L-arginine/NO/cGMP pathway in TMJ kappa-mediated antinociception in both sexes, gonadal hormones do not diminish the activity of this pathway to decrease TMJ kappa-mediated antinociception. Alternatively, they significantly reduce kappa-opioid receptor expression in the trigeminal ganglia6171-34147FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOFAPESP [FAPESP 04/00073-9]2004/00073-

Low doses of 15d-PGJ2 induce osteoblast activity in a PPAR-gamma independent manner

FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOPeroxisome proliferator-activated receptor-gamma (PPAR gamma) regulates both glucose metabolism and bone mass. Evidence suggests that the therapeutic modulation of PPAR gamma with synthetic agonists activity may elicit undesirable effects on bone. However, there is no information regarding its natural agonist 15d-PGJ(2), besides its excellent anti-inflammatory action. In the present study the effects of 15d-PGJ(2) on osteoblastic cells were determined. Osteoblastic cells (MC3T3) were cultured in an osteogenic medium in the presence of 1, 3 or 10 mu M of 15d-PGJ(2) during 21 days and alizarin and Von Kossa staining were employed. The protein expression (type-I collagen, osteonectin, osteopontin, RANKL, osteoprotegerin, HDAC-9c and PPAR-gamma) was evaluated after 3 days in the presence of 15d-PGJ(2) by western blotting and indirect immunofluorescence methods. The production of mineralized extracellular matrix was observed by transmission electron microscopy. After 72 h of culture, the mRNA was extracted for RT-qPCR analysis of RUNX expression. In the presence of all 3 tested 15d-PGJ2 doses, alizarin red and Von kossa staining were positive demonstrating the ability to the osteoblast differentiation. Type-I collagen and osteonectin proteins expression were up-regulated (p < 0.05) after 72 h in the presence of the smaller doses of 15d-PGJ(2). In contrast osteopontin, RANKL and OPG expression did not significantly alter. In the presence of 15d-PGJ(2) it was possible to visualize mineralized nodules in the extracellular matrix confirmed with the increased RUNX mRNA expression. 15d-PGJ(2) at small doses increased the osteoblast activity and the bone-related proteins expression162131138FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOFAPESP [2010/15014-9]CNPq [471305/2009-0]2010/15014-9471305/2009-

15d-PGJ2-loaded in nanocapsules enhance the antinociceptive properties into rat temporomandibular hypernociception

FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOTo verify whether the nanoencapsulation of 15d-PGJ(2) in poly(D,L-lactide-co-glycolide) (PLGA) nanocapsules (15d-PGJ(2)-NC) might potentialize its antinociceptive activity into rats' temporomandibular joint (TMJ). Main methods: Transmission electron microscopy (TEM) and atomic force microscopy (AFM) were used to evaluate the morphology and suspension of the PLGA nanocapsules. Rats were pretreated (15 min) with an intra-TMJ injection of unloaded 15d-PGJ(2) or 15d-PGJ(2)-NC at concentrations of 10, 100 or 1000 pg followed by an ipsilateral intra-TMJ injection of 1.5% formalin. The nociceptive behavioral response was observed during 45 min: animals were then sacrificed and the periarticular tissue was removed for IL-1 beta measurements. TEM and AFM analyses showed that 15d-PGJ(2)-NC is spherical without any aggregates or adhesion confirming that this formulation is a good drug carrier system for 15d-PGJ(2). Pretreatment with 15d-PGJ(2)-NC (100 and 1000 pg/TMJ), but not unloaded 15d-PGJ(2), was found to significantly decrease the release of IL-1 beta cytokine and the animals' nociceptive behavioral response induced by intra-TMJ injection of formalin. The compound 15d-PGJ(2)-NC might be used as a potential antinociceptive and anti-inflammatory agent to treat temporomandibular disorders in clinical practice9023-24944949FAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOFAPESP - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULOCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOFAPESP [2010/15014-9]CNPq [303080/2010-8]2010/15014-9303080/2010-

Low dose of propranolol down-modulates bone resorption by inhibiting inflammation and osteoclast differentiation

CNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOFAPEMIG - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE MINAS GERAISBones are widely innervated, suggesting an important role for the sympathetic regulation of bone metabolism, although there are controversial studies. We investigated the effects of propranolol in a model of experimental periodontal disease. Rats were assigned as follows: animals without ligature; ligated animals receiving vehicle and ligated animals receiving 0.1, 5 or 20 mg.kg-1 propranolol. After 30 days, haemodynamic parameters were measured by cardiac catheterization. Gingival tissues were removed and assessed for IL-1 beta, TNF-alpha and cross-linked carboxyterminal telopeptides of type I collagen (CTX) by ELISA, or intercellular adhesion molecule 1 (ICAM-1), receptor activator of NF-kappa B ligand (RANKL) and osteoprotegerin (OPG) by Western blot analysis. Sections from the mandibles were evaluated for bone resorption. Also, we analysed the ability of propranolol to inhibit osteoclastogenesis in vitro. Propranolol at 0.1 and 5 mg.kg-1 reduced the bone resorption as well as ICAM-1 and RANKL expression. However, only 0.1 mg.kg-1 reduced IL-1 beta, TNF-alpha and CTX levels as well as increased the expression of OPG, but did not alter any of the haemodynamic parameters. Propranolol also suppressed in vitro osteoclast differentiation and resorptive activity by inhibiting the nuclear factor of activated T cells (NFATc) 1 pathway and the expression of tartrate-resistant acid phosphatase (TRAP), cathepsin K and MMP-9. Low doses of propranolol suppress bone resorption by inhibiting RANKL-mediated osteoclastogenesis as well as inflammatory markers without affecting haemodynamic parameters165721402151CNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOFAPEMIG - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE MINAS GERAISCNPQ - CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICOFAPEMIG - FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE MINAS GERAISCNPq [471305/2009-0, 303080/2010-8]PAPE-UNIUBE [2009/001]FAPEMIG [097/09]471305/2009-0; 303080/2010-8097/0