Cristina Gomes de Macedo

Quercetin inhibits inflammatory bone resorption in a mouse periodontitis model.

Periodontitis is a disease that leads to bone destruction and represents the main cause of tooth loss in adults. The development of aggressive periodontitis has been associated with increased inflammatory response that is induced by the presence of a subgingival biofilm containing Aggregatibacter actinomycetemcomitans. The flavonoid quercetin (1) is widespread in vegetables and fruits and exhibits many biological properties for possible medical and clinical applications such as its anti-inflamatory and antioxidant effects. Thus, in the present study, the properties of 1 have been evaluated in bone loss and inflammation using a mouse periodontitis model induced by A. actinomycetemcomitans infection. Subcutaneous treatment with 1 reduced A. actinomycetemcomitans-induced bone loss and IL-1β, TNF-α, IL-17, RANKL, and ICAM-1 production in the gingival tissue without affecting bacterial counts. These results demonstrated that quercetin exhibits protective effects in A. actinomycetemcomitans-induced periodontitis in mice by modulating cytokine and ICAM-1 production.

Secreted osteoclastogenic factor of activated T cells (SOFAT), a novel osteoclast activator, in chronic periodontitis

A novel activated human T cell-secreted cytokine, referred as secreted osteoclastogenic factor of activated T cells (SOFAT), that induce osteoclastogenesis in a RANKL-independent manner was recently described. This study evaluated the role of SOFAT in periodontal tissues and periodontitis. Gingival biopsies were harvested from systemically healthy non-periodontitis (n=15) and chronic periodontitis patients (n=15). The mRNA and protein levels of SOFAT were measured by qPCR and by enzyme-linked immunosorbent assay, respectively. Moreover, RAW 264.7 cells were cultured with SOFAT or Receptor activator of nuclear factor-kB ligand (RANKL) and stained for tartrate-resistant acid phosphatase (TRAP). Also, mice received a palatal injection between the first and second upper molar of SOFAT (100 ng/ml) or saline solution (0.9%). The upper jaw was removed, histologically processed and stained with hematoxilin and eosin to observe the presence of osteoclast-like cells. The mRNA and protein levels of SOFAT were significantly higher in the gingival tissue of the periodontitis group when compared to non-periodontitis one (p<0.05). In addition, SOFAT potently induced TRAP-positive multinucleated cell formation by RAW 264.7 cells as well as induced the formation of osteoclast-like cells in the periodontal ligament in mice. The present study demonstrated that SOFAT may play an important role in periodontitis.

β-Glucans (Saccharomyces cereviseae) Reduce Glucose Levels and Attenuate Alveolar Bone Loss in Diabetic Rats with Periodontal Disease.

The objective of this study was to assess the effects of oral ingestion of β-glucans isolated from Saccharomyces cereviseae on the metabolic profile, expression of gingival inflammatory markers and amount of alveolar bone loss in diabetic rats with periodontal disease. Diabetes mellitus was induced in 48 Wistar rats by intraperitoneal injection of streptozotocin (80 mg/kg). After confirming the diabetes diagnosis, the animals were treated with β-glucans (by gavage) for 28 days. On the 14th day of this period, periodontal disease was induced using a ligature protocol. β-glucans reduced the amount of alveolar bone loss in animals with periodontal disease in both the diabetic and non-diabetic groups (p < 0.05). β-glucans reduced blood glucose, cholesterol and triacylglycerol levels in diabetic animals, both with and without periodontal disease (p < 0.05). Furthermore, treatment with β-glucans reduced the expression of cyclooxygenase-2 and receptor activator of nuclear factor kappa-B ligand and increased osteoprotegerin expression in animals with diabetes and periodontal disease (p < 0.05). It was concluded that treatment with β-glucans has beneficial metabolic and periodontal effects in diabetic rats with periodontal disease.

15-deoxy-Δ12,14-prostaglandin J2 reduces albumin-induced arthritis in temporomandibular joint of rats

The aim of this study was to evaluate the peripheral effect of 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) in albumin-induced arthritis in temporomandibular joint (TMJ) of rats. Antigen-induced arthritis (AIA) was generated in rats with methylated bovine serum albumin (mBSA) diluted in complete Freund׳s adjuvant. Pretreatment with an intra-articular injection of 15d-PGJ2 (100 ng/TMJ) before mBSA intra-articular injection (10 µg/TMJ) (challenge) in immunized rats significantly reduced the albumin-induced arthritis inflammation. The results demonstrated that 15d-PGJ2 was able to inhibit plasma extravasation, leukocyte migration and the release of inflammatory cytokines IL-6, IL-12, IL-18 and the chemokine CINC-1 in the TMJ tissues. In addition, 15d-PGJ2 was able to increase the expression of the anti-adhesive molecule CD55 and the anti-inflammatory cytokine IL-10. Taken together, it is possible to suggest that 15d-PGJ2 inhibit leukocyte infiltration and subsequently inflammatory process, through a shift in the balance of the pro- and anti-adhesive properties. Thus, 15d-PGJ2 might be used as a potential anti-inflammatory drug to treat arthritis-induced inflammation of the temporomandibular joint.

Influence of salivary washout on drug delivery to the oral cavity using coated microneedles: An in vitro evaluation

The objective of this study was to determine whether in buccal tissues, after insertion and removal of coated microneedles, the presence of saliva over the insertion site can lead to loss of the deposited drug, and if saliva can influence in vitro permeation of the drug across the tissue. Microneedles were coated with sulforhodamine (SRD), which was used as a model drug, and inserted in to porcine buccal mucosa in vitro. Fluorescence microscopy was used to study microneedle coating quality and the diffusion of SRD through the mucosa. Permeation experiments were conducted for simulated dynamic or static salivary flow by adding 100μL/h or 100, 200 or 300μL of phosphate buffered saline (PBS) in the donor compartment of the Franz diffusion cells, into which buccal tissue after insertion of SRD-coated microneedles was placed. Microscopy showed that microneedles were uniformly coated with SRD and that SRD was successfully delivered in to the mucosa. Some SRD remained in the tissue even after 24h, despite presence of PBS on top of the coated microneedle insertion site. It was found that salivary washout can result in loss of drug that has been deposited in oral cavity mucosal tissues using coated microneedles, and presence of fluid over the coated microneedle insertion site can increase flux across the tissue. Thus, it is advisable to include salivary flow during in vitro studies related to the use of coated microneedles for drug delivery to the oral cavity in order to not obtain misleading results.

Botulinum toxin type A reduces inflammatory hypernociception induced by arthritis in the temporomadibular joint of rats

Objective This study aimed to investigate the antinociceptive effects of Botulinum toxin type A (BoNT‐A) on persistent inflammatory hypernociception induced by arthritis in the temporomandibular joint (TMJ) of rats. Material and methods Wistar rats were induced to persistent inflammatory hypernociception in the left TMJ. Then, animals were treated with intra‐TMJ injections of BoNT‐A, using doses of 3.5, 7 and 14 U/kg. Saline was used as control group. Behavioral tests were applied to evaluated the effect of BoNT‐A in the inflammatory hypernociception. After that, animals were euthanized and samples from peri‐articular tissues and trigeminal ganglia were obtained for further analyses. Results BoNT‐A reduced the persistent inflammatory hypernociception induced by arthritis in the TMJ of rats. BoNT‐A significantly reduced the peripheral release of the neurotransmitters Substance P and Calcitonin gene related peptide; and the pro‐inflammatory cytokine IL‐1&bgr;. Otherwise, BoNT‐A had no effect in the peripheral release of glutamate and the cytokine TNF‐&agr;. Conclusion These results demonstrate that intra‐articular injection of BoNT‐A reduces the albumin‐induced arthritis persistent hypernociception in TMJ of rats by peripheral inhibition of neuropeptides release. HighlightsBotulinum toxin type A acts reduced behavioral nociceptive response in rats responses in nociceptive animals.Botulinum toxin type A reduces Interleukin 1 beta increasing its antinociceptive effect.Hypernociception is reduced by Botulinum toxin type A injections due to the inhibition of neuropeptides.

Microneedles enhance topical delivery of 15-deoxy-Δ12,14-prostaglandin J2 and reduce nociception in temporomandibular joint of rats

Abstract The pain arising from temporomandibular disorders is often treated with opioids and agents that inhibit the immune response and are associated with substantial adverse effects and long‐term risks. Thus, the development of new therapies that are safer and more effective is of great interest to patients and clinicians. 15‐deoxy‐&Dgr;12,14‐prostaglandin J2 (15d‐PGJ2) is naturally produced in the human body and has anti‐inflammatory properties. We have previously shown in a rat temporomandibular joint (TMJ) model that injection of 15d‐PGJ2 into the rat TMJ can provide antinociceptive relief against a subsequent noxious challenge from formalin injection into the same TMJ. However, intra‐TMJ injections are painful. Thus, to make the treatment patient friendly, this study aimed to evaluate whether the antinociceptive property of 15d‐PGJ2 cream can be enhanced with microneedles (MNs). We found that topical application of 15d‐PGJ2 cream for 15 min directly on the rat TMJ skin did not induce any significant antinociceptive effect. However, if MNs were inserted in the skin for 5 min, removed, and then 15d‐PGJ2 cream was applied, a significant reduction in formalin‐induced nociceptive behavior was observed. This reduction in nociception was comparable to an intra‐TMJ injection of 15d‐PGJ2. A concentration‐dependent effect of 15d‐PGJ2 was observed, with higher concentrations of 15d‐PGJ2 in the cream showing a more durable effect up to 8 h. 15d‐PGJ2 cream associated with MNs also significantly reduced the release of tumor necrosis factor‐&agr; and interleukin‐1 beta, which are pro‐inflammatory cytokines. Our findings suggest that 15d‐PGJ2 cream associated with MNs provides antinociceptive and anti‐inflammatory effect, and can offer a potential patient‐friendly therapeutic option for pain control related to inflammatory disorders of the TMJ. Graphical abstract Figure. No Caption available.

The role of endogenous opioid peptides in the antinociceptive effect of 15-deoxyΔ12,14-prostaglandin J2 in the temporomandibular joint

We have previously demonstrated that peripheral administration of 15d-PGJ2 in the Temporomandibular joint (TMJ) of rats can prevent nociceptor sensitization, mediated by peroxisome proliferator activated receptor-γ (PPAR-γ), and κ- and δ- opioid receptors. However, the mechanism that underlies the signaling of PPAR-γ (upon activation by 15d-PGJ2) to induce antinociception, and how the opioid receptors are activated via 15d-PGJ2 are not fully understood. This study demonstrates that peripheral antinociceptive effect of 15d-PGJ2 is mediated by PPAR-γ expressed in the inflammatory cells of TMJ tissues. Once activated by 15d-PGJ2, PPAR-γ induces the release of β-endorphin and dynorphin, which activates κ- and δ-opioid receptors in primary sensory neurons to induce the antinociceptive effect.

15d-PGJ2-Loaded Solid Lipid Nanoparticles: Physicochemical Characterization and Evaluation of Pharmacological Effects on Inflammation

15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2), a peroxisome proliferator-activated receptor-γ (PPAR-γ) agonist, has physiological properties including pronounced anti-inflammatory activity, though it binds strongly to serum albumin. The use of solid lipid nanoparticles (SLN) can improve therapeutic properties increasing drug efficiency and availability. 15d-PGJ2-SLN was therefore developed and investigated in terms of its immunomodulatory potential. 15d-PGJ2-SLN and unloaded SLN were physicochemically characterized and experiments in vivo were performed. Animals were pretreated with 15d-PGJ2-SLN at concentrations of 3, 10 or 30 μg·kg-1 before inflammatory stimulus with carrageenan (Cg), lipopolysaccharide (LPS) or mBSA (immune response). Interleukins (IL-1β, IL-10 and IL-17) levels were also evaluated in exudates. The 15d-PGJ2-SLN system showed good colloidal parameters and encapsulation efficiency of 96%. The results showed that the formulation was stable for up to 120 days with low hemolytic effects. The 15d-PGJ2-SLN formulation was able to reduce neutrophil migration in three inflammation models tested using low concentrations of 15d-PGJ2. Additionally, 15d-PGJ2-SLN increased IL-10 levels and reduced IL-1β as well as IL-17 in peritoneal fluid. The new 15d-PGJ2-SLN formulation highlights perspectives of a potent anti-inflammatory system using low concentrations of 15d-PGJ2.

Low dose propranolol decreases orthodontic movement

OBJECTIVE Low dose propranolol has previously been demonstrated to suppress bone remodelling. Therefore, its effect on orthodontic movement was tested. DESIGN Rats were assigned as follows (n=5): animals with no orthodontic appliance (G1); the remaining groups were fitted with a Ni-Ti closed-coil spring ligated to the upper left first molar and connected to the incisors using metal and resin and received vehicle only (G2), 0.1mg/kg (G3) or 20mg/kg (G4) of propranolol orally. Cone Beam Computed Tomography was performed using high resolution for image capture. The distance between the first and second upper molars, both with and without the orthodontic appliance, was measured in millimetres. Gingival tissue was harvested and assessed for IL-1β and IL-6 using ELISA and for ICAM-1 and RANKL by Western blotting. RESULTS The orthodontic appliance induced a significant tooth movement in G2 when compared to the animals without an orthodontic appliance (G1) (p<0.05). The animals from G3 showed a significantly reduction in tooth movement (p<0.05) when compared with rats from G2. Animals treated with 20mg/kg of propranolol (G4) showed tooth movement similar to that of G2. The reduced tooth movement observed in the animals treated with 0.1mg/kg of propranolol (G3) occurred due to decreased amounts of IL-1β and IL-6, in addition to lower ICAM-1 and RANKL expression. CONCLUSIONS Low dose propranolol inhibits bone remodelling and orthodontic movement.