Celecoxib treatment dampens LPS-induced periapical bone resorption in a mouse model.

AIM: To evaluate the efficacy of selective and nonselective inhibitors of cyclooxygenase-2 enzymes in the treatment of experimental apical periodontitis induced by bacterial lipopolysaccharide (LPS) in vivo in a mouse model. METHODOLOGY: Thirty-six C57BL/6 mice were used. After access cavity preparation, a solution containing E. coli LPS (1.0 µg µL-1 ) was inoculated into the root canals of the mandibular and maxillary right first molars (n = 72) After 30 days, apical periodontitis was established and the animals were systemically treated with celecoxib, a selective COX-2 inhibitor (15 mg kg-1 ), or indomethacin, a nonselective COX-2 inhibitor (5 mg kg-1 ), for 7 and 14 days. Blocks containing teeth and bone were removed for histopathological and histometric analyses (haematoxylin and eosin), evaluation of osteoclasts numbers (tartrate-resistant acid phosphatase enzyme - TRAP) and immunohistochemistry for RANK, RANKL and OPG. Gene expression was performed using reverse transcription and real-time polymerase chain reaction (qRT-PCR) for RANK, RANKL, OPG, TRAP, MMP-9, cathepsin K and calcitonin receptor. Histopathological, histometric, TRAP, immunohistochemistry and qRT-PCR data were evaluated using Kruskal-Wallis followed by Dunn's test (α = 0.05). RESULTS: Systemic administration of celecoxib for 7 and 14 days prevented periapical bone resorption (P < 0.0001), differently from indomethacin that exacerbated bone resorption at 7 days (P < 0.0001) or exerted no effect at 14 days (P = 0.8488). Celecoxib treatment reduced osteoclast formation in apical periodontitis, regardless of the period of treatment (P < 0.0001 for 7 days and P = 0.026 for 14 days). Administration of celecoxib or indomethacin differentially modulated the expression of genes involved in bone resorption. At 7 days, celecoxib and indomethacin treatment significantly inhibited expression of mRNA for cathepsin K (P = 0.0005 and P = 0.016, respectively) without changing TRAP, MMP-9 and calcitonin receptor gene expression. At 14 days, celecoxib significantly inhibited expression of mRNA for MMP-9 (P < 0.0001) and calcitonin receptor (P = 0.004), whilst indomethacin exerted no effect on MMP-9 (P = 0.216) and calcitonin receptor (P = 0.971) but significantly augmented cathepsin K gene expression (P = 0.001). CONCLUSIONS: The selective COX-2 inhibitor celecoxib reduced osteoclastogenic signalling and activity that dampened bone resorption in LPS-induced apical periodontitis in mice, with greater efficacy than the nonselective inhibitor indomethacin.

Effect of carbodiimide and chlorhexidine on the bond strength longevity of resin cement to root dentine after radiation therapy.

AIM: To evaluate the effect of carbodiimide (EDC) and chlorhexidine (CHX) on the bond strength (BS) of resin cement to root dentine of teeth submitted to radiotherapy. METHODOLOGY: One hundred and twenty extracted maxillary canines were selected and assigned to 2 groups (n = 60): nonirradiated and irradiated (30 cycles of 2 Gy, total 60 Gy). Roots lengths were standardized, and canals were prepared and filled. Post spaces were then prepared, and the samples were redistributed according to dentine treatment (n = 20): saline solution (SF); CHX 2%; or EDC 0.5M. After drying the post space, fibreglass posts were cemented. Cross-sectioned slices were obtained, and in half of the specimens of each subgroup (n = 10), the analysis was performed immediately; the others (n = 10) were stored for 10 months before analyses. The most cervical slice of each third was subjected to a push-out test and failure pattern analysis (n = 10), and the most apical slice submitted to the analysis of the adhesive interface by SEM (n = 5). The bond strength data were submitted to anova and Tukey tests, the adhesive interface adaptation was submitted to Kruskal-Wallis and Dunn's tests, and the Chi-square test was used to evaluate the type of failure. RESULTS: The irradiated specimens had significantly lower bond strength (13.8 ± 4.3) than the nonirradiated (18.1 ± 3.1; P < 0.001). For the irradiated teeth, the bond strengths were significantly lower in the SF and CHX groups (P < 0.001). Also, the bond strengths reduced significantly after 10 months in the SF and CHX groups (P < 0.001). Cohesive failures occurred in dentine for irradiated specimens. Poorer interface adaptation, dentine fractures and microfractures were observed in irradiated specimens, and better adaptation was observed for specimens after EDC treatment. CONCLUSIONS: Radiotherapy was associated with lower bond strength and worse interface adaptation. Dentine treatment with EDC contributed to adhesive interface longevity during the cementation of glass fibre posts in nonirradiated and irradiated teeth.

Immunophenotypic quantification of M1 and M2 macrophage polarization in radicular cysts of primary and permanent teeth.

AIM: To quantify M1 and M2 macrophages in radicular cysts of permanent (n = 14 cases) and primary teeth (n = 15 cases). METHODOLOGY: All patients who attended the School of Dentistry Ribeirão Preto, University of São Paulo with primary teeth or permanent molars that were scheduled for extraction and fulfilled the inclusion criteria: absence of pain; presence/absence of fistulae; extensive coronal destruction due to caries lesions without possibility of restoration; pulp necrosis; radiographically visible apical periodontitis; and no previous treatment, were selected. The radicular cysts were removed and subsequently submitted to histopathologic analysis in order to classify the type of inflammatory infiltrate. In addition, CD68 (M1+, M2+) and CD163 (M1-, M2+) markers were quantified through an immunohistochemistry analysis. The data acquired were submitted to a Mann-Whitney test, with a 5% significance level. RESULTS: The patients had a mean age of 38.6 years and 5.9 years for cysts associated with permanent and primary teeth, respectively. In the histopathological analysis, no significant difference (P = 0.87) was found between radicular cysts in primary and permanent teeth regarding the intensity of the chronic inflammatory infiltrate. A significantly greater prevalence of M2 macrophages (P < 0.05) was observed in the lesions of both permanent and primary teeth, even though both M1 and M2 macrophages were detected. No significant difference (P > 0.05) was found for M1 and M2 macrophages associated with the cysts of primary and permanent teeth. CONCLUSION: M1 and M2 macrophages were present in radicular cysts associated with primary and permanent teeth, with a greater quantity of M2 cells. The immunophenotypic quantification of M1 and M2 macrophage polarization in radicular cysts associated with primary and permanent teeth were similar.

Histopathological, histoenzymological, immunohistochemical and immunofluorescence analysis of tissue response to sealing materials after furcation perforation.

AIM: To evaluate in vivo tissue responses after sealing furcation perforations in dog's teeth with either Biodentine™, mineral trioxide aggregate (MTA) or gutta-percha, by means of histopathological, histoenzymological, immunohistochemical and immunofluorescence analysis. METHODOLOGY: After root canal treatment, perforations were created in the central region of the pulp chamber floor using a round diamond bur and filled with one or other of the materials. The animals were euthanized after 120 days, and the teeth (n = 30) were processed for histopathological analysis of new mineralized tissue formation and collagen fibre reinsertion, immunohistochemical analysis of osteopontin (OPN) and alkaline phosphatase (ALP) and immunofluorescence analysis for bone morphogenetic protein (BMP-2), cementum attachment protein (CAP), bone sialoprotein (BSP), osteocalcin (OCN) and cementum protein1 (CEMP1). Histoenzymology was performed for TRAP activity and osteoclast count. Data were analysed statistically (α = 0.05) using chi-square and Kruskal-Wallis tests. RESULTS: Gutta-percha did not induce mineralized tissue formation. MTA and BiodentineTM formed mineralized tissue in 88% and 92% of specimens, respectively, with no significant difference (P > 0.05). Gutta-percha was associated with scattered collagen fibres parallel to the perforations. Groups treated with MTA or BiodentineTM had partial fibre reinsertion perpendicular to the newly formed mineralized tissue. All materials induced OPN and ALP expression, weakest for gutta-percha and strongest for MTA (P < 0.05). Only MTA induced BMP-2, BSP, OCN, CAP and CEMP1 expression. Osteoclast counts were similar in all groups (P = 0.97). CONCLUSIONS: Mineral trioxide aggregate and BiodentineTM were biocompatible, with formation of mineralized tissue and partial reinsertion of collagen fibres. In addition, the participation of several molecules by which calcium silicate-based materials induce the formation of mineralized tissue were noted, with expression of ALP and OPN mineralization markers, without interference in the number of osteoclasts. Only MTA stimulated the expression of proteins associated with the formation of a cementum-like mineralized tissue.

Longevity of bond strength of resin cements to root dentine after radiation therapy.

AIM: To evaluate the bond strength and adhesive interface between several resin cements and root dentine immediately and 6 months after radiotherapy. METHODOLOGY: Sixty maxillary canines were selected and randomly assigned to two groups (n = 30): one group was not irradiated and the other one was subjected to a cumulative radiation dose of 60 Gy. The teeth were sectioned to obtain roots 16 mm long and the canals were prepared with the Reciproc system (R50) and filled using a lateral condensation technique with an epoxy resin sealer. Each group was divided into three subgroups (n = 10) according to the resin cement used for fibreglass fibre post cementation: RelyX-U200, Panavia-F2.0 and RelyX ARC. The posts were cemented in accordance with the manufacturer's recommendations. Three 1-mm-thick dentine slices were then obtained from each root third. The first two slices in the crown-apex direction of each third were selected for the push-out test. The failure mode after debonding was determined with a stereo microscope. The third slice from each root third was selected for scanning electron microscopy (SEM) analyses to examine the resin cement-dentine interface with 100, 1000, 2000 and 4000× magnification. Bond strength data were analysed by anova and Tukey's test (α = 0.05). RESULTS: Significantly lower bond strength (P < 0.0001) was obtained after irradiation compared to nonirradiated teeth. RelyX-U200 cemented fibre posts had the higher bond strength (15.17 ± 5.89) compared with RelyX ARC (P < 0.001) and Panavia-F2.0 (P < 0.001). The evaluation after 6 months revealed lower bond strength values compared to the immediate values (P < 0.001) for irradiated and nonirradiated teeth. Cohesive failures occurred in the irradiated dentine. SEM revealed fractures, microfractures and fewer collagen fibres in irradiated root dentine. RelyX-U200 and Panavia-F2.0 were associated with a juxtaposed interface of the cement with the radicular dentine in irradiated and nonirradiated teeth, and for RelyX ARC, hybrid layer formation and tags were observed in both irradiated and nonirradiated teeth. CONCLUSION: Radiation was associated with a decrease in the push-out bond strength and with lower resin cement/root dentine interface adaptation. Self-adhesive resin cement was a better alternative for fibre post cementation in teeth subjected to radiation therapy. The bond strength decreased after 6 months.

Influence of therapeutic cancer radiation on the bond strength of an epoxy- or an MTA-based sealer to root dentine.

AIM: To evaluate the influence of radiation on root canal sealer push-out bond strength to dentine and sealer/dentine interface in teeth filled with AH Plus (Dentsply Ind. Com. Ltda, Petrópolis, RJ, Brazil) and MTA Fillapex (Angelus Ind. Prod. Odontológicos S/A, Londrina, PR, Brazil). METHODOLOGY: Thirty-two maxillary canines were selected and randomly assigned to 2 groups (n = 16): one group was not irradiated, and the other was subjected to a cumulative radiation dose of 60 Gy. Root canals were prepared with the Reciproc system (VDW GmbH, Munich, Germany), and each group was divided into 2 subgroups (n = 8) according to the sealer - AH Plus or MTA Fillapex - using the single-cone filling technique. Then, 1-mm-thick dentine slices were obtained from each root third for the push-out test to evaluate sealer bond strength to dentine and for scanning electron microscopy (SEM) to examine the sealer/dentine interface. Failure mode after debonding was determined with a stereomicroscope at ×25 magnification. Bond strength data were analysed by two-way anova with a split-plot design and post hoc Tukey's test (α = 0.05). RESULTS: Significantly lower bond strength (P < 0.0001) was obtained after irradiation (0.71 ± 0.20 versus 0.97 ± 0.29 MPa) and in specimens filled with MTA Fillapex (0.70 ± 0.18 MPa) compared with AH Plus (1.00 ± 0.27 MPa). Percentage of adhesive failures increased after radiation in all root thirds in the teeth filled with AH Plus. SEM revealed more gap-containing regions and fewer tags at the sealer/dentine interface in irradiated specimens, with more tag formation and fewer gaps with AH Plus sealer. CONCLUSIONS: Radiation was associated with a decrease in the push-out bond strength of sealers to intraradicular dentine and formation of more gaps and fewer tags at the sealer/dentine interface regardless of the sealer.

MyD88 knockout mice develop initial enlarged periapical lesions with increased numbers of neutrophils.

AIM: To characterize the formation and progression of experimentally induced periapical lesions in teeth of MyD88 knockout (MyD88 KO) mice compared with wild-type (WT) mice. METHODOLOGY: Periapical lesions were induced in the mandibular first molars of 30 WT and 30 MyD88 KO mice. After 7, 21 and 42 days, the animals were euthanized and the mandibles were subjected to histotechnical processing. Histological sections were stained with haematoxylin and eosin (HE), TRAP histoenzymology, Brown and Brenn staining and immunohistochemistry (RANK, RANKL, OPG). Data were subjected to statistical analysis by the nonparametric Mann-Whitney and Kruskal-Wallis tests and the Dunn post-test, using the SPSS software, version 17.0 (α = 0.05). RESULTS: Regarding the periapical lesion size, the MyD88 KO group had significantly higher values than the WT group in the periods of 7 (P = 0.001) and 21 days (P = 0.05). A larger number of neutrophils in the MyD88 KO group were observed (P = 0.01 at 7 days, P = 0.004 at 21 days and P < 0.001 at 42 days). Regarding the number of osteoclasts, no statistically significant difference was observed between the groups at any of the experimental periods (P = 0.884 at 7 days, P = 0.506 at 21 days and P = 0.211 at 42 days). CONCLUSIONS: In the absence of MyD88, the animals had larger periapical lesions, with a severe inflammatory infiltrate and a significantly larger number of neutrophils.

Analysis of the stomatognathic system functions and mastication muscles in children affected by molar hypomineralization (MH): a cross-sectional study.

PURPOSE: To analyze the functions of the stomatognathic system in children with or without molar-incisor hypomineralization (MIH). METHODS: For this cross-sectional study, 72 children aged 6-12 years were recruited and divided in two groups: with MIH (G1) and without MIH (G2). T-SCAN was used to verify the distribution of occlusal contacts, gnathodynamometer to measure maximum molar bite force, and Iowa Oral Pressure Instrument (IOPI) to assess the strength of facial expression muscles. The t test and paired t test (p ≤ 0.05) were used for statistical comparisons. RESULTS: The molars affected by MIH exhibited lower distribution of occlusal forces (p < 0.001) and lower maximum molar bite force (p < 0.05) compared to the molars in the control group. However, there was no difference between the MIH-affected sides compared to the unaffected side, nor between the molars affected by MIH and their antagonists (p > 0.05). There were no differences in the forces of the facial expression muscles between the groups. CONCLUSIONS: These findings suggest that MIH significantly impacts occlusal force distribution and bite force, but not facial expression musculature.

The treatment of a maxillary lateral incisor with unusual morphology with long-term follow-up.

AIM: To report the treatment of an unusual combination of one dens evaginatus and two dens invaginatus in a single tooth and its healing outcome after 10 years. SUMMARY: The long-term outcome of a maxillary lateral incisor with dens evaginatus combined with two Oehlers type II dens invaginatus and a large periradicular lesion in an 11-year-old female treated endodontically and restoratively is described. The endodontic treatment included intracanal medication with calcium hydroxide and canal filling using a thermoplastic root canal filling technique. The crown was restored with conventional composite resin. During periodic clinical and radiographic follow-up, the patient remained symptom free, and the periradicular region was completely healed, meeting both aesthetic and functional expectations after 10 years. KEY LEARNING POINTS: The co-occurrence of dens evaginatus and two dens invaginatus in the same tooth is an unusual finding that compromises aesthetics and predisposes the patient to dental pulp infection. The complex morphology observed in this case represented both endodontic and restorative challenges.

Association of dental hypersensitivity and anxiety in children with molar-incisor hypomineralisation (MIH).

AIM: The objective of this study was to investigate whether dental hypersensitivity and dental fear were linked to the presence and severity of MIH. METHODS: For this cross-sectional study, 1830 students between the ages of 6 and 12 years were recruited from four randomly selected schools. The Children's Fear Survey Schedule-Dental Subscale questionnaire was used to assess dental anxiety and fear. The children's self-reported dental hypersensitivity resulting from MIH was evaluated using the Wong-Baker Facial Scale and the Visual Analog Scale (VAS). RESULTS: MIH was correlated with tooth hypersensitivity, particularly in severe cases. Dental fear was present in 17.4% of the children with MIH, but it was not associated with dental hypersensitivity, gender, or age. CONCLUSION: No association was found between dental fear and dental hypersensitivity in children with MIH.

Frontonasal dysplasia: oral features, restorative and orthodontic dental treatment in a child.

BACKGROUND: Frontonasal dysplasia is a complex rare malformation, characterised by abnormalities involving the central portion of the face, especially the eyes, nose and forehead. It can manifest independently or associated with other abnormalities as part of some syndromes. CASE REPORT: The purpose of this case report was to describe a 5-year-old patient, diagnosed with frontonasal dysplasia. Among the abnormalities characterised with this disorder were ocular hypertelorism, broad nose tip with median notch, median facial cleft, bifid anterior skull, low set hairline, Poland's syndactyly and ankyloglossia. TREATMENT: Consisted of behavioural management, oral hygiene instruction, prophylaxis, topical fluoride application, extraction of primary teeth, composite resin restorations and sealants in pits and fissures. Preformed metal crowns were also applied to the right and left primary maxillary second molars. FOLLOW-UP: Currently, the patient is 11 years-old in the permanent dentition and therefore was referred for corrective orthodontic and periodontal treatments due to the persistence of gingival retraction of the permanent mandibular right central incisor. CONCLUSION: The treatment in this case was directed to the promotion of oral health and orthodontic corrections, which are of fundamental importance due to medical, physical and social limitations of children affected by this syndrome, hindering healing and rehabilitative treatment. Paediatric dentists should be included in multidisciplinary teams providing care to patients with special needs, improving their quality of life.

Quenching of chlorophyll fluorescence induced by silver nanoparticles.

The interaction between chlorophyll (Chl) and silver nanoparticles (AgNPs) was evaluated by analyzing the optical behavior of Chl molecules surrounded by different concentrations of AgNPs (10, 60, and 100nm of diameter). UV-Vis absorption, steady state and time-resolved fluorescence measurements were performed for Chl in the presence and absence of these nanoparticles. AgNPs strongly suppressed the Chl fluorescence intensity at 678nm. The Stern-Volmer constant (KSV) showed that fluorescence suppression is driven by the dynamic quenching process. In particular, KSV was nanoparticle size-dependent with an exponential decrease as a function of the nanoparticle diameter. Finally, changes in the Chl fluorescence lifetime in the presence of nanoparticles demonstrated that the fluorescence quenching may be induced by the excited electron transfer from the Chl molecules to the metal nanoparticles.