Renato Menezes

Differential Patterns of Receptor Activator of Nuclear Factor Kappa B Ligand/Osteoprotegerin Expression in Human Periapical Granulomas: Possible Association with Progressive or Stable Nature of the Lesions

Receptor activator of nuclear factor kappa B ligand (RANKL) and osteoprotegerin (OPG) are expressed in apical periodontitis, suggesting a role for these molecules during lesion development. However, the profiles of RANKL/OPG expression in periapical lesions remain unknown. In this study we investigated the patterns of RANKL and OPG mRNA expression by real-time polymerase chain reaction in human periapical granulomas (N = 44) and compared them with sites presenting characteristic bone resorbing activity: healthy (n = 14) and orthodontically stretched and compressed periodontal ligament (n = 26), healthy gingiva (n = 24), chronic gingivitis (n = 32), and chronic periodontitis (n = 34) samples. Both RANKL and OPG mRNA expression was higher in periapical granulomas when compared with healthy periodontal ligament. Distinct patterns of RANKL and OPG expression ratio were found in the granulomas and in different physiologic and pathologic conditions, with characteristic bone resorption activity potentially being indicative of the stable or progressive nature of the lesions. Lesions with radiographic image smaller than 5 mm showed higher RANKL/OPG expression than images greater than 5 mm. Periapical granulomas presented heterogeneous patterns of RANKL and OPG expression, ranging from samples with RANKL/OPG ratio similar to that seen in sites with minimal or absent bone resorption to samples with RANKL/OPG expression pattern comparable with active bone resorption sites.

The potential role of suppressors of cytokine signaling in the attenuation of inflammatory reaction and alveolar bone loss associated with apical periodontitis.

Inflammatory cytokines contribute to periapical tissue destruction. Their activity is potentially regulated by suppressors of cytokine signaling (SOCS), which downregulate signal transduction as part of an inhibitory feedback loop. We investigated the expression of the cytokines tumor necrosis factor alpha (TNF-alpha); interleukin (IL)-10 and RANKL; and SOCS-1, -2, and -3 by real-time polymerase chain reaction in 57 periapical granulomas and 38 healthy periapical tissues. Periapical granulomas exhibited significantly higher SOCS-1, -2, and -3, TNF-alpha, IL-10, and RANKL messenger RNA levels when compared with healthy controls. Significant positive correlations were found between SOCS1 and IL-10 and between SOCS3 and IL-10. Significant inverse correlations were observed between SOCS1 and TNF-alpha, SOCS1 and RANKL, and SOCS3 and TNF-alpha. Increased SOCS-1, -2, and -3 messenger RNA levels in periapical granulomas may be related to the downregulation of inflammatory cytokines in these lesions; therefore, SOCS molecules may play a role in the dynamics of periapical granulomas development.

Expression analysis of matrix metalloproteinase-9 in epithelialized and nonepithelialized apical periodontitis lesions

OBJECTIVEnThe objective of this study was to determine the expression of matrix metalloproteinase-9 (MMP-9) in apical periodontitis lesions.nnnSTUDY DESIGNnNineteen epithelialized and 18 nonepithelialized apical periodontitis lesions were collected after periapical surgery. After histological processing, serial sectioning, H&E staining, and microscopic analysis, 10 epithelialized and 10 nonepithelialized lesions were selected for immunohistochemical analysis for MMP-9 and CD 68. At least one third of each specimen collected was frozen at -70 degrees C for further mRNA isolation and reverse transcription into cDNA for real-time-PCR procedures. Geometric averaging of multiple housekeeping genes normalized MMP-9 mRNA expression level.nnnRESULTSnPolymorphonuclear neutrophils, macrophages and lymphocytes presented MMP-9 positive immunostaining in both types of lesions. When present, epithelial cells were also stained. The number and the ratio of MMP-9(+)/total cells were greater in nonepithelialized than epithelialized lesions (P = .0001) presenting a positive correlation to CD68(+)/total cells (P = .045). Both types of lesions presented increased MMP-9 expression (P < .0001) when compared to healthy periapical ligaments. However, no significant differences were observed for MMP-9 mRNA expression between ephithelized and nonephithelized lesions.nnnCONCLUSIONnThe present data suggest the participation of several inflammatory cells, mainly CD68(+) cells, in the MMP-9 expression in apical periodontitis lesions. MMP-9 could be actively enrolled in the extracellular matrix degradation in apical periodontitis lesions.

Tissue response to a membrane of demineralized bovine cortical bone implanted in the subcutaneous tissue of rats

The treatment of persistent bone defects has encouraged the search for proper techniques or bone substitutes. In Dentistry, a common problem in the treatment of periodontal bone defects is the growth of tissues within the lesion, such as the junctional epithelium, which impair regeneration of these tissues. Guided tissue regeneration (GTR), based on the separation of the tissues by means of membranes or barriers, was developed in an attempt to improve periodontal regeneration. The aim of this study was to histologically evaluate the tissue response to a membrane of demineralized bovine cortical bone implanted in the subcutaneous tissue of rats. The study periods were 1, 3, 7, 15, 30 and 60 days after implantation. Analysis of the histological sections demonstrated a moderate to intense inflammatory response at 1 and 3 days, moderate at 7 and 15 days, and almost absent at 30 and 60 days. Resorption of the membrane began 15 days after implantation, and at 60 days only remnants could be detected in some animals. We concluded that the demineralized bovine cortical bone membrane was well tolerated by the tissues and is completely resorbed after 30-60 days by mononuclear cells and multinucleated giant cells, which disappear upon completion of the process.

Microscopic and radiographic analysis of the effect of particle size of demineralized bovine cancellous bone matrix on the repair of bone defects in femurs of rabbits

The bone tissue has a great regenerative potential, with ability to completely restore its structure and original functions. In some situations, though, bone defects cannot be self-repaired, thus requiring the use of grafts for a correct treatment and good prognosis. This work aimed at microscopically analyzing the effect of the particle size of demineralized bovine cancellous bone matrix in micro and macrogranular forms on the repair of bone defects in femurs of rabbits, with blood clot used as control. At 1, 3 and 6 months after implantation of the materials, the animals were killed and the anatomic specimens were removed. A foreign body-type granulomatous reaction containing macrophages and multinucleated giant cells in contact with the implanted particles was observed. These results suggest a failure in demineralization and/or interruption of the antigenic potential during production of the biomaterial. It is concluded that the size of the particles did not influence the evolution of the repair process of bone defects, acting only as bone-filler substances, and that the material implanted should be improved by quality control during production, since it may represent a good alternative for bone graft.

Microscopic analysis of dog dental pulp after pulpotomy and pulp protection with mineral trioxide aggregate and white Portland cement

Considering previous studies on the similarity between the chemical composition of the mineral trioxide aggregate and the Portland cement, the purpose of this study was to investigate the pulp response of dogs teeth after pulpotomy and direct pulp protection with MTA Angelus and white Portland cement. Thirty eight pulp remnants were protected with these materials. One hundred and twenty days after treatment, the animals were sacrificed and the specimens removed and prepared for histological analysis. Both materials demonstrated the same results when used as pulp capping materials, inducing hard tissue bridge formation and maintaining pulp vitality in all specimens. The MTA Angelus and the white Portland cement showed to be effective as pulp protection materials following pulpotomy.

Studies with MMP9 gene promoter polymorphism and nonsyndromic cleft lip and palate.

Ariadne Letra, Rodrigo A. da Silva, Renato Menezes, Ana P. de Souza, Ana L.P.F. de Almeida, Mari C. Sogayar, and Jose M. Granjeiro* Department of Biological Sciences, Bauru Dental School, University of Sao Paulo, Bauru, Sao Paulo, Brazil Oral Biology Program, University of Sagrado Coracao, Bauru, Sao Paulo, Brazil Department of Periodontology, Hospital of Rehabilitation and Craniofacial Anomalies, University of Sao Paulo, Bauru, Sao Paulo, Brazil Institute of Chemistry, University of Sao Paulo, Sao Paulo, Brazil Department of Cellular and Molecular Biology, Fluminense Federal University, Niteroi, Rio de Janeiro, Brazil

Influence of the EDTA, Nd:YAG laser and association of both on the filling of artificial lateral root canals.

This study aimed at evaluating the influence of EDTA, Nd:YAG laser and the combination of both for filling of artificial lateral root canals. Forty-five human mandibular premolars were employed, on which three artificial lateral root canals were prepared by means of a reamer with a similar diameter to a K file #15. The teeth were instrumented through the stepback technique employing Gates Glidden burs at the middle and cervical thirds and manual files at the apical portion, and irrigation with 1% sodium hypochloride. The teeth were divided in three groups: Group 1 -EDTA for 5 minutes; Group 2 -application of Nd:YAG laser at 15 Hz, 100 mJ and 1.5 Watts; and Group 3 - association of both. Roots were filled through the Taggers hybrid technique, radiographed and the radiographs were digitized. Scores were assigned to the filling of the lateral root canals. Statistical analysis revealed no significant differences between the entire groups and also on the analysis of each third.