Paulo Nelson-Filho

Effect of Calcium Hydroxide on Bacterial Endotoxin In Vivo

The aim of this study was the histopathological evaluation of apical and periapical tissues in dog teeth that were submitted to bacterial endotoxin, associated or not with calcium hydroxide. After removal of the pulp from 60 premolars, the teeth were divided into four groups and were filled with bacterial endotoxin (group 1), bacterial endotoxin plus calcium hydroxide (group 2), saline solution (group 3), or had induced periapical lesions with no treatment (group 4). After 30 days, animals were killed and the teeth processed histologically. The inflammatory infiltrate, the thickness of the periodontal ligament, and the presence of resorption areas were similar for groups 1 and 4. Groups 2 and 3 were similar to each other. It can be concluded that the bacterial endotoxin caused a periapical lesion and that calcium hydroxide detoxified the lipopolysaccharides in vivo.

Radiographic Evaluation of the Effect of Endotoxin (LPS) Plus Calcium Hydroxide on Apical and Periapical Tissues of Dogs

The aim of this study was the radiographic evaluation of the apical and periapical region of dog teeth submitted to intracanal bacterial endotoxin (lipopolysaccharide, LPS), associated or not with calcium hydroxide. After removal of the pulp, 60 premolars were divided into four groups and were filled with bacterial endotoxin (group 1), bacterial endotoxin plus calcium hydroxide (group 2), saline solution (group 3), or periapical lesions were induced with no treatment (group 4), for a period of 30 days. Similar periapical lesions were observed in groups 1 and 4. The lamina dura was intact in groups 2 and 3. Bacterial endotoxin (LPS) caused radiographically visible periapical lesions, but when associated with calcium hydroxide, this endotoxin was detoxified.

Bacterial profile in primary teeth with necrotic pulp and periapical lesions

The objective of this study was to evaluate the bacterial profile in root canals of human primary teeth with necrotic pulp and periapical lesions using bacterial culture. A total of 20 primary teeth with necrotic pulp and radiographically visible radiolucent areas in the region of the bone furcation and/or the periapical region were selected. After crown access, 4 sterile absorbent paper points were introduced sequentially into the root canal for collection of material. After 30 s, the paper points were removed and placed in a test tube containing reduced transport fluid (RTF) and were sent for microbiological evaluation. Anaerobic microorganisms were found in 100% of the samples, black-pigmented bacilli in 30%, aerobic microorganisms in 60%, streptococci in 85%, gram-negative aerobic rods in 15% and staphylococci were not quantified. Mutans streptococci were found in 6 root canals (30%), 5 canals with Streptococcus mutans and 1 canal with Streptococcus mutans and Streptococcus sobrinus. It was concluded that in root canals of human primary teeth with necrotic pulp and periapical lesions, the infection is polymicrobial with predominance of anaerobic microorganisms.

Importance of bacterial endotoxin (LPS) in endodontics

New knowledge of the structure and biological activity of endotoxins (LPS) has revolutionized concepts concerning their mechanisms of action and forms of inactivation. Since the 1980s, technological advances in microbiological culture and identification have shown that anaerobic microorganisms, especially Gram-negative, predominate in root canals of teeth with pulp necrosis and radiographically visible chronic periapical lesions. Gram-negative bacteria not only have different factors of virulence and generate sub-products that are toxic to apical and periapical tissues, as also contain endotoxin (LPS) on their cell wall. This is especially important because endotoxin is released during multiplication or bacterial death, causing a series of biological effects that lead to an inflammatory reaction and resorption of mineralized tissues. Thus, due to the role of endotoxin in the pathogenesis of periapical lesions, we reviewed the literature concerning the biological activity of endotoxin and the relevance of its inactivation during treatment of teeth with pulp necrosis and chronic periapical lesion.

Biofilm on the apical region of roots in primary teeth with vital and necrotic pulps with or without radiographically evident apical pathosis

AIM To evaluate, by scanning electron microscopy (SEM), the presence of biofilms on the external surfaces of the apical third of roots of human primary teeth with vital or necrotic pulps with and without radiographically evident periradicular pathosis. METHODOLOGY Eighteen teeth were selected: group I - normal pulp (n = 5), group II - pulp necrosis without radiographic evidence of periapical pathosis (n = 7) and group III - pulp necrosis with well-defined radiographic periapical pathosis (n = 6). After extraction, the teeth were washed with saline and immersed in 0.03 g mL(-1) trypsin solution for 20 min. The teeth were then washed in sodium cacodilate buffer and stored in receptacles containing modified Karnovsky solution. The teeth were sectioned, dehydrated in an ethanol series, critical-point dried with CO(2), sputter coated with gold and the external root surface in the apical third examined by SEM. RESULTS In the teeth of groups I and II, the apical root surfaces were covered by collagen fibres, with no evidence of bacteria (100%). In the teeth of group III, the root apices had no collagen fibres but revealed resorptive areas containing microorganisms (cocci, bacilli, filaments and spirochetes) in all cases (100%). CONCLUSION Microorganisms organized as biofilms on the external root surface (extraradicular infection) were detected in primary teeth with pulp necrosis and radiographically visible periapical pathosis.

Prevalence of microorganisms in root canals of human deciduous teeth with necrotic pulp and chronic periapical lesions

The objective of this study was to evaluate bacterial prevalence in 31 root canals of human deciduous teeth with necrotic pulp and periapical lesions using bacterial culture. After crown access, the material was collected using absorbent paper points for microbiological evaluation and determination of colony forming units (CFU). Anaerobic microorganisms were found in 96.7% of the samples, black-pigmented bacilli in 35.5%, aerobic microorganisms in 93.5%, streptococci in 96.7%, and S. mutans in 48.4%. We concluded that in human deciduous teeth root canals with necrotic pulp and periapical lesions the infection is polymicrobial, with a large number of microorganisms and a predominance of streptococci and anaerobic microorganisms.

In-vivo evaluation of the contamination of Super Slick elastomeric rings by Streptococcus mutans in orthodontic patients

INTRODUCTION We investigated in vivo the contamination by Streptococcus mutans of Super Slick elastomeric rings (TP Orthodontics, LaPorte, Ind), manufactured with Metafasix technology (TP Orthodontics), using microbial culture and scanning electron microscopy (SEM). METHODS Twenty patients undergoing fixed orthodontic appliance therapy were selected. Super Slick elastomeric rings (n = 160) were tied to brackets on the right maxillary premolars or molars and left mandibular premolars or molars. Conventional elastomeric rings (n = 160) were tied to brackets on the contralateral premolars or molars with the same split-mouth design. After a 15-day intraoral period, 75 elastomeric rings of each type were retrieved, submitted to microbiologic processing, and cultured in bacitracin sucrose broth-selective enrichment broth culture media. The number of S mutans colonies or biofilms on the surface of the electrometric rings was counted by using a stereomicroscope. Data were analyzed statistically with the Wilcoxon nonparametric test at the 5% significance level. Four representative rings of each type were chosen for SEM analysis. RESULTS Statistical analysis by the Wilcoxon nonparametric test showed that the Super Slick elastomeric rings had statistically significant greater S mutans contamination than the conventional elastomeric rings (P <.0001). No formation of S mutans colonies or biofilms was observed in the elastomeric rings removed directly from their original packages. SEM micrographs showed fissures on the surface of Super Slick elastomeric rings. No fissures were found on conventional elastomeric rings. When the microbiologic culture was positive, S mutans bacterial biofilm was observed on both types of ligatures. CONCLUSIONS There was no clinical evidence that Super Slick elastomeric rings are effective in reducing bacterial biofilm formation on their surfaces, and a recommendation for their use in orthodontic therapy for that purpose is not justifiable.

Pediatric dental treatment: manifestations of stress in patients, mothers and dental school students

The aim of this study was to compare a group of children with high levels of anxiety during dental treatment who required physical restraint (group 1) to a group of children who collaborated with treatment (group 2). Child stress and its relationship with the manifestations of stress of the mothers and the dental students were assessed. The following psychological evaluation techniques were applied: the Child Stress Scale, the Rutter Parent Scale A2, Dental Fear Survey, and Lipp Stress Symptoms Inventory for Adults. Group 1 presented higher stress indicators, and emotional and behavior difficulties than group 2 (p < or = 0.01). The stress indicators and the dental fear presented by mothers from group 1 were higher than that from group 2 (p < or = 0.01). The dental students who treated the children from both groups presented high stress levels. We conclude that the difficulties experienced by the child and the mother, associated to the students diminished ability to control the situation, may negatively influence the perception that the child has of dental treatment. A set of factors related to the children, mothers or dental students may cause poor collaboration during the dental treatment.

Toll-like Receptor 2 Knockout Mice Showed Increased Periapical Lesion Size and Osteoclast Number

INTRODUCTION The aim of this study was to characterize the formation and progression of experimentally induced periapical lesions in TLR2 knockout (TLR2 KO) mice. METHODS Periapical lesions were induced in molars of 28 wild type (WT) and 27 TLR2 KO mice. After 7, 21, and 42 days, the animals were euthanized, and the mandibles were subjected to histotechnical processing. Hematoxylin-eosin-stained sections were examined under conventional light microscopy for the description of pulpal, apical, and periapical features and under fluorescence microscopy for the determination of the periapical lesion size. The subsequent sections were evaluated by tartrate resistant acid phosphatase histoenzymology (osteoclasts), Brown and Brenn staining (bacteria), and immunohistochemistry (RANK, RANKL, and OPG). Data were analyzed by the Mann-Whitney U and Kruskal-Wallis tests (α = 0.05). RESULTS The WT group showed significant differences (P < .05) in the periapical lesion size and the osteoclast number between 7 and 42 days and between 21 and 42 days. In the TLR2 KO group, significant differences (P < .05) in the periapical lesion size and the osteoclast number were found between 7 days and the other periods. There was a significant difference (P < .05) between the 2 types of animal regarding the periapical lesion size, which was larger in the TLR2 KO animals. No significant differences (P > .05) were found between WT and TLR2 KO mice related to the pulpal, apical, and periapical features; bacteria localization; and immunohistochemical results (except for RANK expression). CONCLUSIONS TLR2 KO animals developed larger periapical lesions with a greater number of osteoclasts, indicating the important role of this receptor in the hosts immune and inflammatory response to root canal and periradicular infection.

Radiation therapy alters microhardness and microstructure of enamel and dentin of permanent human teeth

OBJECTIVES To evaluate, in vitro, the effects of ionizing radiation on the mechanical and micro-morphological properties of enamel and dentin of permanent teeth. METHODS Enamel and dentin microhardness (n=12 hemi-sections) was evaluated at three depths (superficial, middle and deep) prior to (control) and after every 10Gy radiation dose up to a cumulative dose of 60Gy by means of longitudinal microhardness. Data were analyzed using two-way analysis of variance and Tukeys test at a significance level of 5%. Enamel and dentin morphology was assessed by scanning electron microscopy (SEM) for semi-quantitative analysis (n=8 hemi-sections). Data were analyzed using Kruskal-Wallis and Dunns or Fisher exact tests at a significance level of 5%. RESULTS The application of ionizing radiation did not change the overall enamel microhardness, although an increase in superficial enamel microhardness was observed. The micro-morphological analysis of enamel revealed that irradiation did not influence rod structure but interprismatic structure became more evident. Dentin microhardness decreased after 10, 20, 30, 50 and 60Gy cumulative doses (p<0.05) compared with non-irradiated dentin, mainly in the middle portion of the tissue. The micro-morphological analysis revealed fissures in the dentin structure, obliterated dentinal tubules and fragmentation of collagen fibers after 30 and 60Gy cumulative doses. CONCLUSIONS Although ionizing radiation did not affect the enamel microhardness of permanent teeth as a whole, an increase in superficial enamel microhardness was observed. Dentin microhardness decreased after almost all radiation doses compared with the control, with the greatest reduction of microhardness in the middle depth region. The morphological alterations on enamel and dentin structures increased with the increase of the radiation dose, with a more evident interprismatic portion, presence of fissures and obliterated dentinal tubules, and progressive fragmentation of the collagen fibers. CLINICAL SIGNIFICANCE This study shows that irradiation affects microhardness and micro-morphology of enamel and dentin of permanent teeth. The effects of gamma irradiation on dental substrate might contribute to increased risk of radiation tooth decay associated with salivary changes, microbiota shift and high soft and carbohydrate-rich food intake.