Léa Assed Bezerra da Silva

In Vivo Antimicrobial Activity of 2% Chlorhexidine Used as a Root Canal Irrigating Solution

The aim of the present study was to evaluate the in vivo antimicrobial activity of 2% chlorhexidine gluconate (FCFRP-USP) used as a root canal irrigating solution in teeth with pulp necrosis and radiographically visible chronic periapical reactions. Culture techniques and measurement of the inhibition zone were used. Twenty-two root canals of incisors and molars of 12 patients were used. After accessing the canal, the first root canal sample was collected with two sterile paper points that were transferred to a tube containing reduced transport fluid. The root canal was instrumented using chlorhexidine solution. A small sterile cotton pellet was placed at the root canal entrance, and the cavity was sealed with zinc oxide-eugenol cement. The canals were maintained empty for 48 h. Three sterile paper points were then introduced to absorb the root canal fluid (second sample). One paper point was placed on an agar plate inoculated with Micrococcus luteus ATCC 9341 and incubated for 24 h at 37 degrees C, and the other two were submitted to microbiological evaluation. Present in 10 cases at baseline, mutans streptococci was reduced by 100% at the second assessment. Treatment showed an efficiency of 77.78% for anaerobic microorganisms at the second assessment. These data suggest that chlorhexidine prevents microbial activity in vivo with residual effects in the root canal system up to 48 h.

TNF-α Promotes an Odontoblastic Phenotype in Dental Pulp Cells

Dental pulp cells can differentiate toward an odontoblastic phenotype to produce reparative dentin beneath caries lesions. However, the mechanisms involved in pulp cell differentiation under pro-inflammatory stimuli have not been well-explored. Thus, we hypothesized that the pro-inflammatory cytokine tumor necrosis factor-α (TNF-α) could be a mediator involved in dental pulp cell differentiation toward an odontoblastic phenotype. We observed that TNF-α-challenged pulp cells exhibited increased mineralization and early and increased expression of dentin phosphoprotein (DPP), dentin sialoprotein (DSP), dentin matrix protein-1, and osteocalcin during a phase of reduced matrix metalloproteinase (MMP) expression. We investigated whether these events were related and found that p38, a mitogen-activated protein kinase, differentially regulated MMP-1 and DSP/DPP expression and mediated mineralization upon TNF-α treatment. These findings indicate that TNF-α stimulates differentiation of dental pulp cells toward an odontoblastic phenotype via p38, while negatively regulating MMP-1 expression.

Histological evaluation of therapy using a calcium hydroxide dressing for teeth with incompletely formed apices and periapical lesions

Periapical tissue response was evaluated histologically in teeth of 6-month-old dogs with incomplete root formation and periapical lesions in order to compare the effects of two calcium hydroxide-containing pastes prepared with different vehicles on induced root apex formation and repair of the periapical region. A group of teeth with no root canal dressing was used as control. After induction of the periapical lesion, the root canals were instrumented and filled with one of the two pastes (Calen and Calasept), which were renewed monthly for 90 days. At the end of the follow-up period (3 months), the animals were killed. Block sections from the actual areas were dissected free and prepared for histopathological analysis. Both calcium hydroxide pastes helped induce apical sealing and repair of the region, but the Calen paste yielded better results than Calasept. The induced mineralized tissue was predominantly of the cementoid type after the use of both pastes, but the inflammatory infiltrate was significantly less intense with the use of the Calen paste. In the control group, the absence of a root canal dressing impaired the process of periapical repair and yielded no apical root closure in any of the cases analyzed.

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.

EM evaluation of bacterial biofilm and microorganisms on the apical external root surface of human teeth

The aim of this study was to evaluate the presence of bacterial biofilm on the external surface of the root apex in teeth with pulp necrosis, with and without radiographically visible periapical lesions, and in teeth with a vital pulp. Twenty-one teeth were extracted, eight with pulp necrosis and periapical lesions, eight with pulp necrosis without radiographically visible periapical lesions, and five with a vital pulp. The roots were sectioned, and the root apexes (+/- 3 mm) were processed for scanning electron microscope evaluation. The surface of the apical root was evaluated for the presence of microorganisms, root resorption, and biofilm. There were no microorganisms on the apical root surface of either teeth with pulp vitality or with pulp necrosis with no radiographically visible periapical lesions. Microorganisms were always present in teeth with pulp necrosis and radiographically visible periapical lesions. These included cocci, bacilli, and filaments and the presence of an apical biofilm. Apical biofilm is clinically important because microbial biofilms are inherently resistant to antimicrobial agents and cannot be removed by biomechanical preparation alone. This may cause failure of endodontic treatment as a consequence of persistent infection.

Effect of Irrigating Solution and Calcium Hydroxide Root Canal Dressing on the Repair of Apical and Periapical Tissues of Teeth with Periapical Lesion

The objective of this study was to evaluate the apical and periapical repair after endodontic treatment of teeth with pulp necrosis and a chronic periapical lesion in dogs. Seventy-two root canals from four mongrel dogs were submitted to biomechanical preparation, using 5.25% sodium hypochlorite or 2% chlorhexidine digluconate as the irrigating solution. The root canals were subsequently either filled immediately with Sealapex, using active lateral gutta-percha condensation, or a calcium hydroxide root canal dressing was applied for 15 days before filling with Sealapex. After 210 days, the animals were killed by anesthetic overdose, and the obtained histological sections were stained with hematoxylin and eosin for optical microscopic analysis of apical and periapical repair. There was better histological repair in the groups with the root canal dressing (p < 0.05) than the groups with immediate obturation. Comparing the immediate obturation groups, irrigation with chlorhexidine solution resulted in better repair than sodium hypochlorite.

Calcium hydroxide root canal Sealers—Histopathologic evaluation of apical and peripaical repair after endodontic treatment

Eighty root canals of the premolars of 4 dogs, with vital pulp, were instrumented and filled during the same session with the Sealapex, CRCS, Sealer 26, and Apexit sealers. The animals were sacrificed 180 days after root canal filling and their maxillae and mandibles were removed and fixed in 10% formalin. After routine histologic processing, the sections were stained with hematoxylin-eosin and Mallory trichrome. Histopathologic analysis showed that Sealapex was the sealer that best permitted the deposition of mineralized tissue at the apical level and was the only sealer that provided complete sealing (37.5% of cases). With the use of Sealapex, no inflammatory infiltrate occurred and there was no reabsorption of mineralized tissues. In contrast, partial sealing and a moderate inflammatory infiltrate occurred with the use of CRCS. When Apexit and Sealer 26 were used the absence of sealing was frequent and active reabsorption of mineralized tissues occurred in most cases. The inflammatory infiltrate predominating with the use of Apexit was of the severe type, whereas with the use of Sealer 26 the inflammatory infiltrate was mild or absent.

Matrix Metalloproteinase Expression in Teeth with Apical Periodontitis Is Differentially Modulated by the Modality of Root Canal Treatment

INTRODUCTION The objective of this study was to investigate the expression of matrix metalloproteinases (MMPs) in apical periodontitis and during the periapical healing phase after root canal treatment. METHODS Apical periodontitis was induced in dog teeth, and root canal treatment was performed in a single visit or by using an additional calcium hydroxide root canal dressing. One hundred eighty days after treatment the presence of inflammation was examined, and tissues were stained to detect bacteria. Bacterial status was correlated to the degree of tissue organization, and to further investigate molecules involved in this process, tissues were stained for MMP-1, MMP-2, MMP-8, and MMP-9. Data were analyzed by using one-way analysis of variance followed by Tukey test or Kruskal-Wallis followed by Dunn test. RESULTS Teeth with apical periodontitis that had root canal therapy performed in a single visit presented an intense inflammatory cell infiltrate. Periapical tissue was extremely disorganized, and this was correlated with the presence of bacteria. Higher MMP expression was evident, similar to teeth with untreated apical periodontitis. In contrast, teeth with apical periodontitis submitted to root canal treatment with calcium hydroxide presented a lower inflammatory cell infiltrate. This group had moderately organized connective tissue, lower prevalence of bacteria, and lower number of MMP-positive cells, similar to healthy teeth submitted to treatment. CONCLUSIONS Teeth treated with calcium hydroxide root canal dressing exhibited a lower percentage of bacterial contamination, a lower MMP expression, and a more organized extracellular matrix, unlike those treated in a single visit. This suggests that calcium hydroxide might be beneficial in tissue repair processes.

Cone-beam computerized tomographic, radiographic, and histologic evaluation of periapical repair in dogs' post-endodontic treatment.

OBJECTIVE To evaluate the periapical repair after root canal treatment in the teeth of dogs using CT and conventional radiography and to compare these findings with the gold standard microscopic evaluation. STUDY DESIGN The animals were divided into three groups according to endodontic treatment performed: Group 1, single-visit endodontic treatment in teeth without apical periodontitis; Group 2, single-visit endodontic treatment in teeth with apical periodontitis; and Group 3, endodontic treatment in teeth with apical periodontitis using calcium hydroxide as a root canal dressing. Group 4 consisted of teeth with apical periodontitis not submitted to root canal treatment and Group 5 consisted of healthy teeth without periapical disease. Radiographic, tomographic, and microscopic evaluations were performed by blind examiners. At 180 days experimental time, CT and radiographic measurements of periapical disease were compared with the gold standard microscopic measurement using intraclass correlation coefficient. Intergroup comparisons considering different methods of periapical lesions measurement or different clinical protocols of root canal treatment were performed by Kruskal Wallis test followed by Dunn. Integrity of lamina dura, presence of radiolucent areas, and presence of root resorption were analyzed by Fishers exact test. RESULTS There was discontinuity of the lamina dura and CPD in all teeth from Groups 2, 3, and 4 evaluated by tomography and radiography 45 days after CPD induction. Radiographically, 180 days after root canal treatment, there was no periapical lesion in teeth from Groups 1 and 3, different from groups 2 and 4 (p < .05). The highest reduction in the CPD size was observed on Group 3 (p < .05). According to the tomographic results, there was decrease of the size of the CPD on Group 3 but not on Groups 2 or 4. However, in all groups the periapical lesions presented larger mesio-distal extension if compared with radiography, both 45 days after CPD induction and 180 days after root canal treatment. At 180 days, CT measurements were closely related to microscopic results (ICC = 0.95) differently from radiographic evaluation (ICC = 0.86). CONCLUSION CT Scan evaluation of periapical repair following root canal treatment provided similar information than that obtained by microscopic analysis, whereas radiographic evaluation underestimated the size do periapical lesion.

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.