Microbial culture and checkerboard DNA-DNA hybridization assessment of bacteria in root canals of primary teeth pre- and post-endodontic therapy with a calcium hydroxide/chlorhexidine paste.

AIM: To investigate the root canal microbiota of primary teeth with apical periodontitis and the in vivo antimicrobial effects of a calcium hydroxide/chlorhexidine paste used as root canal dressing. DESIGN: Baseline samples were collected from 30 root canals of primary teeth with apical periodontitis. Then, the root canals were filled with a calcium hydroxide paste containing 1% chlorhexidine for 14 days and the second bacteriologic samples were taken prior to root canal filling. Samples were submitted to microbiologic culture procedure to detect root canal bacteria and processed for checkerboard DNA-DNA hybridization. RESULTS: Baseline microbial culture revealed high prevalence and cfu number of anaerobic, black-pigmented bacteroides, Streptococcus, and aerobic microorganisms. Following root canal dressing, the overall number of cfu was dramatically diminished compared to initial contamination (P <0.05), although prevalence did not change (P > 0.05). Of 35 probes used for checkerboard DNA-DNA hybridization, 31 (88.57%) were present at baseline, and following root canal dressing, the number of positive probes reduced to 13 (37.14%). Similarly, the number of bacterial cells diminished folowing application of calcium hydroxide/chlorhexidine root canal dressing (P = 0.006). CONCLUSION: Apical periodontitis is caused by a polymicrobial infection, and a calcium hydroxide/chlorhexidine paste is effective in reducing the number of bacteria inside root canals when applied as a root canal dressing.

Ex vivo accuracy of an apex locator using digital signal processing in primary teeth.

PURPOSE: The purpose of this study was to evaluate ex vivo the accuracy an electronic apex locator during root canal length determination in primary molars. METHODS: One calibrated examiner determined the root canal length in 15 primary molars (total=34 root canals) with different stages of root resorption. Root canal length was measured both visually with the placement of a K-file 1 mm short of the apical foramen or the apical resorption bevel, and electronically using an electronic apex locator (Digital Signal Processing). Data were analyzed statistically using the intraclass correlation (ICC) test. RESULTS: Comparing the actual and electronic root canal length measurements in the primary teeth showed a high correlation (ICC=0.95). CONCLUSIONS: The Digital Signal Processing apex locator is useful and accurate for apex foramen location during root canal length measurement in primary molars.

Arrest of root formation in a permanent maxillary central incisor subsequent to trauma and pulp necrosis to the primary predecessor.

This paper reports a case in which a previous traumatic injury at the age of 2 and pulp necrosis to a primary incisor resulted in a rare injury to the permanent successor tooth. The radiographic examination at the age of 9 showed the arrest of root formation of the permanent maxillary right central incisor, which did not erupt. Tooth 11 was extracted and a functional removable space maintainer was prepared. At the age of 17, the patient received an anterior fixed prosthesis for re-establishment of the esthetics, phonetics and deglutition.

Root canal adhesive filling in dogs' teeth with or without coronal restoration: a histopathological evaluation.

The purpose of this study was to evaluate in vivo the response of the periradicular tissues after endodontic treatment and root filling with Epiphany/Resilon (Penntron Clinical Technologies, LLC, Wallingford, CT) or gutta percha and new Sealapex (SybronEndo, Glendora, CA) in dogs' teeth with or without coronal restoration. Teeth without coronal restorations were used to assess the influence of continuous exposure to the oral environment on the periradicular tissues. Sixty root canals with vital pulps in three dogs were instrumented and obturated in a single session and randomly assigned to one of four groups as follows. group 1: root canal filling with Epiphany/Resilon with coronal restoration, group 2: root canal filling with Sealapex sealer and gutta percha with restoration, group 3: root canal filling with Epiphany/Resilon without restoration, and group 4: root canal filling with Sealapex sealer and gutta percha without coronal restoration. After 90 days, the animals were euthanized, and the maxillas and mandibles were removed and submitted for histologic processing. Longitudinal sections were obtained and stained with hematoxylin and eosin, Mallory's trichrome, and Brown and Brenn stains and examined under light microscopy. There were significant differences found between the four groups (p < 0.05). The results showed that roots canals filled with Epiphany/Resilon, with coronal restoration, had significantly less periradicular inflammation than roots canals filled with gutta percha and Sealapex, with coronal restoration (p = 0.021). No significant difference was observed in the intensity of inflammation between roots canals filled with Epiphany/Resilon with no restoration and roots filled with gutta percha and Sealapex with restoration (p = 0.269). Roots canals filled with gutta percha and Sealapex sealer without coronal restoration showed the greatest degree of periradicular inflammation.

Scanning electron microscopy of the apical structure of human teeth.

The objective of this research was to evaluate, by scanning electron microscopy, the apical structure of extracted human permanent teeth with different degrees of pulp and periapical pathology. A total of 25 teeth were extracted: 5 teeth with vital pulp (group I); 10 teeth with pulp necrosis without radiographically visible periapical lesion (group II); 10 teeth with pulp necrosis with radiographically visible periapical lesion (group III). The root apex was sectioned and processed for scanning electron microscopy. In groups I and II, fibers covered the root cementum and there was no cementum resorption or microorganisms. There were areas of cementum resorption in group III with microorganisms on the root apex surface (biofilm) and no fibers. The authors conclude that the presence of chronic periapical lesions causes severe changes in the apical structure with a destruction of fibers and different degrees of cementum resorption forming lacunae in which bacterial biofilm persisted.

Residual antibacterial activity of chlorhexidine digluconate and camphorated p-monochlorophenol in calcium hydroxide-based root canal dressings.

The purpose of this study was to evaluate the residual antibacterial activity of several calcium hydroxide [Ca(OH)2]-based pastes, placed in root canals of dogs' teeth with induced chronic periapical lesions. Root canals were instrumented with the ProFile rotary system and filled with 4 pastes: G1 (n=16): Ca(OH)2 paste + anesthetic solution; G2 (n=20): Calen paste + camphorated p-monochlorophenol (CMCP); G3 (n=18): Calen; and G4 (n=18): Ca(OH)2 paste + 2% chlorhexidine digluconate. After 21 days, the pastes were removed with size 60 K-files and placed on Petri plates with agar inoculated with Micrococcus luteus ATCC 9341. Pastes that were not placed into root canals served as control. After pre-diffusion, incubation and optimization, the inhibition zones of bacterial growth were measured and analyzed by Mann-Whitney U test at 5% significance level. All pastes showed residual antibacterial activity. The control samples had larger halos (p<0.05). The mean residual antibacterial activity halos in G1, G2, G3 and G4 were 7.6; 10.4; 17.7 and 21.4 mm, respectively. The zones of bacterial growth of G4 were significantly larger than those of G1 and G2 (p<0.05). In conclusion, regardless of the vehicle and antiseptic, all Ca(OH)2-based pastes showed different degrees of measurable residual antibacterial activity. Furthermore, unlike CMCP, chlorhexidine increased significantly the antibacterial activity of Ca(OH)2.

Apical Periodontitis Healing Following Treatment is Impacted by Root Canal Sealer Composition: An in Vivo and in Vitro Investigation

Abstract Objective: To evaluate the periapical healing following root canal treatment in teeth with apical periodontitis (in vivo) and the cytotoxic potential of root canal sealers in vitro. Material and Methods: Apical periodontitis was induced in 60 dogs' teeth and root canals were filled with Sealapex (40 roots), EndoREZ (40 roots), intracanal dressing (20 roots), or left untreated (20 roots). After 30 and 90 days, histopathological analyses were made. In vitro, J774.1 macrophages were stimulated with root canal sealers extracts, cytotoxicity was assessed using lactate dehydrogenase assay, and qRT-PCR was used to analyze TNF-α gene expression. Results: In vivo, smaller apical periodontitis and lower inflammatory cell infiltrate were found in teeth treated with Sealapex compared to EndoREZ. In vitro, EndoREZ was cytotoxic and induced TNF-α gene expression by macrophages differently from Sealapex. Conclusion: Sealapex allowed improved tissue repair following root canal treatment in teeth with apical periodontitis compared to EndoREZ. Synthesis of TNF-α induced by LPS was enhanced by EndoREZ, whereas Sealapex prevented pro-inflammatory gene expression (AU).

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.

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.

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.

Influence of root canal dressings and sealers on repair of apical periodontitis after endodontic treatment.

OBJECTIVE: The objective of this study was to evaluate the histopathologic response of periapical tissues after root canal treatment of necrotic dog teeth with chronic apical periodontitis by using 2 calcium hydroxide-based root canal dressings and 2 root canal sealers. STUDY DESIGN: Seventy-eight root canals were instrumented by using 5.25% sodium hypochlorite as the irrigating solution, after which a calcium hydroxide paste (Calen/PMCC or Calasept) was placed for 30 days as a dressing. The root canals were then filled by using cold lateral gutta-percha condensation and an endodontic sealer (Sealapex or AH Plus). After 360 days, the animals were killed by anesthetic overdose; then, the teeth were histologically prepared, sectioned, and stained with hematoxylin and eosin for optical microscopic analysis of apical and periapical tissue repair. RESULTS: Statistical analysis showed that the poorest histopathologic results were observed in the Calasept/AH Plus group and that the Sealapex sealer overall resulted in better apical repair than the AH Plus sealer. The histopathologic results of Calen/PMCC paste with both AH Plus and Sealapex and Calasept paste with only Sealapex were statistically similar but were different from the results of Calasept with AH Plus. CONCLUSIONS: The results of this study in the dog showed differences in apical and periapical tissue repair of teeth with chronic apical periodontitis by using 2 calcium hydroxide root canal dressings and 2 sealers. More research is necessary to determine the best combination of dressings and sealers.

Histological study of the effect of some irrigating solutions on bacterial endotoxin in dogs.

The aim of this study was to evaluate, histopathologically, the effectiveness of mechanical preparation of root canals using different irrigating solutions in dog teeth filled with LPS after pulpectomy. A total of 120 root canals of 6 mongrel dogs were filled with a solution of LPS after pulpectomy. The irrigating solutions used were saline, 1, 2.5, and 5% sodium hypochlorite, and 2% chlorhexidine. No irrigation was used in the control group. The animals were sacrificed after 60 days and the teeth were fixed and demineralized. Subsequently, serial 6-microm sections were stained with hematoxylin and eosin and Mallory's trichrome for histopathological analysis and Brown-Brenn for verification of bacterial contamination. Analysis showed that the inflammatory infiltrate was statistically less intense in the groups in which the root canals were irrigated with 5% sodium hypochlorite and 2% chlorhexidine. However, none of the irrigating solutions completely inactivated the harmful effects of LPS. Mechanical preparation associated with different irrigating solutions did not completely inactivate LPS.

Calcium hydroxide root canal dressing. Histopathological evaluation of periapical repair at different time periods.

The objective of this study was to evaluate periapical and apical repair using calcium hydroxide root canal dressings for different lengths of times in teeth with induced chronic periapical lesions. A total of 61 root canals of maxillary and mandibular premolars from 4 dogs were used. After mechanical preparation of the root canals using the crown-down technique, and 5.25% NaOCl as irrigating solution, the apical foramen was enlarged in all cases. A calcium hydroxide root canal dressing was applied. The control group did not receive a root canal dressing. The animals were killed at 7, 15 or 30 days. After histological preparation, serial sections were stained with hematoxylin-eosin and Mallory's trichrome. The best histopathological results occurred at 15 and 30 days, and the worst results occurred at 7 days and in the control group.

Apical and periapical repair of dogs' teeth with periapical lesions after endodontic treatment with different root canal sealers.

The aim of this study was to evaluate the apical and periapical repair after root canal treatment of dogs' teeth with pulp necrosis and chronic periapical lesion using different root canal sealers. After periapical lesion induction, forty-four root canals of 3 dogs were submitted to biomechanical preparation using 5.25% sodium hypochlorite as an irrigating solution. A calcium hydroxide dressing (Calen PMCC) was applied for 15 days and the root canals were filled using the lateral condensation technique with gutta-percha points and Sealapex, AH Plus or Sealer Plus for sealing. After 180 days, the animals were sacrificed by anesthetic overdose and the obtained histological sections were stained with hematoxylin-eosin for optical microscopic analysis of the apical and periapical repair. The groups filled with Sealapex and AH Plus had better histological repair (p < 0.05) than the group filled with Sealer Plus, that had unsatisfactory results.

Pulpal and periapical response of dogs' teeth after pulpotomy and use of enamel matrix derivative as a capping agent

Objective: To evaluate the pulpal and periapical response of dogs' teeth after pulpotomy and use of Enamel Matrix Derivative (EMD) as capping agent. Methods: Pulpotomy was performed in 40 teeth from 4 dogs and the remaining pulp tissue was recovered with the following materials: Groups I and IV: EMD (Emdogain®); Groups II and V: calcium hydroxide; Groups III and VI: zinc oxide and eugenol cement. After 7 days (Groups I, II, and III) and 70 days (Groups IV, V, and VI), the animals were euthanized and the teeth were removed and processed for histological analysis; were analyzed using Kruskal-Wallis test followed by Dunn test or Mann Whitney test ( =0.05). Results: 7-day period: in Group I, it was observed a mild to moderate inflammatory infiltrate and intense vascular proliferation while Group II presented a mild inflammatory infiltrate and an intact pulp tissue (p < 0.05). Group III presented a moderate to severe inflammatory infiltrate. 70- day period: Groups IV and VI showed no dentin bridge formation, the remaining pulp tissue presented necrotic areas with inflammatory cells in the periapical region and bone and cementum resorption (p > 0.05). In Group V, there was dentin bridge formation, absence of inflammation and absence of mineralized tissue resorption (p < 0.05). Conclusion: EMD as a capping material after pulpotomy did not show either satisfactory tissue response or capacity of inducing deposition of mineralized tissue in dental pulp.

Objetivo: Avaliar a resposta pulpar e periapical de dentes de cães após pulpotomia e uso de Derivados de Matriz de Esmalte (EMD) como agente de capeamento. Métodos: A pulpotomia foi realizada em 40 dentes de 4 cães e o tecido pulpar remanescente foi recuperado com os seguintes materiais: Grupos I e IV: EMD (Emdogain®); Grupos II e V: hidróxido de cálcio; Grupos III e VI: cimento de óxido de zinco e eugenol. Após 7 dias (Grupos I, II e III) e 70 dias (Grupos IV, V e VI), os animais foram eutanásiados e os dentes foram removidos e processados para análise histológica; foram analisados pelo teste de Kruskal-Wallis, seguido pelo teste de Dunn ou Mann Whitney ( = 0,05). Resultados: Período de 7 dias: no Grupo I, observou-se infiltrado inflamatório leve a moderado e intensa proliferação vascular, enquanto o Grupo II apresentou infiltrado inflamatório leve e tecido pulpar intacto (p <0,05). O grupo III apresentou infiltrado inflamatório moderado a grave. Período de 70 dias: os grupos IV e VI mostraram, na formação da ponte de dentina, que o tecido pulpar remanescente apresentava áreas necróticas com células inflamatórias na região periapical, reabsorção óssea e de cemento (p> 0,05). No Grupo V, houve formação de ponte de dentina, ausência de inflamação e ausência de reabsorção tecidual mineralizada (p <0,05). Conclusão: O EMD como material de cobertura após pulpotomia não mostrou resposta tecidual satisfatória ou capacidade de induzir deposição de tecido mineralizado na polpa dentária.

Subcutaneous connective tissue response to primary root canal filling materials.

This study evaluated the response of the subcutaneous connective tissue of BALB/c mice to root filling materials indicated for primary teeth: zinc oxide/eugenol cement (ZOE), Calen paste thickened with zinc oxide (Calen/ZO) and Sealapex sealer. The mice (n=102) received polyethylene tube implants with the materials, thereby forming 11 groups, as follows: I, II, III: Calen/ZO for 7, 21 and 63 days, respectively; IV, V, VI: Sealapex for 7, 21 and 63 days, respectively; VII, VIII, IX: ZOE for 7, 21 and 63 days, respectively; X and XI: empty tube for 7 and 21 days, respectively. The biopsied tissues were submitted to histological analysis (descriptive analysis and semi-quantitative analysis using a scoring system for collagen fiber formation, tissue thickness and inflammatory infiltrate). A quantitative analysis was performed by measuring the area and thickness of the granulomatous reactionary tissue (GRT). Data were analyzed by Kruskal-Wallis, ANOVA and Tukey's post-hoc tests (?=0.05). There was no significant difference (p>0.05) among the materials with respect to collagen fiber formation or GRT thickness. However, Calen/ZO produced the least severe inflammatory infiltrate (p<0.05). The area of the GRT was significantly smaller (p<0.05) for Calen/ZO and Sealapex. In conclusion, Calen/ZO presented the best tissue reaction, followed by Sealapex and ZOE.

Inhibition of sodium hypochlorite antimicrobial activity in the presence of bovine serum albumin.

INTRODUCTION: This study investigated the inhibition of the antimicrobial activity of sodium hypochlorite (NaOCl) by bovine serum albumin (BSA). The killing of Enterococcus faecalis, Candida albicans, Staphylococcus epidermidis, and Escherichia coli by NaOCl in concentrations from 2% to 0.03% was measured in the presence of BSA in concentrations between 6.7% and 0.1%. METHODS: NaOCl, BSA, and microorganism suspensions were mixed, and, after 30 seconds, 6 minutes, and 30 minutes, samples were taken and NaOCl was inactivated by 5% sodium thiosulphate. The microbes were incubated in tryptic soy broth broth for up to 7 days for the detection of growth. RESULTS: All microorganisms were killed within 30 seconds by 0.03% NaOCl when BSA was not present. High concentrations of BSA significantly reduced the antimicrobial activity of NaOCl against the four species. CONCLUSIONS: The inhibition of sodium hypochlorite by BSA was directly dependent on their quantitative relationships. The result partly explains the poorer performance in vivo of NaOCl as compared to in vitro experiments.

Electronic working length determination in primary teeth by ProPex and Digital Signal Processing.

The purpose of this study was to evaluate the accuracy of electronic apex locators Digital Signal Processing (DSP) and ProPex, for root canal length determination in primary teeth. Fifteen primary molars (a total of 34 root canals) were divided into two groups: Group I - without physiological resorption (n = 16); and Group II - with physiological resorption (n = 18). The length of each canal was measured by introducing a file until its tip was visible and then it was retracted 1 mm. For electronic measurement, the devices were set to 1 mm short of the apical resorption. The data were analysed statistically using the intraclass correlation coefficient (ICC). Results showed that the ICC was high for both electronic apex locators in all situations - with (ICC: DSP = 0.82 and Propex = 0.89) or without resorption (ICC: DSP = 0.92 and Propex = 0.90). Both apex locators were extremely accurate in determining the working length in primary teeth, both with or without physiological resorption.

Congenital eruption cyst: a case report.

Congenital pathologies are those existing at or dating from birth. Occurrence of congenital cystic lesions in the oral cavity is uncommon in neonates. Eruption cyst (EC) is listed among these unusual lesions. It occurs within the mucosa overlying teeth that are about to erupt and, according to the current World Health Organization (WHO) classification of epithelial cysts of the jaws, EC is a separate entity. This paper presents a case of congenital EC successfully managed by close monitoring of the lesion, without any surgical procedure or tooth extraction. Eruption of the teeth involved, primary central incisors, occurred at the fourth month of age. During this time neither the child nor mother had any complication such as pain on sucking, refusal to feed, airway obstruction, or aspiration of fluids or teeth.

Histopathological evaluation of root canal filling materials for primary teeth.

This study aimed to assess the response of apical and periapical tissues of dogs' teeth after root canal filling with different materials. Forty roots from dogs' premolars were prepared biomechanically and assigned to 4 groups filled with: Group I: commercial calcium hydroxide and polyethylene glycol-based paste (Calen) thickened with zinc oxide; Group II: paste composed of iodoform, Rifocort and camphorated paramonochlorophenol; Group III: zinc oxide-eugenol cement; Group IV: sterile saline. After 30 days, the samples were subjected to histological processing. The histopathological findings revealed that in Groups I and IV the apical and periapical regions exhibited normal appearance, with large number of fibers and cells and no resorption of mineralized tissues. In Group II, mild inflammatory infiltrate and mild edema were observed, with discrete fibrogenesis and bone resorption. Group III showed altered periapical region and thickened periodontal ligament with presence of inflammatory cells and edema. It may be concluded that the Calen paste thickened with zinc oxide yielded the best tissue response, being the most indicated material for root canal filling of primary teeth with pulp vitality.