Effect of a calcium hydroxide/chlorhexidine paste as intracanal dressing in human primary teeth with necrotic pulp against Porphyromonas gingivalis and Enterococcus faecalis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Antimicrobial activity of triantibiotic paste, 2% chlorhexidine gel, and calcium hydroxide on an intraoral-infected dentin biofilm model

Introduction: The purpose of this study was to evaluate the antimicrobial activity of calcium hydroxide, 2% chlorhexidine gel, and triantibiotic paste (ie, metronidazole, minocycline, and ciprofloxacin) by using an intraorally infected dentin biofilm model. Methods: Forty bovine dentin specimens were infected intraorally using a removable orthodontic device in order to induce the biofilm colonization of the dentin. Then, the samples were treated with the medications for 7 days. Saline solution was used as the control. Two evaluations were performed: immediately after the elimination of the medication and after incubation in brain-heart infusion medium for 24 hours. The Live/Dead technique (Invitrogen, Eugene, OR) and a confocal microscope were used to obtain the percentage of live cells. Nonparametric statistical tests were performed to show differences in the percentage of live cells among the groups (P < .05). Results: Calcium hydroxide and 2% chlorhexidine gel did not show statistical differences in the immediate evaluation. However, after application of the brain-heart infusion medium for 24 hours, 2% gel chlorhexidine showed a statistically lesser percentage of live cells in comparison with calcium hydroxide. The triantibiotic paste significantly showed a lower percentage of live cells in comparison with the 2% chlorhexidine gel and calcium hydroxide groups in the immediate and secondary (after 24 hours) evaluations. Conclusions: The triantibiotic paste was most effective at killing the bacteria in the biofilms on the intraorally infected dentin model in comparison with 2% chlorhexidine gel and calcium hydroxide

Subcutaneous tissue response of isogenic mice to calcium hydroxide-based pastes with chlorhexidine

This study was evaluated the response of subcutaneous connective tissue of isogenic mice to calcium hydroxide-based pastes with chlorhexidine digluconate (CHX). Seventy isogenic male BALB/c mice aged 6-8 weeks and weighing 15-20 g were randomly assigned to 8 groups. The animals received polyethylene tube implants as follows: Groups I, II, and III (n=10) - Calen® paste mixed with 0.4% CHX (experimental paste; Calen/CHX) for 7, 21, and 63 days, respectively; Groups IV, V, and VI (n=10) - UltraCal™ paste mixed with 2% CHX (experimental paste supplied by Ultradent Products Inc.; Ultracal/CHX) for 7, 21, and 63 days, respectively; and Groups VII and VIII (n=5): empty tube for 7 and 21 days, respectively. At the end of the experimental periods, the implants were removed together with the surrounding tissues (skin and subcutaneous connective tissue). The biopsied tissues were subjected to routine processing for histological analysis. Using a descriptive analysis and a four-point (0-3) scoring system, the following criteria were considered for qualitative and quantitative analysis of the tissue around the implanted materials: collagen fiber formation, tissue thickness and inflammatory infiltrate. A quantitative analysis was performed by measuring the thickness (µm), area (µm²) and perimeter (µm) of the reactionary granulomatous tissue formed at the tube ends. Data were analyzed statistically by the Kruskal-Wallis test and Dunn's post-test (α=0.05). Calen/CHX showed biocompatibility with the subcutaneous and reactionary tissues, with areas of discrete fibrosis and normal conjunctive fibrous tissue, though without statistically significant difference (p>0.05) from the control groups. In Groups I to III, there was a predominance of score 1, while in Groups IV to VI scores 2 and 3 predominated for all analyzed parameters. UltraCal/CHX, on the other hand, induced the formation of an inflammatory infiltrate and abundant exudate, suggesting a persistent residual aggression from the material, even 63 days after implant placement. In conclusion, the Calen paste mixed with 0.4% CHX allowed an adequate tissue response, whereas the UltraCal paste mixed with 2% CHX showed unsatisfactory results.

The effectiveness of mineral trioxide aggregate, calcium hydroxide and formocresol for pulpotomies in primary teeth

Aim To compare the effectiveness of mineral trioxide aggregate (MTA), calcium hydroxide (CH) and formocresol (FC) as pulp dressing agents in carious primary teeth. Methodology Forty-five primary mandibular molars with dental caries in 23 children [AUTHOR QUERY: How many children?] between 5 and 9 years old were treated by a conventional pulpotomy technique. The teeth were randomly assigned to the experimental ( CH or MTA) or control ( FC) groups. After coronal pulp removal and haemostasis, remaining pulp tissue was covered with MTA paste or CH powder in the experimental groups. In the control group, diluted FC was placed with a cotton pellet over the pulp tissue for 5 min and removed; the pulp tissue was then covered with zinc oxide-eugenol (ZOE) paste. All teeth were restored with reinforced ZOE base and resin modified glass-ionomer cement. Clinical and radiographic successes and failures were recorded at 3, 6, 12, 18 and 24 month follow-up. Results Forty-three teeth were available for follow-up. In the FC and MTA groups, 100% of the available teeth were clinically and radiographically successful at all follow-up appointments; dentine bridge formation could be detected in 29% of the teeth treated with MTA. In the CH group, 64% of the teeth presented clinical and radiographic failures detected throughout the follow-up period, and internal resorption was a frequent radiographic finding. Conclusions Mineral trioxide aggregate was superior to CH and equally as effective as FC as a pulpotomy dressing in primary mandibular molars. Internal resorption was the most common radiographic finding up to 24 month after pulpotomies performed with CH.

Cytotoxic effects and pulpal response caused by a mineral trioxide aggregate formulation and calcium hydroxide

Purpose: To evaluate: the in vivo pulpal response after pulpotomy with different capping agents. In addition, the in vitro cytotoxic effects of both materials were assessed by applying them on culture of pulp cells. Methods: For the in vivo test, the coronal pulp of 28 teeth of dogs was mechanically removed and the root pulps were capped with the following dental materials: Group 1: Pro-Root NITA (PRMTA); and Group 2 (control): calcium hydroxide saline paste (CH). After 60 days, the animals were sacrificed and the teeth processed for histological analysis. In the in vitro test, experimental extracts obtained from both capping agents were applied on the cultured MDPC-23 odontoblast-like cells. Results: In the root pulps capped with PRMTA or CH, coagulation necrosis partially replaced by dystrophic calcification as well as tubular dentin matrix laid down by elongated pulp cells was observed. None or mild inflammatory response occurred beneath the capped pulpal wound. Regarding the pulpal response, PRMTA and CH presented no statistical difference. However, the teeth capped CH presented greater healthy pulp loss which resulted in convex shape of the hard barrier than PRMTA. When applied on the cultured cells, it was demonstrated that PRMTA and CH solutions decreased the cell metabolic activity by 9.9% and 29.4%, respectively. CH caused higher cytotoxic effects to the MDPC-23 cells as well as deeper healthy pulp tissue loss than PRMTA. However, similar sequence of healing occurred after pulpotomy with both dental materials.

Periapical tissue reactions to calcium hydroxide and MTA after external root resorption as a sequela of delayed tooth replantation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Analysis of the healing process in delayed tooth replantation after root canal filling with calcium hydroxide, Sealapex and Endofill: a microscopic study in rats

The major concern in the therapeutics of tooth replantation refers to the occurrence of root resorption and different approaches have been proposed to prevent or treat these complications. The purpose of this study was to evaluate tissue response to delayed replantation of anterior rat teeth treated endodontically using calcium hydroxide, Sealapex, and Endofill without the placement of gutta-percha cones. Thirty rats had their right upper incisor extracted and maintained in dry storage for 60 min. After removal of the dental papilla, enamel organ, pulp tissue, and periodontal ligament remnants, the teeth were immersed in 2% sodium fluoride phosphate acidulated, pH 5.5, for 10 min. The root canals were dried with absorbent paper points and the teeth were assigned to three groups (n = 10) according to the filling material. Group I - calcium hydroxide and propyleneglycol paste, Group II - Sealapex, and Group III - Endofill. The sockets were irrigated with saline and the teeth were replanted. Replacement resorption, inflammatory resorption and ankylosis were observed in all groups. Although the occurrence of inflammatory resorption was less frequent in Group I, there were no statistically significant differences among the groups. It may be concluded that compared to the paste, filling the root canals with Sealapex and Endofill sealers without the placement of gutta-percha cones did not provide better results.

Cytotoxic effects and pulpal response caused by a mineral trioxide aggregate formulation and calcium hydroxide

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Release and diffusion of hydroxyl ion from calcium hydroxide-based medicaments

The release and diffusion of hydroxyl ions (OH-) of calcium hydroxide (Ca(OH)2)-based intracanal medications may be affected by the association with other substances. The aim of this study was to evaluate the diffusion of OH- ions through root dentin by the medications: G1, Ca(OH)2/saline; G2, Calen; G3, Calen/camphorated p-monochlorophenol (CMCP); and G4, Calen/0.4% chlorhexidine (CHX). Root canals from bovine teeth were prepared in a standardized manner. A cavity until dentin was prepared in the middle third of the root surface of each specimen. The external surface of the root was made impermeable using a layer of adhesive, except the prepared cavity. The root canals were filled with different medications, and teeth were individually stored in flasks containing 10 ml distilled water at 37 degrees C. The water pH was measured at 1, 3, 7, 14, 21, 30, and 60 days. Data obtained were subjected to anova and Tukeys tests. Increase in pH was observed at 3 days for Calen/CHX and from 7 to 14 days for the other mixtures. Calen paste promoted pH increase up to 21 days. Calen/CMCP had the highest pH up to 21 days, and all groups had similar results at 30 days. At 60 days, the greatest pH values were observed for Calen/CMCP and Calen alone. All different formulations of Ca(OH)2-based medications tested release hydroxyl ion that can diffuse through the dentin.

Effect of rotary or manual instrumentation, with or without a calcium hydroxide/1% chlorhexidine intracanal dressing, on the healing of experimentally induced chronic periapical lesions

Objective. To evaluate the healing of experimentally induced chronic periapical lesions in dogs at 30, 75, and 120 days after root canal instrumentation with rotary NiTi files or manual K-files, with or without a calcium hydroxide/1% chlorhexidine paste intracanal dressing.Study design. The second, third, and fourth mandibular premolars and the second and third maxillary premolars of 5 dogs (12 to 18 months of age, weighing 8 to 15 kg) were selected for treatment (a total of 82 root canals). After pulp removal, the root canals were left exposed to the oral cavity for 7 days to allow microbial contamination, after which the root canals were sealed with ZOE cement until periapical lesions were confirmed with radiography. Group I and II teeth were instrumented with manual K-files using the crown-down technique. In group III and IV teeth, NiTi rotary files were used. The apical delta was perforated by using #20 to #30 K-files at the length of the tooth, thus creating a standardized apical opening. The apical stop was enlarged to size 70, with 2.5% sodium hypochlorite irrigation at each file change. Teeth in groups II and IV were dressed with calcium hydroxide (Ca(OH)(2))/1% chlorhexidine (CHX) paste for 15 days before root filling. Group I and III teeth did not receive an intracanal dressing. The access openings of the teeth were permanently restored with silver amalgam condensed on a glass ionomer cement base. Pairs of standardized periapical radiographs were taken at the beginning of the treatment (0 days) and at 30, 75, and 120 days after filling.Results. There was no significant difference in the rate of radiographic healing of the periapical lesions between manual and rotary instrumentation. Radiographs taken at 120 days showed that the treatment with Ca(OH)(2)/1% CHX paste resulted in a significant reduction in mean size of the periapical lesions in comparison to single-session treatment. These findings were also true for histologic observations.Conclusion. The findings support the hypothesis that, regardless of the instrumentation technique (manual or rotary), the use of an intracanal dressing is important in the endodontic treatment of dog's teeth with experimentally induced chronic periapical lesions.

Vehicle influence on calcium hydroxide pastes diffusion in human and bovine teeth

The purpose of this study was to compare the pH and calcium ion liberation after use of calcium hydroxide pastes with different paste vehicles in human or bovine teeth. Ninety-two single-rooted human and bovine roots were used. The roots were instrumented and an external cavity preparation was performed. The roots were divided in to human and bovine groups. Each group was subdivided into four subgroups (SB) according to the vehicle:SB1, detergent; SB2, saline; SB3, polyethylenoglycol + camphorated paramonochlorophenol (Calen PMCC) and SB4, polyethylenoglycol + furacyn paramonochlorophenol (FPMC). Specimens were immersed into saline solution at 37 degrees C and after 7 and 14 days pH and calcium ion measurements were made. The results were analyzed by ANOVA and Tukey tests (P < 0.05). There was no statistical difference between bovine and human teeth in the pH analysis (P < 0.05), but bovine teeth provided larger calcium ion liberation than human teeth. Calen PMCC was statistically more effective for pH increase and calcium ion liberation in all analyses, followed by FPMC and saline. Detergent showed the lowest pH alterations and calcium ion liberation. The period of 14 days showed more calcium ionic liberation than the 7-day period.

Connective tissue responses to calcium hydroxide-based root canal medicaments

Aim: The objective of the present study was to evaluate the tissue inflammatory response induced by calcium hydroxide pastes, with or without paramonochlorophenol and camphor. Methodology: Isogenic BALB/c mice were inoculated into the subcutaneous tissue with either 0.1 mL of a suspension of Calen, Calen with camphorated paramonochlorophenol, Calen with paramonochlorophenol, Calasept paste or phosphate-buffered saline (control). After 6, 12 and 24 h and 2, 3, 5, 7 and 15 days, three animals in each group were sacrificed and the excised lesions processed for histopathological evaluation of the inflammatory response. Events monitored and graded included the assessment of vascular congestion, oedema, haemorrhage, inflammatory infiltrate, necrosis and tissue repair. Results: The pastes induced an inflammatory response at every observation period, although the intensity, duration and extension of inflammation varied. Calen paste always produced an initial short-term inflammatory response whilst the other pastes produced extended reactions. All pastes allowed repair to take place by the end of the experimental period, although the speed of this process varied between the materials. Calen presented the best biocompatibility; the phenolic compound caused greater tissue response, which was even more severe in the absence of camphor. Calasept paste was damaging and the repair process slower. Conclusions: All calcium hydroxide formulations caused an inflammatory response. The severity and longevity of the responses varied between pastes as a result of the various antiseptic agents. Although irritating, repair was apparent with all formulations.

Effect of rotary instrumentation and of the association of calcium hydroxide and chlorhexidine on the antisepsis of the root canal system in dogs

This study aimed at evaluating the antisepsis of the root canal system (RCS) and periapical region (PR) provided by rotary instrumentation associated with chlorhexidine + calcium hydroxide as intracanal medicament. Chronic periapical lesions were induced in 26 pre-molar roots in two dogs. After microbiological sampling, automatic instrumentation using the Profile system and irrigation with 5.25% sodium hypochlorite solution, with a final rinse of 14.3% EDTA followed by profuse irrigation with physiological saline were carried out in 18 root canals. After drying the canals, a paste based on calcium hydroxide associated with a 2% chlorhexidine digluconate solution was placed inside them. After 21 days, the medication was removed, leaving the root canals empty and coronally sealed. After 96 hours, a final microbiological sample was obtained, followed by histomicrobiological processing by the Brown & Brenn method. Eight untreated root canals represented the control group (C-G). Based on the Mann-Whitney test at a confidence level of 5% (p < 0.05), the procedures of antisepsis used offered significant efficacy (p < 0.05) resulting in 100.0% of the canals free of microorganisms. In the C-G, an elevated incidence of various microbial morphotypes was confirmed in all sites of the RCS, with the presence of microbial colonies in the periapical region. In contrast, the experimental group showed a similar pattern of infection in the RCS, although less intense and a reduced level of periapical infection (p < 0.05). It was concluded that adequate instrumentation followed by the application of calcium hydroxide + chlorhexidine offered significant elimination of microorganisms.

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 pmonochlorophenol (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.

Antimicrobial activity of endodontic sealers based on calcium hydroxide and MTA.

The aim of this study was to evaluate the antimicrobial activity of a new root canal sealer containing calcium hydroxide (Acroseal) and the root canal sealer based on MTA (Endo CPM Sealer), in comparison with traditional sealers (Sealapex, Sealer 26 and Intrafill) and white MTA-Angelus, against five different microorganism strains. The materials and their components were evaluated after manipulation, employing the agar diffusion method. A base layer was made using Müller-Hinton agar (MH) and wells were made by removing agar. The materials were placed into the wells immediately after manipulation. The microorganisms used were: Micrococcus luteus (ATCC9341), Staphylococcus aureus (ATCC6538), Pseudomonas aeruginosa (ATCC27853), Candida albicans (ATCC 10231), and Enterococcus faecalis (ATCC 10541). The plates were kept at room temperature for 2 h for prediffusion and then incubated at 37 degrees C for 24 h. The results showed that Sealapex and its base paste, Sealer 26 and its powder, Endo CPM Sealer and its powder, white MTA and its powder all presented antimicrobial activity against all strains. Intrafill and its liquid presented antimicrobial activity against all strains except P. aeruginosa and Acroseal was effective only against M. luteus and S. aureus.