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An in vitro study of coronal leakage after intraradicular preparation of cast-dowel space.

STATEMENT OF PROBLEM: Coronal leakage can produce contamination of periapical tissues, resulting in endodontic failure. PURPOSE: The purpose of this in vitro study was to evaluate the ability of 2 sealers to prevent coronal leakage in canals filled with gutta-percha and prepared for cast dowels but without coronal sealing. MATERIAL AND METHODS: The crowns of 60 extracted single-rooted teeth were amputated. The root canals were prepared corono-apically and filled with gutta-percha cones and 1 of 2 different endodontic sealers: a resin-based sealer (AH Plus) and a calcium hydroxide-based sealer (Sealapex). Specimens were then stored in water for 7 days to allow the sealers to set. The specimens were prepared in 1 of 2 ways: no preparation for cast dowel or preparation of cast-dowel space (n=15). External surfaces of the roots were sealed with cyanoacrylate cement. The teeth were thermal cycled at 5 degrees and 55 degrees C in water baths (dwell time=30 seconds) for 500 cycles. Specimens were then submerged in 2% methylene blue colorant for 24 hours. Microleakage was measured according to the percentage of area stained with the colorant. Effects of each factor (cast-dowel preparation and type of sealant) on microleakage were analyzed by the Student t test (alpha=.05). RESULTS: The AH Plus and Sealapex sealers with cast-dowel preparation resulted in significantly (P<.001) more leakage compared to sealers with no dowel preparation. CONCLUSION: Cast dowel-space preparation had a negative influence on the sealing ability of the remnant root-canal filling material.

Calcium Hydroxide↗

Postoperative pain after endodontic retreatment: single- versus two-visit treatment.

OBJECTIVE: The purpose of this clinical study was to determine the effect of 1- or 2-visit root canal treatment on the postoperative pain in the retreatment cases. STUDY DESIGN: Two hundred eighteen cases that required retreatment were included in the study. Obturated and unfilled canal space and the status of periapical tissues were evaluated according to the PAI index. The patients were subcategorized in regard to the presence or the absence of preoperative pain. Approximately half of each category was treated in 1 appointment. After removing the previous root canal obturation materials and biomechanic preparation of root canals, the teeth in the 1-visit group were obturated at the first appointment by using AH 26 sealer and laterally compacted gutta-percha, and those in the 2-visit group were medicated with calcium hydroxide-chlorhexidine combination and then closed with a temporary filling material. One week after the initial appointment, patients were asked about the occurrence of postoperative pain. The level of discomfort was rated as no pain, mild pain, moderate pain, or severe pain (flare-up). Data were statistically analyzed using the chi-squared and Fischer exact tests. RESULTS: Eight patients from the 1-visit group and 2 patients from the 2-visit group had flare-ups. There was a statistical difference between the groups (P <.05). Two-visit root canal treatment was more effective in completely eliminating pain than 1-visit treatment of previously symptomatic teeth (P <.05). CONCLUSIONS: Two-visit endodontic treatment with intracanal medication was found to be effective in reducing postoperative pain of previously symptomatic teeth and decreased the number of flare-ups in all retreatment cases.

Bismuth↗

Effects of intracanal carbon dioxide laser irradiation on cultured human fibroblasts.

OBJECTIVE: The purpose of this study was to evaluate, under various conditions, the damage that might occur to viable cells after CO2 laser irradiation of the root canal. STUDY DESIGN: A laser tip was placed within the root canal of extracted human teeth that were positioned above cultured fibroblasts (NB cells), and then irradiation was applied. The irradiation modes used were either pulse (40 pps) or super-pulse (151 pps). The laser energy was set at 32 J in both modes, and the tip of the laser beam was positioned to a point 2 mm from the fibroblasts. RESULTS: When the pulse mode irradiation was applied, there was significant difference (P < .05) in the percentage of viable cells between teeth with closed apical foramen (1-mm wall between root canal and root apex) and the #15 group (No. 15 K-file used), #30 group (No. 30 K-file), or #50 group (No. 50 K-file). The difference between the #30 group and the #50 group (P < .05) was also significant. The larger the diameter of the apical foramen became, the lower was the percentage of viable cells. CONCLUSION: If a CO2 laser irradiates a root canal system enlarged to within 1 mm of the root canal length, little damage to the periapical tissues would be expected to occur.

Carbon Dioxide↗

In vitro study of the effect of sodium hypochlorite and glutaraldehyde on substrate adherence capacity of macrophages.

The purpose of this study was to investigate the in vitro effect of two irrigation solutions used in endodontics (5.25% sodium hypochlorite and 1% glutaraldehyde) on substrate adherence capacity of macrophages to determine if these substances can alter macrophage function. Inflammatory macrophages were obtained from Wistar rats and resuspended in RPMI-1640 medium. Substrate adherence capacity assays were carried out in Eppendorf tubes for 15 min of incubation at 37 degrees C in a humidified atmosphere of 5% CO2. The adherence index was calculated. Results showed that both sodium hypochlorite and glutaraldehyde significantly decreased the substrate adherence capacity of inflammatory macrophages. To take into account that adhesion is the first step in the phagocytic process of macrophages and in antigen presentation, sodium hypochlorite and glutaraldehyde could inhibit macrophage function and reduce inflammatory reactions in periapical tissues when they are used in root-canal therapy.

Animals↗

In vitro effect of the resin component bisphenol A on substrate adherence capacity of macrophages.

This study was design to investigate the "in vitro" effect of bisphenol A (BPA), a component of resin used in dentistry, on viability, and substrate adherence capacity of macrophages. Peritoneal macrophages were obtained from Wistar rats and resuspended in RPMI-1640 medium. Viability was determined by trypan blue exclusion. As a test of macrophage adhesion, the adherence capacity of macrophages to a plastic surface was determined and the adherence index was calculated. Assays were conducted in Eppendorf tubes for 60 min of incubation at 37 degrees C in a humidified atmosphere of 5% CO2 in air. BPA did not alter significantly macrophage viability at concentrations as high as 10(-5) M, but BPA decreased in a dose-dependent manner the adherence index of rat peritoneal macrophages. Control peritoneal macrophages showed an adherence index = 81.5 +/- 7.9%. In the presence of 10(-8) M BPA, the Al of macrophages decreased to 41.4 +/- 12.2% (p < 0.05). Higher BPA concentrations (10(-7) to 10(-5) M) also caused a significant inhibition of the adherence index. Half-maximal inhibition (IC50) was obtained at 4.92 +/- 0.39 x 10(-6) M BPA. The in vitro study shows that the resin component BPA can alter macrophage adhesion. Taking into account that adhesion is the first step in the phagocytic process of macrophages and in antigen presentation, BPA could inhibit macrophage function and modulate immune and inflammatory responses in dental pulp and periapical tissues.

Analysis of Variance↗

Calcium hydroxide inhibits substrate adherence capacity of macrophages.

The purpose of this study was to investigate the effect of calcium hydroxide on substrate adherence capacity of rat inflammatory macrophages to determine if calcium hydroxide can alter macrophage function. Inflammatory macrophages were obtained from Wistar rats and resuspended in RPMI-1640 medium. Substrate adherence capacity assays were carried out in Eppendorf tubes for 15 min of incubation at 37 degrees C in a humidified atmosphere of 5% CO2. The adherence index (AI) was calculated. Results showed that calcium hydroxide decreased substrate adherence capacity of inflammatory macrophages in a time and dose-dependent manner. The lowest calcium hydroxide concentration that caused a significant inhibition of AI was 1 mM (p < 0.05), and the concentration of calcium hydroxide that caused half-maximal inhibition (IC50) was 1.54 mM (p < 0.01). We conclude that calcium hydroxide decreased substrate adherence capacity of macrophages. When adhesion as the first step in the phagocytic process and in antigen presentation is taken into account, calcium hydroxide could inhibit macrophage function and reduce inflammatory reactions in periapical tissues or in dental pulp when it is used in root-canals therapy or in direct pulp capping and pulpotomy, respectively. Moreover, this effect could explain, at least in part, the mineralized tissue-inducing property of calcium hydroxide.

Animals↗

In vitro effect of parachlorophenol and camphorated parachlorophenol on macrophages.

The purpose of this study was to investigate the "in vitro" effect of parachlorophenol and camphorated parachlorophenol, used in endodontics for the disinfection of root canals, on the substrate adherence capacity of macrophages. Inflammatory macrophages were obtained from Wistar rats and resuspended in RPMI-1640 medium. As a test of macrophage phagocytic function, the adherence capacity of macrophages to a plastic surface was determined. Assays were conducted in Eppendorf tubes for 15 min of incubation at 37 degrees C in a humidified atmosphere of 5% CO2. The adherence index was calculated. Results showed that parachlorophenol and camphorated parachlorophenol significantly decreased the substrate adherence capacity of inflammatory macrophages. Taking into account that adhesion is the first step in the phagocytic process of macrophages and in antigen presentation, parachlorophenol and camphorated parachlorophenol could inhibit macrophage function and modulate immune and inflammatory reactions in periapical tissues.

Animals↗

Antimicrobial activity of 2.5% sodium hypochlorite and 0.2% chlorhexidine gluconate separately and combined, as endodontic irrigants.

Sodium hypochlorite irrigant is known to be toxic to periapical tissues. Chlorhexidine gluconate, a safer and effective antimicrobial irrigant, is not known to dissolve pulpal tissues. To obtain their optimal properties, their combined action within the root canal was evaluated. Ten single rooted nonvital anterior teeth were irrigated using either 2.5% sodium hypochlorite alone, 0.2% chlorhexidine gluconate alone, 2.5% sodium hypochlorite and 0.2% chlorhexidine gluconate combined within the root canal, or 0.9% saline, respectively. Microbiological samples for culture and Gram's staining were taken before and proceeding irrigation. This study indicates that the use of sodium hypochlorite and chlorhexidine gluconate combined within the root canal resulted in the greatest percentage reduction of postirrigant positive cultures. This may be due to formation of "chlorhexidine chloride," which increases the ionizing capacity of the chlorhexidine molecule. This reduction was significant compared to use of sodium hypochlorite alone but not significant compared to use of chlorhexidine gluconate alone.

Anti-Infective Agents, Local↗

Effect of dentin bonding agents on the secretion of inflammatory mediators from macrophages.

Dentin bonding agents (DBAs) have been proposed as substitutes for amalgam as root-end filling materials. The current study tested the hypothesis that certain components of DBAs could alter the secretion of cytokines from macrophages. Such alteration would likely be undesirable for healing of the periapical tissues. Human THP-1 macrophages were exposed to 2-hydroxyethyl methacrylate, 4-methacryloxyethyl trimelliate anhydride, bisphenol-gycidylmethacrylate, and urethane dimethacrylate. The secretion of interleukin-1 beta (IL-1 beta) and tumor necrosis factor-alpha (TNF-alpha) were measured with or without challenge by lipopolysaccharide (LPS). Results showed that all DBA components completely suppressed LPS-induced IL-1 beta and TNF-alpha secretion at concentrations that suppressed mitochondrial activity by 50%. In addition, 4-methacryloxyethyl trimelliate anhydride induced secretion of IL-1 beta, but not TNF-alpha, without the LPS challenge. These results indicate that DBA components may alter normal macrophage-directed inflammatory responses if the macrophages are exposed to sufficiently high concentrations of these components.

Acrylic Resins↗

Comparative effects of two endodontic irrigants, chlorhexidine digluconate and sodium hypochlorite, on macrophage adhesion to plastic surfaces.

This study was designed to compare the "in vitro" effect of chlorhexidine digluconate, proposed as a new irrigant solution, with sodium hypochlorite, the currently irrigant solution used to disinfect the root canal system before obturation of the canal, on substrate adherence capacity of macrophages. Inflammatory macrophages were obtained from Wistar rats and resuspended in RPMI-1640 medium. The adherence capacity of macrophages to a plastic surface was determined. Assays were conducted in Eppendorf tubes for 15 min of incubation at 37 degrees C in an humidified atmosphere of 5% CO2 in air. The adherence index was calculated. Chlorhexidine digluconate inhibited substrate adherence capacity of macrophages in all conditions tested. Half-maximal inhibition (IC50) was observed at 1:5.6 dilution. Chlorhexidine digluconate was less potent than 5.25% sodium hypochlorite (IC50 = 1:24.1 dilution) in inhibiting substrate adherence capacity of macrophages. Taking into account that substrate adherence is the first step in the phagocytic process of macrophages and in antigen presentation, chlorhexidine digluconate could inhibit macrophage function and modulate inflammatory reactions at the level of inflamed periapical tissues.

Animals↗

Effect in vitro of Tifell (formocresol-eugenol) on macrophage adhesion.

The purpose of this study was to investigate the effect in vitro on macrophage adhesion of Tifell (formocresoleugenol), used as an intracanal medicament and in therapeutic pulpotomies in primary teeth, Macrophages were obtained from Wistar rats. As a test of macrophage phagocytic function the adherence capacity of macrophages to a plastic surface was determined. Assays were carried out in Eppendorf tubes after 15 min of incubation at 37 degrees C in a humidified atmosphere of 5% carbon dioxide. The adherence index (AI) was calculated. Results showed that Tifell decreased significantly (P < 0.05) the AI of macrophages. Half maximal inhibition of AI was obtained at 1:334.5 Tifell dilution (AI = 27.75; P < 0.05). Taking into account that substrate adherence is the first step in the phagocytic action of macrophages and in antigen presentation, Tifell could inhibit macrophage function and modulate immune and inflammatory responses in dental pulp and periapical tissues.

Analysis of Variance↗

Comparison between a conventional technique and two bone regeneration techniques in periradicular surgery.

AIM: The aim was to demonstrate the efficacy of two materials for bone regeneration during periradicular surgery and their effects on the healing of periapical tissues. METHODOLOGY: Twenty-eight patients (30 surgical sites) were selected and distributed into three groups:group A, conventional technique; group B, conventional technique plus nonbioabsorbable GoreTex R Augmentation membrane; and group C, conventional technique plus the same membrane placed over a synthetic bioactive resorbable graft of a hydroxylapatite (OsteoGen) product in the bony defect. Clinical and radiological evaluations were completed immediately prior to surgery, a week later and every 3 months after surgery up to 12 months. Two histological evaluations were performed (at the beginning and at 12 months). RESULTS: The results showed complete clinical and radiographic healing (eight cases) for group C, with histologic evidence of trabecular bone in all cases. For group B, six out of nine cases showed complete radiographic healing, incomplete in one case and uncertain in two cases, with histologic evidence of trabecular bone in five out of eight cases, scar tissue in one case and granuloma in two cases. For group A, there was complete radiographic healing in four out of nine cases, incomplete in four and unsuccessful in one case, with evidence of granuloma in four out of eight, scar tissue in two cases and trabecular bone in two cases. CONCLUSIONS: It was concluded that the conventional technique was less predictable in its healing response during the 12 months of this study. The use of bone regeneration materials, such as nonbioabsorbablemembranes and resorbable hydroxylapatite improved the predictability of clinical, radiographic and histological healing.

Absorbable Implants↗

Host collagen signal induces antigen I/II adhesin and invasin gene expression in oral Streptococcus gordonii.

Microbial interactions with host molecules, and programmed responses to host environmental stimuli, are critical for colonization and initiation of pathogenesis. Bacteria of the genus Streptococcus are primary colonizers of the human mouth. They express multiple cell-surface adhesins that bind salivary components and other oral bacteria and enable the development of polymicrobial biofilms associated with tooth decay and periodontal disease. However, the mechanisms by which streptococci invade dentine to infect the tooth pulp and periapical tissues are poorly understood. Here we show that production of the antigen I/II (AgI/II) family polypeptide adhesin and invasin SspA in Streptococcus gordonii is specifically upregulated in response to a collagen type I signal, minimally the tri-peptide Gly-Pro-Xaa (where Xaa is hydroxyproline or alanine). Increased AgI/II polypeptide expression promotes bacterial adhesion and extended growth of streptococcal cell chains along collagen type I fibrils that are characteristically found within dentinal tubules. These observations define a new model of host matrix signal-induced tissue penetration by bacteria and open the way for novel therapy opportunities for oral invasive diseases.

Adhesins, Bacterial↗

In vivo resistance of coronally induced bacterial ingress by an experimental glass ionomer cement root canal sealer.

The resistance of an experimental sealer (KT-308) to bacterial ingress was assessed in six beagle dogs. In four mandibular premolars per dog, canals were prepared, filled with condensed gutta-percha and either KT-308 or Roth 801 cement (n = 24 roots), and the pulp chambers inoculated with plaque. Two additional premolars per dog were similarly root-filled, but not inoculated (n = 12 and 11, respectively). One incisor per dog was inoculated, but not root-filled (n = 6). Dogs were terminated after 6 months, and jaw blocks were retrieved and processed for light microscopic examination of the periapical tissues. Inflammation about the inoculated roots was significantly lower (p < 0.03) for KT-308 (17%) than Roth 801 cement (46%). Inflammation about the noninoculated roots did not differ significantly between KT-308 (8%) and Roth 801 cement (36%). This study demonstrated a better functional efficacy of KT-308 than of Roth 801 cement, and validated this in vivo model for assessment of root filling materials.

Animals↗

Secretion of tumor necrosis factor-alpha by mouse peritoneal macrophages in the presence of dental sealers, sealapex and endomethasone.

After filling root canals, the healing process depends on the chemical composition or physical-chemical properties of the material used, among other factors. All root canal sealers, whether solid or plastic, are foreign matter for the body if they remain in permanent contact with apical and periapical tissues. As a result, the first organic reaction that occurs is an attempt to phagocytize the material. During phagocytosis, macrophages release a large number of cell mediators into the area, among which are cytokines that are essential in intercellular communication and in many physiological and pathophysiological processes. One of these cytokines is tumor necrosis factor-alpha (TNF-alpha), which acts through links to specific receptors on the cell membrane initiating a cascade of events leading to induction, activation, or inhibition of numerous cytokine-regulated genes in the cell nucleus. The release of TNF-alpha in a cell culture of mouse peritoneal macrophages incubated with three concentrations (25, 50, and 100 mg/ml) of two endodontic sealers was measured. The solutions containing the calcium hydroxide-based root canal sealer (Sealapex) released fewer units of TNF-alpha than solutions containing the zinc oxide and eugenol-based sealer (Endomethasone).

Animals↗

Induction of vascular endothelial growth factor gene expression by proinflammatory cytokines in human pulp and gingival fibroblasts.

Vascular endothelial growth factor (VEGF) enhances the permeability of blood vessels, which is an important vascular change observed during inflammatory processes. The purpose of this in vitro study was to investigate the effect of proinflammatory cytokines on the expression of VEGF mRNA gene in human pulp and gingival fibroblasts. Interlukin-1alpha (IL-1alpha) and tumor necrosis factor-alpha (TNF-alpha) were used to evaluate VEGF mRNA gene expression in human pulp and gingival fibroblasts. The levels of mRNAs were measured by quantitative reverse-transcriptase polymerase chain reaction analysis. Both IL-1alpha and TNF-alpha induced significantly high levels of VEGF mRNA gene expression in human pulp and gingival fibroblasts (p < 0.05). In addition, TNF-alpha was found to be more effective in the induction of VEGF mRNA gene expression in pulp than gingival fibroblasts (p < 0.05). Moreover, IL-1alpha was found to be more effective in the induction of VEGF mRNA gene expression than TNF-alpha in gingival fibroblast cultures (p < 0.05). These results indicate that proinflammatory cytokines can induce VEGF mRNA gene expression, and such an effect may partially contribute to the destruction of pulpal and periapical tissues through promoting expansion of the vascular network coincident to progression of the inflammation.

Analysis of Variance↗

Pulp response to, and cariogenicity of, a strain of Streptococcus mutans.

Gnotobiotic rats mono-infected with Streptococcus mutans (NCTC 10919) were maintained on a high sucrose diet in an isolator. When they were 8 weeks old the pulps of their maxillary first and second molars were mechanically exposed and left open to the oral environment. The rats were sacrificed in groups after 7 and 28 days. Thirty-seven teeth were available for study in the 7-day group, and 30 teeth in the 28-day group. The test S. mutans was non-cariogenic. After 7 days, necrosis of the superficial layers of the pulp was noted; however, after 28 days pulpal necrosis was much more extensive and calcific repair was observed in only one-third of the teeth examined. The microorganism did not provoke inflammation in the pulp or periapical tissues. This is the second strain of S. mutans tested in this manner. These results are very similar to those reported previously for S. mutans (NCTC 10449) and strengthens the view that organisms of the S. mutans group have a low pathogenicity for the pulp.

Animals↗