Antimicrobial efficacy of a human ß-defensin-3 peptide using an Enterococcus faecalis dentine infection model.

AIM: To assess the antibacterial efficacy of a human ß-defensin-3 (HBD3) peptide against Enterococcus faecalis biofilms. METHODOLOGY: Standardized human dentine blocks were infected with E. faecalis ATCC 29212 for 3 weeks. Aqueous calcium hydroxide paste (n = 12, CH), a 2% chlorhexidine gel (n = 12, CHX), an HBD3 peptide gel (n = 12) and saline (n = 12) were tested as experimental groups. A mismatched peptide gel group (n = 12, MP) and sterilized but noninoculated block group (n = 12) were included as controls. After 1 week of medication, the dentinal samples at the depth of 200 and 400 µm were collected from medicated canal lumens. Bacterial growth was assessed by spectrophotometric analysis of optical density (OD) after 72 h of incubation. Statistical analysis was performed with repeated-measures anova and Tukey's post hoc test. RESULTS: The HBD3 group was associated with significantly lower OD values (P < 0.05) than the CH or CHX groups at both depths. The CH group did not differ significantly from MP or Saline group at either depth (P > 0.05). There was no significant difference (P > 0.05) in the OD values of the inner (200 µm) and outer (400 µm) dentinal samples for any group. CONCLUSIONS: The HBD3 peptide inhibited the growth of E. faecalis biofilms in infected dentine blocks.

Minimum-intensity projection for in-depth morphology study of mesiobuccal root.

OBJECTIVE: This micro-computed tomography (MCT) study investigated the utility of thin-slab minimum-intensity projection (TS-MinIP) technique as an adjunct to 3-dimensional (3D) modeling for in-depth morphology study. STUDY DESIGN: One hundred one extracted maxillary first molars were scanned for microtomographic analysis (SkyScan). Two-dimensional TS-MinIP and 3D images of mesiobuccal (MB) roots were produced and analyzed to record the number and configurations of the canals, the incidence and location of accessory canals, loop, and intercanal connections, and number of foramina. RESULTS: Multiple-canal MB roots were present in 76.2%, and all of the roots had intercanal communications. Weine type III configuration was the most common in the multiple-canal roots. Accessory canals were found in 78.2% of the roots. Configurations that were nonclassifiable were found in 10.9% of the MB roots. CONCLUSIONS: MB root canal anatomy was complex, and MinIP may serve as an adjunct to 3D modeling for in-depth morphology study.

CONTROVERSIES IN ENDODONTICS.

Diseases of the dental pulp often have an infectious origin, and treatments are aimed to control infections of the root canal system. Endodontic treatment principles originally evolved on the basis of trial and error, and only in recent decades have scientific methods been adopted to support clinical strategies. Yet, relevant research on the disease processes, their diagnoses, and efficient treatment are rare in the endodontic literature. Hence, the advancement of biologically based knowledge significant to clinical endodontics has been slow. Therefore, many differences of opinion still prevail in this field of dentistry. This review highlights and analyzes the background of some of the more heavily debated issues in recent years. Specifically, it deals with disagreements regarding the clinical management of pulpal exposures by caries in the adult dentition, definitions of success and failure of endodontic therapy, and causes of and measures to control infections of the root canal system. Clearly, a most apparent gap in the published endodontic literature is the lack of randomized clinical trials that address the more significant controversial matters relating to the management of pulpal wounds, medication, and the number of appointments required for the treatment of infected root canals. However, trials in endodontics require extremely long follow-up periods if valid conclusions are to be generated. Therefore, it is not to be expected that there will be rapid solutions to these issues in the foreseeable future.

Enamel matrix derivative prolongs primary osteoblast growth.

Enamel matrix derivative (EMD) secreted by cells of the epithelial root sheath plays an important role in cementogenesis and periodontal tissue formation. The mechanisms by which EMD influences cell function are not known. The purpose of this study was to determine the effect of EMD on cell growth of primary mouse osteoblasts. Osteoblasts were digested from 6- to 8-day-old mouse calvaria and plated into 6-well cell culture plates at an initial density of 5000 cells/cm2. After 24-h incubation with Dulbecco's modified eagle medium (DMEM) supplemented with 10% fetal bovine serum, cells were incubated in three different groups of media: DMEM only as control, DMEM with 25 microg/ml EMD, and DMEM with 100 microg/ml EMD. At days 3, 7, 10, and 14, the total cell number per well was calculated, and cell morphology was examined. At each observation period the number of cells in the EMD groups was significantly greater (ANOVA, p < 0.01) than that in the control group. EMD had a greater effect on osteoblast survivor in the higher concentration than in the lower concentration. Furthermore normal morphology of the primary osteoblasts was maintained in the EMD groups. These results suggest that EMD prolongs primary osteoblast growth and may have an effect on osteoblasts during periodontal regeneration.

A prognostic model for assessment of the outcome of endodontic treatment: Effect of biologic and diagnostic variables.

OBJECTIVE: Many biological variables, endodontic treatment factors, and restorative considerations have been suggested in the literature to affect the outcome of endodontic treatment. However, few attempts have been made recently to study these variables further. The purpose of this study was to identify the biologic and endodontic treatment-associated variables that are most predictive of treatment outcome for conventional endodontic therapy and to determine the magnitude of risk these variables pose on the outcome. STUDY DESIGN: The population of this historical prospective cohort study comprised a total of 200 teeth with 441 root canals. Diagnostic and treatment information was abstracted from the original patient records. An endodontic follow-up examination was conducted 4 +/- 0.5 years after obturation. Each tooth/root was analyzed according to 3 indices of periradicular status at 2 time points. The main outcome measure was the presence of apical periodontitis. The criteria used for evaluation of the outcome were modified from Strindberg. Data were subjected to univariate and multivariate analysis. Logistic regression models were fit by using various clinical measures to determine which combination of biologic and treatment-associated factors best predicted treatment outcome. RESULTS: The preoperative pulp diagnosis, the periapical diagnosis, the preoperative periapical radiolucency size, and the sex of the patients were revealed, by means of univariate analysis, to exert a significant influence on endodontic treatment outcome (P <.05). In the logistic regression model, the strongest effect on postoperative healing was the presence and magnitude of preoperative apical periodontitis. In the presence of this variable, no other factor contributed value to the prediction. The correct prediction of this model was 74.7% (P <.05). CONCLUSION: The major biologic factors influencing the outcome of endodontic treatment appear to be the extent of microbiological insult to the pulp and periapical tissue, as reflected by the periapical diagnosis and the magnitude of periapical pathosis.

Effects of enamel matrix derivative on gene expression of primary osteoblasts.

OBJECTIVE: The aim of this study was to investigate the effects of enamel matrix derivative (EMD) on gene expression of collagen alpha1 (I), osteocalcin, prostaglandin G/H synthase 2 (PGHS-2), interleukin-6, and insulin-like growth factor I in primary mouse osteoblasts. STUDY DESIGN: Primary osteoblasts were digested from 6- to 8-day-old mouse calvaria. Cells were divided into 4 groups and cultured for 24, 48, and 72 hours with a serum-free modified Eagle medium as negative control, modified Eagle medium with 25 microg/mL EMD, modified Eagle medium with 100 microg/mL EMD, and modified Eagle medium plus 10% fetal bovine serum as positive control. Gene expression was determined by Northern blot and reverse transcription polymerase chain reaction analysis. RESULT: EMD enhanced collagen I, interleukin-6, and PGHS-2 expression and did not stimulate the expression of osteocalcin and IGF-I. CONCLUSION: These results indicate that EMD might regulate certain gene expression during periodontal tissue regeneration.

A comparison of stainless steel and nickel-titanium H-type instruments of identical design: torsional and bending tests.

OBJECTIVE: For a true comparison of clinically relevant qualities of instruments manufactured from different alloys, the instrument must be identical in design. The purpose of this study was to perform mechanical tests on stainless steel and nickel-titanium instruments of identical design. STUDY DESIGN: Identical experimental endodontic files of H-type with double helix were fabricated from stainless steel and nickel-titanium alloys. Torsional and bending tests were carried out according to American National Standards Institute/American Dental Association specification No. 58 and International Standards Organization No. 3630/1. Changes in instruments during the testing were also analyzed with scanning electron microscope technique. RESULTS: The results indicated that the torsional moment of the stainless steel files was significantly higher than for the nickel-titanium files (P <.001), whereas the mean angular deflection for the nickel-titanium files was significantly higher. The mean bending moment for the nickel-titanium files was significantly lower (P <.001) than the mean value for the stainless steel files. The scanning electron microscope demonstrated that the fracture surfaces of the stainless steel files were brittle, whereas the nickel-titanium files had a ductile fracture. CONCLUSIONS: When the design of endodontic instruments of different alloys is identical, the nickel-titanium files are more flexible in bending. However, nickel-titanium files require less force to deform to fracture.

Reaction of bony tissue to implanted silver glass ionomer and a reinforced zinc oxide-eugenol cement.

OBJECTIVE: To evaluate the tissue responses to implants of Ketac Silver and Super EBA cement in the guinea pig mandible. STUDY DESIGN: Sixteen guinea pigs were used for 2 experimental periods of 4 and 12 weeks. Both materials were placed in Teflon applicators and implanted into surgically prepared sites in the mandible. A histologic examination for reaction to the material occurred after the animals were killed and processed. RESULTS: After 4 weeks, minimal inflammatory reactions were observed in Ketac Silver implants, whereas the Super EBA implants showed minimal to moderate inflammation. Localized foreign body reaction was present in areas of fragmented small particles of Ketac Silver. At 12-weeks observation, no inflammatory reactions were present around either material. Direct bone contact was observed in 1 Ketac Silver implant. CONCLUSIONS: Ketac Silver and Super EBA cement elicited mild reactions under the conditions of this model. From a biologic point of view, these 2 materials may offer equal utility in endodontic surgery.

Adhesion of human osteoblasts on root-end filling materials.

Adhesion of human osteoblasts to root-end filling materials (mineral trioxide aggregate (MTA), IRM, composite, and amalgam) was observed by scanning electron microscopy. Root-end filling materials were inserted into 96-well flat-bottomed plates and condensed to disks of approximately 1 mm thick and the same diameter as the wells. After the disks were set, they were placed in the bottom of Nunc four-well culture plates at one disk per well. Then human osteoblasts were seeded into the wells at 1.5 x 10(5) cells per well. After 1 day in culture the disks of root-end filling materials along with cells grown on their surface were examined with a scanning electron microscopy. Results showed that osteoblasts attached and spread on MTA and composite by forming a monolayer. Osteoblasts also attached on amalgam, but with few cells spreading. In the presence of IRM, osteoblasts appeared rounded with no spreading. These results indicate that osteoblasts have a favorable response to MTA and composite resin compared with IRM and amalgam.

Shaping ability of three ProFile rotary instrumentation techniques in simulated resin root canals.

The aim of this study was to compare the shaping ability of three ProFile rotary instrumentation techniques and a conventional step-back method in simulated root canals. Prevalence of canal aberrations, change in working length, and preparation time were measured. A total of 48 composite images were made from pre- and postcanal scanned images using Corel Photopaint 8.0 and then the amount of coronal substance the instruments removed was also calculated two-dimensionally on digitized images with the Brain C software to compare the enlarging efficiency. There were no significant differences between the three rotary groups in preparation time, change in working length, and the incidence of aberrations (p > 0.05). The amount of coronal substance the instruments removed in the ProFile .04 taper group was significantly smaller than the other three groups (p < 0.05).

Permeability changes of dentine treated with titanium tetrafluoride.

OBJECTIVES: The aim of this research was to evaluate the effect of titanium tetrafluoride on dentine permeability in comparison with sodium fluoride and acidulated phosphate fluoride to determine their contribution to acid resistance of dentine. METHODS: Thirty-two dentine discs were prepared from extracted mandibular molars. The hydraulic conductance of all discs was measured before and after the formation of smear layer. The discs were then randomly divided into five experimental and one-control groups. Coronal surfaces of dentine discs were treated with either fluoride solutions of NaF, APF, 1% TiF4, 0.5% TiF4 and 0.1% TiF4 or de-ionised water. Following the measurement of hydraulic conductance, treated surfaces were subjected to 25% citric acid application. Then, final permeability measurements were made. The data were statistically analysed using ANOVA and Tukey's HSD multiple comparisons. RESULTS: While smear layer formation considerably reduced dentinal permeability of dentine discs, fluoride or de-ionised water application to smeared surfaces did not cause any significant change in hydraulic conductance (p > 0.05). After citric acid application, control and NaF groups showed higher permeability values than 1%, 0.5% and 0.1% TiF4 and APF groups (p < 0.01). CONCLUSION: Clinical use of acidic solutions of titanium tetrafluoride in dentine cavities may be considered since smeared dentine surfaces are modified to a stabile and acid-resistant state.

Cytotoxic evaluation of root-end filling materials in cultures of human osteoblast-like cells and periodontal ligament cells.

The cytotoxicity of three root-end filling materials (amalgam, IRM, and Super-EBA) was evaluated in cultures of human periodontal ligament cells and human osteoblast-like cells. Ten-millimeter-long plastic test tubes were filled with 3 mm of freshly mixed root-end filling materials at one end (1.5 mm diameter). The opposite end was sealed and attached by heat to a 35-mm cell culture dish. Human periodontal ligament cells and human osteoblast-like cells were seeded in the dishes. The size of cell-free zones around the root-end filling materials and the total cell number per dish were calculated after 3 and 7 days. Empty test tubes used as controls did not influence the growth and distribution of the cultured cells. Cell density increased in all groups in the test period. Amalgam had a larger cell-free zone, compared with IRM and Super-EBA and showed a reduction in total cell number per dish for both tested cell types. IRM and Super-EBA also had a cell-free inhibition zone for both cell types, but no significant reduction in total cell number per dish. This study showed that amalgam had a higher cell toxicity to human periodontal ligament cells and human osteoblast-like cells than IRM and Super-EBA.

Antifungal effects of sodium hypochlorite and chlorhexidine in root canals.

The purpose of this study was to evaluate the antifungal properties of 0.12% chlorhexidine, 1% NaOCl, and 5% NaOCl. Root sections were enlarged and the smear layer was removed in half of the specimens. The specimens were fixed in the wells of tissue culture plates. Each root canal was dispensed with an inoculum of Candida albicans. After 10 days, the root sections were treated with 3 ml of either disinfectant solution for 1 min, 5 min, 30 min, and 1 h. Then, root sections were incubated in test tubes having Sabouraud's Dextrose Broth at 37 degrees C for 24 h. In the presence of the smear layer, antifungal activity was observed only in 1-h treatment groups for all solutions. However, in the absence of the smear layer, 5% NaOCl alone started to show antifungal activity after 30 min. The antimicrobial effectiveness of irrigating solutions should be re-evaluated, particularly in patients predisposed to oral candidiasis.

Morphologic effects on L929 fibroblasts of titanium tetrafluoride application.

OBJECTIVE: The aim of this study was to investigate the effect of titanium tetrafluoride solution on L929 fibroblasts by scanning electron microscopy. Titanium tetrafluoride was then compared with sodium fluoride and acidulated phosphate fluoride. STUDY DESIGN: Cells were treated with fluoride solutions for 1 minute either directly, through a filter membrane with a pore size of 0.4-micron, or indirectly, through dentin disks; they were then investigated at an electron microscopic level. RESULTS: Fluoride application on smeared dentin disks showed fewer cytotoxic effects on fibroblasts than application on nonsmeared dentin disks. Acidulated phosphate fluoride and titanium tetrafluoride appeared to be more cytotoxic than sodium fluoride. Because all fluoride solutions used in this study contained the same fluoride concentration, pH was considered to be the main factor causing the higher toxicity. CONCLUSION: Because these solutions demonstrated toxicity in vitro, they must be further evaluated under in vivo conditions to ascertain their clinical safety.

The role of integrin beta 1 in human dental pulp cell adhesion on laminin and fibronectin.

OBJECTIVE: This study is to identify the expression of integrin beta 1 in human dental pulp cells and the role of integrin beta 1 in pulp cell adhesion on extracellular matrix protein laminin and fibronectin. STUDY DESIGN: Immunoblot detection of integrin beta 1 in human pulp cells was with the use of monoclonal anti-beta 1 antibody. Dental pulp cell adhesion assay on extracellular matrix protein laminin and fibronectin and blocking cell adhesion was performed with monoclonal anti-beta 1 antibody. RESULT: Integrin beta 1 was identified in human dental pulp cells. Pulp cells adhered and spread on both laminin and fibronectin. Monoclonal anti-beta 1 antibody inhibited human dental pulp cells adhesion on laminin but not on fibronectin. CONCLUSIONS: Integrin beta 1 was expressed on human dental pulp cells and mediated cell adhesion on laminin. Human dental pulp cells also adhered on fibronectin but the adhesion was not regulated by beta 1 integrin.

Integrin expression in human dental pulp cells and their role in cell attachment on extracellular matrix proteins.

Integrins are a family of heterodimeric glycoproteins consisting of alpha and beta subunits that noncovalently interact to form cell surface adhesion receptors. The objective of this study was to identify integrins in human dental pulp cells and determine their role in human dental pulp cell attachment to the biological active molecules, laminin and fibronectin. Integrin expression was studied by immunoblot and immunoprecipitation using monoclonal integrin antibodies. The role of integrin in human dental pulp cell adhesion on laminin and fibronectin was determined by inhibition of cell adhesion with those antibodies. This study found human dental pulp cells expressed alpha 1, alpha 3, alpha 5, alpha 6, alpha v, and beta 1 integrin subunits. The adhesion of human dental pulp cells to laminin and fibronectin was not inhibited by monoclonal antibody to any subunit, except that anti-beta 1 antibody inhibited pulp cells adhesion on laminin. These data provide information for further studying the role of integrins in dental pulp cell biological function.