Root canal instruments are poorly standardized.

In an earlier study we demonstrated that root canal instruments vary greatly in their effectiveness in removing substrates. To further analyze the reason for this variation of results, the dimensions of the instruments used were considered of interest as their matching effectiveness may be size dependent. Nine different brands of Hedstrom files, seven brands of K files, and four brands of special files were examined. The measurements were done in a computerized measuring microscope and the data were compared with earlier reported data on machining. Although the dimensions of the instruments varied greatly, no correlation could be found between the machining and dimensions. Only one brand of files was completely within allowable size range.

Dentin permeability to bacterial proteins in vitro.

Passage of bacterial components through dentin is a subject of recent research with in vitro as well as with in vivo models. Diffusive transport of Porphyromonas gingivalis ATCC 33277 proteins has been demonstrated by the authors with an in vitro setup closely simulating the pulp chamber. The purpose of this investigation was to study the filtration of these proteins through dentin, measure possible concentration changes resulting from the filtering, and elaborate on the physical aspects of the binding process. The hydraulic conductance (Lp) of 10 dentin specimens was determined in three experiments using standard procedures: initially with phosphate-buffered saline, subsequently with a P. gingivalis ATCC 33277 suspension (200 micrograms/ml of protein), and finally with phosphate-buffered saline. The results showed significant, nonpermanent, alterations of the Lps of the dentin disks, and variable retention of the bacterial proteins among samples. A positive correlation between the bacterial protein retention and reduction of Lps was also demonstrated. The reduction of the Lps of the dentin samples was attributed to microstructural changes of the dentin, while the bacterial protein retention was considered to take place either on the surface of the disk or intratubularly. The intratubular interactions in the present experiment seemed to have a mechanical rather than a chemical basis.

Comparison of apical leakage in root canals obturated with various gutta percha techniques using a dye vacuum tracing method.

In this study, residual apical microlumina remaining after obturation with five different gutta-percha obturation methods were traced with an efficient vacuum method. Seventy-nine human maxillary anterior teeth were instrumented and divided into five groups. The groups were obturated with single-cone techniques, lateral condensation, vertical condensation, Thermafil, and Ultrafil techniques. All specimens were immersed in a vacuum flask containing 2% methylene blue dye solution. The air was evacuated with a vacuum pump to an absolute pressure of 75 torr for 30 min. The roots were sectioned longitudinally and the linear extent of dye penetration was measured. Group mean linear dye penetration values were: single-cone technique, 1.55 +/- 0.95 mm; lateral condensation, 2.25 +/- 1.46 mm; vertical condensation, 2.61 +/- 2.61 mm; Thermafil 1.41 +/- 1.16 mm; and Ultrafil 3.51 +/- 4.70 mm. There were no statistically significant differences between the gutta-percha obturation methods. The clinical implications of these findings are discussed.

In vitro cytotoxicity of root canal filling materials: 1. Gutta-percha.

Gutta-percha (GP) has been the most widely used root canal filling material because of its well-known low toxicity. The inertness of GP, however, was challenged recently. The purpose of this study was to evaluate the toxicity of marketed endodontic GP using the radiochromium release test. Fourteen commercially available and three experimental GP brands were tested. Raw GP, zinc oxide, and barium sulfate, which were considered major components of GP points, and zinc ions were also evaluated. The material was spread to cover the bottom of testing wells after being dissolved in chloroform or warmed. A labeled suspension of L929 cells was added to the wells. After incubation at 37 degrees C for 4 and 24 h, extracellular radiochromium in the culture medium was measured and calculated in percentage of the total intracellular label. Spontaneous release of radiochromium was used as control and the results were considered to be within normal limits either at 4 or 24 h. All chloroform-dissolved GP showed low toxicity at 4 h, whereas warmed GP showed statistically significant differences at 4 h. Both dissolved and warmed GP were toxic at 24 h. The raw materials and barium sulfate were not toxic, whereas zinc oxide and zinc ions showed marked toxicity. All GP points tested were toxic at longer observation periods, and the toxicity was attributed to leakage of zinc ions into the fluids.

Machining efficiency of endodontic K files and Hedstrom files.

Methods for the objective evaluation of the machining property of endodontic files were described earlier. These methods were used in this investigation to assess the characteristics of machining of 7 different brands of K files and 10 brands of Hedstrom files. The results show that there are large variations in amounts of material removed by the files. These variations are large between brands of instruments as well as between files of the same brand. Thus, among the 510 files evaluated, the file with the highest machining quality was more than 40 times as effective as the least effective instrument. The highest values among K files were obtained with files manufactured by Maillefer followed by Antaeos, Healthco, JS Dental, Brasseler, and Miltex. Among the Hedstrom files, the most efficient brands were Antaeos and Zipperer followed by Healthco, Hygenic, JS Dental, Miltex, Maillefer, Union Broach, and the least efficient Brasseler. The files did not wear noticeably during these testing procedures. These results clearly indicate that the provider needs more information from the manufacturer in regard to machining and cutting in order to intelligently select instruments. It is suggested that procedures be agreed upon to evaluate machining and cutting of root canal files in order to provide the endodontist with better information about instruments.

Machining efficiency of endodontic files: a new methodology.

The ability of endodontic files to effectively remove hard tissues is difficult to assess as results from published investigations often vary considerably. An analysis is presented of errors occurring during efficiency assessments. It was found that the machining property varies around the circumference of a single file as well as between various files of the same brand. A new method is proposed for the assessment of the quantity and quality of the machining properties of an endodontic file.

Machining efficiency of Flex-R, K-Flex, Trio-Cut, and S Files.

Many root canal instruments with several different designs have been introduced during recent years. This article evaluates the machining properties of four such instruments: the Flex-R, K-Flex, Trio-Cut, and S file. There are large variations in the machining properties of these instruments. The results are comparable to findings when evaluating conventional root canal files which show that the Hedstrom file is much more efficient when considering removal of material than the K file. The Flex-R file, K-Flex file, and Trio-Cut file are similar in design. The two former brands perform in a similar fashion, while the Trio-Cut file is less efficient than any of the evaluated K files. The S file performs as a Hedstrom file which can be expected from its similarity in design to that instrument.

Biological evaluation of collagen gels containing calcium hydroxide and hydroxyapatite.

Dentinal chips and different biocompatible materials have been used in the past as apical plugs to enhance healing and simplify obturation. The purpose of this study was to evaluate bone reactions to implants of hydroxyapatite, collagen, and calcium hydroxide, alone or in different combinations. The materials were implanted bilaterally in the mandibles of 36 guinea pigs and the reactions were compared histologically over a period of 16 wk. No major inflammatory reactions were observed in any of the implant combinations. Hydroxyapatite was not resorbed over the examination periods, but calcium hydroxide and collagen implants were partially or totally resorbed and replaced by bony tissue. The results indicate that cylinders which may be useful as apical plugs can be performed and will heal with minimal biological reactions.

Toxicity of sonicated extracts of Bacteroides gingivalis on human pulpal cells and L929 cells in vitro.

Human pulpal fibroblasts and L929 cells were treated with sonicated extracts of two strains of Bacteroides gingivalis (W83 and ATCC 33277). The cell reaction was evaluated by monitoring cell growth and DNA synthesis. Light and scanning electron microscopic analysis were used to evaluate morphological changes of the cells. Extracts from both bacterial strains exerted a growth inhibitory effect on the cells. The pulpal cells were more sensitive than L929 cells. The ATCC 33277 strain of B. gingivalis was more cytotoxic than the W83 strain. Pulpal cells appeared to be markedly affected on the microscopic level. The diffusion of these toxic bacterial by-products, through dentin to the pulp, may account for pulpal cell damage that contributes to the initiation of pulpal pathosis.

Indirect longitudinal cytotoxicity of root canal sealers on L929 cells and human periodontal ligament fibroblasts.

The cytotoxicity of two root canal sealers was evaluated in vitro. The powder components of both sealers, mainly zinc, were the same. The liquid for one sealer, Canals, was clove oil (included eugenol in more than 80%) and other materials. For the other, Canals-N, the liquid was composed of higher fatty acids and glycol. The experiments included two cell lines, heteroploid L929 mouse fibroblasts and diploid human periodontal ligament fibroblasts. Cytotoxicity was assessed using the radiochromium release method with 4-h exposure time. The assay involved using insert chambers in multiwell arrays to produce indirect contact of materials with the cell monolayer at a controlled distance of approximately 1 mm. This model also allowed for the longitudinal study of the same material sample to assess time-dependent changes in toxicity. Freshly mixed Canals was highly toxic (p < 0.01) to both cell lines. On and after 24 h of setting no toxicity was detected. At no time could cytotoxicity be observed when experimenting with Canals-N. These results indicate that both materials have a low content of water diffusible toxic components. Substituting eugenol can further decrease the toxicity of the sealer.

Root canal dentinal tubule disinfection.

Dentinal tubules of the root canal walls of human teeth were infected in vitro with a known bacterial isolate. The roots were exposed to either calcium hydroxide or iodine potassium-iodide for various periods of time and the viability of microorganisms was determined by incubation of entire root samples in a culture medium. The effects of the two agents on microbial viability were evaluated and compared. Iodine potassium-iodide disinfected dentin effectively. In contrast, bacteria remained viable in the dentin after relatively extended periods of calcium hydroxide treatment.

Silver glass ionomer cement as a retrograde filling material: a study in vitro.

Comparison was made of the apical microleakage of retrograde fillings with amalgam and with silver glass ionomer cements using a modified dye penetration method. Forty instrumented human teeth were divided into four groups. Each group was characterized by a different retrograde filling material or technique: Group 1--silver glass ionomer (SGI); group 2--SGI with previous acid wash of the cavity; group 3--SGI in a previously acid washed cavity, protected with varnish; and group 4--zinc-free amalgam. A paper cone for dye adsorption analysis was placed in contact with retrofill and the main canal was obturated with thermoplasticized gutta-percha. All teeth were coated, immersed in methylene blue dye, and evacuated to assure complete penetration of the dye in any possible void. Later the teeth were cleared and evaluated under stereomicroscope. Statistical analysis showed that group 1 had the least microleakage of all. In addition, the biocompatibility of the SGI and varnish was compared with that of amalgam using the 51Cr release evaluation method. The results showed that SGI and varnish were less cytotoxic than amalgam. It was concluded that SGI cement can be considered an alternative retrograde filling material.

The endodontic file is a disposable instrument.

Most cutting and machining tools wear when properly operated on intended substrates. Little is known about endodontic files when machining on human dentin. It was shown earlier that the efficiency of files was very variable when evaluated on Plexiglas substrates. The present study of seven different brands and types of files was undertaken to understand their wear better when machining human dentin. It was shown that all files evaluated rapidly deteriorated when machining dentin. This decline in efficiency was significant but different within, as well as, among brands. It was suggested that endodontic files be disposable.

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.

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.

Reduced cytotoxicity of a root canal sealer through eugenol substitution.

The cytotoxicity of two zinc oxide root canal sealers was investigated in vitro. The sealers were freshly mixed and set for 24 and 168 h. The sealers had identical powders but different liquid components. One (Canals) used eugenol, while the other (Canals-N) used fatty acids. L929 cells were incubated for 4 and 24 h in direct contact with the materials or with an eluate of the materials. The toxicity was evaluated using the radiochromium release assay. In the direct exposure assay, both sealers were cytotoxic when freshly prepared or after 24 h of setting. After 1 wk of setting, Canals was still toxic, while Canals-N was not significantly different from the control in the 4-h assay. In the elution assay the materials showed very low cytotoxicity. Only the eluate from freshly prepared Canals was clearly cytotoxic after 24 h. The liquid of Canals-N was clearly less cytotoxic than liquid from Canals. The results showed that the cytotoxicity of a root canal sealer can be reduced by replacing eugenol with fatty acids.

Endodontic anesthesia in mandibular molars: a clinical study.

Sixty-one mandibular molar teeth with clinically manifest pulpitis, which required endodontic therapy, were studied. Twenty-seven subjects received standard inferior alveolar nerve block (IANB) with 2% lidocaine HCI with 1:100,000 epinephrine and 34 subjects received IANB with 3% mepivacaine with no vasoconstrictor. Pulpal anesthesia was assessed with dichlorodifluormethane (DDM). Subjects who gave a positive response to DDM were given a periodontal ligament injection with 2% lidocaine with 1:100,000 epinephrine. This study showed that 3% mepivacaine HCI is as effective as 2% lidocaine HCI in achieving pulpal anesthesia in mandibular molars with IANB. Of a total 61 IANB with lip anesthesia, 23 subjects required periodontal ligament injection to achieve a negative response to DDM. It was concluded that lip anesthesia is not a reliable indicator of pulpal anesthesia. The use of DDM is a reliable method of determining true pulpal anesthesia.

An in vitro method for longitudinal evaluation of toxicity of endodontic sealers.

An in vitro method for longitudinal evaluation of root canal sealers was developed and applied. A newly introduced cell culture chamber was used to evaluate the cytotoxic effects of test samples immediately after mixing and for an extended period of time thereafter. A ranking of the test materials, based on their cytotoxicity, was allowed by the method.

Influence of entrapped air on the accuracy of leakage studies using dye penetration methods.

Entrapped air can inhibit dye penetration resulting in failure to demonstrate existing voids. A new approach to study dye penetration was developed in which the entrapped air was evacuated before the dye was introduced. This method was compared with the regular technique of passive dye diffusion. Uniformly prepared root canal specimens were filled with AH26. Two different sizes of standardized voids were created in the specimens. Specimens were either immersed passively in 2% methylene blue for a week or placed in a chamber with 100 mtorr absolute pressure, after which the dye was introduced while the vacuum was maintained. Leakage was measured stereomicroscopically. Controls with no voids showed insignificant external leakage. Passive dye penetration resulted in incomplete filling of the voids regardless of size of the voids, whereas vacuum dye delivery resulted in complete filling of the voids. Results of this study suggest that entrapped air produces artifacts in passive immersion technique; therefore, the sample should be evacuated prior to the dye introduction in order to demonstrate the full extent of void in future dye penetration studies.

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.