Is it really penetration? Part 2. Locomotion of Enterococcus faecalis cells within dentinal tubules of bovine teeth.

OBJECTIVE: The aim of the present vitro study was to examine the question whether devitalized Enterococcus faecalis (E. faecalis) cells can migrate into dentinal tubules and if that process takes place in a time-dependent manner. DESIGN: Sixty bovine root canals were incubated with devitalized and vital streptomycin-resistant E. faecalis strains after root canal enlargement (size 80, taper .02) with 3% NaOCl solution. Incubation times 7, 14, 21, 28, 35, and 42 days. Samples were processed for analysis by scanning electron microscopy (SEM) and DAPI (4',6-diamidino-2-phenylindole) staining. The penetration depth was calculated with the measurement tool of the Axio Vision program (Zeiss, Jena, Germany). Statistical analysis was performed by Kruskal-Wallis (α = 0.05) and Mann-Whitney U test (p < 0.05). RESULTS: Devitalized E. faecalis strains were able to migrate into dentinal tubules. The total number and penetration depth of devitalized E. faecalis cells was lower compared to the vital suspension of E. faecalis. It was noted, that bacterial penetration was not common to all of the dentinal tubules in the vital E. faecalis control and especially in the devitalized control. The migration took place in a time-dependent migration characteristic. CONCLUSIONS: Devitalized E. faecalis cells are still able to migrate into the dentinal tubules due to possible electrokinetic and osmotic processes. Thereby, increased exposure times lead to a time-dependent penetration characteristic. CLINICAL RELEVANCE: Since devitalized bacteria can migrate as well into dentinal tubules, the presence of bacteria within dentinal tubules cannot be interpreted as a failure of tested preparation regimens.

Development of a Function-Integrative Sleeve for Medical Applications.

Function-integrative textiles bear the potential for a variety of applications in the medical field. Recent clinical investigations suggest that the application of a function-integrative fabric could have a positive impact on currently applied diagnostic procedures of a specific type of tumour. In this context, the fabric should enable local warming of a patient's upper extremity as well as blood flow measurement. Existing solutions comprise a warming a warming system but lack a measuring apparatus for blood flow determination. With regard to the quality of results of current diagnostic procedures, the local warming of the patients' upper extremity and the simultaneous determination of the blood flow plateau are crucial. In the present paper, the development process of a function-integrative sleeve is introduced. Besides the development of an adaptable sleeve-design, the manufacturing process of an integrated warming system was also addressed. Furthermore, the identification of crucial physiological effects, using a Laser Doppler Perfusion Monitor, is introduced. During testing of the function-integrative sleeve, modulation of the desired physiological effects was observed. The results support the initial assumptions and dictate further investigations on increasing user-friendliness and cost-efficiency during adjusting and determining the physiological effects in the course of tumour diagnosis.

Design and Testing of a Co-Rotating Vibration Excitation System.

A vibration excitation system (VES) in a form of an active coupling is proposed, designed and manufactured. The system is equipped with a set of piezoelectric stack actuators uniformly distributed around the rotor axis and positioned parallel to each other. The actuator arrangement allows an axial displacement of the coupling halves as well as their rotation about any transverse axis. Through the application of the VES an aimed vibration excitation is realised in a co-rotating coordinate system, which enables a non-invasive and precise modal analysis of rotating components. As an example, the VES is applied for the characterisation of the structural dynamic behaviour of a generic steel rotor at different rotational speeds. The first results are promising for both stationary and rotating conditions.

Determination of a Representative and 3D-Printable Root Canal Geometry for Endodontic Investigations and Pre-Clinical Endodontic Training-An Ex Vivo Study.

Models of artificial root canals are used in several fields of endodontic investigations and pre-clinical endodontic training. They allow the physical testing of dental treatments, the operating of instruments used and the interaction between these instruments and the tissues. Currently, a large number of different artificial root canal models exist whose geometry is created either on the basis of selected natural root canal systems or to represent individual geometrical properties. Currently, only a few geometric properties such as the root canal curvature or the endodontic working width are taken into consideration when generating these models. To improve the representational capability of the artificial root canal models, the aim of the current study is therefore to generate an artificial root canal based on the statistical evaluation of selected natural root canals. Here, the approach introduced by Kucher for determining the geometry of a root canal model is used, which is based on the measurement and statistical evaluation of the root canal center line's curvatures and their cross-sectional dimensions. Using the example of unbranched distal root canals of mandibular molars (n = 29), an artificial root canal model representing the mean length, curvature, torsion and cross-sectional dimensions of these teeth could be derived.

Efficacy of Endodontic Disinfection Protocols in an E. faecalis Biofilm Model-Using DAPI Staining and SEM.

The aim of this study was to investigate the antimicrobial efficacy of different disinfection protocols in a novel Enterococcus faecalis biofilm model based on a visualization method and to evaluate the potential alteration of dentinal surface. A total of 120 extracted human premolars were allocated to 6 groups with different irrigation protocols. The assessment of the effectiveness of each protocol and the alteration of dentinal surface were visualized by using SEM and fluorescence microscopy (DAPI). A dense E. faecalis biofilm with a penetration depth of 289 µm (medial part of the root canal) and 93 µm (apical part) validated that the biofilm model had been successfully implemented. A significant difference between the 3% NaOCl groups and all the other groups in both observed parts of the root canal (p < 0.05) was detected. However, the SEM analysis revealed that the dentinal surface in the 3% NaOCl groups was severely altered. The established biofilm model and the visualization method based on DAPI are appropriate for bacterial quantification and evaluation of the depth effect of different disinfection protocols in the root canal system. The combination of 3% NaOCl with 20% EDTA or MTAD with PUI allows the decontamination of deeper dentine zones within the root canal but simultaneously alters the dentinal surface.

Analysis of a Film Forming Process through Coupled Image Correlation and Infrared Thermography.

The aim of the present investigation was to determine the dependence of the material and process parameters of the bending process of thermoplastic films. In this context, parameter combinations leading to high resulting forming ratios were identified. To measure the relevant parameters within the hot bending process, a coupled evaluation of infrared thermography (IRT) and deformation measurement using digital image correlation (DIC) was performed. The coupled measurement enables the identification of the actual mechanically stressed bending area of the film as a result of the bending process. This allows for the specification of the local forming temperatures required for the desired forming ratios. Furthermore, the mechanical and thermal strain along the defined measuring sections and their deviation in individual tests as well as the effect of thermal strain on process control on a larger scale were determined. Based on the results, a process window was defined for the film materials investigated, which will serve as a starting point for future efforts to develop a continuous manufacturing process.

Material Selection Process for Acoustic and Vibration Applications Using the Example of a Plate Resonator.

In this work, a new method for selecting suitable materials is presented. This method has a high potential for a variety of engineering applications, such as the design of sound-absorbing and vibration-loaded structures, where a large number of different requirements have to be met. The method is based on the derivation of functional dependencies of selected material parameters. These dependencies can be used in parameter studies to consider parameter combinations that lie in the range of real existing and targeted material groups. This allows the parameter space to be reduced, the calculation to be accelerated, and suitable materials to be (pre-)selected for the respective application, which contributes to a more target-oriented design. The method is applied to the example of a plate resonator. For this purpose, a semi-analytical model is implemented to calculate the transmission loss as well as the reflected and dissipated sound power of plate silencers, taking into account the influence of flow velocity and fluid temperature on the performance of plate silencers.

Quantification of Bacterial DNA from Infected Human Root Canals Using qPCR and DAPI after Disinfection with Established and Novel Irrigation Protocols.

The removal of bacterial infections within the root canal system is still a challenge. Therefore, the cleansing effect of established and new irrigation-protocols (IP) containing silver diamine fluoride (SDF) 3.8% on the whole root canal system was analyzed using quantitative PCR (qPCR) and 4',6-diamidino-phenylindole-(DAPI)-staining. Extracted human premolars were instrumented up to F2 (ProTaper Gold) under NaCl 0.9% irrigation and incubated with Enterococcus faecalis for 42 days. Subsequently, different ultrasonically agitated IP were applied to the roots: control (no irrigation), 1. NaOCl 3%, EDTA 20%, CHX 2%, 2. NaOCl 3%, EDTA 20%, 3. NaOCl 3%, EDTA 20%, SDF 3.8%, 4. SDF 3.8%, and 5. NaCl 0.9%. One half of the root was investigated fluorescent-microscopically with DAPI. The other half was grinded in a cryogenic mill and the bacterial DNA was quantified with qPCR. The qPCR results showed a statistically significant reduction of bacteria after the application of IP 1, 2, and 3 compared to the control group. While IP 4 lead to a bacterial reduction which was not significant, IP 5 showed no reduction. These data corresponded with DAPI staining. With qPCR a new molecular-biological method for the investigation of the complete root canal system was implemented. The novel IP 3 had an equally good cleansing effect as the already established IP.

An Automated Measurement Method for the Endodontic Working Width of Lower Molars by Means of Parametric Models Using Cone-beam Computed Tomographcy and Micro-Computed Tomography.

INTRODUCTION: A new method for the approximation of the root canal's cross-sectional shape and its working width using cone-beam computed tomographic (CBCT) or micro-computed tomographic (micro-CT) imaging was introduced. METHODS: Scanned data from 29 extracted human mandibular first and second molar distal root canals without instrumentation were reconstructed and analyzed with a self-developed measurement algorithm. The 3-dimensional volume models were sliced perpendicular to the vertical axis. Using different 2-dimensional parametric models, the contour of each root canal slice was approximated and used to determine the canal's cross-sectional dimensions. The measurements of minor width, major width, and the root canal's conicity were statistically analyzed using analysis of variance. RESULTS: The measured minor and major widths of the investigated root canals were significantly higher (probability value P < .05) when evaluated by CBCT images than the results obtained from micro-CT data. Both dimensions increased starting from the apical foramen (P < .01). The narrowest measured canal widths were 0.19-0.24 mm for CBCT imaging and 0.09-0.21 mm for micro-CT imaging in the apical part. The maximum values for conicity were between 13% and 17% in the cervical third. CONCLUSIONS: The 3-dimensional imaging data from CBCT and micro-CT imaging enabled a valuable anatomic assessment of the root canal's cross-sectional working width along the canal up to the physiological foramen in order to determine an adequate apical diameter as well as the correct measured taper in the cervical and medial part.

Mapping of the Micro-Mechanical Properties of Human Root Dentin by Means of Microindentation.

The extensive knowledge of root dentin's mechanical properties is necessary for the prediction of microstructural alterations and the teeth's deformations as well as their fracture behavior. Standardized microindentation tests were applied to apical, medial, and cervical root sections of a mandibular human first molar to determine the spatial distribution of the hard tissue's properties (indentation modulus, indentation hardness, Martens hardness, indentation creep). Using an indentation mapping approach, the inhomogeneity of mechanical properties in longitudinal as well as in transversal directions were measured. As a result, the tooth showed strongly inhomogeneous material properties, which depended on the longitudinal and transversal positions. In the transversal cutting planes of the cervical, medial, apical sections, the properties showed a comparable distribution. A statistical evaluation revealed an indentation modulus between 12.2 GPa and 17.8 GPa, indentation hardness between 0.4 GPa and 0.64 GPa and an indentation creep between 8.6% and 10.7%. The established standardized method is a starting point for further investigations concerning the intensive description of the inhomogeneous mechanical properties of human dentin and other types of dentin.

Sound Transmission Loss of a Sandwich Plate with Adjustable Core Layer Thickness.

Compressible Constrained Layer Damping (CCLD) is a novel, semi-active, lightweight-compatible solution for vibration mitigation based on the well-known constrained layer damping principle. The sandwich-like CCLD set-up consists of a base structure, a constraining plate, and a compressible open-cell foam core in between, enabling the adjustment of the structure's vibration behaviour by changing the core compression using different actuation pressures. The aim of the contribution is to show to what degree, and in which frequency range the acoustic behaviour can be tuned using CCLD. Therefore, the sound transmission loss (TL), as an important vibro-acoustic index, is determined in an acoustic window test stand at different actuation pressures covering a frequency range from 0.5 to 5 kHz. The different actuation pressures applied cause a variation of the core layer thickness (from 0.9 d0 to 0.3 d0), but the resulting changes of the stiffness and damping of the overall structure have no significant influence on the TL up to approximately 1 kHz for the analysed CCLD design. Between 1 kHz and 5 kHz, however, the TL can be influenced considerably well by the actuation pressure applied, due to a damping-dominated behaviour around the critical frequency.

Quantification of Bacterial Colonization in Dental Hard Tissues Using Optimized Molecular Biological Methods.

Bacterial infections of root canals and the surrounding dental hard tissue are still a challenge due to biofilm formation as well as the complex root canal anatomy. However, current methods for analyzing biofilm formation, bacterial colonization of root canals and dental hard tissue [e.g., scanning electron microscopy, confocal laser scanning microscopy (CLSM) or determination of colony forming units (CFU)] are time-consuming and only offer a selective qualitative or semi-quantitative analysis. The aim of the present study is the establishment of optimized molecular biological methods for DNA-isolation and quantification of bacterial colonization via quantitative PCR (qPCR) from dental hard tissue. Root canals of human premolars were colonized with Enterococcus faecalis. For isolation of DNA, teeth were then grinded with a cryo mill. Since the hard tissues dentin and especially enamel belong to the hardest materials in the human organism, the isolation of bacterial DNA from root dentin is very challenging. Therefore, treatment steps for the isolation of DNA from grinded teeth were systematically analyzed to allow improved recovery of bacterial DNA from dental hard tissues. Starting with the disintegration of the peptidoglycan-layer of bacterial cells, different lysozyme solutions were tested for efficacy. Furthermore, incubation times and concentrations of chelating agents such as EDTA were optimized. These solutions are crucial for the disintegration of teeth and hence improve the accessibility of bacterial DNA. The final step was the determination of prior bacterial colonization of each root canal as determined by qPCR and comparing the results to alternative methods such as CFU. As a result of this study, optimized procedures for bacterial DNA-isolation from teeth were established, which result in an increased recovery rate of bacterial DNA. This method allows a non-selective and straightforward procedure to quantify bacterial colonization from dental hard tissue. It can be easily adapted for other study types such as microbiome studies and for comparable tissues like bones.

Continuous Measurement of Three-Dimensional Root Canal Curvature Using Cone-Beam Computed and Micro-Computed Tomography: A Comparative Study.

The knowledge of root canal curvature is crucial regarding the prevention of ledge formation, root perforation and the possibility of endodontic instruments' fracture during endodontic treatments. Therefore, a quantification method of the root canal curvature as well as the applicability of diagnostically relevant tomographic three-dimensional (3D) imaging data is necessary. Hereby, cone-beam computed tomography (CBCT) images and micro-computed tomography (µCT) data of distal root canals were analysed concerning the continuous three-dimensional curvature of human mandibular molars ( n = 50). The curvature of the canal's three-dimensional centre line was determined by evaluating the tomographic images. The centroids of each root canal slice were identified and approximated by spline curves to obtain the centre line and therefore, its curvature. Comparing the results evaluated from CBCT and µCT images, minimum radii of curvature of 2.6 mm and 2.1 mm were determined, respectively. The observation of the centre line demonstrated the requirement of the three-dimensional imaging data from CBCT and µCT for a reliable curvature analysis. Conclusively, the evaluation of CBCT and µCT images results in comparable radii of curvature. Thus, the application of the introduced method in combination with CBCT applied to patient cases could offer an important preliminary diagnostical step to prevent endodontic treatment complications.

Effects of Endodontic Irrigants on Material and Surface Properties of Biocompatible Thermoplastics.

Passive irrigation is an efficient method for a successful endodontic treatment. During sonic activation biocompatible polymer tips are used to activate irrigants. Compared to ultrasonic activation with metallic tips, polymer tips have the advantage of a reduced risk of fracture and minimise dentine damage. Hence, two polymers, polyether ether ketones (PEEK) and polyamide (PA6), were identified for the manufacturing of novel irrigation tips. The chemical resistance against the irrigants ethylenediaminetetraacetic acid (EDTA) 20%, chlorhexidine gluconate (CHX) 2% and sodium hypochlorite (NaOCl) 5.25% was analysed. Using microindentation, the change of hardness, elasticity, surface roughness and appearance of the polymers was determined. PA6 had a high absorption of irrigant compared to PEEK. PEEK was resistant to the investigated irrigants and showed no significant alteration of surface and mechanical properties, whereas PA6 slightly increased its hardness, elastic modulus and surface roughness during long-term exposure at 37 °C. However, PA6 tips seem to be a promising disposable product due to the material's high deformability and low manufacturing costs. Particularly with regard to structural-dynamic properties and high chemical resistance, PEEK can be considered as a material for reusable irrigation tips.

An Approach for a Mathematical Description of Human Root Canals by Means of Elementary Parameters.

INTRODUCTION: Root canal geometry is an important factor for instrumentation and preparation of the canals. Curvature, length, shape, and ramifications need to be evaluated in advance to enhance the success of the treatment. Therefore, the present study aimed to design and realize a method for analyzing the geometric characteristics of human root canals. METHODS: Two extracted human lower molars were radiographed in the occlusal direction using micro-computed tomographic imaging. The 3-dimensional geometry of the root canals, calculated by a self-implemented image evaluation algorithm, was described by 3 different mathematical models: the elliptical model, the 1-circle model, and the 3-circle model. RESULTS: The different applied mathematical models obtained similar geometric properties depending on the parametric model used. Considering more complex root canals, the differences of the results increase because of the different adaptability and the better approximation of the geometry. CONCLUSIONS: With the presented approach, it is possible to estimate and compare the geometry of natural root canals. Therefore, the deviation of the canal can be assessed, which is important for the choice of taper of root canal instruments. Root canals with a nearly elliptical cross section are reasonably approximated by the elliptical model, whereas the 3-circle model obtains a good agreement for curved shapes.

Is it really penetration? Locomotion of devitalized Enterococcus faecalis cells within dentinal tubules of bovine teeth.

OBJECTIVE: The aim of the present study was to evaluate the penetration characteristics of devitalized and vital E. faecalis cells into root dentinal tubules. DESIGN: Thirteen root canals were incubated with devitalized (4days, 7days, 14days, 28days) and vital (28days) E. faecalis strains (streptomycin-resistant strains) after root canal enlargement (size 80, taper 0.02) with 3 % NaOCl solution. The smear layer was intentionally removed with 20 % EDTA before inoculation. Samples were processed for analysis by scanning electron microscopy (SEM) and DAPI (4',6-diamidino-2-phenylindole) staining. DAPI was conducted for fluorescence microscopic visualization of the bacterial penetration into dentinal tubules. The penetration depth was calculated with the measurement tool of the Axio Vision program (Zeiss, Jena, Germany). RESULTS: Devitalized E. faecalis strains were able to penetrate into dentinal tubules of the root canal. Apikal penetration depths of the devitalized cells were 100.67µm±26.54µm after 7days, 230.67µm±111.5µm after 14days and 266.5µm±92.63µm after 28days of incubation. The total number and penetration depth of E. faecalis cells was lower compared to a vital suspension of E. faecalis (1002.45µm) after 28days. It was noted that bacterial penetration was not common to all of the dentinal tubules in the vital E. faecalis control and especially in the devitalized control. CONCLUSIONS: Increased exposure times of devitalized bacteria into root canals lead to an increased number of penetrated dentinal tubules as well as to a deeper penetration.