Biomechanical characterization of a fibrinogen-blood hydrogel for human dental pulp regeneration.

In dental practice, Regenerative Endodontic Treatment (RET) is applied as an alternative to classical endodontic treatments of immature necrotic teeth. This procedure, also known as dental pulp revitalization, relies on the formation of a blood clot inside the root canal leading to the formation of a reparative vascularized tissue similar to dental pulp, which would provide vitality to the affected tooth. Despite the benefit of this technique, it lacks reproducibility due to the fast degradation and poor mechanical properties of blood clots. This work presents a method for constructing a fibrinogen-blood hydrogel that mimics the viscoelastic properties of human dental pulp while preserving the biological properties of blood for application in RET. By varying the blood and fibrinogen concentrations, gels with different biomechanical and biological properties were obtained. Rheology and atomic force microscopy (AFM) were combined to study the viscoelastic properties. AFM was used to evaluate the elasticity of human dental pulp. The degradation and swelling rates were assessed by measuring weight changes. The biomimetic properties of the gels were demonstrated by studying the cell survival and proliferation of dental pulp cells (DPCs) for 14 days. The formation of an extracellular matrix (ECM) was assessed by multiphoton microscopy (MPM). The angiogenic potential was evaluated by an ex vivo aortic ring assay, in which the endothelial cells were observed by histological staining after migration. The results show that the Fbg-blood gel prepared with 9 mg ml-1 fibrinogen and 50% blood of the Fbg solution volume has similar elasticity to human dental pulp and adequate degradation and swelling rates. It also allows cell survival and ECM secretion and enhances endothelial cell migration and formation of neovessel-like structures.

In vitro, ex vivo, and in vivo models for dental pulp regeneration.

Based on the concept of tissue engineering (Cells-Scaffold-Bioactive molecules), regenerative endodontics appeared as a new notion for dental endodontic treatment. Its approaches aim to preserve dental pulp vitality (pulp capping) or to regenerate a vascularized pulp-like tissue inside necrotic root canals by cell homing. To improve the methods of tissue engineering for pulp regeneration, numerous studies using in vitro, ex vivo, and in vivo models have been performed. This review explores the evolution of laboratory models used in such studies and classifies them according to different criteria. It starts from the initial two-dimensional in vitro models that allowed characterization of stem cell behavior, through 3D culture matrices combined with dental tissue and finally arrives at the more challenging ex vivo and in vivo models. The travel which follows the elaboration of such models reveals the difficulty in establishing reproducible laboratory models for dental pulp regeneration. The development of well-established protocols and new laboratory ex vivo and in vivo models in the field of pulp regeneration would lead to consistent results, reduction of animal experimentation, and facilitation of the translation to clinical practice.

Concordance study between regular face-to-face dental diagnosis and dental telediagnosis using fluorescence.

BACKGROUND: Teledentistry consultations are an effective way to increase access to care. Whether it be for a screening, referral or even an adapted treatment plan for a certain number of patients whose access to care is complicated, demonstrating the reliability of remote consultations is essential in allowing the technique to become generalised. AIM: This study aimed to determine if teledentistry consultations using fluorescence are of the same quality as regular consultations in the diagnosis of caries. METHODS: Patients were seen in consultation in the dental care centre at the Montpellier University Hospital (France) and in the centre at Kyushu Dental University Hospital (Japan). The protocol was broken down into three parts: the regular consultation, the recording of videos with the Soprocare camera and the remote consultation. The regular consultation and the remote consultation were blinded and carried out by two different dentists. The recording of videos was carried out by a third dentist. The carious diagnosis was based on the International Caries Detection and Assessment System: a clinical rating system for the detection and assessment of caries. RESULTS: One hundred and ninety-five patients met the predefined inclusion criteria. Most patients had at least one surface at stage 3 or higher (73%) with a higher proportion amongst French patients (81% compared to 66%). However, they had good dental hygiene, given that dental hygiene was only deemed unsatisfactory for 10.8% (19% for French patients and 2% for Japanese patients). The odontogram (presence/absence of each tooth) seemed to be correctly identified during the remote consultation (reinterpretation). Out of the 195 patients, 168 (86.2%) were identified without error. CONCLUSIONS: Teledentistry consultations can represent acceptable diagnostic performance with regard to the detection of dental caries. The Soprocare camera enables an early diagnosis of carious lesions with optimal efficiency. Several areas still need to be improved, however, so that the use of the camera during remote consultations is as coherent and effective as possible, especially with regard to the organisational aspects of remote consultations.

Performance of Fluorescence-based Systems in Early Caries Detection: A Public Health Issue.

AIM: Modern clinical caries management involves early stage caries diagnosis and should fit with dental health policy. The objective of this study was to achieve early caries detection in enamel and dentine with a laser-based system (DIAGNOdent™ pen) first and secondary with a new fluorescence intra-oral camera (Soprolife®). A visual inspection with a loupe was used as control. MATERIALS AND METHODS: Following the consolidated standards of reporting trials recommendations, 628 occlusal fissures were included for analysis. RESULTS: The sensitivity and specificity of both devices varied depending on the cutoff threshold of the caries score, and the ROC curve showed higher values for the Soprolife® than for DIAGNOdent™ pen. The values of the area under the curve decreased from 0.81 (Soprolife® in daylight) to 0.79 (Soprolife® in fluorescent mode) and 0.67 for DIAGNOdent™ pen. DIAGNOdent™ pen reproducibility (intra and inter-investigator) showed a wide dispersion, with many values scattered beyond the confidence limits (±2 SD), and the weighted kappa coefficient, which was quite low (0.58), confirmed this tendency. CONCLUSION: Caries prevalence in terms of public health policy is of interest and caries detection increased significantly when using an fluorescence-based intra-oral camera. CLINICAL SIGNIFICANCE: The clinical significance of these findings is that fluorescence could help improve caries diagnosis, reduce clinical misinterpretations, and finally benefit the patients. How to cite this article: Terrer E, Slimani A, Giraudeau N, et al. Performance of Fluorescence-based Systems in Early Caries Detection: A Public Health Issue. J Contemp Dent Pract 2019;20(10):1126-1132.

A Five-year Comparative Study of Perceived Stress Among Dental Students at Two European Faculties.

AIM: To compare the levels, causes and the impact of stress from cohort of fifthyear dental students in the Faculties of Dental Medicine in Plovdiv, Bulgaria and Montpellier, France. MATERIALS AND METHODS: The questionnaire we used comprised the modified version of the 56-item Dental Environment Stress (DES) questionnaire. It was completed by fifth-year undergraduate dental students in both faculties in 2011 and 2016, totaling 335 dental students. A five-point Likert scale was used to record the responses from the subjects. Statistical analysis was performed with STATA version 14.1 (StataCorp, College Station, USA) specialized software. RESULTS: The mean age of studied population was not significantly different between 2011 and 2016 (p=0.08). The common stress value per student was 1.74 (CI: 1.64; 1.84, range: 0.12; 4.50). It was higher in Plovdiv (1.79±0.81) than in Montpellier (1.63±0.66); it was statistically higher for girls (1.94±0.70) than for boys (1.49±0.77), and decreased between 2011 and 2016. A significant overall effect of the common mean stress was influenced by the town (p=0.008), year of study (p=0.003), gender (p=0.0001) and accommodation (p=0.01) of the student. CONCLUSION: Academicals and clinical factors of stress are prevailing stressors during education. The challenge is now to decrease the influence of both, without altering the quality of dental education. Regular discussions and debriefs about the clinical cases before and after clinical activity. Alternative methods of education and examination, such as interactive methods, could be considered in order to reduce the stress of theoretical exams.

Multiphoton Microscopy for Caries Detection with ICDAS Classification.

Dentin carious lesion is a dynamic process that involves demineralization and collagen denaturation. Collagen type I is the major protein in dentin and it has been investigated based on its optical properties. Multiphoton microscopy (MPM) is a nonlinear imaging technique that reveals the caries process using the collagen two-photon excitation fluorescence (2PEF) and its second-harmonic generation (SHG). Combining the histological and the International Caries Detection and Assessment System (ICDAS) classifications with nonlinear optical spectroscopy (NLOS), 2PEF and SHG intensities of enamel and dentin were highly altered during the caries process. It has been proven that the ratio SHG/2PEF is a relevant indicator of the organic matrix denaturation [Terrer et al.: J Dent Res 2016; 96: 574-579]. In the present study, a series of measurable signals is made to detect early stages of carious lesion according to the ICDAS classification and to explore the relationship between these measures and the ICDAS scale. Comparison of the efficiency of nonlinear optical signals for caries detection with the ICDAS classification is essential to evaluate their potential for clinical application. In our study, the use of the NLOS measured by MPM allowed us to monitor a quantitative parameter (SHG/2PEF ratio) according to the dentin carious lesion state (ICDAS and histological examination). Three coherent new groups were defined (ICDAS 0/1; ICDAS 2/3; ICDAS 4/5/6), where the carious process can be clearly described with a statistically significant decrease of the SHG/2PEF ratio.

Confocal Raman mapping of collagen cross-link and crystallinity of human dentin-enamel junction.

The separation zone between enamel and dentin [dentin-enamel junction (DEJ)] with different properties in biomechanical composition has an important role in preventing crack propagation from enamel to dentin. The understanding of the chemical structure (inorganic and organic components), physical properties, and chemical composition of the human DEJ could benefit biomimetic materials in dentistry. Spatial distribution of calcium phosphate crystallinity and the collagen crosslinks near DEJ were studied using confocal Raman microscopy and calculated by different methods. To obtain collagen crosslinking, the ratio of two peaks 1660 cm-1 over 1690 cm-1 (amide I bands) is calculated. For crystallinity, the inverse full-width at half maximum of phosphate peak at 960 cm-1, and the ratio of two Raman peaks of phosphate at 960/950 cm-1 is provided. In conclusion, the study of chemical and physical properties of DEJ provides many benefits in the biomaterial field to improve the synthesis of dental materials in respect to the natural properties of human teeth. Confocal Raman microscopy as a powerful tool provides the molecular structure to identify the changes along DEJ and can be expanded for other mineralized tissues.

Strategies For Immobilization Of Bioactive Organic Molecules On Titanium Implant Surfaces - A Review.

Numerous approaches have been used to improve the tissue-implant interface of titanium (Ti) and titanium alloy (Ti6Al4V). They all aim at increasing cell migration and attachment to the metal, preventing unspecific protein adsorption and improving post-implantation healing process. Promising methods for titanium and titanium alloy surface modification are based on the immobilization of biologically active organic molecules. New and interesting biochemical approaches to such surface modification include layer-by-layer deposition of polyelectrolyte films, phage display-selected surface binding peptides and self-assembled DNA monolayer systems. The present review summarizes the scientific information about these methods, which are at in vitro or in vivo development stages, and hopes to promote their future application in dental implantology and in oral and maxillofacial surgery.

Root canal hydrophobization by dentinal silanization: improvement of silicon-based endodontic treatment tightness.

A new strategy to improve silicon-based endodontic treatment tightness by dentine hydrophobization is presented in this work: root dentine was silanized to obtain a hydrophobic dentine-sealer interface that limits fluid penetration. This strategy was based on the grafting of aliphatic carbon chains on the dentine through a silanization with the silane end groups [octadecyltrichlorosilane (OTS) and octadecyltriethoxysilane]. Dentine surface was previously pretreated, applying ethylenediaminetetraacetic acid and sodium hypochlorite, to expose hydroxyl groups of collagen for the silane grafting. Collagen fibers exposure after pretreatment was visible with scanning electron microscopy, and Fourier transform infrared (FTIR) spectroscopy showed their correct exposition for the silanization (amide I and II, with 1630, 1580, and 1538 cm⁻¹ peaks corresponding to the vibration of C=O and C--N bonds). The grafting of aliphatic carbon chains was confirmed by FTIR (peaks at 2952 and 2923 cm⁻¹ corresponding to the stretching of C--H bonds) and by the increasing of the water contact angle. The most efficient hydrophobization was obtained with OTS in ethyl acetate, with a water contact angle turning from 51° to 109°. Gas and liquid permeability tests showed an increased seal tightness after silanization: the mean gas and water flows dropped from 2.02 × 10⁻8 to 1.62 × 10⁻8 mol s⁻¹ and from 10.8 × 10⁻³ to 5.4 × 10⁻³ µL min⁻¹, respectively. These results show clear evidences to turn hydrophilic dentine surface into a hydrophobic surface that may improve endodontic sealing.

Functional mapping of human sound and carious enamel and dentin with Raman spectroscopy.

The goals of this trial were, first, to produce a Raman mapping of decay and sound dentin samples, through accurate analysis of the Raman band spectra variations of mineral and organic components. The second goal was to confirm the correlation between the Raman signal and the signal of a fluorescent camera, by assaying the concentration of pentosidine and natural collagen fluorescent crosslink using reverse phase high-pressure liquid chromatography. The first correlation assumed a possible relationship between the signal observed with the camera and Raman spectroscopy. The second correlation assumed an association with the Maillard reaction. Absence of a correlation for this trial was that no association could be found between Raman spectra characteristics, fluorescence variation and the HPLC assay. Our results void this absence.

In vitro investigation of fluorescence of carious dentin observed with a Soprolife® camera.

OBJECTIVES: Our aim was to determine the origin of the red fluorescence of carious dentine observed with the Soprolife® camera. METHODS: We conducted in vitro studies to evaluate the origin of the red fluorescence using acids and matrix metalloproteinase (MMP) to mimic caries and methylglycoxal (MGO) to evaluate the effect of glycation reactions on the red fluorescence. In every step of these models, we detected the changes of dentin photonic response with Soprolife® in daylight mode and in treatment mode. A Raman spectroscopy analysis was performed to determine the variations of the dentin organic during the in vitro caries processes. Raman microscopy was performed to identify change in the collagen matrix of dentine. RESULTS: The red fluorescence observed in carious dentine using a Soprolife® camera corresponds to the brownish color observed using daylight. Demineralization using nitric acid induces a loss of the green fluorescence of dentine. The red fluorescence of carious dentine is resistant to acid treatment. Immersion of demineralized dentine in MGO induces a change of color from white to orange-red. This indicates that the Maillard reaction contributes to lesion coloration. Immersion of demineralized dentine in an MMP-1 solution followed by MGO treatment results in a similar red fluorescence. Raman microspectroscopy analysis reveals accumulation of AGE's product in red-colored dentine. CONCLUSIONS: Our results provide important information on the origin of the fluorescence variation of dentine observed with the Soprolife® camera. We demonstrate that the red fluorescence of carious dentine is linked to the accumulation of Advanced Glycation End products (AGE). CLINICAL RELEVANCE: The study provides a new biological basis for the red fluorescence of carious dentine and reinforces the importance of the Soprolife® camera in caries diagnostics.

Multiphoton imaging of the dentine-enamel junction.

Multiphoton microscopy has been used to reveal structural details of dentine and enamel at the dentin-enamel junction (DEJ) based on their 2-photon excited fluorescence (2PEF) emission and second harmonic generation (SHG). In dentine tubule 2PEF intensity varies due to protein content variation. Intertubular dentin produces both SHG and 2PEF signals. Tubules are surrounded by a thin circular zone with a lower SHG signal than the bulk dentine and the presence of collagen fibers perpendicular to the tubule longitudinal axis is indicated by strong SHG responses. The DEJ appears as a low intensity line on the 2PEF images and this was never previously reported. The SHG signal is completely absent for enamel and aprismatic enamel shows a homogeneous low 2PEF signal contrary to prismatic enamel. The SHG intensity of mantle dentine is increasing from the dentine-enamel junction in the first 12 µm indicating a progressive presence of fibrillar collagen and corresponding to the more external part of mantle dentine where matrix metallo-proteases accumulate. The high information content of multiphoton images confirms the huge potential of this method to investigate tooth structures in physiological and pathological conditions.

Assessment of leakage at the implant-abutment connection using a new gas flow method.

PURPOSE: The aim of this study was to evaluate, with a new gas flow technique, leakage at the implant/abutment junction in systems with four different connections. MATERIALS AND METHODS: Five Branemark System, five One Morse, five Intra-lock System, and five Ankylos Plus implants and abutments were used. A hole was drilled in the apex of each implant to allow gas to flow through the connection from negative to atmospheric pressure. The gas flow was calculated (slope of pressure decrease, in hPa.s-1). Each connection was tested after both manual and key tightening. Statistical analysis was performed on a generalized linear model with repeated measurements. The significance level was set at α=.05. RESULTS: A global significant difference was observed between the various systems (P=.0001). After manual tightening, gas leakage was (Ln[hPa.s-1], means±standard deviations): One Morse: 0.20 (±1.70); Branemark System: -4.56 (±2.61); Intra-lock: -4.31 (±4.17); Ankylos Plus; -7.59 (±0.76). After key tightening, mean values were: One Morse: -2.51 (±2.72); Branemark System: -7.23 (±1.01); Intra-lock: -7.76 (±0.50); Ankylos Plus; -7.73 (±0.62). CONCLUSION: This study confirms that gas flow is an appropriate method to assess connection leakage. Ankylos Plus connection leakage was very low when the assembly was tightened manually. Among conical connection systems, low (Ankylos Plus) and high (One Morse) leakage was observed. This gas flow study suggests, therefore, that connection design is not the most important parameter for implant/abutment connection leakage.

SVSVGMKPSPRP: a broad range adhesion peptide.

BACKGROUND: A combinatorial phage display approach was previously used to evolve a 12-mer peptide (SVSVGMKPSPRP) with the highest affinity for different semiconductor surfaces. The discovery of the multiple occurrences of the SVSVGMKPSPRP sequence in an all-against-all basic local alignment search tool search of PepBank sequences was unexpected, and a Google search using the peptide sequence recovered 58 results concerning 12 patents and 16 scientific publications. The number of patent and articles indicates that the peptide is perhaps a broad range adhesion peptide. METHODS: To evaluate peptide properties, we conducted a study to investigate peptide adhesion on different inorganic substrates by mass spectrometry and atomic force microscopy for gold, carbon nanotubes, cobalt, chrome alloy, titanium, and titanium alloy substrates. RESULTS: Our results showed that the peptide has a great potential as a linker to functionalize metallic surfaces if specificity is not a key factor. This peptide is not specific to a particular metal surface, but it is a good linker for the functionalization of a wide range of metallic materials. CONCLUSION: The fact that this peptide has the potential to adsorb on a large set of inorganic surfaces suggests novel promising directions for further investigation. Affinity determination of SVSVGMKPSPRP peptide would be an important issue for eventual commercial uses.

Perceived sources of stress in fifth year dental students--a comparative study.

INTRODUCTION: The term "stress" refers to the effect of certain external factors (physical or mental) on the individual's physical and psychological health. Stress is very high in dental medicine. It may well be generated by the process of training in dentistry. During their training dental students express high levels of stress and considerable stress-related symptoms such as physical problems, depression, obsessive-compulsive and personality disorders. The perceived stress can be dependent on socio-cultural factors. The type of personality, gender, emotional intelligence and other individual characteristics may also influence the stress effects. AIM: To determine the main sources of stress in students during their dental training and compare the level of stress fifth-year dental students experience in the Faculty of Dental Medicine, Plovdiv, Bulgaria and the Faculty of Dentistry in Montpellier, France. METHODS: We recruited in the study fifth-year students from the Faculty of Dental Medicine in Plovdiv, Bulgaria and the Faculty of Dentistry in Montpellier, France. The modified Dental Environment Stress (DES) questionnaire was used to assess the effect of the various stressors. The questionnaire survey was conducted between February and March 2008. RESULTS: The living arrangements domain indicates higher levels of stress in Bulgarian students compared to their French counterparts with significant statistical difference for all factors. In personal factors the highest levels of stress were found with respect to the reduced holidays during school year and financial problems. Exams were the highest stressor in academic work domain and for the survey as a whole. In educational environment factors the stressors with the highest significance were the interactions with patients, academic environment (for French students) and risk of cross transmitted infections (for Bulgarian students). The high stress during clinical work is related to completing the assigned clinical workload quota and the shortage of time for clinical work. CONCLUSIONS: The students from both faculties report the highest mean levels of stress are related to examinations and clinical factors. Bulgarian students experienced higher mean levels of stress compared to their French colleges. The differences in mean stress levels are associated with socio-cultural factors and variations in educational curriculums at both dental faculties.

[Dental pulp stem cells: characteristics, cryopreservation and therapeutic potentialities]. / Les cellules souches de la pulpe dentaire : caractéristiques, cryopréservation et potentialités thérapeutiques.

Stem cells discovery and their potential have led to the emergence of new forms of therapy with the development of bio-engineering cell and tissue methods underlying future medicine. The availability of stem cells and their preservation thus become an issue for everyone's health. Among the different sources of stem cells, those in the dental pulp have the advantage of being pluripotent, they can be cryopreserved and stored for long periods without losing their multiplication and differentiation capacities and finally they are easily accessible. The wisdom or natal teeth extracted for medical reasons are an opportunity for everyone to preserve stem cells for an autologous use. Biobanks authorized and specialized in the preparation and storage of pulp stem cells provide access to autologous regenerative medicine of tomorrow.

Molecular structural analysis of carious lesions using micro-Raman spectroscopy.

In clinical situations carious dentine tissues can be discriminated by most caries fluorescence detection tools, including a new fluorescence intra-oral camera. The objectives of this study were: (i) to analyze the Raman spectra of sound, carious, and demineralized dentine, (ii) to compare this spectral analysis with the fluorescence variation observed when using a fluorescence camera, and (iii) to evaluate the involvement of the Maillard reaction in the fluorescence variations. The first positive hypothesis tested was that the fluorescence of carious dentine obtained using a fluorescence camera and the Raman spectra variation were closely related. The second was that the variation of fluorescence could be linked with the Maillard reaction. Sound dentine, sound dentine demineralized in aqueous nitric acid solution, carious soft dentine, sound dentine demineralized in lactic acid solution, sound dentine demineralized in aqueous nitric acid solution and immersed in methylglycoxal solution, and sound dentine demineralized in aqueous nitric acid solution and immersed in methylglycoxal and glucose solutions, were studied using micro-Raman spectroscopy. Modifications in the band ratio of amide, phosphate, and carbonate were observed in the decayed and demineralized groups compared with the sound dentine group. The results indicate that a close relationship exists between the Maillard reaction and fluorescence variation.

Development of a new quantitative gas permeability method for dental implant-abutment connection tightness assessment.

BACKGROUND: Most dental implant systems are presently made of two pieces: the implant itself and the abutment. The connection tightness between those two pieces is a key point to prevent bacterial proliferation, tissue inflammation and bone loss. The leak has been previously estimated by microbial, color tracer and endotoxin percolation. METHODS: A new nitrogen flow technique was developed for implant-abutment connection leakage measurement, adapted from a recent, sensitive, reproducible and quantitative method used to assess endodontic sealing. RESULTS: The results show very significant differences between various sealing and screwing conditions. The remaining flow was lower after key screwing compared to hand screwing (p = 0.03) and remained different from the negative test (p = 0.0004). The method reproducibility was very good, with a coefficient of variation of 1.29%. CONCLUSIONS: Therefore, the presented new gas flow method appears to be a simple and robust method to compare different implant systems. It allows successive measures without disconnecting the abutment from the implant and should in particular be used to assess the behavior of the connection before and after mechanical stress.

Modeling colorant leakage techniques: application to endodontics.

OBJECTIVES: Our aim was to improve the comprehension of in vitro tracer leakage studies and to determine in which conditions such studies can be reliable. We aimed to develop different theoretical models to describe either an initially dry or a wet interface (slit) between sealer and dentin. METHODOLOGY: Equations based on physical laws were derived to model theoretically in vitro tracer penetration. For the dry interfaces, atmospheric, hydrostatic, tracer gravimetric, capillary and internal air pressures were considered as the underlying forces that control tracer penetration. For wet interfaces, the laws of diffusion were used to model colorant penetration. RESULTS: In both cases penetration is influenced by the width of the interface and by the size of the colorant. Calculations for dry conditions have shown that penetration is quick, mainly driven by the capillary pressure, and the penetration increases as the width of the interface diminishes. Dentinal tubules and the extent of their interconnection modify the penetration depth. For wet conditions, tracer size is the main factor controlling the penetration length and speed (the bigger the tracer, the slower the penetration). SIGNIFICANCE: Our model calculations demonstrate that tracer penetration studies have to be performed under strict experimental conditions. Dry and wet interfaces are two extreme cases with very different tracer penetration modes. In vitro colorant penetration tests should be performed in both of these conditions avoiding cases where the slit contains both air and water. Theses models can be adapted to other dental situations as well.

Gas permeability: a new quantitative method to assess endodontic leakage.

AIM: The aim of this study was to evaluate a new endodontic leakage measurement method. MATERIALS AND METHODS: Permeability was assessed measuring the gas flow passing through the root. Positive and negative tests were carried out to assess the validity of the method. We used glass capillaries for calibration (diameters of 15, 30, 40, 50 and 75 microm). The applicability of the method was assessed with human teeth using three sealing methods: GuttaFlow (GF) and a single cone; Pulp Canal Sealer (PCS) and a single cone; PCS and system B. RESULTS: This method demonstrated to be highly reproducible as the standard deviation was approximately 1% on average with glass capillaries. Significantly higher leakage (p<0.05) was recorded for endodontic treatment with GF and single cone compared to PCS and single cone and PCS with system B. CONCLUSION: Gas permeability is quantitative, sensitive, non-destructive and reproducible and seems appropriate for endodontic tests. It would participate in the indirect comprehension of leakage phenomena.