The effects of ageing on the biomechanical properties of root dentine and fracture.

OBJECTIVES: Knowledge of the mechanical behaviour of root dentine can facilitate better understanding of spontaneous vertical root fracture (VRF), an age-related disease initiated mainly at the root apex. We tested the hypothesis that the biomechanical properties of root dentine change with ageing. METHODS: Sixteen human premolars were divided into "old" (17-30 years) and "young" (50-80 years) groups. The elastic modulus, nano-hardness, micro-hardness, elemental contents, tubular density/area of root dentine in cervical, middle and apical root regions were evaluated using atomic force microscopy-based nano-indentation, Knoop indentation, scanning electron microscopy and energy dispersive X-ray spectroscopy, respectively. RESULTS: The apical dentine showed a lower nano-hardness, a lower elastic modulus, a lower calcium content, a lower calcium-to-phosphorus ratio and a smaller tubular density/area than the cervical dentine in both age groups, whereas spatial differences in micro-hardness were observed only in old roots. Compared with young dentine, old dentine showed a greater hardness, a higher elastic modulus, a greater mineral content and a smaller tubular size in the cervical portion, whereas the age-induced changes in tubular density were insignificant. Finite element analysis revealed that due to its higher elastic modulus, old apical dentine has a higher stress level than young dentine. CONCLUSIONS: The intrinsic material properties of root dentine have spatial variations, and they are altered by ageing. The higher stress level in old apical dentine may be one reason, if not the most important one, why spontaneous VRFs are more likely to occur in the elderly population.

Dental caries and chemical analyses in reconstruction of diet, health and hygienic behaviour in the Middle Euphrates valley (Syria).

OBJECTIVES: The aim of this study was to use two methods, biological and chemical, to examine changes in diet and health in individuals from the Middle Euphrates valley (Syria). We determined the frequency distribution of dental caries. Chemical analyses were concerned with the presence of elements such as strontium, barium, calcium and stable carbon isotopes ((13)C/(14)C). We chose three consecutive periods: Late Roman (2nd-4th century AD), Islamic (600-1200 AD) and Modern Islamic (1850-1950 AD). MATERIALS AND METHODS: We analysed the dental remains of 145 individuals, with a total of 2530 teeth. We used visual research (magnifying glass/sharp dental probe) and radiography. The frequencies of caries were calculated on the basis of the proportional correction factor of Erdal and Duyar. We chose 39 permanent second molars for chemical analyses. RESULTS: The frequency of carious lesions was similar in all three periods (6-8%). In the Modern Islamic and Islamic periods, occlusal surfaces were infected with caries most often, while the cemento-enamel junction (CEJ) and approximal surfaces were affected to a lesser degree. However, in the Late Roman period, the CEJ and approximal surfaces showed caries most frequently, in contrast to occlusal surfaces, which seldom showed signs of caries. Chemical analyses showed lower Sr/Ca ratios and Observed Ratio index values for the Modern Islamic and higher values for the Islamic and Late Roman periods. Mean stable isotope (δ(13)C) analyses demonstrated that the Modern Islamic period was strongly divergent from the other periods. DISCUSSION: These data suggest a similar socio-economic status during the Late Roman and Islamic periods. The diet of the population living in the Late Roman and Islamic periods contained a larger number of products containing strontium than calcium. In the modern population these proportions have been reversed.

Fracture behavior of human molars.

Despite the durability of human teeth, which are able to withstand repeated loading while maintaining form and function, they are still susceptible to fracture. We focus here on longitudinal fracture in molar teeth-channel-like cracks that run along the enamel sidewall of the tooth between the gum line (cemento-enamel junction-CEJ) and the occlusal surface. Such fractures can often be painful and necessitate costly restorative work. The following study describes fracture experiments made on molar teeth of humans in which the molars are placed under axial compressive load using a hard indenting plate in order to induce longitudinal cracks in the enamel. Observed damage modes include fractures originating in the occlusal region ('radial-median cracks') and fractures emanating from the margin of the enamel in the region of the CEJ ('margin cracks'), as well as 'spalling' of enamel (the linking of longitudinal cracks). The loading conditions that govern fracture behavior in enamel are reported and observations made of the evolution of fracture as the load is increased. Relatively low loads were required to induce observable crack initiation-approximately 100 N for radial-median cracks and 200 N for margin cracks-both of which are less than the reported maximum biting force on a single molar tooth of several hundred Newtons. Unstable crack growth was observed to take place soon after and occurred at loads lower than those calculated by the current fracture models. Multiple cracks were observed on a single cusp, their interactions influencing crack growth behavior. The majority of the teeth tested in this study were noted to exhibit margin cracks prior to compression testing, which were apparently formed during the functional lifetime of the tooth. Such teeth were still able to withstand additional loading prior to catastrophic fracture, highlighting the remarkable damage containment capabilities of the natural tooth structure.

Chemical characterization of etched dentin in non-carious cervical lesions.

PURPOSE: Bonding to non-carious cervical lesion (NCCL) sclerotic dentin that involves acid etching continues to be a challenging problem due to its altered chemical structure. In the present study, the objective was to investigate the chemical response of NCCL sclerotic dentin to the different acid etching times. MATERIALS AND METHODS: Extracted human premolars affected with NCCLs were selected, and a cavity matching the natural lesion with respect to size and location was prepared on the lingual surface of each tooth to serve as the control. The dentin surfaces were treated for 15 s and 30 s using 37% phosphoric acid and then analyzed by Raman microspectroscopic mapping/imaging. RESULTS: NCCL dentin substrates had dramatic effects on the chemical profile of dentin demineralization. The spectral comparison showed that the demineralized layer generated by the acid treatment was highly irregular in terms of depth and mineral component retained, especially when NCCL sclerotic dentin was etched for 15 s. When the etching time was increased to 30 s, the demineralization of NCCL sclerotic dentin was more effective and comparable to the nonsclerotic control that was treated for 15 s. Different etching times affected the depth, degree, and profile of the dentin demineralization. CONCLUSION: The shorter etching time (ie, 15 s) might not be adequate for NCCL sclerotic dentin. However, the longer etching time (ie, 30 s) would induce much deeper demineralized dentin for nonsclerotic substrates. Thus, although extended etching times can be used to remove the hypermineralized layer, further studies are required to analyze the impact this might have on the dentin bonding.

Mineral density of hypomineralised enamel.

OBJECTIVES: To characterize molar-incisor hypomineralisation (MIH) defects of different severities quantitatively and qualitatively using X-ray microtomography (XMT) and to measure the range of reduction in mineral density (MD) of MIH enamel compared with the normal range. METHODS: Ten sound teeth and ten MIH teeth were scanned using a commercial XMT system. Four hydroxyapatite phantoms of different densities were used as calibration standards with each scan. A calibration equation derived from the phantoms with each tooth was used for MD calibration. MD was traced from the cementum-enamel junction (CEJ) to the cusp tip and from the dentine-enamel junction (DEJ) to the outer enamel surface. RESULTS: In sound teeth, MD increased from CEJ to cusp/incisal tip, while in MIH teeth MD dropped from the CEJ to the occlusal region, then increased again at the cusp tip. MD was highest midway between DEJ and outer enamel in sound teeth. In MIH, enamel showed normal thickness and MD was highest near the DEJ and then decreased towards the outer enamel. MD of MIH enamel was on average about 19% lower than sound enamel. The MIH defects seemed to follow the incremental lines of enamel formation. CONCLUSIONS: MIH defects are hypomineralised defects of different severities that follow the natural incremental lines of enamel formation. Cuspal areas are usually only mildly affected and cervical enamel always appears to be sound.

Structure, chemical composition and mechanical properties of coronal cementum in human deciduous molars.

OBJECTIVES: It was hypothesized that the coronal cementum containing collagen forms a weak junction with enamel unlike the well integrated DEJ and CDJ. METHODS: The hypothesis was investigated in two parts: (1) evaluate the structure, chemical composition and mechanical properties of coronal cementum and its junction with enamel using scanning electron microscopy, micro-X-ray computed tomography, and atomic force microscopy. The chemical composition and mechanical properties were determined by evaluating the spatial variations of inorganic (PO(4)(3-)nu(1) mode at 960 cm(-1)) and organic (C-H deformation at 1452 cm(-1); C-H stretch at 2940 cm(-1)) contents using Raman microspectroscopy and elastic modulus and hardness values using nanoindentation. (2) Estimate the strength and evaluate the microstructure of coronal cementum interface with enamel using SEM and MicroXCT. RESULTS AND CONCLUSIONS: Coronal cementum is heterogeneous because it is a combination of laminar acellular afibrillar cementum and acellular extrinsic fiber cementum with relatively higher organic content. It integrates micromechanically via a scallop-like weak interface with enamel unlike the biomechanically efficient DEJ and CDJ and is continuous with primary root cementum. A single tooth could exhibit all three types of cementum enamel junctions; an overlap, butt and a gap depending on the sectioning plane. The elastic modulus of coronal cementum (11.0+/-5.8 GPa) is significantly lower (p<0.05; Student's t-test with 95% confidence interval) than primary cementum (15.8+/-5.3 GPa).

Chemical profile of the dentin substrate in non-carious cervical lesions.

OBJECTIVE: The molecular structural nature of the dentin substrate in non-carious cervical lesions (NCCLs) is poorly understood. This investigation characterized the chemical structure including inhomogeneities, composition, mineral crystallinity, collagen organization of normal dentin and affected dentin substrates within NCCLs using Raman microspectroscopic mapping/imaging. MATERIALS AND METHODS: Three extracted human pre-molars affected with NCCLs were selected and cavities matching the natural lesion with respect to size and location were prepared on the lingual/palatal surface of each tooth to serve as controls. The specimens were sectioned to expose the gingival and occlusal margins of the NCCLs and the control cavities. Micro-Raman spectra and imaging were acquired at 1.5 microm spatial resolution at positions perpendicular to the lesion surfaces. RESULTS: The Raman spectra and imaging comparisons showed the distinct compositional and structural alterations in mineral and matrix components of NCCL affected dentin. A heterogeneous hyper-mineralized layer, with characteristic features such as high phosphate/low carbonate content, high degree of crystallinity and partially denatured collagen were revealed in affected dentin substrate of NCCLs. SIGNIFICANCE: Generating Raman images based on different strategies from the same data set provides a powerful means to study the structural alterations within heterogeneous dental tissues. Direct overlay of the images indicated that the changes in chemical structure and composition are synchronized. Further studies are required to understand the role that these alterations play in response to acid etching and bonding to these clinically relevant substrates.

Tooth morphology and characteristics of non-carious cervical lesions.

OBJECTIVES: Two 3-dimensional models, one of a lower second premolar and one of a lower central incisor were used to investigate effects of load on the location and magnitude of cervical strains. METHODS: Point loads of 100 N were applied to the model premolar (at the cusp tip, parallel and 45 degrees to the long axis of the tooth) and incisor (at the mid-buccal point on incisal edge, parallel and 45 degrees to the long axis of the tooth). Outputs were presented in the form of tensile-strain contours and vector plots. RESULTS: Strains were concentrated near the cementoenamel junction (CEJ) regardless of load direction, and oblique loading showed higher tensile strains, opposite to the point of loading, than vertical loading for both models. For the premolar model, regardless of loading direction, strains were concentrated at the mid- buccal CEJ. In contrast, under vertical loading on the incisor model, tensile strains were concentrated at the line-angle of CEJ. From oblique loads, vector plots of both models showed tensile vectors in vertical directions while vertical loads showed tensile vectors in horizontal directions. CONCLUSION: Mechanisms of non-carious cervical lesions (NCCLs) may in part be due the changing orientation of tensile strains as well as their magnitude. Stress concentration at the CEJ related well to the common location of clinical NCCLs. The crown-root morphology may have an influence on the initial location of non-carious cervical lesions.

The effect of adjacent dentine blocks on the demineralisation and remineralisation of enamel in vitro.

The aim was to investigate interactions between enamel and dentine at low pH under conditions simulating those at the enamel-dentine junction. Sound enamel blocks were demineralised in acid-gel systems, at pH 4.6, either in isolation, next to one, or in the middle of two, abutting dentine blocks. The gels were initially infinitely undersaturated with respect to enamel. In a second study, enamel blocks containing pre-formed lesions were placed in acid-gel systems, at pH 5.0, either in isolation or next to dentine blocks. The systems were initially either partially or infinitely undersaturated. In the partially saturated systems, calcium and phosphate concentrations were representative of plaque fluid. In the first study, demineralisation of enamel next to one dentine block was reduced in inverse proportion to the distance from the dentine. Demineralisation of enamel between two dentine blocks was retarded markedly across the whole block. In the second study, in the partially saturated systems, enamel lesions next to dentine blocks remineralised, whereas those in isolation demineralised further. We suggest that diffusion of dissolved dentine mineral over the enamel in the infinitely undersaturated system was sufficient to reduce undersaturation, thus retarding demineralisation, and that in the partially saturated systems, dentine dissolution together with the added calcium phosphate caused remineralisation of enamel lesions. Fluoride released from dissolving dentine may have augmented these effects. Different rates of demineralisation in enamel and dentine, or enamel remineralisation with concurrent dentine demineralisation, enabled by differences in their solubilities, could help explain the progression of so-called 'hidden caries'.

Raman spectroscopic study of noncarious cervical lesions.

The surface of noncarious cervical lesions (NCCLs) consists of sclerosed dentin. This type of dentin may affect the ability of adhesive restorative materials to bond well to its surface, but little information exists on the chemical nature of this dentin surface and how it may be affected during acidic treatment. The inorganic part of normal dentin and dentin from NCCLs before and after acid conditioning with phosphoric acid or polyacrylic acid was investigated. Ten premolars with NCCLs and four human third molars (control) were used. Replicas of NCCLs were examined using scanning electron microscopy (SEM). Surfaces and longitudinal sections of four NCCLs and control dentin discs were analyzed using Raman spectroscopy. The discs and NCCLs were sectioned, and treated with 35% phosphoric acid or 20% polyacrylic acid/3% aluminum chloride, and Raman spectra obtained. The area under phosphate nu1 of the dentin spectrum was computed to obtain a ratio with the area under the second-order spectrum of a silicon phonon comparative standard. Mean phosphate nu1 and silicon phonon ratios from normal dentin and NCCLs were compared using a linear model with repeated measurements and Tukey's pairwise tests. Mean ratios from different locations of the NCCLs were compared using one-way analysis of variance (ANOVA) and Tukey's pairwise tests. SEM micrographs of NCCL surfaces showed variation from relatively smooth with no dentinal tubule openings to surfaces with occluded tubules. The mean phosphate nu1 and silicon phonon ratios for NCCLs were higher than those of normal dentin in all treatment groups (P < 0.05). Ratios from the untreated specimens were higher than those of the polyacrylic acid-treated specimens, and those for the phosphoric acid-treated group were the lowest (P < 0.05). The ratios obtained for the surfaces of NCCLs were higher than those halfway towards the pulp, and those adjacent to the pulp were the lowest (P < 0.05).

Spectroscopic imaging of mineral maturation in bovine dentin.

Dentin is a useful model for the study of mineral maturation. Using Fourier Transform Infrared Imaging (FTIRI), we characterized distinct regions in developing dentin at 7- micro m spatial resolution. Mineral-to-matrix ratio and crystallinity in bovine dentin from cervical and incisal parts of 3rd-trimester fetal compared with one-year-old incisor crowns showed that virtually all maturation stages in dentin could be spectroscopically isolated and analyzed. In the fetal incisors, mantle and circumpulpal dentin presented distinct patterns of mineral maturation. Gradients in both mineral properties examined were observed at the mineralization front and at the dentino-enamel junction.

Effect of calcium hydroxide as a supplementary barrier in the radicular penetration of hydrogen peroxide during intracoronal bleaching in vitro.

AIM: To examine pH changes in the cervical external root surface, when calcium hydroxide was used as a supplementary barrier to the protective base material during intracoronal bleaching. METHODOLOGY: Twenty-eight single-rooted human premolars extracted for orthodontic reasons were instrumented with K-Flex files, obturated with gutta-percha and subjected to thermocatalytic bleaching. The teeth were divided into four groups. In group A, a glass-ionomer cement barrier was placed at the cemento-enamel junction (CEJ) level and in group C, the barrier was placed 1 mm apical to the CEJ. In groups B and D, Ca(OH)2 was placed in contact and apical to the glass-ionomer cement at the CEJ and 1 mm apical to the CEJ, respectively. The teeth were placed in vials containing distilled water and the pH values of the medium surrounding the teeth were recorded after 1, 2, 4, 10 and 15 days, following renewal of the medium. RESULTS: The pH in the medium became acidic in all groups. No statistically significant differences existed between groups for all the experimental days (P = 0.790). CONCLUSION: The placement of Ca(OH)2 as a supplementary barrier during intracoronal bleaching did not have a significant effect in reversing the acidic pH created at the external root surface in vitro. Its potential effect during these procedures in vivo needs to be further investigated.

A Raman spectroscopic investigation of dentin and enamel structures modified by lactic acid.

Confocal Raman microspectrometry allows a thorough molecular analysis of mineralised dental tissues. The output information is provided in the form of curves representing the intensity of the signal according to the frequency, and its mathematical exploitation permits all sorts of comparative and quantitative analyses. By this process, we investigated the in vitro action of lactic acid on enamel and dentin from human permanent teeth. Modifications due to the acidic attack essentially concern phosphate grouping PO(4)(3-), which represents the mineral phase in enamel and dentin (hydroxyapatite): on Raman spectra, changes in intensity of the PO(4)(3-) band are linked to the type of dentin, to its anatomical location, and to the age of the subject. The variability of the dentinal chemical structure was confirmed by a quantitative statistical analysis, revealing a significant spectral difference between coronal and root dentin.

Fluoride distribution in sound and carious root tissues of human teeth.

Proton probe analysis has been used to provide for the first time quantitative F concentration data in carious root tissues from subjects consuming water containing 1 ppm F. In small lesions at the neck of the tooth with minimal tissue loss the F concentration was significantly higher at the outer lesion edge than at the outer edge of adjacent sound root tissue. In one sample with high F values the lesion edge had 19,000 ppm F and the adjacent sound root surface 5,400 ppm F microg F/g apatite). In large lesions with extensive cavitation F was again concentrated in the outer edge of the lesion and was significantly higher (1,800-4,100 ppm) than in adjacent sound inner dentine (190-290 ppm). Fluoride concentrations varied markedly along the outer edge of both normal and carious root tissues. Fluoride increase at the lesion edge is not an effect of tissue shrinkage but probably a result of remineralisation events during caries. This additional F may be expected to increase tissue resistance to further acid attacks.

Immunodetection of enamel- and cementum-related (bone) proteins at the enamel-free area and cervical portion of the tooth in rat molars.

Enamel and dentin at the cervical portion of the tooth are frequently covered by a collagen-free matrix referred to as acellular afibrillar cementum (AAC). It is believed that AAC deposition occurs when the enamel organ is displaced or disrupted, and mesenchymal cells from the dental follicle gain access to the tooth surface, differentiate into cementoblasts, and secrete noncollagenous proteins typically found in collagen-based mineralized tissues. A similar thin layer of mineralized matrix is found at the enamel-free area (EFA) of rodent molars, but in this case the matrix is covered by inner enamel epithelium (IEE) throughout development. We have, therefore, used this site as a paradigm to test the hypothesis that typical mesenchymal matrix proteins can also be found in association with epithelial cells. To this end, we have analyzed the presence and distribution of enamel- and cementum-related matrix proteins at the EFA and at the cervical portion of the tooth. Rat mandibular molars were processed for colloidal gold immunolabeling with antibodies to amelogenins, bone sialoprotein (BSP), osteopontin (OPN), osteocalcin (OC), and dentin sialoprotein (DSP), and the plasma proteins alpha 2 HS-glycoprotein and albumin. The EFA matrix was immunoreactive for amelogenins as well as for BSP, OPN, OC, and alpha 2 HS-glycoprotein, but not for albumin and DSP. The AAC was, similar to the EFA matrix, labeled for BSP, OPN, OC, and alpha 2 HS-glycoprotein. These data show for the first time that the EFA matrix is comprised of a mixture of enamel- and cementum-related proteins, a situation that parallels the distribution of matrix constituents at the cervical portion of the tooth. Since the EFA matrix is deposited on top of the mineralized dentin, and since the enamel organ seals off the forming matrix, it is concluded that EFA cells are responsible for the production of these proteins. Consistent with previous reports showing that epithelial cells can produce both BSP and OPN in some circumstances, the data also suggest that AAC may be deposited by cells of epithelial origin. Furthermore, they lend support to the possibility that cells derived from Hertwig's epithelial root sheath may likewise be capable of producing cementum matrix proteins.

Role of cementoenamel junction on the radicular penetration of 30% hydrogen peroxide during intracoronal bleaching in vitro.

Intracoronal bleaching of nonvital, teeth with 30% hydrogen peroxide is occasionally associated with external cervical root resorption. The exact mechanism by which bleaching induced root resorption occurs is not yet fully understood. The relationship of cementum to the enamel at the cementoenamel junction may have clinical significance. Seventeen single rooted human mandibular premolars extracted atraumatically for orthodontic reasons were used. The radicular hydrogen peroxide penetration in each tooth was measured in vitro by an indirect colorimetric method. Thereafter, the teeth were examined with a scanning electron microscope to determine the type of the cementoenamel junction. It was found that the radicular penetration of 30% hydrogen peroxide was related to the type of cementoenamel junction.