Type IV collagen is a novel DEJ biomarker that is reduced by radiotherapy.

The dental basement membrane (BM) is composed of collagen types IV, VI, VII, and XVII, fibronectin, and laminin and plays an inductive role in epithelial-mesenchymal interactions during tooth development. The BM is degraded and removed during later-stage tooth morphogenesis; however, its original position defines the location of the dentin-enamel junction (DEJ) in mature teeth. We recently demonstrated that type VII collagen is a novel component of the inner enamel organic matrix layer contiguous with the DEJ. Since it is frequently co-expressed with and forms functional complexes with type VII collagen, we hypothesized that type IV collagen should also be localized to the DEJ in mature human teeth. To identify collagen IV, we first evaluated defect-free erupted teeth from various donors. To investigate a possible stabilizing role, we also evaluated extracted teeth exposed to high-dose radiotherapy--teeth that manifest post-radiotherapy DEJ instability. We now show that type IV collagen is a component within the morphological DEJ of posterior and anterior teeth from individuals aged 18 to 80 yr. Confocal microscopy revealed that immunostained type IV collagen was restricted to the 5- to 10-µm-wide optical DEJ, while collagenase treatment or previous in vivo tooth-level exposure to > 60 Gray irradiation severely reduced immunoreactivity. This assignment was confirmed by Western blotting with whole-tooth crown and enamel extracts. Without reduction, type IV collagen contained macromolecular α-chains of 225 and 250 kDa. Compositionally, our results identify type IV collagen as the first macromolecular biomarker of the morphological DEJ of mature teeth. Given its network structure and propensity to stabilize the dermal-epidermal junction, we propose that a collagen-IV-enriched DEJ may, in part, explain its well-known fracture toughness, crack propagation resistance, and stability. In contrast, loss of type IV collagen may represent a biochemical rationale for the DEJ instability observed following oral cancer radiotherapy.

Extracts of irradiated mature human tooth crowns contain MMP-20 protein and activity.

OBJECTIVES: We recently demonstrated a significant correlation between enamel delamination and tooth-level radiation dose in oral cancer patients. Since radiation can induce the synthesis and activation of matrix metalloproteinases, we hypothesized that irradiated teeth may contain active matrix metalloproteinases. MATERIALS AND METHODS: Extracted teeth from oral cancer patients treated with radiotherapy and from healthy subjects were compared. Extracted mature third molars from healthy subjects were irradiated in vitro and/or incubated for 0-6 months at 37°C. All teeth were then pulverized, extracted, and extracts subjected to proteomic and enzymatic analyses. RESULTS: Screening of irradiated crown extracts using mass spectrometry identified MMP-20 (enamelysin) which is expressed developmentally in dentine and enamel but believed to be removed prior to tooth eruption. MMP-20 was composed of catalytically active forms at Mr=43, 41, 24 and 22kDa and was immunolocalized predominantly to the morphological dentine enamel junction. The proportion of different sized MMP-20 forms changed with incubation and irradiation. While the pattern was not altered directly by irradiation of healthy teeth with 70Gy, subsequent incubation at 37°C for 3-6 months with or without prior irradiation caused the proportion of Mr=24-22kDa MMP-20 bands to increase dramatically. Extracts of teeth from oral cancer patients who received >70Gy radiation also contained relatively more 24 and 22kDa MMP-20 than those of healthy age-related teeth. CONCLUSION: MMP-20 is a radiation-resistant component of mature tooth crowns enriched in the dentine-enamel. We speculate that MMP-20 catalyzed degradation of organic matrix at this site could lead to enamel delamination associated with oral cancer radiotherapy.

FTIR and SEM analysis of CO2 laser irradiated human enamel.

OBJECTIVES: Considering the enamel chemical structure, especially carbonate band, which has a major role in the caries prevention, the objective of the present study was to assess the chemical alterations on the enamel irradiated with CO(2) laser by means of FTIR spectroscopy and SEM analysis. DESIGN: The enamel surfaces were analysed on a spectrometer for acquisition of the absorption spectrum relative to the chemical composition of the control sample. The irradiation was conducted with a 10.6-µm CO(2) laser (0.55W, 660W/cm(2)). The carbonate absorption band at 1600-1291cm(-1) as well as the water absorption band at 3793-2652cm(-1) was measured in each sample after the irradiation. The water band was measured again 24-h after the irradiation. The band area of each chemical compound was delimited, the background was subtracted, and the area under each band was integrated. Each area was normalized by the phosphate band (1190-702cm(-1)). RESULTS: There was a statistically significant decrease (p<0.05) in the water content after irradiation (control: 0.184±0.04; irradiated: 0.078±0.026), which increased again after rehydration (0.145±0.038). The carbonate/phosphate ratio was measured initially (0.112±0.029) and its reduction after irradiation indicated the carbonate loss (0.088±0.014) (p<0.05). CONCLUSION: The 10.6-µm CO(2) laser irradiation diminishes the carbonate and water contents in the enamel after irradiation.

Impact of pelleted diets with different mineral compositions on the crown size of mandibular cheek teeth and mandibular relative density in degus (Octodon degus).

The impact of pelleted diets with different mineral compositions on the crown size of the mandibular cheek teeth, as well as the mandibular bone and cheek teeth density, in degus (Octodon degus) was investigated. A total of 28 animals were randomly divided into four groups and fed different calcium or phosphorous dietary content, in the presence or absence of UV light, for 14 months. Dental radiographs and CT images of the head were taken, and the crown size of premolar and molar teeth was recorded. Apical and coronal crown elongation of all cheek teeth and significantly smaller relative cheek teeth and mandibular densities were recorded in degus fed a high-phosphorus diet with an improper calcium:phosphorus ratio. A diet with a calcium:phosphorus ratio of 1:1 was also responsible for the rapid development of dental disease with subsequent severe health impairment.

Effects of chlorhexidine and fluoride on irradiated enamel and dentin.

The effectiveness of mouthwash protocols in preventing gamma irradiation therapy damage to the ultimate tensile strength (UTS) of enamel and dentin is unknown. It was hypothesized that the use of chlorhexidine and fluoride mouthwash would maintain the UTS of dental structures. One hundred and twenty teeth were divided into 2 groups: irradiated (subjected to 60 Gy of gamma irradiation in daily increments of 2 Gy) and non-irradiated. They were then subdivided into 2 mouthwash protocols used 3 times per day: 0.12% chlorhexidine, 0.05% sodium fluoride, and control group (n = 10). The specimens were evaluated by microtensile testing. The results of the Tukey test (p < 0.05) indicated that the gamma irradiation therapy significantly reduced the UTS of the enamel, crown, and root dentin. Macromolecular alterations were suggested by optical retardation data in dentin. Structural alterations, in both substrates, were detected by scanning electron microscopy analysis. Mouthwash with 0.12% chlorhexidine partially prevented the damage to the mechanical properties of the irradiated crown dentin, whereas the 0.05% sodium-fluoride-irradiated enamel showed UTS similar to that of non-irradiated enamel.

Fluctuating molar asymmetry in relation to environmental radioactivity.

OBJECTIVE: Fluctuating dental asymmetry is considered a sensitive indicator of environmental stress. We used fluctuating asymmetry of teeth in Greek children to assess the effect of the Chernobyl accident to the Greek population, which received relatively large radiation exposure compared to other European countries. DESIGN: Sixty dental casts were divided into two groups according to date of birth prior to or after the Chernobyl accident. The intercuspal distances of the mandibular first permanent molars were measured with digital calipers. We followed the data analysis procedure proposed by Palmer and Strobeck (2003). RESULTS: Fluctuating asymmetry was found at levels above measurement error in all traits except for the buccal measurement. Overall, there were no significant differences in the fluctuating asymmetry between the two groups, after removal of outliers. CONCLUSIONS: These results suggest that environmental radioactivity from the Chernobyl incident did not significantly affect the symmetry of lower permanent molar intercuspal distances in the specific population, even though increased levels of radionuclides have been reported in teeth and other tissues.

Relationship between laser fluorescence and enamel hypomineralisation.

OBJECTIVES: To study the relation between the mechanical properties of hypomineralised enamel, and its laser fluorescence (LF). METHODS: Five extracted teeth with molar-incisor hypomineralisation (MIH) were sectioned longitudinally through the defects and polished to prepare the hypomineralised enamel for testing. Hardness (H) and elastic modulus (E) of enamel were measured using nanoindentation. Measurement recording started from the cervicoenamel junction and proceeded occlusally in increments of 200 microm. Laser fluorescence readings were taken along the same line and at the same sites using a DIAGNOdent pen. RESULTS: H, E, and LF readings from cervical enamel were within the expected range for normal sound enamel. After log transformation of the H and E measurements to allow for linear correlation analysis, there was a significant and moderately strong inverse correlation between LF and H or E samples (r (between specimens)=0.59, r (between specimens)=0.39, respectively; p<0.001). CONCLUSIONS: This study shows that, in the absence of dental caries, increased DIAGNOdent readings can indicate enamel hypomineralisation. While the increased LF readings in carious enamel are thought to be related to the presence of caries bacterial metabolites, the increased readings in hypomineralised enamel may be related to proteins in the hypomineralised enamel and/or light scattering by the inhomogeneous enamel.

Effect of sterilization by gamma radiation on nano-mechanical properties of teeth.

OBJECTIVES: Extracted teeth used in dental research need to be considered infective and hence be sterilized without the materials' properties being altered. This study examined the effect of gamma radiation on the nano-mechanical properties of dentin and enamel of extracted human third molars. METHODS: Whole teeth were sterilized using gamma radiation doses of 7 kGy and 35 kGy, respectively; teeth of the control group were not treated with gamma radiation. Crowns were sectioned occlusally and polished. Elastic modulus and hardness were tested using atomic force microscopy with nano-indentations under wet conditions. RESULTS: The authors found no significant dose-response relationship in elastic modulus or hardness in either dentin or enamel. SIGNIFICANCE: Nano-indentation is a common technique for the determination of local mechanical properties in biological hard tissues. Gamma radiation is an efficient way to sterilize extracted teeth while alteration of dentin and enamel mechanical properties are minimized.

Hard tissue ablation with a spray-assisted mid-IR laser.

The objective of this study was to understand the dominant mechanism(s) for dental enamel ablation with the application of water spray. A free-running Er,Cr:YSGG (yttrium, scandium, gallium, garnet) laser was used to ablate human enamel tissue at various radiant exposures. During dental ablation, distilled water was sprayed on the sample surface, and these results were compared to ablation without a spray (dry ablation). In order to identify dominant ablation mechanisms, transient acoustic waves were compared to ablation thresholds and the volume of material removed. The ablation profile and depth were measured using optical coherence tomography (OCT). Irregular surface modification, charring and peripheral cracks were associated with dry ablation, whereas craters for spray samples were relatively clean without thermal damage. In spite of a 60% higher ablation threshold for spray associated irradiations owing to water absorption, acoustic peak pressures were six times higher and ablation volume was up to a factor of 2 larger compared to dry ablation. The enhanced pressure and ablation performance of the spray-assisted process was the result of rapid water vaporization, material ejection with recoil stress, interstitial water explosion and possibly liquid-jet formation. With water cooling and abrasive/disruptive mechanical effects, the spray ablation can be a safe and efficient modality for dental treatment.

Disturbed root development of permanent teeth after pediatric stem cell transplantation. Dental root development after SCT.

BACKGROUND: Deficient dental root development has been reported after conventional pediatric anticancer therapy, but less information is available on stem cell transplantation (SCT) recipients. METHODS: Root-crown (R/C) ratios of fully developed permanent teeth were assessed from panoramic radiographs of 52 SCT recipients, who were treated when they were age < 10 years. Using standard deviation scores (SDSs), the authors compared the R/C ratios to the corresponding tooth and gender-specific values in a healthy population. The percentage of affected R/C ratios per individual was examined in a subgroup of 39 (SG39) patients with advanced tooth development. The effects of total body irradiation (TBI) and SCT age on the R/C ratios were studied in TBI and high-dose chemotherapy (HDC = non-TBI) groups and in 3 age groups (< or = 3.0 years, 3.1-5.0 years, > or = 5.1 years). RESULTS: Per individual, 77% of the fully developed permanent teeth were affected in SG39. At the tooth level, in 77% of the 945 teeth studied (52 patients), the R/C ratios were outside +/-2 SDSs. More teeth were affected in the TBI (85%) than in the non-TBI (55%) group (P < 0.001). The teeth of the patients who were ages 3.1-5.0 years old at SCT presented with the most severe aberrations of the R/C ratio (mean SDS = -4.4) whereas the teeth of the youngest (age < or = 3.0 years) and the oldest (age > or = 5.1 years) patients were equally affected (mean SDSs = -3.1 and -3.0, respectively). CONCLUSIONS: Disturbances of dental root growth always followed pediatric SCT. HDC alone intensely harmed root growth but TBI further increased the adverse effects that were most extensive in the patients 3.1-5.0 years at SCT. These sequelae should be taken into account during the lifelong dental follow-up to minimize the clinical consequences of dental injuries.

An in vitro L-band electron paramagnetic resonance study of highly irradiated whole teeth.

Regarding in vivo L-band dosimetry with human teeth, a number of preliminary experiments were carried out that were linked to the resonators response and the relative contribution of enamel to the EPR signal intensity of irradiated whole teeth. The sensitivity of the extended loop resonator varies in the antenna plane, but this variation tends to vanish when the sample is moved away from this plane. When the loop antenna is placed just above the highly irradiated molar, around 88% of the dosimetric signal is due to the crown enamel. The sensitivity inside a birdcage cavity is approximately equal over the volume of a molar; only 30% of the molar's total dosimetric signal results from enamel. Some decrease in the intensity of the dosimetric signal from enamel is observed after irradiation. At room temperature, the signal is reduced by about 20% within 90 days and approaches a plateau with a time constant of about 35 days.

Thin layer alpha-Al2O3:C beta dosemeters for the assessment of current dose rate in teeth due to 90Sr intake, and comparison with electron paramagnetic resonance dosimetry.

The use of thin-layer alpha-Al2O3:C thermoluminescent detectors (TLDs) for the assessment of current beta dose rate in human teeth due to 90Sr intake is investigated. The teeth used in this study were collected from members of the Techa river population who were exposed to radiation as a result of releases of the Mayak plutonium production facilities (Southern Urals-Russia) between 1949 and 1956. The beta dose rates from different parts of the tooth (enamel, crown dentine, and root) were determined by storing the detectors over the samples in a shielded environment. The cumulative dose measured by electron paramagnetic resonance (EPR) in different dental tissues is found to be proportional to current dose rate obtained from alpha-Al2O3:C thermoluminescence dosemeters. The retention of 90Sr in various parts of the teeth is discussed.

Using ultrasound transmission velocity to analyse the mechanical properties of teeth after in vitro, in situ, and in vivo irradiation.

Ultrasound transmission velocity (UTV) in isotropic material as a measure for the modulus of elasticity was correlated to mechanical properties. Changes in micromechanical properties of radiated teeth and influence of the oral cavity were to be evaluated nondestructively. UTV was measured in extracted teeth after 36 Gy and 62 Gy of in situ (enorally, with no contact to the oral cavity) and in vitro irradiation. Relative to controls, teeth subjected to 62 Gy in vivo showed higher UTV values for dentine and enamel. Sound teeth irradiated with 60 Gy in situ also showed higher UTV values for enamel, whereas dentine values were not significantly different from those of control. The mechanical properties of teeth irradiated in vitro were affected only after high experimental doses of up to 500 Gy. The difference between in vivo and in vitro mechanical properties may be due to radioxerostomia-induced damages as well as the status of dentine vitality. This supports the concept of direct radiation-induced damage in synergy with radioxerostomia-induced caries.

Impaired dentofacial development after radiotherapy of a non-Hodgkin lymphoma: report of case.

Since the advances in therapy of childhood malignancies have improved life expectancy attention is now increasingly focused on the long-term effects of antineoplastic therapy. Developmental abnormalities due to antineoplastic therapy have been claimed to preferentially occur in children treated before the age of six years. This report of a case demonstrates severe developmental disturbances following radiotherapy of a cervical non-Hodgkin lymphoma at the age of eight years. The morphological changes included microdontia, root shortening, blunting and thinning as well as mandibular hypoplasia.

Effects of Nd: and Ho:yttrium-aluminium-garnet lasers on human dentine fluid flow and dental pulp-chamber temperature in vitro.

Dentine specimens were prepared from freshly extracted third molars and initial permeability measured. Each specimen was subjected to Nd:yttrium-aluminium-garnet (YAG) (1.06 or 1.32 microns wavelength) or Ho:YAG (2.10 microns wavelength) laser energy while temperatures in the pulp chambers were recorded. Permeability was again measured and the surfaces examined by scanning electron microscopy. Six crown segments were used for each laser variable and eight permeability measurements were taken before and eight after laser exposure, while temperature was recorded during treatment. All wavelengths reduced permeability but temperature rises were high enough to have caused pulpal damage, indicating that shorter treatment times and lower power settings may be necessary if used in vivo.