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Objective: To evaluate the effect of four different photoactivation protocols (according to "photoactivated faces" - mesial/distal, cervical/incisal or center - and "photoactivation time" - 6-3 s) of a high-power photo activator (Valo Cordless ® -Ultradent) on the shear bond strength (SBS) between metal brackets and dental enamel and on the degree of conversion (DC) of an orthodontic resin., Materials and Methods: 40 bovine incisor crowns were randomly assigned to 4 groups (n = 10). The brackets were bonded with Transbond XT ® resin using 4 protocols according to the "photoactivation protocol" factor (which was subdivided into photoactivated faces and photoactivation time): V3C = 3 s + center; V6C = 6 s + center; V3M3D = 3 s on mesial + 3 s on distal; V3C3I = 3 s on cervical + 3 s on incisal. All the samples were stored for 4 months (water,37ºC) and then subjected to a SBS test (100KgF,1 mm/min). 40 resin discs were made to evaluate the monomer degree of conversion. Data from the SBS and DC were assessed by One-way ANOVA and Tukey's test (5%). Bond failures were analyzed according to the Adhesive Remnant Index (ARI) and evaluated by the Kruskal-Wallis test (5%)., Results: There was a statistically significant difference (p = 0.008) in the One-way ANOVA result for SBS values between all groups, but the protocols showed statistically similar results (p ≥ 0.05-Tukey's tests) concerning the photoactivated faces (V6C, V3M3D and V3C3I) and photoactivation time (V3C and V6C) factors individually. There was no statistically significant difference (p ≥ 0.05) in the One-way ANOVA result for DC values., Conclusion: The SBS and DC values will vary depending on the protocol applied., Clinical Relevance: It is possible to maintain the bracket fixation quality with the use of a high-power LED photo activator associated with a shorter photoactivation time. However, it is assumed that not all types of protocols that might be applied will provide quality bonding, such as V3C, V3M3D and V3C3I, which may - depending on the SBS and DC values - affect the final treatment time, due to brackets debonding, or increase of possibility of damage to dental enamel during bracket removal. Clinical studies are suggested to confirm the hypotheses of this research., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

This study aimed to assess the in vitro effects of re-irradiation on enamel and dentin properties, simulating head and neck cancer radiotherapy retreatment. Forty-five human permanent molars were classified into five groups: non-irradiated; irradiated 60 Gy, and re-irradiated with doses of 30, 40, and 50 Gy. Raman spectroscopy, scanning electron microscopy (SEM), and energy dispersive x-ray spectroscopy (EDS) were employed for analysis. Raman spectroscopy assessed intensity, spectral area, and specific peaks comparatively. Statistical analysis involved Kolmogorov-Smirnov and One-Way ANOVA tests, with Tukey's post-test (significance level set at 5%). Significant changes in irradiated, non-irradiated, and re-irradiated enamel peaks were observed, including phosphate (438 nm), hydroxyapatite (582 nm), phosphate (960 nm), and carbonate (1070 nm) (p < 0.05). Re-irradiation affected the entire tooth (p > 0.05), leading to interprismatic region degradation, enamel prism destruction, and hydroxyapatite crystal damage. Dentin exhibited tubule obliteration, crack formation, and progressive collagen fiber fragmentation. EDX revealed increased oxygen percentage and decreased phosphorus and calcium post-reirradiation. It is concluded that chemical and morphological changes in irradiated permanent teeth were dose-dependent, exacerbated by re-irradiation, causing substantial damage in enamel and dentin., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Objectives: This in vitro study evaluated the effect of different colouring solutions and primer systems used in the bonding of brackets on enamel colour change and bond strength., Materials and Methods: 120 premolar teeth were divided into four main groups; brackets were bonded with 37% orthophosphoric acid + Transbond XT Primer in Group 1, 3 M Single Bond Universal in Group 2, Transbond Plus SEP in Group 3, and G-Premio Bond in Group 4. Each group was divided into three subgroups, and the teeth were placed in a cup containing coffee and tea mixture, in a cup containing cola and in distilled water. A bond strength test was applied to all teeth. Colour measurements of all teeth were performed at 2 different times: before bonding and after the bond strength test., Results: The average bond strength of the 37% orthophosphoric acid group was higher than that of the other groups. The effect of primer and solution groups on colour change was statistically significant (p = 0.001 and p = 0.023, respectively)., Conclusions: In this study, the bond strength was clinically sufficient in all primer groups. The highest colour change was observed when the tea-coffee solution and Transbond Plus SEP primer were used., Clinical Relevance: This study has identified enamel discoloration and bond strength from different colouring solutions and primer systems used for bonding braces, which can be used to inform clinicians and patients to achieve better treatment results., (© 2024. The Author(s).)

Objectives: The current research on single-nucleotide polymorphism (SNP) mutation sites at different positions of the FAM83H gene and their phenotypic changes leading to amelogenesis imperfecta (AI) is inconsistent. We identified a previously reported heterozygous nonsense mutation c.1192C>T (p.Q398*) in the FAM83H gene and conducted a comprehensive analysis of the dental ultrastructure and chemical composition changes induced by this mutation. Additionally, we predicted the protein feature affected by this mutation site. The aim was to further deepen our understanding of the diversity of AI caused by different mutation sites in the FAM83H gene., Methods: Whole-exome sequencing (WES) and Sanger sequencing were used to confirm the mutation sites. Physical features of the patient's teeth were investigated using various methods including cone beam computer tomography (CBCT), scanning electron microscopy (SEM), contact profilometry (roughness measurement), and a nanomechanical tester (nanoindentation measurement). The protein features of wild-type and mutant FAM83H were predicted using bioinformatics methods., Results: One previously discovered FAM83H heterozygous nonsense mutation c.1192C>T (p.Q398*) was detected in the patient. SEM revealed inconsistent dentinal tubules, and EDS showed that calcium and phosphorus were lower in the patient's dentin but higher in the enamel compared to the control tooth. Roughness measurements showed that AI patients' teeth had rougher occlusal surfaces than those of the control tooth. Nanoindentation measurements showed that the enamel and dentin hardness values of the AI patients' teeth were both significantly reduced compared to those of the control tooth. Compared to the wild-type FAM83H protein, the mutant FAM83H protein shows alterations in stability, hydrophobicity, secondary structure, and tertiary structure. These changes could underlie functional differences and AI phenotype variations caused by this mutation site., Conclusions: This study expands the understanding of the effects of FAM83H mutations on tooth structure., Clinical Relevance: Our study enhances our understanding of the genetic basis of AI and may contribute to improved diagnostics and personalized treatment strategies for patients with FAM83H-related AI., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Objectives: The aim of this in vitro study was to evaluate the role of highly fluoridated dentifrice on remineralization characteristics of lowly and highly pre-demineralized enamel artificial caries lesions., Methods: Bovine enamel specimens were prepared (pH 4.95; 21 days) and discriminated in either lowly [L] or highly [H] pre-demineralized artificial caries lesions. Specimens with a mean ΔZ baseline,L (95% CI) of 5120 (4995; 5245) vol.% × μm and a mean ΔZ baseline,H of 8187 (8036; 8339) vol.% × μm were selected and randomly allocated to 12 groups (n = 20). Treatments during pH-cycling (28 days; 6 × 60 min demineralization/day) were brushing 2×/day with fluoride-free (0 ppm F - [L 0 /H 0 ]), 1100 ppm F - [L 1100 /H 1100 ], 2800 ppm F - [L 2800 /H 2800 ], 5000 ppm F - [L 5000 /H 5000 ], 5000 ppm F -  + glycerin [L 5000 + glycerin /H 5000 + glycerin ], and 5000 ppm F -  + TCP [L 5000 + TCP /H 5000 + TCP ] containing dentifrices. Dentifrice slurries were prepared with deionized water (1:3wt/wt). After cycling specimens presenting lesion surface loss were discarded and for the remaining 202 specimens, transversal microradiographic (TMR) analyses (ΔZ pH-cycle /LD pH-cycle ) were performed again. Changes in mineral loss (ΔΔZ = ΔZ baseline - ΔZ pH-cycle ) and lesion depth (ΔLD = LD baseline - LD pH-cycle ) were calculated., Results: Significant differences for ΔΔZ could be found between L 0 , L 1100 , and L 5000 as well as H 0 , H 1100 , and H 2800 /H 5000 (p ≤ 0.01; ANCOVA). Except for 0 ppm F - , higher ΔΔZ could be found in highly compared with lowly demineralized specimens (p ≤ 0.004; ANCOVA). After pH-cycling, a second lesion front could only be observed in H 5000 and H 5000 + TCP . The correlation between ΔΔZ and F - was moderate for lowly and highly demineralized lesions (r L  = 0.591; p L  < 0.001; r H  = 0.746; p H  < 0.001), indicating a fluoride dose response for both., Conclusion: For both baseline substrate conditions, a dose response for fluoride could be revealed., Clinical Significance: Remineralization characteristics of enamel directly depended on baseline mineral loss.

Objectives: The purpose of this study was to examine the enamel bond durability of universal adhesives in the self-etch mode under 2-year water storage and thermal cycling conditions., Materials and Methods: Three commercially available universal adhesives and a gold standard two-step self-etch adhesive were used. Ten specimens of bovine enamel were prepared per test group, and shear bond strength (SBS) was measured to determine the bonding durability after thermal cycling (TC) or long-term water storage (WS). The bonded specimens were divided into three groups: (1) specimens subjected to TC, where the bonded specimens were stored in 37 °C distilled water for 24 h before being subjected to 3000, 10,000, 20,000 or 30,000 TC; (2) specimens stored in 37 °C distilled water for 3 months, 6 months, 1 year or 2 year; and (3) specimens stored in 37 °C distilled water for 24 h, serving as a baseline., Results: The two-step self-etch adhesive showed significantly higher SBS than the universal adhesives tested, regardless of the type of degradation method. All universal adhesives showed no significant enamel SBS reductions in TC and WS, when compared to baseline and the other degradation conditions., Conclusions: Compared to the bond strengths obtained with the two-step self-etch adhesive, significantly lower bond strengths were obtained with universal adhesives. However, the enamel bond durability of universal adhesives was relatively stable under both degradation conditions tested., Clinical Relevance: The present data indicate that the enamel bond durability of universal adhesives in the self-etch mode might be sufficient for clinical use.

Objectives: The morphology and chemical composition of enamel submitted to different sterilization methods was studied., Methods: X-ray photoelectron spectroscopy (XPS), field emission gun scanning electron microscopy (FEG-SEM), and energy-dispersive X-ray spectroscopy (EDS) were performed to evaluate 50 bovine enamel specimens sterilized using four methods: microwaving (MI), gamma irradiation (GI), ethylene oxide (EO), and steam autoclave (SA). Non-sterilized specimens were used as control., Results: XPS indicated that the concentration of P (phosphorus), CO 3 (carbonate), and CO 3 /P was not changed in all groups. GI produced no significant change on elemental composition. SA produced the major decrease in calcium (Ca), Ca/P ratio, and increase in N (nitrogen). MI was found to decrease Ca, Ca/P ratio and O (oxygen), and increase in C (carbon) and N. EO produced decrease in Ca and O with increased C concentration. FEG-SEM revealed surface and in-depth morphological changes on SA specimens. Minor surface alterations were observed for EO and for MI groups, and no alteration was observed on GI group. EDS indicated no difference on elemental composition of enamel bulk among groups., Conclusions: SA produced mineral loss and morphological alterations on surface and in depth. MI and EO sterilization caused mineral loss showing only slight alteration on enamel surface. GI sterilization preserves the morphological characteristics of enamel. The sterilization methods could be classified from lower to high damage as GI < MI < EO < SA., Clinical Relevance: This is a comprehensive comparative study where different methods for enamel sterilization were investigated in terms of chemical changes. The results presented here may help researchers to choose the most appropriate method for their research setting and purpose.

Objectives: This study tested the effect of enamel salivary exposure time prior to an acid challenge (30 min, 1, 2, or 12 h) and type of intraoral appliance (palatal or mandibular) on initial erosion., Methods: After initial surface hardness evaluation, enamel blocks (n = 340) were randomly divided into groups and volunteers (n = 20). The control group was not exposed to saliva previously to the erosive challenge. The volunteers wore palatal and mandibular appliances simultaneously. After salivary exposure, the blocks were subjected to acid exposure by immersion in hydrochloric acid (0.01 M, pH 2.3) for 30 s. Then, the enamel surface hardness was evaluated. Data were analyzed using ANOVA, Kruskal-Wallis and Tukey's test (p < 0.05)., Results: No difference was observed on percent surface hardness change (% SHC) in the enamel blocks between the types of intraoral appliances. Exposure to saliva for 30 min and 1 h promoted similar enamel resistance to the erosive attack, which was similar to the control group for both appliances. Blocks exposed to saliva for 2 h showed less hardness loss when compared to 30 min. Keeping the blocks in saliva during 12-h overnight resulted in similar percentage of enamel hardness loss compared to 2 h., Conclusions: A 2-hour in situ exposure to saliva is adequate to promote partial protection against initial erosive lesions, independently of the type of intraoral appliance used., Clinical Significance: This finding will help researchers in the development of erosion studies, which will provide information for dentists to offer a better treatment for erosion.

Objectives: This study aimed to evaluate the ultramorphological characteristics of tooth-resin interfaces and the bond strength (BS) of multimode adhesive systems to enamel and dentin., Methods: Multimode adhesives (Scotchbond Universal (SBU) and All-Bond Universal) were tested in both self-etch and etch-and-rinse modes and compared to control groups (Optibond FL and Clearfil SE Bond (CSB)). Adhesives were applied to human molars and composite blocks were incrementally built up. Teeth were sectioned to obtain specimens for microtensile BS and TEM analysis. Specimens were tested after storage for either 24 h or 1 year. SEM analyses were performed to classify the failure pattern of beam specimens after BS testing., Results: Etching increased the enamel BS of multimode adhesives; however, BS decreased after storage for 1 year. No significant differences in dentin BS were noted between multimode and control in either evaluation period. Storage for 1 year only reduced the dentin BS for SBU in self-etch mode. TEM analysis identified hybridization and interaction zones in dentin and enamel for all adhesives. Silver impregnation was detected on dentin-resin interfaces after storage of specimens for 1 year only with the SBU and CSB., Conclusions: Storage for 1 year reduced enamel BS when adhesives are applied on etched surface; however, BS of multimode adhesives did not differ from those of the control group. In dentin, no significant difference was noted between the multimode and control group adhesives, regardless of etching mode., Clinical Relevance: In general, multimode adhesives showed similar behavior when compared to traditional adhesive techniques. Multimode adhesives are one-step self-etching adhesives that can also be used after enamel/dentin phosphoric acid etching, but each product may work better in specific conditions.

Objectives: The present study explored the histological and chemical effects of infantile thiamine deficiency (ITD) on enamel development through the examination of exfoliated deciduous teeth from a patient who had been fed during his first year of life with a thiamine-deficient milk substitute., Materials and Methods: Ground sections derived from six exfoliated primary teeth were examined. Slices from a light microscope were photographed for histological analysis. We calculated the time when the amelogenesis insults occurred, and the data were cross-examined with the patient's medical history. We then measured the enamel content of calcium, phosphate, oxygen, carbon, and magnesium on two lines from the dentino-enamel junction (DEJ) to the outer surface using an energy dispersive X-ray spectrometer., Results: Carbon (organic matter) concentration in postnatal enamel was 2.37 times higher in ITD, phosphate levels were lower, and magnesium and calcium levels tended to be higher in ITD teeth., Conclusion: Chemical and histological analysis enabled us to confirm that thiamine deficiency in infancy impaired postnatal amelogenesis and resulted in less calcified enamel with a higher level of organic matter. Higher postnatal enamel carbon and magnesium concentration found in ITD may derive from either impaired mineralization caused by disturbed cellular metabolism or indirect damage to the ameloblasts due to the physical condition. Ca/P mean ratio in ITD teeth was higher than the mean ratio in the control displaying a damaged mineralization process., Clinical Relevance: This is probably the first description of infantile thiamine deficiency effect on amelogenesis resulting in less calcified enamel.

Objectives: This study evaluated the effects of radiotherapy on the composition of deciduous teeth enamel using micro-energy-dispersive X-ray fluorescence and Fourier transform Raman spectroscopy before and after a pH cycling process., Materials and Methods: Ten deciduous molars were sectioned and divided into two groups (n = 10). The radiotherapy group (RT) was irradiated with 54 Gy at 2 Gy/day, 5 days per week for 5 weeks and 2 days, and the normal group (N) was not irradiated. The RT group was evaluated before radiotherapy (RTb), after radiotherapy (RTa), and after radiotherapy and pH cycling (RTc). The normal group was evaluated before (N) and after pH cycling (Nc). The weight percentage (wt%) of calcium (Ca), phosphorus (P), and organic content; the Ca/P ratio; and the integrated area of the Raman bands relative to the organic, carbonate, and phosphate contents were also evaluated., Results: The exclusive use of RT reduced the organic content of the enamel (p = 0.000). The RTc group exhibited a decrease in P wt% (p = 0.016), an increase in the Ca/P ratio (p = 0.000), and a reduction in the integrated area of the phosphate band (p = 0.046). An increase in the Ca/P ratio (p = 0.000) and a reduction in the areas of the carbonate and phosphate bands were found in the RTb/RTc treatments., Conclusions: RT application at a therapeutic dose reduced the organic content of the deciduous enamel., Clinical Relevance: Preventive measures should be included in the patient treatment protocol because of RT-induced chemical changes to the deciduous enamel.

Waterborne radioactive releases into the Techa River from the Mayak Production Association in Russia during 1949-1956 resulted in significant doses to about 30,000 persons who lived in downstream settlements. The residents were exposed to internal and external radiation. Two methods for reconstruction of the external dose are considered in this paper, electron paramagnetic resonance (EPR) measurements of teeth, and fluorescence in situ hybridization (FISH) measurements of chromosome translocations in circulating lymphocytes. The main issue in the application of the EPR and FISH methods for reconstruction of the external dose for the Techa Riverside residents was strontium radioisotopes incorporated in teeth and bones that act as a source of confounding local exposures. In order to estimate and subtract doses from incorporated (89,90)Sr, the EPR and FISH assays were supported by measurements of (90)Sr-body burdens and estimates of (90)Sr concentrations in dental tissues by the luminescence method. The resulting dose estimates derived from EPR to FISH measurements for residents of the upper Techa River were found to be consistent: The mean values vary from 510 to 550 mGy for the villages located close to the site of radioactive release to 130-160 mGy for the more distant villages. The upper bound of individual estimates for both methods is equal to 2.2-2.3 Gy. The EPR- and FISH-based dose estimates were compared with the doses calculated for the donors using the most recent Techa River Dosimetry System (TRDS). The TRDS external dose assessments are based on the data on contamination of the Techa River floodplain, simulation of air kerma above the contaminated soil, age-dependent lifestyles and individual residence histories. For correct comparison, TRDS-based doses were calculated from two sources: external exposure from the contaminated environment and internal exposure from (137)Cs incorporated in donors' soft tissues. It is shown here that the TRDS-based absorbed doses in tooth enamel and muscle are in agreement with EPR- and FISH-based estimates within uncertainty bounds. Basically, this agreement between the estimates has confirmed the validity of external doses calculated with the TRDS.

Objective: The objective of this study was to analyse the failure mode of adhesive interfaces by comparing OCT and scanning electron microscope (SEM) analysis of class V restoration margins located on enamel and dentin., Materials and Methods: Three groups were tested that differed in the application of a 3-step etch-and-rinse adhesive system (OptiBond FL) prior to cavity filling with restorative composite resin (Clearfil AP-X). After tooth restoration and polishing, the samples were loaded in a fatigue machine, and adhesive interfaces were evaluated with OCT and SEM., Results: Important and complementary information could be obtained with OCT analysis in respect to how marginal defects can propagate inside the cavity, compromising the restoration's long-term performance. A self-etching effect was observed with OptiBond FL due to the presence of an acidic primer (GPDM) within its composition. Our results could show that areas of bonding and gaps coexisted within the same restoration., Conclusions: When marginal imperfections, or non-continuous margins, were detected by SEM, also imperfections beneath the surface could be observed at the tooth-restoration interface with OCT. Restoration loss occurred above the borderline of 50% of marginal gaps on enamel and dentin., Clinical Relevance: Marginal discrepancies of adhesive restorations can propagate inside the cavity and lead to restoration loss.

Objectives: This study evaluated the in vitro enamel remineralization capacity of experimental composite resins containing sodium trimetaphosphate (TMP) combined or not with fluoride (F)., Materials and Methods: Bovine enamel slabs were selected upon analysis of initial surface hardness (SH1) and after induction of artificial carious lesions (SH2). Experimental resins were as follows: resin C (control—no sodium fluoride (NaF) or TMP), resin F (with 1.6% NaF), resin TMP (with 14.1% TMP), and resin TMP/F (with NaF and TMP). Resin samples were made and attached to enamel slabs (n = 12 slabs per material). Those specimens (resin/enamel slab) were subjected to pH cycling to promote remineralization, and then final surface hardness (SH3) was measured to calculate the percentage of surface hardness recovery (%SH). The integrated recovery of subsurface hardness (ΔKHN) and F concentration in enamel were also determined. Data was analyzed by ANOVA and Student-Newman-Keuls test (p < 0.05)., Results: Resins F and TMP/F showed similar SH3 values (p = 0.478) and %SH (p = 0.336) and differed significantly from the other resins (p < 0.001). Considering ΔKHN values, resin TMP/F presented the lowest area of lesion (p < 0.001). The presence of F on enamel was different among the fluoridated resins (p = 0.042), but higher than in the other resins (p < 0.001)., Conclusions: The addition of TMP to a fluoridated composite resin enhanced its capacity for remineralization of enamel in vitro., Clinical Relevance: The combination of two agents with action on enamel favored remineralization, suggesting that composite resins containing sodium trimetaphosphate and fluoride could be indicated for clinical procedures in situations with higher cariogenic challenges.

Objectives: Bone replacement grafting materials play an important role in regenerative dentistry. Despite a large array of tested bone-grafting materials, little information is available comparing the effects of bone graft density on in vitro cell behavior. Therefore, the aim of the present study is to compare the effects of cells seeded on bone grafts at low and high density in vitro for osteoblast adhesion, proliferation, and differentiation., Materials and Methods: The response of osteoblasts to the presence of a growth factor (enamel matrix derivative, (EMD)) in combination with low (8 mg per well) or high (100 mg per well) bone grafts (BG; natural bone mineral, Bio-Oss®) density, was studied and compared for osteoblast cell adhesion, proliferation, and differentiation as assessed by real-time PCR. Standard tissue culture plastic was used as a control with and without EMD., Results: The present study demonstrates that in vitro testing of bone-grafting materials is largely influenced by bone graft seeding density. Osteoblast adhesion was up to 50 % lower when cells were seeded on high-density BG when compared to low-density BG and control tissue culture plastic. Furthermore, proliferation was affected in a similar manner whereby cell proliferation on high-density BG (100 mg/well) was significantly increased when compared to that on low-density BG (8 mg/well). In contrast, cell differentiation was significantly increased on high-density BG as assessed by real-time PCR for markers collagen 1 (Col 1), alkaline phosphatase (ALP), and osteocalcin (OC) as well as alizarin red staining. The effects of EMD on osteoblast adhesion, proliferation, and differentiation further demonstrated that the bone graft seeding density largely controls in vitro results. EMD significantly increased cell attachment only on high-density BG, whereas EMD was able to further stimulate cell proliferation and differentiation of osteoblasts on control culture plastic and low-density BG when compared to high-density BG., Conclusion: The results from the present study demonstrate that the in vitro conditions largely influence cell behavior of osteoblasts seeded on bone grafts and in vitro testing., Clinical Relevance: These results also illustrate the necessity for careful selection of bone graft seeding density to optimize in vitro testing and provide the clinician with a more accurate description of the osteopromotive potential of bone grafts.

A systematic investigation, based on highly spectrally resolved Raman spectroscopy, was undertaken to research the efficacy of vibrational assessments in locating chemical and crystallographic fingerprints for the characterization of dental caries and the early detection of non-cavitated carious lesions. Raman results published by other authors have indicated possible approaches for this method. However, they conspicuously lacked physical insight at the molecular scale and, thus, the rigor necessary to prove the efficacy of this spectroscopy method. After solving basic physical challenges in a companion paper, we apply them here in the form of newly developed Raman algorithms for practical dental research. Relevant differences in mineral crystallite (average) orientation and texture distribution were revealed for diseased enamel at different stages compared with healthy mineralized enamel. Clear spectroscopy features could be directly translated in terms of a rigorous and quantitative classification of crystallography and chemical characteristics of diseased enamel structures. The Raman procedure enabled us to trace back otherwise invisible characteristics in early caries, in the translucent zone (i.e., the advancing front of the disease) and in the body of lesion of cavitated caries.

The Raman spectroscopic method has quantitatively been applied to the analysis of local crystallographic orientation in both single-crystal hydroxyapatite and human teeth. Raman selection rules for all the vibrational modes of the hexagonal structure were expanded into explicit functions of Euler angles in space and six Raman tensor elements (RTE). A theoretical treatment has also been put forward according to the orientation distribution function (ODF) formalism, which allows one to resolve the statistical orientation patterns of the nm-sized hydroxyapatite crystallite comprised in the Raman microprobe. Close-form solutions could be obtained for the Euler angles and their statistical distributions resolved with respect to the direction of the average texture axis. Polarized Raman spectra from single-crystalline hydroxyapatite and textured polycrystalline (teeth enamel) samples were compared, and a validation of the proposed Raman method could be obtained through confirming the agreement between RTE values obtained from different samples.

The aim of this pilot study was to evaluate possible doses in teeth received by workers of a uranium processing plant, in excess to the natural background dose. For this, the electron paramagnetic resonance dosimetry method was applied. Absorbed doses in teeth from the workers were compared with those measured in teeth from the Stepnogorsk city population and a control pool population from Astana city. The measured tooth samples were extracted according to medical indications. In total, 32 tooth enamel samples were analyzed, 5 from Astana city, Kazakhstan (control population), 21 from the residents of Stepnogorsk city (180 km from Astana city), and 6 from the workers of a uranium processing plant. The estimated doses in tooth enamel from the uranium processing plant workers were not significantly different to those measured in enamel from the control population. In teeth from the workers, the maximum dose in excess to background dose was 33 mGy. In two teeth from residents of Stepnogorsk city, however, somewhat larger doses were measured. The results of this pilot study encourage further investigations in an effort to receiving a final conclusion on the exposure situation of the uranium processing plant workers and the residents of Stepnogorsk city.

Objectives: This in situ/ex vivo study analysed the anti-erosive/abrasive effect of TiF4 and NaF varnish and solution on enamel wear., Materials and Methods: Twelve subjects took part in this study which was performed in three periods (phases) with the duration of 5 days each. Each two human enamel specimens per subject were pretreated with experimental NaF varnish or solution (phase A), experimental-TiF4 varnish or solution (phase B) and placebo varnish or untreated control (phase C). The specimens were worn in palatal appliances; one enamel specimen, from each treatment, was subjected to erosion (ERO; cola soft drink, 4 × 90 s/day), and the other specimen was subjected to erosion plus abrasion (ERO + ABR; tooth brushing, 2 × 10 s/day). The tooth wear was quantified by a contact profilometer (micrometre) and analysed using two-way repeated measures ANOVA and Bonferroni's test (n = 12 subjects, p < 0.05)., Results: All fluoride varnishes and solutions reduced the enamel wear (around 25 %) significantly compared to the control and placebo varnish. There were no significant differences among the fluoride formulations and between the conditions ERO and ERO + ABR., Conclusions: Therefore, it can be concluded that TiF4 has the same protective potential as NaF formulations to reduce human enamel wear under this experimental in situ model., Clinical Significance: In vitro studies have indicated a better anti-erosive/abrasive effect of TiF4 compared to NaF varnish. The present in situ study does not support the previous findings. Therefore, any of the tested professional fluoride varnishes in principle could be able to partially reduce enamel wear.

Objectives: Recent studies suggest that a combination of enamel matrix derivative (EMD) with grafting material may improve periodontal wound healing/regeneration. Newly developed calcium phosphate (CaP) ceramics have been demonstrated a viable synthetic replacement option for bone grafting filler materials., Aims: This study aims to test the ability for EMD to adsorb to the surface of CaP particles and to determine the effect of EMD on downstream cellular pathways such as adhesion, proliferation, and differentiation of primary human osteoblasts and periodontal ligament (PDL) cells., Materials and Methods: EMD was adsorbed onto CaP particles and analyzed for protein adsorption patterns via scanning electron microscopy and high-resolution immunocytochemistry with an anti-EMD antibody. Cell attachment and cell proliferation were quantified using CellTiter 96 One Solution Cell Assay (MTS). Cell differentiation was analyzed using real-time PCR for genes encoding Runx2, alkaline phosphatase, osteocalcin, and collagen1α1, and mineralization was assessed using alizarin red staining., Results: Analysis of cell attachment revealed significantly higher number of cells attached to EMD-adsorbed CaP particles when compared to control and blood-adsorbed samples. EMD also significantly increased cell proliferation at 3 and 5 days post-seeding. Moreover, there were significantly higher mRNA levels of osteoblast differentiation markers including collagen1α1, alkaline phosphatase, and osteocalcin in osteoblasts and PDL cells cultured on EMD-adsorbed CaP particles at various time points., Conclusion: The present study suggests that the addition of EMD to CaP grafting particles may influence periodontal regeneration by stimulating PDL cell and osteoblast attachment, proliferation, and differentiation. Future in vivo and clinical studies are required to confirm these findings., Clinical Relevance: The combination of EMD and CaP may represent an option for regenerative periodontal therapy in advanced intrabony defects.

Objective: The aim of the study was to evaluate the impact of flow velocity under laminar flow conditions of different acidic solutions on enamel erosion., Material and Methods: A total of 240 bovine enamel specimens were prepared and allocated to 30 groups (n = 8 each). Samples of 18 groups were superfused in a flow chamber system with laminar flow behavior using 1 ml of citric acid or hydrochloric acid (HCl) of pH 2.0, 2.6 or 3.0. Flow rates in the sample chamber were adjusted to 10, 60 or 100 μl/min. To simulate turbulent flow behavior, samples of six groups were immersed in 1 ml of the respective solution, which was vortexed (15 min, 600 rpm). For simulating non-agitated conditions, specimens of the remaining six groups were immersed in 1 ml of the respective solution without stirring. Calcium in the solutions, released from the enamel samples, was determined using Arsenazo III method., Results: For acidic solutions of pH 2.6 and 3.0, erosive potential of citric acid was equivalent to that of HCl at a flow of 100 μl/min. The same observation was made for the samples subjected to turbulent conditions at pH 3. At all other conditions, citric acid induced a significantly higher calcium loss than HCl., Conclusion: It is concluded that under slow laminar flow conditions, flow rate variations lead to higher erosive impact of citric acid compared to hydrochloric acid at pH 2.0, but not at pH ≥ 2.6 and increasing laminar flow or turbulent conditions., Clinical Relevance: Erosive enamel dissolution under laminar flow conditions is a complex issue influenced by flow rate and acidic substrate.

Following a radiological or nuclear emergency event, there is a need for quick and reliable dose estimations of potentially exposed people. In situations where dosimeters are not readily available, the dose estimations must be carried out using alternative methods. In the present study, the optically stimulated luminescence (OSL) properties of tooth enamel and different dental repair materials have been examined. Specimens of the materials were exposed to gamma and beta radiation in different types of liquid environments to mimic the actual irradiation situation in the mouth. Measurements were taken using a Risø TL/OSL reader, and irradiations were made using a (90)Sr/(90)Y source and a linear accelerator (6 MV photons). Results show that the OSL signal from tooth enamel decreases substantially when the enamel is kept in a wet environment. Thus, tooth enamel is not reliable for retrospective dose assessment without further studies of the phenomenon. Dental repair materials, on the other hand, do not exhibit the same effect when exposed to liquids. In addition, dose-response and fading measurements of the dental repair materials show promising results, making these materials highly interesting for retrospective dosimetry. The minimum detectable dose for the dental repair materials has been estimated to be 20-185 mGy.

This is the first detailed report about the collar enamel of the teeth of Polypterus senegalus. We have examined the fine structure of the collar enamel and enamel organ of Polypterus during amelogenesis by light and transmission electron microscopy. An immunohistochemical analysis with an antibody against bovine amelogenin, an antiserum against porcine amelogenin and region-specific antibodies or antiserum against the C-terminus, middle region and N-terminus of porcine amelogenin has also been performed to examine the collar enamel matrix present in these teeth. Their ameloblasts contain fully developed Golgi apparatus, rough endoplasmic reticulum and secretory granules. During collar enamel formation, an amorphous fine enamel matrix containing no collagen fibrils is found between the dentin and ameloblast layers. In non-demineralized sections, the collar enamel (500 nm to 1 μm thick) is distinguishable from dentin, because of its higher density and differences in the arrangement of its crystals. The fine structural features of collar enamel in Polypterus are similar to those of tooth enamel in Lepisosteus (gars), coelacanths, lungfish and amphibians. The enamel matrix shows intense immunoreactivity to the antibody and antiserum against mammalian amelogenins and to the middleregion- and C-terminal-specific anti-amelogenin antibodies. These findings suggest that the proteins in the enamel of Polypterus contain domains that closely resemble those of bovine and porcine amelogenins. The enamel matrix, which exhibits positive immunoreactivity to mammalian amelogenins, extends to the cap enameloid surface, implying that amelogenin-like proteins are secreted by ameloblasts as a thin matrix layer that covers the cap enameloid after enameloid maturation.

The aim of the present study was to evaluate the remineralization potential of five dentifrices with different fluoride concentrations. Initial caries lesions were created in 72 cylindrical enamel blocks from deciduous teeth. The specimens were randomly distributed among six experimental groups corresponding to six experimental periods. Each of the six volunteers carried two deciduous enamel specimens fixed in an intraoral appliance for a period of 4 weeks. They brushed their teeth and the enamel blocks at least two times a day with dentifrices containing 0 ppm (period 1), 250 ppm (period 2), and 500 ppm fluoride (period 3), respectively. A second group of volunteers (n = 6) used dentifrices with a fluoride content of 0 ppm (period 4), 1,000 ppm (period 5), or 1,500 ppm (period 6). At the end of the respective period, the mineral content was determined by transversal microradiography (TMR). The use of dentifrices containing 500 ppm fluoride (38% MR), 1,000 ppm fluoride (42% MR), and 1,500 ppm fluoride (42% MR) resulted in a statistically significant higher mineral recovery compared to the control group (0 ppm fluoride). Mineral recovery was similar after use of dentifrices containing 0 and 250 ppm fluoride (24%; 25%). It is concluded that it is possible to remineralize initial carious lesions in deciduous enamel in a similar way as it has been described for enamel of permanent teeth.

The objective of this research was to investigate nanoindentation-induced residual stresses in human enamel using Raman microspectroscopy and establish if this approach can be used as a stress meter. Healthy human premolars and sintered hydroxyapatite samples were embedded, cut, and the surfaces were polished finely with a 0.05 microm polishing paste before Berkovich and spherical indentations were made with a force of 100 mN. Spectra were collected using a Renishaw Raman InVia reflex microscope equipped with an air-cooled charge-coupled device (CCD) camera. Sample excitation was achieved using either an argon ion laser emitting at 514.5-nm or a NIR diode laser emitting at 830-nm. The residual micro stresses within and surrounding the indentation impressions were monitored by mapping the position of the nu(1)(PO(4)) band of (crystalline) hydroxyapatite. The Raman maps coincided well with the optical micrographs of the samples. Despite the presence of a fluorescence background from the organic component of human enamel, spectra collected using 514.5-nm excitation exhibited more significant shifts in the position of the nu(1)(PO(4)) band than spectra collected using 830-nm excitation. This implies that the former excitation may be a more appropriate excitation for stress detection. It was concluded that Raman microspectroscopy provides a novel high-resolution and non-destructive method for exploring the role of microstructure on the residual stress distribution within natural biocomposites.

The demineralization of enamel that is associated with early caries formation affects the optical properties of the enamel. Polarized Raman spectroscopy and optical coherence tomography have been used to detect these changes and potentially offer a means to detect and monitor early caries development. The total optical attenuation coefficient as measured by optical coherence tomography and the polarization anisotropy of the Raman peak arising from the symmetric nu(1) vibration of PO4(3-) at approximately 959 cm(-1) have been demonstrated as being sensitive markers of early caries. This ex vivo study on extracted human teeth demonstrates that these measurements can be made with reasonable precision with concomitantly good repeatability and reproducibility in sound enamel. Such reliability is crucial for these techniques to have a practical clinical value.

The aim of this study was to investigate the effect of fluoridated milk on enamel lesion formation as assessed by laser fluorescence (LF). The material consisted of 18 extracted premolar teeth that were cut in mesial-distal direction and pairwise assigned to either test or control samples in an experimental caries model. The teeth were exposed to a low-pH 5% cellulose gel for 4 h, 5 days per week immediately followed by a 4-h period in either fluoridated (5 ppm, test) or nonfluoridated milk (control). In the meantime, the specimens were stored in pooled human-stimulated whole saliva in room temperature. All teeth were examined by visual inspection with a magnifying glass and by LF readings (DIAGNOdent) at baseline and after 2 and 4 weeks. The baseline LF readings ranged from 3 to 7 with a mean value of 5.6+/-0.9. The mean values increased with time in both groups but the increase was more marked in the control teeth, 8.7+/-2.3 vs 12.8+/-3.3 after 4 weeks, this difference being statistically significant (p<0.01). The visual examination could not distinguish between the test or control samples after 2 and 4 weeks, respectively. The findings indicated that fluoride added to milk may to some extent counteract enamel lesion formation as assessed by LF in an experimental caries model.

A sensitive, quantitative method for investigating changes in enamel mineralization of specimens subjected to in vitro or in situ experimentation is presented. The fluorescence-detecting instrument integrates a Xenon arc light source and an object positioning stage, which makes it particularly suitable for the nondestructive assessment of demineralized or remineralized enamel. We demonstrate the ability of in vitro quantitative light-induced fluorescence (QLF) to quantify changes in mineralization of bovine enamel discs that had been exposed in vitro to a demineralizing gel (n=36) or biofilm-mediated demineralization challenges (n=10), or were carried in situ by three volunteers during a 10-day experiment (n=12). Further experiments show the technique's value for monitoring the extent of remineralization in 36 specimens exposed in vitro to oral multispecies biofilms and document the repeatability of in vitro QLF measurements (n=10) under standardized assay conditions. The validity of the method is illustrated by comparison with transversal microradiography (TMR), the invasive current gold standard for assessing experimental changes in enamel mineralization. Ten discs with 22 measurement areas for comparison demonstrated a positive correlation between TMR and QLF (r=0.82). Filling a technological gap, this QLF system is a promising tool to assay in vitro nondestructively localized changes in mineralization of enamel specimens.

The aim of this in vivo study was to assess the association between caries prevalence and changes in mineralization measured with a colorimeter (Color Compare CC 400, JENOPTIK, Jena, Germany). After a clinical examination (defs/DMFS, initial caries lesions), an area of a deciduous tooth was etched in each of the 35 children (psi = 8.11+/-2.41 years) with 37% phosphoric acid gel for 1 min. Immediately after, this demineralization was stained with 2% aqueous methylene blue and the red/green/blue spectrum measured with a colorimeter. Twenty-four hours later, the remineralization of this area was measured following the same staining procedure. Color measurements were clearly reduced after 24 h, indicating remineralization, and they correlated highly with the age of the children (Spearman correlation coefficient r = -0.48, p = 0.004). Correlations between the number of initial caries lesions in the deciduous and permanent dentition and color measurements after demineralization were statistically significant (r = 0.41 and 0.37, p = 0.02 and 0.045, respectively). The difference between the first and second measurements correlated significantly with the number of initial caries lesions in the permanent dentition (r = 0.42, p = 0.02). The values after artificial demineralization correlated with the number of initial lesions for the permanent (r = 0.368, p = 0.045) and deciduous (r = 0.408, p = 0.015) dentition. This resistance to artificial demineralization had stronger correlation coefficients with the caries incidence than the caries experience and initial lesions, which are considered to be the most valid caries predictors. In conclusion, these data suggest that the degree of demineralization after etching and its changes with time could be associated with caries parameters. Its use in prospective clinical trials on caries activity could be a successful approach.

The Mayak worker cohort is one of the major sources of information on health risks due to protracted exposures to plutonium and external ionizing radiation. Electron paramagnetic resonance (EPR) measurements in tooth enamel in combination with personal dose monitoring can help to improve external dose assessment for this cohort. Here, the occupational lifetime external exposure was evaluated individually for 44 nuclear workers of three plants of the Mayak Production Association by EPR measurements of absorbed doses in collected tooth enamel samples. Analysis included consideration of individual background doses in enamel and dose conversion coefficients specific for photon spectra at selected work areas. As a control, background doses were assessed for various age groups by EPR measurements on teeth from non-occupationally exposed Ozyorsk residents. Differences in occupational lifetime doses estimated from the film badges and from enamel for the Mayak workers were found to depend on the type of film badge and the selected plant. For those who worked at the radiochemical processing plant and who were monitored with IFK film badges, the dose was on average 570 mGy larger than estimated from the EPR measurements. However, the average difference was found to be only -4 and 6 mGy for those who were monitored with IFKU film badges and worked at the reactor and the isotope production plant respectively. The discrepancies observed in the dose estimates are attributed to a bias in film badge evaluation.

In the present study naturally loose deciduous molars were investigated. The feasibility of separating enamel from small size molars was analysed. EPR spectrum parameters of whole molars and separated enamel only were evaluated before and after laboratory irradiation. The Electron paramagnetic resonance (EPR) signal amplitudes of CO (2) (-) and native signals were determined by spectrum deconvolution, as a function of radiation dose in the range 0.1-10 Gy. A detection threshold of absorbed dose from deciduous molars of 198 and 21 mGy is estimated for massive (that contained both enamel and dentine) and grainy samples (that contained enamel only), respectively. The elimination of the organic material from the massive deciduous samples reduced the mean bias dose for the native signal from 90+/-18 to 34+/-13 mGy. A decay of the background signal within 2 weeks after irradiation was found, while the dosimetric signal was stable before and after the irradiation process. The presented results suggest deciduous teeth to be suitable for retrospective dose assessment. To get reasonable dose estimates, however, any organic material must be eliminated, and the measurements should be performed 2 weeks after the chemical and mechanical preparation and the irradiation process are done.

We have recently identified a protein, RP59, in bone marrow cells and young osteoblasts, in cells involved in bone repair and in young erythroblasts and megakaryocytes. Here, we report immunohistochemical data at the light- and electron-microscope level indicating that RP59 is also present in newly secreted tooth enamel of the rat and in ameloblasts, the formative cells. In enamel matrix, RP59 was located proximal to secretory ameloblasts only, i.e. in newly secreted material. Distal enamel and enamel in association with maturation stage ameloblasts were unlabelled. Secretory ameloblasts contained RP59 in the matrix-proximal region including Tomes' processes, post-secretory ameloblasts in the cell-matrix interface. Western blotting of proteins from tooth germs identified RP59 as a band at 90 kD, co-migrating with RP59 from bone marrow and spleen. Antisera versus a chemically synthesised RP59 peptide and versus a bacteria-synthesised protein fragment reacted in the same manner. In situ hybridisation of tooth tissue revealed RP59 RNA specifically in ameloblasts. The reverse transcription/polymerase chain reaction method identified tooth RNA coding for RP59. Sequence analysis indicated that RP59 RNA from tooth and marrow had the same sequence. An internal sequence motif was found in rat RP59 resembling a signal implicated in secretion of the chicken "engrailed" gene product. The findings indicate that RP59 is a genuine product of ameloblasts and that it is secreted in the course of enamel formation together with other matrix components.

The particle-induced X-ray emission (PIXE) of thick biomineral targets provides pertinent surface analysis, but if good reference materials are missing then complementary approaches are required to handle the matrix effects. This is illustrated by our results from qualitative and semiquantitative analysis of biomaterials and calcified tissues in which PIXE usually detected up to 20 elements with Z > 14 per sample, many at trace levels. Relative concentrations allow the classification of dental composites according to the mean Z and by multivariate statistics. In femur bones from streptozotocin-induced diabetic rats, trace element changes showed high individual variability but correlated to each other, and multivariate statistics improved discrimination of abnormal pathology. Changes on the in vitro demineralization of dental enamel suggested that a dissolution of Ca compounds in the outermost layer results in the uncovering of deeper layers containing higher trace element levels. Thus, in spite of significant limitations, standardless PIXE analysis of thick biomineral samples together with proper additional procedures can provide relevant information in biomedical research.

Human tooth enamel provides a nearly permanent and chronological record of an individual's nutritional status and anthropogenic trace metal exposure during development; it might thus provide an excellent bio archive. We investigated the micro-spatial distribution of trace metals (Cu, Fe, Mg, Sr, Pb, and Zn) in 196 x 339 microm2 raster pattern areas (6.6 x 10(4) microm2) in a deciduous tooth using laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS). Ablated areas include prenatal and postnatal enamel, the neonatal line, the dentine-enamel junction (DEJ), dentine, and the dentine-pulp junction. Topographic variations in the surface elemental distribution of lead, zinc, strontium, and iron intensities in a deciduous tooth revealed heterogeneous distribution within and among regions. 43Ca normalized elemental intensities showed the following order: Sr>Mg>>Zn>Pb>Fe>Cu. Elevated zinc and lead levels were present in the dental pulp region and at the neonatal line. This study demonstrates the ability of LA-ICP-MS to provide unique elemental distribution information in micro spatial areas of dental hard tissues. Elemental distribution plots could be useful in decoding nutrition and pollution information embedded in their bio apatite structure.

The development of dentin and of enamel share a common starting locus: the dentinoenamel junction (DEJ). In this study the relationship between enamel and dentin crystals has been investigated in order to highlight the guiding or modulating role of the previously mineralized dentin layer during enamel formation. Observations were made with a high-resolution electron microscope and, after digitalization, image-analysis software was used to obtain digital diffractograms of individual crystals. In general no direct epitaxial growth of enamel crystals onto dentin crystals could be demonstrated. The absence of direct contact between the two kinds of crystals and the presence of amorphous areas within enamel particles at the junction with dentin crystals were always noted. Only in a few cases was the relationship between enamel and dentin crystals observed, which suggested a preorganization of the enamel matrix influenced by the dentin surface structure. This could be explained either by the existence of a proteinaceous continuum between enamel and dentin or by the orientation of enamel proteins by dentin crystals.

The development of interglobular dentine in the first upper and lower molars of Wistar rats aged 3, 7, 14, 21, 42 days was examined histochemically using a lectin, succinyl wheat germ agglutinin (sWGA), which is specific for N-acetyl-D-glucosamine. sWGA stained the interglobular dentine, predentin and Golgi area of odontoblasts. Interglobular dentine was not formed in the first molars of 3-day rats, but appeared in those of 7-day rats near the enamel-free area. In 14-day rats, interglobular dentine was present in most areas of the coronal dentine except the cervical area. At the interface between dentine and predentin, numerous sWGA-negative calcospherites were seen, suggesting that the interglobular dentine is formed actively there. In 21-day rats, the interlobular dentine was more numerous than in 14-day rats. Interglobular dentine was present in the cervical root dentine as well as in the coronal dentine, including the cervical area. The distribution of interglobular dentine in 42-day rats was similar to that in 21-day rats, but fluorescence of sWGA binding was less intense in the former. Because the development of interglobular dentine appeared to be time and position specific its relation to the stages of ameloblasts was analysed. Thin enamel matrix was formed at cusps in molars of 3-day rats and thickness of enamel matrix increased in 7-day rats. In these teeth, the ameloblasts were at the differentiating or secretory stage. The Golgi area and Tomes' processes of the secretory ameloblasts, the cells of intermediate layer and the enamel matrix were weakly positive with sWGA. The epithelial cells at the enamel-free area were also stained with sWGA. In 14-day rats, most of the ameloblasts in the first maxillary molars were at the maturative stage except in the cervical area, where the ameloblasts were at the transitional stage. sWGA stained the distal border and the Golgi area of the maturative ameloblasts as well as the cells of the papillary layer. The distal border of the maturative ameloblasts appeared either thick or thin, suggesting a ruffle-end and smooth-end of the cells. Ameloblasts were absent in the first molars of 21-day rats and the cervical part of the enamel was covered with the stratified epithelium like that of 42-day rats. The present study has demonstrated that interglobular dentine contains sWGA-binding glycoconjugates and the formation of the interglobular dentine is largely associated with the enamel maturation. These results suggest that matrix-to-cell interaction is important for the development of interglobular dentine.

During 1949-1956, about 76 x 10(6)m3 of radioactive liquid waste containing a total activity of 10(17) Bq was discharged into the Techa River by the first Russian industrial nuclear facility Mayak. As a consequence, the population living in the river valley received considerable internal and external radiation doses. The results of a first application of electron paramagnetic resonance (EPR) of tooth enamel for a retrospective individual dose evaluation of the residents of the Techa riverside are presented. Three main contributions to the dose absorbed in tooth enamel have been considered: external exposure mainly from the Techa River sediments, internal exposure mainly due to 90Sr; and background radiation including all other sources of exposure except the Techa River. The teeth of 86 inhabitants of the town Kamensk-Uralskii were analysed to determine the age-dependent contribution of the background radiation to the enamel dose. For 22 residents of the middle and lower Techa riverside, measurements of the 90Sr whole-body content and EPR measurements of the absorbed dose in enamel were used to establish a correlation between these two quantities. Finally, absorbed doses in the enamel of five residents of the upper Techa riverside were determined by EPR method. Contributions of the background radiation and the internal 90Sr contamination were subtracted to determine the external exposure of the residents.

Protostylid pits on 50 lower third molars were studied. The depth and width of the pits, the thickness of the enamel at the pit bottom and the concavity of the dentino-enamel junction under the pit were measured by light microscopy. The pit content was examined by scanning electron microscopy and X-ray spectral microanalysis. The results show that the morphometric variables of protostylid pits are similar to those of occlusal fissures and pits. The protostylid pits were seen to contain globular formations, belonging mainly to dental calculus; organic rod-shaped bodies, which could represent remnants of the enamel organ--or more probably--foreign bodies, were frequently observed. The study suggests that the development of the most common form of protostylid is basically similar to that of an occlusal cusp, with variations attributable to the low expression of the protostylid and particularities of cusp development in the lateral tooth wall.

During epithelial-mesenchymal interactions associated with mammalian tooth development, epithelially-derived and mesenchymally-derived extracellular matrix molecules form a discrete dentine-enamel junction. The developmental and molecular processes required to form this junction are not known. To address this problem we designed studies to test the hypothesis that ectodermally-derived epithelial cells synthesize and secrete enamel proteins which function to nucleate and regulate the growth of enamel calcium phosphate crystals. Initial enamel crystals were detected separate from the adjacent dentine. Electron-microprobe analyses revealed that early enamel crystals were octacalciumphosphate or tricalciumphosphate rather than hydroxyapatite. Thereafter, enamel crystals became confluent with the adjacent, albeit significantly smaller hydroxyapatite crystals associated with mineralized dentine. Therefore, we interpret our data to indicate that de novo enamel crystal nucleation and growth are independent from the mineralization processes characterized for dentine. We further argue that gene expression of enamel protein appears to have a constitutive function during early enamel formation and that supramolecular aggregates of amelogenin and enamelin provide the microenvironment for the nucleation and crystal growth of the initial enamel matrix.

A characteristic layer containing numerous fibrils is associated with the basement membrane of the inner enamel epithelium during the early stages of odontogenesis. However, its nature is not well understood. In this study, the layer was examined with high-resolution electron microscopy and immuno-histochemical staining. Tooth germs of monkeys (Macaca fuscata) were studied and each fibril in the layer was found to be a tubular structure, 8-9 nm in width, resembling a "basotubule", the tubular structure previously observed in various basement membranes. The space between the fibrils was filled with a network formed by irregular anastomosing strands with an average thickness of 4 nm; these strands resembled the "cords" forming the network in the lamina densa of basement membranes. After immunoperoxidase staining, fine threads immunoreactive for laminin staining were seen winding along the strands of the network, and 1.5-nm wide filaments, immunoreactive for type IV collagen, took the form of a network arrangement. The 5-nm-wide ribbon-like structures associated with the strands were identified as heparan sulfate proteoglycan by immunostaining. These results are similar to those obtained for the cord network of the lamina densa. The "fibrillar layer" therefore represents a highly specialized lamina fibroreticularis of the basement membrane of the inner enamel epithelium, and rich in basotubules.

Immunohistochemistry was used to study the ontogeny of GH receptor/binding protein (GHR/BP) and IGF-I from the 13-day-old embryo (E13) to the E19 rat fetus in the developing incisor and molar. Analysis of serial sections revealed diffuse staining of GHR/BP and IGF-I at the bud and early cap stages within both the mesenchyme of the dental papilla and the ectodermal-derived enamel organ. Just before transition to the cap stage, immunoreactivity of GHR/BP and IGF-I increased in the epithelial bud and extended to the condensed dental mesenchyme. At the cap stage, the dental epithelium showed an intense expression of GHR/BP and IGF-I, whereas the dental mesenchymal cells showed very weak staining. The inner enamel epithelium and the outer enamel epithelium were positive for both GHR/BP and IGF-I in the bell stage. Differentiating ameloblasts, odontoblasts and the secretory ameloblasts and odontoblasts continued to express GHR/BP and IGF-I in incisors. These findings support the premise that growth hormone and IGF-I may play a role in embryonic tooth development by regulating the epithelial-mesenchymal interactions that influence events in growth and cytodifferentiation.

Enamel proteins were extracted from the newly formed layer of immature porcine enamel, and the 25 kDa amelogenin, 89 kDa enamelin and 13-17 kDa nonamelogenins were purified. Specific antisera were raised against these proteins. Antibodies specific to the C-terminal region (residues 149-173) of the 25 kDa amelogenin were generated by absorption of the anti-25 kDa amelogenin serum with 20 kDa amelogenin, which contains residues 1-148 of the antigen. Immunoelectro-transfer blotting of the extracted porcine enamel proteins showed that the anti-25 kDa amelogenin serum recognized the 25 kDa and other low and high molecular weight amelogenins. The C-terminal specific anti-25 kDa amelogenin serum reacted only with amelogenins having molecular weights over 23 kDa. The anti-89 kDa enamelin serum recognized the 89 kDa enamelin and lower molecular weight proteins, but neither the amelogenins nor the 13-17 kDa nonamelogenins. The antiserum against the 13-17 kDa nonamelogenins showed no cross reactivity to the 89 kDa enamelin, but recognized higher molecular weight nonamelogenins. In immunohistochemical preparations of the porcine tooth germs, the 25 kDa amelogenin-like immunoreactivity over immature enamel decreased in a gradient from the enamel surface to the middle layer. In the inner layer immunoreactivity was concentrated over the prism sheaths. The C-terminal specific 25 kDa amelogenin-like immunoreactivity was intense at the outer layer of immature enamel and decreased sharply toward the middle layer. Prism sheaths were intensely stained by the antiserum to the 13-17 kDa nonamelogenins.(ABSTRACT TRUNCATED AT 250 WORDS)