Lead contents in the surface enamel of primary and permanent teeth, whole blood, serum, and saliva of 6- to 8-year-old children.

This study aimed to evaluate associations of Pb-enamel values determined in two successive enamel microbiopsies taken from a primary and a permanent tooth, and to assess how Pb-enamel correlates with Pb-blood and Pb-serum as well as whole (Pb-whole-saliva), submandibular/sublingual (Pb-sub-saliva) and parotid saliva (Pb-parotid-saliva). The study population included 444 children aged 6 to 8years attending 4 government schools in the district of Campos Eliseos, in Ribeirao Preto, Sao Paulo State, Brazil. Whole blood, serum, parotid, submandibular/sublingual ("sub-saliva"), and whole saliva were collected in trace element-free tubes. Two successive microbiopsies were taken from the surface of a primary and a permanent tooth of each child. Lead concentrations were determined by inductively coupled plasma mass spectrometry (ICP-MS). There was a significant correlation between primary and permanent teeth in terms of Pb-enamel, for both the first and the second microbiopsies. When the median Pb-enamel values were compared between the two genders using only the highest 10th percentile Pb-enamel, there was a significant difference between girls and boys for both primary (474.2 vs 910.0µg/g, respectively; p=0.02) and permanent teeth (739.5 vs 1325µg/g, respectively; p=0.04). There were no significant correlations between Pb-enamel and Pb-blood or between Pb-enamel and lead in the 3 different salivas. However, there was a statistically significant correlation between Pb-enamel and Pb-serum when only the highest 10th percentile Pb-enamel was analyzed (r=0.57 and p=0.0002 for primary teeth; r=0.56 and p<0.0001, for permanent teeth). In conclusion, our findings have demonstrated a significant correlation between Pb-enamel found in primary and permanent teeth, as well as a significant correlation between Pb-serum and lead in primary and permanent tooth enamel. Boys presented higher Pb-enamel than girls. Our study also suggests that Pb-enamel has no correlation with Pb-blood or with lead in the 3 different salivas.

Whole blood, serum, and saliva lead concentrations in 6- to 8-year-old children.

With increasing evidence of adverse health effects of lower lead levels (below 10microg/dL in whole blood), studies on novel internal dose biomarkers are needed. This study aimed at: (1) assessing the lead exposure by measuring this element in whole blood (Pb-blood), serum (Pb-serum), and 3 different types of saliva: whole (Pb-whole-saliva), submandibular/sublingual (Pb-sub-saliva) and parotid saliva (Pb-parotid-saliva); (2) correlating Pb-blood with Pb-serum; and (3) relating Pb-blood and Pb-serum with the lead found in the different salivas. The study population included 444 children aged 6 to 8 years attending 4 government schools in the district of Campos Eliseos, in Ribeirao Preto, Sao Paulo State, Brazil. Whole blood, serum, parotid, submandibular/sublingual ("sub-saliva"), and whole saliva were collected in trace element-free tubes. Lead concentrations were determined by inductively coupled plasma-mass spectrometry (ICP-MS). Median Pb-blood and Pb-serum were 2.1microg/dL and 0.4microg/L, respectively. Ten percent of the children had Pb-blood between 4.0 and 9.4microg/dL. Boys showed higher Pb-blood than girls (2.3 vs. 2.0microg/dL, p<0.0003). Lead concentrations in whole, sub, and parotid saliva were 1.7, 1.4, and 1.3microg/L, respectively. No significant correlations were found between Pb-blood and Pb-serum, between Pb-blood and lead in the 3 different salivas, or between Pb-serum and Pb-whole-saliva or Pb-sub-saliva. However, there was a weak statistically significant correlation between Pb-serum and Pb-parotid-saliva. In conclusion, the mean Pb-blood concentration was 2.4microg/dL, with 10% of the children exhibiting Pb-blood between 4.0 and 9.4microg/dL. Boys presented higher Pb-blood. Results suggest that Pb-blood has no correlation with Pb-serum or lead in the 3 different salivas collected here. A weak but statistically significant correlation between Pb-serum and Pb-parotid-saliva was found. Our results also point to the need for carrying out more studies on sources of exposure and lead levels in children in Brazil.

Analysis of enamel microbiopsies in shed primary teeth by Scanning Electron Microscopy (SEM) and Polarizing Microscopy (PM).

The aims of this study were 1) to verify how close to the theoretically presumed areas are the areas of enamel microbiopsies carried out in vivo or in exfoliated teeth; 2) to test whether the etching solution penetrates beyond the tape borders; 3) to test whether the etching solution demineralizes the enamel in depth. 24 shed upper primary central incisors were randomly divided into two groups: the Rehydrated Teeth Group and the Dry Teeth Group. An enamel microbiopsy was performed, and the enamel microbiopsies were then analyzed by Scanning Electron Microscopy (SEM) and Polarizing Microscopy (PM). Quantitative birefringence measurements were performed. The "true" etched area was determined by measuring the etched enamel using the NIH Image analysis program. Enamel birefringence was compared using the paired t test. There was a statistically significant difference when the etched areas in the Rehydrated teeth were compared with those of the Dry teeth (p=0.04). The etched areas varied from -11.6% to 73.5% of the presumed area in the Rehydrated teeth, and from 6.6% to 61.3% in the Dry teeth. The mean percentage of variation in each group could be used as a correction factor for the etched area. Analysis of PM pictures shows no evidence of in-depth enamel demineralization by the etching solution. No statistically significant differences in enamel birefringence were observed between values underneath and outside the microbiopsy area in the same tooth, showing that no mineral loss occurred below the enamel superficial layer. Our data showed no evidence of in-depth enamel demineralization by the etching solution used in the enamel microbiopsy proposed for primary enamel. This study also showed a variation in the measured diameter of the enamel microbiopsy in nineteen teeth out of twenty four, indicating that in most cases the etching solution penetrated beyond the tape borders.

Lead in saliva from lead-exposed and unexposed children.

INTRODUCTION: Whole blood is used for diagnosis of lead exposure. A non-invasive method to obtain samples for the biomonitoring of lead contamination has become a necessity. This study 1) compares the lead content in whole saliva samples (Pb-saliva) of children from a city with no reported lead contamination (Ribeirão Preto, São Paulo State, Brazil) and children of a region notoriously contaminated with lead (Bauru, São Paulo State, Brazil), and 2) correlates Pb-saliva with the lead content in the enamel microbiopsy samples (Pb-enamel) in the case of these two populations. METHODS: From a population of our previous study that had included 247 children (4- to 6-year-old) from Ribeirão Preto, and 26 children from Bauru, Pb-saliva was analyzed in 125 children from Ribeirão Preto and 19 children from Bauru by inductively coupled plasma mass spectrometry (ICPMS). To correlate Pb-saliva with Pb-enamel, we used Pb-enamel data obtained in our previous study. The Mann-Whitney test was employed to compare the Pb-saliva data of the two cities. Pb-saliva and Pb-enamel values were then Log10 transformed to normalize data, and Pb-saliva and Pb-enamel were correlated using Pearson's correlation coefficient. RESULTS: Median Pb-saliva from the Ribeirão Preto population (1.64 microg/L) and the Bauru population (5.85 microg/L) were statistically different (p<0.0001). Pearson's correlation coefficient for Log10 Pb-saliva versus Log10 Pb-enamel was 0.15 (p=0.08) for Ribeirão Preto and 0.38 (p=0.11) for Bauru. CONCLUSIONS: A clear relationship between Pb-saliva and environmental contamination by lead is shown. Further studies on Pb-saliva should be undertaken to elucidate the usefulness of saliva as a biomarker of lead exposure, particularly in children.

Lead contents in the surface enamel of deciduous teeth sampled in vivo from children in uncontaminated and in lead-contaminated areas.

This study aimed to: (1) measure lead contents in the surface enamel of two populations consisting of 4-6-year-old children, one from an apparently uncontaminated area (Ribeirão Preto, São Paulo State, SP, Brazil, n=247) and the other from an area notoriously contaminated with lead (Bauru, São Paulo State, Brazil, n=26); (2) compare biopsy depths between the two populations; (3) correlate biopsy depth with lead content; (4) stratify samples according to biopsy depth to compare lead contents in samples from similar biopsy depths. A surface enamel acid-etch microbiopsy was performed in vivo on a single upper deciduous incisor for each sample. Lead was measured by graphite furnace atomic absorption spectrometry (GFAAS) while phosphorus was measured colorimetrically to establish biopsy depth. Samples from both populations were classified into categories of similar biopsy depths based on biopsy depth quartiles. Median lead contents were statistically different between the Ribeirão Preto population (206 microg/g, range: 5-1399 microg/g) and the Bauru population (786 microg/g, range: 320-4711 microg/g) (p<0.001); however, biopsy depth did not differ between the Ribeirão Preto (3.9 microm, Standard Deviation, SD=0.9) and Bauru (3.8 microm, SD=0.9) populations (p=0.7940). Pearson's correlation coefficient for biopsy depths versus log10 lead values was -0.29 for Ribeirão Preto and -0.18 for Bauru. Lead contents were statistically different between the two populations for all quartiles of biopsy depth. These findings suggest that lead accumulated in the surface enamel of deciduous teeth is linked to the environment in which people reside, indicating that this tissue should be further explored as an accessible biomarker of lead exposure.