A novel electron-microscopic method for measurement of mineral content in enamel lesions

Arch Oral Biol. 2018 Oct:94:10-15. doi: 10.1016/j.archoralbio.2018.06.013. Epub 2018 Jun 13.

Abstract

Objective: To assess Scanning Electron Microscopy in Back-Scattered Emission mode (BSE-SEM) for measurement of lesion mineral content as a function of depth. Direct comparison is made with Transverse Micro-Radiography (TMR) and Surface Micro-Hardness (SMH) on carious and erosive lesions.

Design: Caries lesions prepared from sound bovine enamel at 37 °C and pH 4.6 in unsaturated (7d) or part-saturated (8d, 4.1 mM Ca2+, 8 mM Pi) lactic acid /methyl cellulose gel system, followed by TMR analysis. Erosive lesions prepared from sound bovine enamel (1% citric acid, pH3.8, room temperature) for 5, 10, 15 or 20 min at n = 10 per treatment group. SMH readings (Vickers diamond, 1.9 N, 20 s) were taken from acid-treated and reference areas of each sample. BSE-SEM performed on polished cross-sections of lesioned samples (Jeol JSM6490LV SEM; high vacuum, 10 keV beam voltage, magnification x500 with constant working distance of 10 mm). Under identical SEM conditions, polished standards i.e. MgF2, alumina, Mg, Al and Si provided a calibration plot of BSE-SEM signal vs. atomic number (z¯). Mineral content vs. depth plots were derived from the cross-sectional BSE-SEM data.

Results: Cross-sectional BSE-SEM images clearly differentiate between caries and erosive lesions. Comparison of caries lesion mineral loss from BSE-SEM with TMR data showed good correlation (R2 = 0.98). Similarly, comparison of BSE-SEM data from erosive lesions showed good correlation (R2 = 0.99) with hardness loss data from SMH.

Conclusion: BSE-SEM provides a relatively rapid and cost-effective method for the assessment of mineral content in demineralised tooth enamel and is applicable to both caries and erosive lesions.

Keywords: Caries; Demineralisation; Erosion; Radiography; Remineralisation; Scanning electron microscopy.

MeSH terms

  • Animals
  • Cattle
  • Cost-Benefit Analysis
  • Dental Caries / diagnostic imaging*
  • Dental Caries / pathology*
  • Dental Enamel / chemistry*
  • Dental Enamel / drug effects
  • Dental Enamel / ultrastructure*
  • Hardness
  • Hydrogen-Ion Concentration
  • Incisor / diagnostic imaging
  • Incisor / pathology
  • Lactic Acid / adverse effects
  • Microscopy, Electron, Scanning / methods*
  • Minerals / analysis*
  • Radiography, Dental / methods
  • Sensitivity and Specificity
  • Surface Properties
  • Tooth Demineralization / pathology
  • Tooth Erosion / diagnostic imaging
  • Tooth Erosion / pathology
  • Tooth Remineralization / methods

Substances

  • Minerals
  • Lactic Acid