The objective of this in vitro study was to quantify the amount of mineral change in demineralized dentin at pH 5.5 after the application of dental varnishes containing fluoride with casein phosphopeptide-amorphous calcium phosphate, fluoride and bioglass, or fluoride alone.

Methods and materials::

A total of 12 extracted human sound mandibular premolar root samples were coated with an acid-resistant varnish, leaving a 2 × 3 mm window at the outer root surface. These root specimens were then randomly divided into four groups and separately subjected to the demineralizing cycle at a pH of 4.8 for five days to create artificial caries-like lesions in dentin. Subsequently, each sample was imaged using quantitative x-ray microtomography (XMT) at a 15-μm voxel size. Each test group then received one of the following treatments: dental varnish containing casein phosphopeptide-amorphous calcium phosphate and fluoride (CPP-ACP, MI varnish, GC Europe), bioglass and fluoride (BGA, Experimental, Dentsply Sirona), or fluoride alone (NUPRO, Dentsply Sirona), as well as a control group, which received no treatment. These samples were kept in deionized water for 12 hours. The thin layer of varnish was then removed. All samples including the nonvarnish group were subjected to the second demineralizing cycle at pH 5.5 for five days. The final XMT imaging was then carried out following the second demineralizing cycle. XMT scan was also carried out to varnish samples at 25 μm voxel size. The change in mineral concentration in the demineralized teeth was assessed using both qualitative and quantitative image analysis.


There was an increase in radiopacity in the subtracted images of all varnish groups; a significant increase in mineral content, 12% for the CPP-ACP and fluoride ( p≤0.05 and p≤0.001), 25% BGA ( p≤0.001), and 104% fluoride alone varnish ( p≤0.001). There was an increase in the size of radiolucency in the lesion area with a significant decrease in mineral content in the nonvarnish group, 10% ( p≤0.05 and p≤0.001).


There was encouraging evidence of a remineralization effect following the application of dental varnish on dentin and also an observed resistance to demineralization during the acidic challenge in all cases. However, a dental varnish containing fluoride alone appeared to have a much greater effect on dentin remineralization when compared with CPP-ACP with fluoride and bioglass with fluoride.