Quantitative evaluation of remineralizing potential of three agents on artificially demineralized human enamel using scanning electron microscopy imaging and energy‑dispersive analytical X‑ray element analysis: An in vitro study

Nitin Khanduri, Deepak Kurup, Malay Mitra


Background: The aim of this study is to quantitatively evaluate the remineralization potential
of three remineralizing systems as follows: fluoride, casein phosphopeptide‑amorphous calcium
phosphate (CPP‑ACP), and CPP‑ACP with fluoride, under scanning electron microscope with
energy‑dispersive X‑ray analysis.
Materials and Methods: In this in vitro study A total of 40 enamel specimens were prepared
from the buccal or lingual surfaces of human premolars extracted for orthodontic reason.
Specimens were then placed in demineralizing solution for 96 h, to produce artificial caries‑like
lesion. Calcium and phosphate weight percentage of demineralized specimens was measured.
Specimens were divided into four groups as follows: (a) control, (b) CPP‑ACP, (c) CPP‑ACP
with fluoride, and (d) fluoride varnish. Except for the control group, the entire specimens were
subjected to remineralization using respective remineralizing agents of their groups. The prepared
specimens were assessed for calcium and phosphate weight percentage using scanning electron
microscopy‑energy dispersive X‑ray spectroscopy. One way analysis of variance (ANOVA), followed
by Tukey’s test, was performed with the help of critical difference (CD) or least significant difference
at 5% and 1% level of significance. P ≤ 0.05 was taken to be statistically significant and P < 0.001
as statistically highly significant.
Results: The mean weight percentage of calcium and phosphorus of specimens treated with
CPP‑amorphous calcium phosphate nanocomplexes plus fluoride (ACPF) was significantly higher
than other groups.
Conclusion: All the groups showed statistically significant remineralization. However, because of
added benefit of fluoride, CPP‑ACPF showed statistically significant amount of remineralization
than CPP‑ACP.


Casein phosphopeptide‑amorphous calcium phosphate, demineralization, remineralization

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