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Updated: Jul 9, 2026

Microhardness Measurements on Tooth and Alveolar Bone in Rodent Oral Disease Models
Published on: April 26, 2024
Change in surface hardness of enamel by a cola drink and a CPP-ACP paste
D Tantbirojn1, A Huang, M D Ericson
1Department of Restorative Sciences, University of Minnesota, Minneapolis, USA. tant0002@umn.edu
Objectives:
This in vitro study used surface microhardness to evaluate whether a paste containing casein phosphopeptide amorphous calcium phosphate (CPP-ACP) can reharden tooth enamel softened by a cola drink, and how different saliva-substitute solutions affect the enamel hardness.
Methods:
Twenty-four bovine incisors, each tooth consisting of treatment and control halves, were immersed in a cola drink (Coke) for 8 min, then placed under a 0.4 mL/min drip with various saliva-substitute solutions. The saliva-substitute solutions were: saliva-like solution (SLS) with 1 ppm fluoride, SLS without fluoride, and Biotene mouthwash. CPP-ACP paste was applied to the treatment halves for 3 min at 0, 8, 24, and 36 h. Knoop microhardness measurements were performed at baseline, after the cola drink immersion, and after 24 and 48 h contact with saliva-substitute solution.
Results:
Enamel hardness significantly decreased after immersion in cola drink (ANOVA, p<0.05). After contact with saliva-like solutions for 48 h, those treated with CPP-ACP paste were significantly harder than those untreated regardless of the presence of 1 ppm fluoride in the saliva-like solution (ANOVA, p<0.05). Biotene mouthwash significantly softened the enamel surface (ANOVA, p<0.05). Two-way ANOVA showed significant effects of the CPP-ACP paste application and types of saliva-substitute solutions on the changes in surface hardness of the softened enamel at a significance level of 0.05.
Conclusion:
The application of CPP-ACP paste with continuous replenishment of saliva-like solution for 48 h significantly hardened enamel softened by a cola drink. Biotene mouthwash softened enamel surface after 48 h contact.
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