# Assessing the Impact of Varied Dark Chocolate Concentrations on Enamel and Dentine Microhardness
## 3 Results
### Comparision of Enamel and Dentin Microhardness between Pairs of Groups
In addition, it contains unsaturated free fatty acids like oleic and linoleic acids, which are antibacterial for Streptococcus mutans [32]. The larger molecular weight of polyphenic molecules present in cocoa bean husk extract possesses significant antiglycostransferase activity. The cariostatic effects of the cocoa bean husk are due to these physiologically active substances [32]. Streptococcus mutans is recognized as the primary contributor to dental caries in humans. It produces three types of glucosyltransferase(GTF), namely GTFB, GTFC, and GTFD. These enzymes are responsible for synthesizing glucans from sucrose, resulting in an adhesive and water-insoluble substance that firmly adheres to the tooth surface [32]. The adhesive glucan, where acid accumulation takes place, plays a role in the formation of dental plaque, leading to localized demineralization of the tooth enamel surface [33]. Previous studies investigating the mechanism of the anticariogenic properties of theobromine have revealed the development of a medium with an apatite structure that promotes the rehardening of the tooth surface through remineralization [34]. In this study, enamel and dentine surface microhardness assessments were conducted utilizing a digital micro-Vickers hardness tester (Laryee, Beijing, China) under (0.24 N load for 15 s) before and after immersion of the specimens in the selected concentration of dark chocolate and using artificial saliva as a control group. The findings of the current investigation indicated that there is an increase in enamel and dentin hardness after immersion of the prepared specimen in different concentrations of dark chocolate and even in artificial saliva. Therefore, the current study's null hypothesis was rejected. The possible explanation for this effect is attributed to the composition of artificial saliva, specifically the inorganic components, primarily calcium and phosphate. No significant difference was observed between human saliva and artificial saliva containing mucin, suggesting that it can serve as a suitable substitute for human saliva in in vitro cycling studies focused on remineralization and hardness assessment [35]. Also in the major composition of dark chocolate are minerals like iron, magnesium, zinc, copper, potassium, selenium, phosphorus, and manganese, which are the essential minerals for strengthening tooth structure, according to certain previous studies. Theobromine increased the hardness of the enamel, and the inorganic minerals were directly related to enhancing the hardness of the enamel [4,6]. That agrees with our study. In the present investigation, 70% dark chocolate was employed, characterized by a composition of 70% cocoa along with cocoa butter and sugar. Likewise, 85% dark chocolate was utilized, incorporating 85% cocoa, cocoa butter, and sugar, while 100% dark chocolate consisted solely of cocoa and butter without the addition of sugar. The study revealed a notable correlation between the increasing concentration of dark chocolate and a significant enhancement in the hardness of enamel and dentin, attributable to the heightened cocoa content. This study specifically examined the authentic impact of dark chocolate on enamel and dentin hardness without resorting to demineralization processes, demonstrating a favorable influence on dental structures. Particularly noteworthy is the observed increase in theobromine and mineral content with escalating chocolate concentration. Prior research has primarily focused on the impact of theobromine post-demineralization; however, a dearth of literature exists regarding the direct examination of the inherent effects of dark chocolate on enamel and dentin. According to the studies, theobromine outperformed fluorides in safeguarding teeth. In a laboratory experiment, researchers evaluated the effectiveness of a ginger, honey, and chocolate mixture in remineralizing the initial enamel caries lesions.