## 4 Discussion
Documented evidence shows that the presence of biofilm is a risk factor for periodontitis, which generates inflammatory reactions [51]. Furthermore, the presence of periodontitis and the attendant inflammation has been associated with diseases like endothelial dysfunction and coronary heart disease [52]. Therefore, the treatment of biofilm associated dental diseases is an important health approach. Moreover, a recent report confirms the link between pathogenic oral bacteria and the progression of periodontal-induced biofilm [25]. Visualization with confocal imaging confirmed concentration-dependent dispersion and loose colonies with mangiferin treatment in this study. These findings indicate that mangiferin potentially disturbed the process of adherence in early _S. mutans_ colonizers, thereby interrupting biofilm formation. The studies of [PERSON] et al. [38] and [PERSON] et al. [26] thus support this outcome and this could mean an anti-adherentproperty. The behavior of _S. mutans_ to aggregate and adhere contributes to the pathogenesis of dental plaque. If not controlled, will eventually lead to dental caries and associated diseases of the oral cavity [10]. In addition, biofilm analysis on the tooth showed that viable CFU were significantly reduced. Meaning that microbial milieu favourable to colonization has been altered by mangiferin. Evidently, from this present study, the synthesis of water-soluble and water insoluble glucans were reduced in the presence of mangiferin. These effects were only achieved in the concentration of 125-1000 m of mangiferin. Reported studies show that _S. mutans_ utilizes water soluble glucan to interact with salivary proteins in the oral cavity, whereas the synthesized water insoluble glucan is used to form stable dental plaques with increased adherence that attract other microbes [39; 53]. Other natural products like EGCC and Ethyl gallate have similarly been reported to suppress the propensity of _S. mutans_ to synthesize both water-soluble and water-insoluble glucans [39; 53]. In our study, mangiferin suppressed biofilm formation on the tooth surface by attenuating the very element that enables adhesion. Furthermore, to evaluate the effect of mangiferin on gene expression of glucansucrase virulence elements used by _S. mutans_ to form and maintain functional biofilms. The elucidation of these genes and their presence provided more information about the mode of action of mangiferin. Here, we demonstrated that mangiferin significantly downregulated the expression of _gtfB_, _gtfC_, and _gtfD_ virulence genes involved in biofilm formation. The inhibition compromised their ability to express their full virulence attributes. Evidence indicates that reduced expressions of these genes proactively decreased the production of extracellular polysaccharide, bacterial aggregation, and consequently biofilm formation [39]. In addition, mangiferin was found to reduce the expression of glucan binding protein (_gbpB_). It is noted to be responsible for mediating microbial cell adhesion and plays an active role in the maturation of biofilm [54]. Our finding is consistent with the report of [PERSON] et al. [50] who documented the reduction of the _gbpB_ virulent gene involved in biofilm formation using a modified natural compound. Another essential element critical for dental plaque development, used by _S. mutans_ to establish persistent biofilm, is the competence stimulating peptide (CSP) gene _comDE_[55]. The _comDE_ gene is involved in the quorum-sensing system, regulating the expressions and competence of glucanotransferase enzymes. It also modulates the oral microbial environmental adaptation of bacteria involved in biofilm formation [56]. In our present investigation, we report that mangiferin suppressed _comDE_ gene expression. [PERSON] et al. [10] demonstrated that the suppression of _comDE_ expression is critical for plaque formation, affecting cell wall integrity and the loss of adherence. Reports indicate that _S.