In Vitro Anticariogenic Effects of Drymocallis rupestris Extracts and Their Quality Evaluation by HPLC-DAD-MS3 Analysis

Michał Tomczyk, Małgorzata Pleszczyńska, Adrian Wiater, Sebastian Granica
2013 Molecules  
In this study, for the first time, we investigated in vitro inhibitory effects of Drymocallis rupestris extracts and their subfractions obtained with solvents of different polarity (aqueous, 50% ethanolic, diethyl ether, ethyl acetate and n-butanolic) against bacterial viability and caries virulence factors of Streptococcus spp. strains. The diethyl ether subfraction (PRU2) showed bacteriostatic and bactericidal activity against mutans streptococci, with minimum inhibitory concentrations (MICs)
more » ... ncentrations (MICs) in the range of 0.75-1.5 mg/mL and minimum bactericidal concentrations (MBCs) in the range of 1.5-3 mg/mL. Furthermore, PRU2 inhibited biofilm formation by Streptococci in a dose-dependent manner. It was also found that all five D. rupestris preparations exhibited diverse inhibitory effects on de novo synthesis of water-insoluble and water-soluble α-D-glucans by glucosyltransferases of the mutans group streptococci. The phytochemical profile of investigated samples was determined by spectrophotometric and chromatographic (HPLC-DAD-MS 3 ) methods. The high polyphenol (total phenol, phenolic acids, tannins, proantocyanidins, and flavonoids) contents were found which correlated with anticariogenic activity of the analyzed samples. The results demonstrate that D. rupestris extracts and their subfractions could become useful supplements for pharmaceutical products as a new anticariogenic agent in a wide OPEN ACCESS Molecules 2013, 18 9118 range of oral care products. Further studies are necessary to clarify which phytoconstituents of D. rupestris are responsible for anticaries properties. Introduction Rock cinquefoil [Drymocallis rupestris (L.) Soják (syn. Potentilla rupestris L.; Potentilla corsica Sieber ex Lehm) Rosaceae] is chiefly found on soils derived from base-rich, igneous rocks. On the European continent, D. rupestris is usually found in mountain areas on rock and scrubby, rocky slopes which are often steep and south-facing [1,2]. However, little is known about chemical composition of extracts prepared from D. rupestris. The plant has only been reported to contain 2-pyrone-4,6dicarboxylic acid as well as long-chain polyprenyl alcohols. More recently, the presence of flavonoids and carboxylic acids in the aerial parts of rock cinquefoil has also been described [3] [4] [5] . Considering the traditional use of the Potentilla L. species [6] , it is justified to investigate its biological properties in the dental field. Dental caries is a common chronic, social disease, and therefore is an important issue not only for dentists, but also researchers from other fields of science. There is a whole range of methods for preventing or reducing tooth decay. These include, among the others: proper oral hygiene (brushing, toothpaste, dental floss, and mouthwash); the use of unfermentable sugar substitutes [7]; addition of fluoride to drinking water, salt, milk or toothpaste [8]; the use of antimicrobial agents that inhibit the development of dental plaque [9]; sealing of occlusal pits and fissures of caries-susceptible teeth [10] ; the induction of immune responses [11] , and replacement therapy [12] . However, the problem of caries still remains valid. Hence searching for alternative prevention methods and products for oral diseases that are safe, effective and economical is needed. One of them is the use of plant-derived products [13, 14] . The purpose of this study was to evaluate the effects of extracts and their subfractions prepared by the partitioning of aqueous methanolic extract with solvents of different polarity against cariogenic Streptococcus spp. strains. Furthermore, their inhibitory effects on de novo synthesis of water-soluble and water-insoluble α-glucans by glucosyltransferases of mutans streptococci and artificial dental plaque formation were examined. The phytochemical profile of the investigated samples was also determined. Results and Discussion Determination of Polyphenolic Compounds In order to roughly estimate the total polyphenol content (TPC), and that of related polyphenolic compounds like phenolic acids (TPA), flavonoids (TFC), tannins (TTC), proanthocyanidins (TPDC) in the aerial parts of D. rupestris (Table 1) were measured by different analytical methods preceeding the HPLC-MS study. The results obtained in the present study showed that all the analyzed extracts and subfractions contain relatively huge quantities of polyphenolic compounds. Both, the diethyl ether subfraction (PRU2) and the ethyl acetate subfraction (PRU3) showed the highest values of the total Sample Availability: Samples of the extracts and subfractions are available from the authors (M.T.).
doi:10.3390/molecules18089117 pmid:23903184 fatcat:7ckr3rngznhqnlhfluezbm6yqq