S. Mutansin Research Paper

Effect of chalepensin on S. mutans growth
Fifty microliters of a S. mutans suspension (1 x 103 bacteria/ml) in BHI medium (Remel, Lenexa, KS) were transferred to flat-bottomed 96-well plates (Corning Incorporated, Corning, NY) containing serial dilutions (1:2) of 50 μl of chalepensin, or 1 μg/ml tetracycline, chalepensin-free vehicle (vehicle was similarly processed as with chalepensin extraction, but without plant material), and culture medium controls, and incubated for 6 h at 37°C. Next, MTT was added to all wells (0.5 mg/ml in saline solution, final concentration) and microplate cultures were incubated for 4 additional hours. Next, 50 μl of extraction buffer (Gomez-Flores et al., 1995) were added to all wells, microplates incubated (Yamato IC600 incubator) for 16 h at 37°C, and optical densities read at 570 nm (microplate reader, Beckman Coulter, Inc., Fullerton, CA) (Gomez-Flores et al., 1995). For colony forming units (CFU) determination, 1:10,000 dilutions from treatment and control wells, as explained above, were seeded on BHI agar plates (Becton Dickinson, Mexico, D.F.) and incubated for 24 h at 37°C, after which CFU were counted (ULB-100,
Plants such as Glycyrrhiza glabra, Allium sativum, Aloe vera, Physalis angulata, Annona senegalensis, Dryopteris crassirhizoma, Quercus infectoria, Englerophytum magalismontanum, Euclea natalensis, Solanum panduriforme, Rosmarinus officinalis Linn., Baeckea frutescens, and Parinari curatellifolia were demonstrated to have antibacterial activity against S. mutans (Bachrach et al., 2011; Ban et al., 2012; Fani and Kohanteb, 2012; Hwang et al., 2004; More et al., 2008). In addition, substances such as linoleic, linolenic, oleanolic, betulinic acids, betulin, and beta-sitosterol glucoside, among others, were reported to suppress adherence of S. mutans in-vitro (Wu,