JCDP Anticaries10.5005/jp-journals-10024-2361 Efficacy of Liquorice Lollipop ORIGINAL RESEARCH Anticaries Efficacy of Liquorice Lollipop: Anex vivo Study 1Gauri Krishnakumar, 2Dhananjaya Gaviappa, 3Shwetha Guruswamy ABSTRACT Source of support: Nil Aims: To investigate the anti-bacterial efficacy of liquorice Conflict of interest: None lollipop and also to investigate the remineralizing potential of the extract. INTRODUCTION Materials and methods: An interventional study was carried out on 20 volunteers aged between 18 and 21 years. Antibacterial Dental caries still remains the most common dental efficacy was investigated by evaluating the bacterial counts problem to affect individuals across the globe. With the of salivary Streptococcus mutans and Lactobacilli before and gradual change in lifestyle trends and food habits, adults after the consumption of liquorice lollipops for 10 days while the and especially children are becoming more susceptible remineralization potential was studied with scanning electron to caries. Thus, there is a pressing need to develop new microscopy (SEM) analysis of the enamel block embedded on a Hawley’s appliance worn by volunteers for 14 days after natural and innovative antimicrobial agents for the consumption of the liquorice lollipop. treatment of dental caries.1,2 Herbal extracts in the form of leaves, fruits, flowers, seeds, roots, and barks have Results: A reduction in the bacterial counts of S. mutans which was statistically significant was observed. However, been studied by researchers and have shown to possess Lactobacillus growth was not observed before or after use of anti-inflammatory, antibiotic, analgesic, and antinocicep- liquorice lollipop. No structural or morphological changes were tive properties. Validation of the uses of these different observed on the enamel surface through SEM imaging. therapeutic modalities is particularly important to help Conclusion: Thus, liquorice lollipop was effective in reducing serve as a potent, safer, and cost-effective solution to S. mutans but did not show to possess a remineralizing potential. dental diseases.3 Clinical significance: The unique approach of a lollipop delivery Root extracts like ginger, garlic, turmeric, and system to reduce S. mutans count in children is also observed liquorice (mulethi) are traditionally used in India and have to be palatable for children. This raises hopes of a simple and been known to have an antibacterial potential. The anti- effective way to deliver a targeted intervention to young children bacterial efficacy of liquorice has been studied by various who are at risk for dental caries. researchers, but the effective mode of administration of Keywords: Adolescent, Antibacterial, Anticarious, Herbal, the extract by which the optimal antibacterial potential Liquorice, Remineralizing, Scanning electron microscope can be attained has not been established.4 Literature analysis. reveals that liquorice has an inhibitory effect on S. mutans How to cite this article: Krishnakumar G, Gaviappa D, and the modes of administration reported are either as Guruswamy S. Anticaries Efficacy of Liquorice Lollipop: An ex mouthwashes or through a lollipop delivery system.5,6 vivo Study. J Contemp Dent Pract 2018;19(8):937-942. The novel approach of incorporation of the liquorice extract with active compound Glycyrrhizol A into a lol- lipop delivery system had been devised by Hu et al,7 and 1-3 Department of Pedodontics and Preventive Dentistry, Faculty a reduction in salivary S. mutans levels postconsumption of Dental Sciences, M. S. Ramaiah University of Applied Sciences, Bengaluru, Karnataka, India of the liquorice lollipops has been observed and it has also been reported that the mode of delivery was well Corresponding Author: Gauri Krishnakumar, Department of Pedodontics and Preventive Dentistry, Faculty of Dental accepted by children. Sciences, M. S. Ramaiah University of Applied Sciences The potential of liquorice lollipop has been proven Bengaluru, Karnataka, India, Phone: +919650232720, e-mail: but has not yet been investigated in the Indian scenario. [email protected] Further, the remineralizing potential of liquorice, if The Journal of Contemporary Dental Practice, August 2018;19(8):937-942 937 Gauri Krishnakumar et al any, has also not been investigated. Thus, the scope of Assessment of Remineralizing Potential this study is to investigate the antibacterial efficacy of Twenty premolars extracted for orthodontic purpose were liquorice lollipops among volunteers and to investigate obtained from the Oral Surgery Department of the same if the extract has a remineralizing potential. The hypoth- institution. The radicular part of each tooth was removed esis is that there is no difference in the bacterial counts and the coronal part was then sectioned along the long of salivary S. mutans and Lactobacilli before and after the axis of the tooth into two portions of buccal and lingual consumption of liquorice, and there is no remineralizing using a high-speed diamond-tipped disk. Further, the tooth effect of liquorice on the enamel surface. was sectioned into three sections of occlusal, middle, and cervical third using the high-speed diamond-tipped disk. MATERIALS AND METHODS Four enamel specimens were prepared from each tooth and The study was carried out in the Department of only the buccal sections were selected for the study (Fig. 1). Pedodontics and Preventive Dentistry, M. S. Ramaiah Each enamel block was standardized to 3 × 2 mm in size. University of Applied Sciences, Bengaluru, Karnataka, The prepared enamel blocks were further subjected India, from March 2017 to September 2017. The volunteers to demineralization by suspending each section of the were selected from the undergraduate students of the enamel block into a glass bottle containing 20 mL of institution and were in the age group of 18 to 21 years. demineralization solution for 72 hours in an incubator at The exclusion criteria included adolescents who were a temperature of 35°C.8 The composition of the deminer- on any antibiotic regimen for the past 3 months, under- alizing solution used was CaCl2 = 2.2 mM, NaH2PO4 = going any orthodontic treatment, and if suffering from 2.2 mM, lactic acid = 0.05 M, fluoride = 0.2 ppm. Solution any systemic disease or condition. All volunteers of the was adjusted with 50% NaOH to a pH of 4.5.9 The pH was study had signed consent for the same. The sample size recorded each time with litmus paper before subjecting determination was done in accordance with a previous the enamel blocks to demineralization. study by Hu et al,7 power of 80%, error of 5%, dropout Maxillary and mandibular alginate impressions were of 10%, and confidence level of 5%. recorded in order to fabricate a Hawley’s appliance for each The liquorice lollipops were procured from the subject. Once the Hawley’s appliances were fabricated, a LOLOZTM Company who had fabricated the lollipops to window measuring 5 × 5 mm was prepared in the palatal contain 7 to 15 mg of the active compound, Glycyrrhizol A. aspect of the acrylic. The demineralized enamel blocks The genotoxicity and cytotoxicity of this liquorice lollipop were incorporated into this prepared window with cold had been earlier studied7 and is reported to be safe for cure acrylic (Fig. 2). The incorporated appliances were ster- children and adults. The recommended regimen is twice ilized by 3% ethylene oxide gas sterilizer. The subjects were daily for 10 days for an antibacterial effect. instructed to wear the appliance while consumption and 20 minutes postconsumption of the lollipop. When not Assessment of Antibacterial Efficacy worn, the Hawley’s appliance was stored in a plastic con- tainer. The liquorice lollipops were consumed for 14 days. Unstimulated saliva (2–2.5 mL) was collected for the Post the treatment regimen, the enamel blocks were sub- purpose of evaluating the antibacterial efficacy of jected to SEM analysis for evaluation of remineralization. liquorice extract. The samples were collected between 8 and 10 am in accordance with the circadian rhythm, and at least 1 hour after the consumption of any food or drink. The collected samples were transported to the laboratory within 3 to 4 hours for processing and estimation of the bacterial counts of S. mutans and Lactobacillus. Saliva samples were inoculated into mitis salivarius bacitracin (MSB agar) and Rogosa agar for specific bacterial culturing and identification of colony characteristics, and counting was done by digital colony counter. Saliva collection was performed twice during the study period. Saliva was first recorded at baseline before start of the liquorice regimen to evaluate the salivary bacterial status of the subject. The liquorice regimen was then followed for a period of 10 days. Similar procedure was performed again to evaluate the salivary bacterial status posttreatment. Fig. 1: Enamel block preparation 938 JCDP Anticaries Efficacy of Liquorice Lollipop Fig. 2: LOLOZ liquorice lollipops used in the study Fig. 3: Hawley’s appliance with the incorporated enamel blocks ETHICAL AND HUMANE CONSIDERATIONS The study was approved by the ethical review board University Ethics Committee for Human Trials of the university and is in accordance with the Declaration of Helsinki. The commercially produced liquorice lollipop was procured from LOLOZTM 601 SW 2nd Ave, Portland, OR (Fig. 3). Data were subjected to statistical analysis performed using Statistical Package for the Social Sciences software version 20.0. Paired Student’s