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Betulin inhibits cariogenic properties of Streptococcus mutans by targeting vicRK and gtf genes

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Abstract

Streptococcus mutans, a multivirulent pathogen is considered the primary etiological agent in dental caries. Development of antibiotic resistance in the pathogen has created a need for novel antagonistic agents which can control the virulence of the organism and reduce resistance development. The present study demonstrates the in vitro anti-virulence potential of betulin (lup-20(29)-ene-3β,28-diol), an abundantly available plant triterpenoid against S. mutans UA159. Betulin exhibited significant dose dependent antibiofilm activity without affecting bacterial viability. At 240 µg/ml (biofilm inhibitory concentration), betulin inhibited biofilm formation and adherence to smooth glass surfaces by 93 and 71 % respectively. It reduced water insoluble glucan synthesis by 89 %, in conjunction with down regulation of gtfBC genes. Microscopic analysis confirmed the disruption in biofilm architecture and decreased exopolysaccharide production. Acidogenicity and aciduricity, key virulence factors responsible for carious lesions, were also notably affected. The induced auto-aggregation of cells upon treatment could be due to the down regulation of vicK. Results of gene expression analysis demonstrated significant down-regulation of virulence genes upon betulin treatment. Furthermore, the nontoxic effect of betulin on peripheral blood mononuclear cells even after 72 h treatment makes it a strong candidate for assessing its suitability to be used as a therapeutic agent.

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Acknowledgments

The authors sincerely acknowledge Dr. K. Padima Devi and Mr. T. Rajavel of the Department of Biotechnology, Alagappa University for their help in performing cytotoxicity assay. The authors thank the Department of Biotechnology, Government of India for providing Bioinformatics Infrastructure Facility (Grant No. BT/BI/25/012/2012 (BIF)). The instrumentation facility provided by Department of Science and Technology, Government of India through PURSE [Grant No.SR/S9Z-23/2010/42(G)] & FIST (Grant No.SR-FST/LSI-087/2008) and University Grants Commission, New Delhi through SAP-DRS1 [Grant No.F.3-28/2011(SAP-II)] is gratefully acknowledged. Financial support provided to Dharmaprakash Viszwapriya in the form of Senior Research Fellowship (File. No. 09/688(0017)/2011-EMR-I) by Council of Scientific and Industrial Research (CSIR), New Delhi is thankfully acknowledged.

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  1. Department of Biotechnology, Alagappa University, II Floor, Science Campus, Karaikudi, Tamil Nadu, 630 004, India

    Dharmaprakash Viszwapriya, Ganapathy Ashwinkumar Subramenium, Solai Radhika & Shunmugiah Karutha Pandian

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  1. Dharmaprakash Viszwapriya
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  2. Ganapathy Ashwinkumar Subramenium
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Correspondence to Shunmugiah Karutha Pandian.

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Viszwapriya, D., Subramenium, G.A., Radhika, S. et al. Betulin inhibits cariogenic properties of Streptococcus mutans by targeting vicRK and gtf genes. Antonie van Leeuwenhoek 110, 153–165 (2017). https://doi.org/10.1007/s10482-016-0785-3

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