Research Article | Volume: 16, Issue: 7, July, 2026

Comparative Evaluation of Ultrasonic-activated Herbal and Inorganic Salt Irrigants Against Enterococcus faecalis Biofilms in Endodontic Disinfection: An In Vitro Study

Hemalatha Hiremath,  Anand C Patil,  Tarasha Gupta,  Vijay M Kumbar,  and Sayali Manjarekar   

Hemalatha Hiremath1, Anand C Patil1, Tarasha Gupta1, Vijay M Kumbar2, and Sayali Manjarekar2

1Department of Conservative Dentistry and Endodontics, KLE V. K. Institute of Dental Sciences, KLE Academy of Higher Education and Research (KAHER), Belagavi, Karnataka, India.

2Dr Prabhakar Kore’s Basic Science Research Laboratory, KLE Academy of Higher Education and Research (KAHER), Belagavi, Karnataka, India.

Open Access   

Published:  May 02, 2026

DOI: 10.1177/22313354261438283
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Abstract

Elimination of Enterococcus faecalis (E. faecalis) biofilms from the root canal system is a challenging issue in endodontics. The objective of this in vitro study was to evaluate the antibacterial efficacy of ultrasonic-activated herbal irrigants (black walnut [BW] and wormwood) and inorganic salts (alum and sodium chloride [NaCl]) against E. faecalis biofilms, compared with conventional irrigation. E. faecalis (MTCC 439) biofilms were grown on microtiter plates and treated with the test irrigants with and without ultrasonic activation for 5 and 10 minutes. Antibacterial efficacy was determined by Minimum Biofilm Eradication Concentration (MBEC) analysis using crystal violet staining and optical density measurement, and fluorescence microscopy using acridine orange and ethidium bromide staining to distinguish live and dead cells. All irrigants showed increased antibacterial activity when combined with ultrasonic activation (p < .05), with BW showing the highest biofilm reduction, followed by wormwood, alum, and NaCl. A 10-minute exposure time caused significantly greater biofilm disruption than 5 minutes. Ultrasonic activation dramatically enhances the antibacterial potential of herbal and inorganic irrigants, with BW, wormwood, and alum showing promise as adjuncts to conventional endodontic disinfection protocols.


Keyword:     Alum biofilm black walnut Enterococcus faecalis fluorescence microscopy herbal irrigants Inorganic salt MBEC NaCl ultrasonic irrigation wormwood

Citation:

Hiremath H, Patil AC, Gupta T, Kumbar VM, Manjarekar S. Comparative Evaluation of Ultrasonic-activated Herbal and Inorganic Salt Irrigants Against Enterococcus faecalis Biofilms in Endodontic Disinfection: An In Vitro Study. J Appl Pharm Sci. 2026;16(7):308-317. https://doi.org/10.1177/22313354261438283

Copyright: © The Author(s). This is an open-access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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