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. 2021 Nov 10;13(11):1907. doi: 10.3390/pharmaceutics13111907

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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(A) Representation of sample collection from infected dentin of patient. (B) Sample transferred to brain heart infusion (BHI) broth with 20% glycerol. (C) Separation of crown from the root of bovine tooth using a straight handpiece with a diamond disc. (D) Crown attached to a circular sample cutter with the buccal side facing upwards. (E) Tooth cut with a trephine drill. (F) Specimen inserted into a metal matrix for polishing. (G) Bovine specimen prepared. (H) Specimen positioned in the metal wire and submerged in BHI broth to form the biofilms. (I) Biofilm formed on bovine specimen. (J) Irradiation of biofilms with LED at 660 nm. (K) Colonies formed on BHI agar. (L) Scanning electron microscope. (M) Lactic dehydrogenase LDH UV K014 (Bioclin) to determine the acid production.