Following root-end resection and filling bacterial colonization may lead to inflammatory reactions at the periapical tissues; the viability of the colonized bacteria may be affected by the type of root-end filling material.

Objectives: This work assesses different methods to interfere with Enterococcus faecalis biofilms formed on human dentin slabs.Methods: First, methods are presented that select for small molecule inhibitors of biofilm targets using multi-well polystyrene biofilm plates. Next, we establish methodologies to study and interfere with biofilm formation on a medically relevant model, whereby biofilms are grown on human root dentin slabs.Results: Non-conventional D-amino acid (D-Leucine) can efficiently disperse biofilms formed on dentin slabs without disturbing planktonic growth. Cation chelators interfere with biofilm formation on dentin slabs and polystyrene surfaces, and modestly impact planktonic growth. Strikingly, sodium hypochlorite, the treatment conventionally used to decontaminate infected root canal systems, was extremely toxic to planktonic bacteria, but did not eradicate biofilm cells. Instead, it induced a viable but non-culturable state in biofilm cells when grown on dentin slabs.Conclusion: Sodium hypochlorite may contribute to bacterial persistence. A combination of the methods described here can greatly contribute to the development of biofilm inhibitors and therapies to treat Enterococcus faecalis infections formed in the root canal system.

The combination of ethylenediaminetetraacetic acid (EDTA) and sodium hypochlorite (NaOCl) has been advocated as an effective irrigation methodology to remove organic and inorganic matter in root canal therapy. Yet, it was suggested that both solutions might lead to structural changes of the dentinal wall surface, depending on the order of application which might affect sealer mechanical retention. This study aims to evaluate the effect of different irrigating protocols on dentin surface roughness using quantitative 3D surface texture analysis. Data stems from 150 human root dentin sections, divided into five groups, each prepared according to one of the following protocols: Negative control; 17% EDTA; 17% EDTA followed by 5.25% NaOCl; 5.25% NaOCl; and 5.25% NaOCl followed by 17% EDTA. Each dentin sample was examined for its three-dimensional surface texture using a high-resolution confocal disc-scanning measuring system. EDTA 17% and the combined EDTA 17% with NaOCl 5.25% showed considerably higher roughness properties compared to the control and to NaOCl 5.25% alone. However, the irrigation sequence did not affect the dentin roughness properties. Therefore, mechanical retention is probably not dependent upon the selection of irrigation protocol sequence.

In this study, we evaluated the invasion of bacteria into the dentinal tubules of retrofilled extracted human teeth, and the influence of different fluorescently labeled retrograde filling materials on the bacterial invasion and viability, by means of confocal laser scanning microscopy (CLSM). The root apices of extracted teeth were cut, prepared, and filled retrogradely using either intermediate restorative material (IRM), mineral trioxide aggregate (MTA), or Biodentine. The roots were filled with Enterococcus faecalis bacteria from their coronal part for 21 days. Then, 3-mm-long apical segments were cut to get root axial slices, and the bacteria were fluorescently stained and evaluated by CLSM. Bacterial penetration into the dentinal tubules favored the bucco-lingual directions. The filling materials penetrated up to 957 µm into the tubuli, and the bacteria, up to 1480 µm (means: 130 and 167 μm, respectively). Biodentine fillings penetrated less and the associated bacteria penetrated deeper into the tubuli compared to MTA or IRM (p = 0.004). Deeper filling penetration was associated with shallower penetration of both dead and live, or live alone, bacteria (p = 0.015). In conclusion, the current study enables better understanding of the microbiological–pathological course after endodontic surgical procedures. It was found that even with retrograde fillings, bacteria invade deep into the dental tubules, where deeper filling penetration prevents deeper penetration of the bacteria and adversely affects the viability of the bacteria.