Bacterial colonization in the apical part of extracted human teeth following root-end resection and filling: a confocal laser scanning microscopy study

Objectives: The purpose of this study was to evaluate Enterococcus faecalis colonization at the apical part of root canals following root-end resection and filling using confocal laser scanning microscopy (CLSM). Materials and methods: The apical 3-mm root-ends of 55 extracted single rooted human teeth were resected, and 3-mm retrograde cavities were prepared and filled using either mineral trioxide aggregate (MTA), intermediate restorative material (IRM), or Biodentine (n = 10 each); 25 teeth served as controls. The roots were placed in an experimental model, sterilized, and coronally filled with E. faecalis bacterial suspension for 21 days. Then, the apical 3-mm segments were cut to get two slabs (coronal and apical). The slabs were stained using LIVE/DEAD BacLight Bacterial Viability Kit and evaluated using CLSM. Results: The fluorescence-stained areas were larger in the bucco-lingual directions compared with the mesio-distal directions (p < 0.05). The mean and maximal depths of bacterial colonization into the dentinal tubules were 755 and 1643 μm, respectively, with no differences between the root-end filling materials (p > 0.05). However, more live bacteria were found in the MTA group in comparison to IRM and Biodentine groups (p < 0.05). Conclusions: CLSM can be used to histologically demonstrate bacterial root-end colonization following root-end filling. This colonization at the filling-dentine interfaces and deeper into the dentinal tubules may be inhomogeneous, favoring the bucco-lingual aspects of the root. Clinical relevance: 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.