The aim of this study was to evaluate bacterial nanocellulose (BNC) membranes incorporated with antimicrobial agents regarding cytotoxicity in fibroblasts of the periodontal ligament (PDLF), antimicrobial activity, and inhibition of multispecies biofilm formation.

The tested BNC membranes were BNC + 1% clindamycin (BNC/CLI); BNC + 0.12% chlorhexidine (BNC/CHX); BNC + nitric oxide (BNC/NO); and conventional BNC (BNC; control). After PDLF culture, the BNC membranes were positioned in the wells and maintained for 24 hours. Cell viability was then evaluated using the MTS calorimetric test. Antimicrobial activity against Enterococcus faecalis, Actinomyces naeslundii, and Streptococcus sanguinis (S. sanguinis) was evaluated using the agar diffusion test. To assess the antibiofilm activity, BNC membranes were exposed for 24 hours to the mixed culture. After sonicating the BNC membranes to remove the remaining biofilm and plating the suspension on agar, the number of colony-forming units (CFU)/mL was determined. Data were analyzed by 1-way analysis of variance and the Tukey, Kruskal-Wallis, and Dunn tests (α = 5%).

PDLF metabolic activity after contact with BNC/CHX, BNC/CLI, and BNC/NO was 35%, 61% and 97%, respectively, compared to BNC. BNC/NO showed biocompatibility similar to that of BNC (p = 0.78). BNC/CLI showed the largest inhibition halos, and was superior to the other BNC membranes against S. sanguinis (p < 0.05). The experimental BNC membranes inhibited biofilm formation, with about a 3-fold log CFU reduction compared to BNC (p < 0.05).

BNC/NO showed excellent biocompatibility and inhibited multispecies biofilm formation, similarly to BNC/CLI and BNC/CHX.

The aim of this study was to evaluate discoloration of teeth undergoing regenerative endodontic procedures (REPs) using blood clot or platelet-rich fibrin (PRF) as the scaffolds and different calcium silicate-based materials as the intracanal coronal barriers in an ex vivo model.

Forty-eight bovine incisors were prepared and disinfected using 1 mg/mL double antibiotic paste (DAP). The specimens were then randomly divided into 2 groups (n = 24) according to the scaffolds (blood or PRF). After placement of scaffolds each group was divided into 2 subgroups (n = 12) according to the intracanal coronal barriers (ProRoot MTA or Biodentine). The pulp chamber walls were sealed with dentin bonding agent before placement of DAP and before placement of scaffolds. The color changes (∆E) were measured at different steps. The data were analyzed using 2-way analysis of variance.

Coronal discoloration induced by DAP was not clinically perceptible (ΔE ≤ 3.3). Regarding the type of the scaffold, coronal discoloration was significantly higher in blood groups compared with PRF groups at the end of REP and after 1 month (p < 0.05). However, no significant difference was found between PRF and blood clot after 6 months (p > 0.05). Considering the type of intracanal coronal barrier, no significant difference existed between ProRoot MTA and Biodentine (p > 0.05).

With sealing the dentinal tubules of pulp chamber with a dentin bonding agent and application of DAP as an intracanal medicament, coronal color change of the teeth following the use of PRF and blood sealed with either ProRoot MTA or Biodentine was not different at 6-month follow-up.

The biocompatibility of two experimental scaffolds for potential use in revascularization or pulp regeneration was evaluated.

One resilient lyophilized collagen scaffold (COLL), releasing metronidazole and clindamycin, was compared to an experimental injectable poly(lactic-co-glycolic) acid scaffold (PLGA), releasing clindamycin. Human dental pulp stem cells (hDPSCs) were seeded at densities of 1.0 × 10⁴, 2.5 × 10⁴, and 5.0 × 10⁴. The cells were investigated by light microscopy (cell morphology), MTT assay (cell proliferation) and a cytokine (IL-8) ELISA test (biocompatibility).

Under microscope, the morphology of cells coincubated for 7 days with the scaffolds appeared healthy with COLL. Cells in contact with PLGA showed signs of degeneration and apoptosis. MTT assay showed that at 5.0 × 10⁴ hDPSCs, COLL demonstrated significantly higher cell proliferation rates than cells in media only (control, p < 0.01) or cells co-incubated with PLGA (p < 0.01). In ELISA test, no significant differences were observed between cells with media only and COLL at 1, 3, and 6 days. Cells incubated with PLGA expressed significantly higher IL-8 than the control at all time points (p < 0.01) and compared to COLL after 1 and 3 days (p < 0.01).

The COLL showed superior biocompatibility and thus may be suitable for endodontic regeneration purposes.