Restor Dent Endod.  2021 May;46(2):e20. 10.5395/rde.2021.46.e20.

A novel antimicrobial-containing nanocellulose scaffold for regenerative endodontics

• Kichler V
• Teixeira LS
• Prado MM
• Colla G
• Schuldt DPV
• Coelho BS
• Porto LM
• Almeida Jd

• KMID: 2548064
• DOI: http://doi.org/10.5395/rde.2021.46.e20

Abstract

Objectives
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.
Materials and Methods
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%).
Results
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).
Conclusions
BNC/NO showed excellent biocompatibility and inhibited multispecies biofilm formation, similarly to BNC/CLI and BNC/CHX.

Keyword

Anti-infective agents; Biofilms; Regenerative endodontics; Tissue scaffolds

Figure

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  Figure 1 Percentage of periodontal ligament fibroblast (PDLF) metabolic activity after contact with the different membranes.BNC, bacterial nanocellulose; NO, nitric oxide; CLI, clindamycin; CHX, chlorhexidine.*Indicate significant differences compared to conventional BNC (control).

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  Figure 2 Mean colony-forming unit (CFU) values (log CFU/membrane) present in the multispecies biofilm that adhered to the different experimental bacterial nanocelluloses (BNCs).NO, nitric oxide; CLI, clindamycin; CHX, chlorhexidine.*Indicate significant differences compared to control (conventional BNC).

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  Figure 3 Dense and homogeneous bacterial biofilm covering the membrane surface in samples of the control group (conventional bacterial nanocellulose [BNC]) (magnification ×2,000 and ×4,000).

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  Figure 4 Isolated bacterial cells and small bacterial clusters, of varying sizes, irregularly distributed on the surface of the bacterial nanocellulose (BNC)/nitric oxide (NO) (A, B), BNC/clindamycin (CLI) (C, D), and BNC/chlorhexidine (CHX) (E, F) membranes (magnification ×2,000 and ×4,000).

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