Estudos têm investigado o uso de scaffolds alternativos para Procedimentos Endodônticos Regenerativos (PERs), uma vez que ainda existem limitações técnicas significativas nos materiais atualmente empregados. O objetivo deste estudo in vitro foi avaliar a atividade antibiofilme de scaffolds tridimensionais de nanocelulose bacteriana (BNC) incorporados com 0,12% de digluconato de clorexidina (BNC/CHX), 1% de amoxicilina (BNC/AMOX) e 1% de clindamicina (BNC/CLI). Scaffolds de BNC sem agente antimicrobiano compuseram o grupo controle (BNC/C). Os scaffolds de BNC foram imersos em uma cultura multiespécies de E. faecalis, A. naeslundii e S. sanguinis, servindo como substrato para o crescimento do biofilme. Após 24 horas, 7 e 15 dias, determinou-se o valor médio de unidades formadoras de colônia (UFCs). A formação de biofilme multiespécies nos scaffolds de BNC foi avaliada por Microscopia Eletrônica de Varredura (MEV) e por Microscopia Confocal de Varredura a Laser (CLSM). Para o teste de viabilidade celular do biofilme, o valor médio de UFC/mL de cada amostra foi determinado e os dados foram normalizados pela transformação logaritmo de base 10 (log₁₀) de cada valor de UFC/mL. Os dados foram analisados pelos testes de ANOVA de dois fatores, seguida do teste de comparações múltiplas post-hoc de Bonferroni (α=5%). No período de 24h, o BNC/AMOX apresentou valores médios de UFCs significativamente menores do que BNC/CHX, BNC/CLI e BNC/C (p<0,05). Os valores médios de UFCs aumentaram ao longo dos períodos experimentais, sem diferença significativa em relação aos demais grupos (p>0,05). A análise por MEV e CLSM revelou, nos períodos iniciais, um número menor de microrganismos nos grupos experimentais em comparação ao grupo controle. Scaffolds de BNC contendo 1% de amoxicilina e 1% de clindamicina demonstraram maior atividade antimicrobiana contra o biofilme multiespécies nos períodos iniciais de incubação.
Autoria
person Fernanda Keil Kressin
schoolDepartment of Dentistry, Endodontics Division, University of Southern Santa Catarina, Palhoça, Santa Catarina, BrazilUniversity of Southern Santa CatarinaBrazilPalhoça, Santa Catarina, BrazilDepartment of Dentistry, Endodontics Division, University of Southern Santa Catarina, Palhoça, Santa Catarina, Brazil
person Isabela Aparecida Stahelin
schoolDepartment of Dentistry, Endodontics Division, University of Southern Santa Catarina, Palhoça, Santa Catarina, BrazilUniversity of Southern Santa CatarinaBrazilPalhoça, Santa Catarina, BrazilDepartment of Dentistry, Endodontics Division, University of Southern Santa Catarina, Palhoça, Santa Catarina, Brazil
person Gilmar da Rosa Souza Jr
schoolDepartment of Dentistry, Endodontics Division, University of Southern Santa Catarina, Palhoça, Santa Catarina, BrazilUniversity of Southern Santa CatarinaBrazilPalhoça, Santa Catarina, BrazilDepartment of Dentistry, Endodontics Division, University of Southern Santa Catarina, Palhoça, Santa Catarina, Brazil
person Karina Cesca
schoolDepartment of Chemical Engineering, Federal University of Santa Catarina, Florianópolis, Santa Catarina, BrazilFederal University of Santa CatarinaBrazilFlorianópolis, Santa Catarina, BrazilDepartment of Chemical Engineering, Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil
person Ricardo Ruiz Mazzon
schoolDepartment of Microbiology, Immunology and Parasitology, Federal University of Santa Catarina, Florianópolis, Santa Catarina, BrazilFederal University of Santa CatarinaBrazilFlorianópolis, Santa Catarina, BrazilDepartment of Microbiology, Immunology and Parasitology, Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil
person Rayssa Sabino-Silva
schoolDepartment of Dentistry, Endodontics Division, Health Sciences Center, Federal University of Santa Catarina, Florianópolis, Santa Catarina, BrazilFederal University of Santa CatarinaBrazilFlorianópolis, Santa Catarina, BrazilDepartment of Dentistry, Endodontics Division, Health Sciences Center, Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil
person Taynara Santos Goulart
schoolDepartment of Dentistry, Endodontics Division, Health Sciences Center, Federal University of Santa Catarina, Florianópolis, Santa Catarina, BrazilFederal University of Santa CatarinaBrazilFlorianópolis, Santa Catarina, BrazilDepartment of Dentistry, Endodontics Division, Health Sciences Center, Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil
person Cleonice da Silveira Teixeira
schoolDepartment of Dentistry, Endodontics Division, Health Sciences Center, Federal University of Santa Catarina, Florianópolis, Santa Catarina, BrazilFederal University of Santa CatarinaBrazilFlorianópolis, Santa Catarina, BrazilDepartment of Dentistry, Endodontics Division, Health Sciences Center, Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil
person Lucas da Fonseca Roberti Garcia
schoolDepartment of Dentistry, Endodontics Division, Health Sciences Center, Federal University of Santa Catarina, Florianópolis, Santa Catarina, BrazilFederal University of Santa CatarinaBrazilFlorianópolis, Santa Catarina, BrazilDepartment of Dentistry, Endodontics Division, Health Sciences Center, Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil
person Josiane de Almeida
schoolDepartment of Dentistry, Endodontics Division, University of Southern Santa Catarina, Palhoça, Santa Catarina, BrazilUniversity of Southern Santa CatarinaBrazilPalhoça, Santa Catarina, BrazilDepartment of Dentistry, Endodontics Division, University of Southern Santa Catarina, Palhoça, Santa Catarina, Brazil
Correspondence: Dr. Lucas da Fonseca Roberti Garcia - Departamento de Odontologia - Endodontia, Centro de Ciências da Saúde - Universidade Federal de Santa Catarina - Campus Reitor João David Ferreira Lima - Zip Code: 88040-900 - Florianópolis - Santa Catarina - Brazil. Phone +55 (48) 3721-5841. e-mail address:
drlucas.garcia@gmail.com
drlucas.garcia@gmail.com
Conflict of interest
The authors deny any conflicts of interest related to this study.
Responsible editor
Manoel Damião de Sousa-Neto
SCIMAGO INSTITUTIONS RANKINGS
Department of Dentistry, Endodontics Division, University of Southern Santa Catarina, Palhoça, Santa Catarina, BrazilUniversity of Southern Santa CatarinaBrazilPalhoça, Santa Catarina, BrazilDepartment of Dentistry, Endodontics Division, University of Southern Santa Catarina, Palhoça, Santa Catarina, Brazil
Department of Chemical Engineering, Federal University of Santa Catarina, Florianópolis, Santa Catarina, BrazilFederal University of Santa CatarinaBrazilFlorianópolis, Santa Catarina, BrazilDepartment of Chemical Engineering, Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil
Department of Microbiology, Immunology and Parasitology, Federal University of Santa Catarina, Florianópolis, Santa Catarina, BrazilFederal University of Santa CatarinaBrazilFlorianópolis, Santa Catarina, BrazilDepartment of Microbiology, Immunology and Parasitology, Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil
Department of Dentistry, Endodontics Division, Health Sciences Center, Federal University of Santa Catarina, Florianópolis, Santa Catarina, BrazilFederal University of Santa CatarinaBrazilFlorianópolis, Santa Catarina, BrazilDepartment of Dentistry, Endodontics Division, Health Sciences Center, Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil
Figuras | Tabelas
imageopen_in_new

imageFigure 1 Representative SEM images of the different BNC scaffolds after the experimental periods of incubation (24 hours, 7 and 15 days - left to right). BNC/C (A-C) Regular, homogeneous, and denser multispecies biofilm, with the extracellular matrix layer (*) covering the entire nanofiber network surface, regardless of the period of analysis. BNC/CHX (D and E) Isolated bacterial cells and small bacterial clusters were distributed on the surface of the nanofiber network in the initial periods of analysis. (F) Dense and homogeneous multispecies biofilm covering the nanofiber network surface in the 15-day period. BNC/CLI (G) Prevalence of rod-shaped microorganisms, characteristic of A. naeslundii, in the 24-hour period. (H and I) Note the presence of cocci-type microorganisms adhered to the scaffolds incorporated with clindamycin over the course of the experimental periods. BNC/AMOX (J) No microorganisms in the 24-hour period. (K and L) Note a more complex formation, with greater aggregation of microorganisms over the course of the experimental periods. Magnification of 5,000×. open_in_new

imageFigure 2 Representative CLSM images of the different BNC scaffolds after the experimental periods of incubation (24 hours, 7, and 15 days - left to right). Green colour (SYTO 9) indicates viable microbial cells, and red color (propidium iodide) indicates non-viable cells. BNC/C (A-C) shows substantial proportions of viable microorganisms adhered to the nanofiber network surface, particularly at the initial experimental periods, serving as the reference for comparison with the experimental groups. Compared with BNC/C, BNC/CHX (D-F), BNC/CLI (G-I), and BNC/AMOX (J-L) showed reduced viable microorganism counts at 24 hours and 7 days. At the 15-day period, a greater proportion of non-viable microorganisms (red) adhered to the BNC scaffolds was observed across all groups, regardless of the experimental condition. All bars represent 50 µm. open_in_new

table_chartTable 1
Mean values (log10) and standard deviation (SD) of CFUs/mL corresponding to viable bacteria adhered to the BCN scaffolds incorporated with the different antimicrobial agents at 24 hours, 7 days, and 15 days.
| Group | Log CFUs/mL ± SD | ||
|---|---|---|---|
| 24 hours | 7 days | 15 days | |
| BNC/C | 7.35 ± 0.22 a,A | 8.08 ± 0.04 ab,A | 8.63 ± 0.2 b,A |
| BNC/CHX | 7.04 ± 0.22 a,A | 8.08 ± 0.03 b,A | 8.49 ± 0.23 b,A |
| BNC/CLIN | 6.05 ± 0.21 a,B | 7.85 ± 0.04 a,B | 8.33 ± 0.24 b,A |
| BNC/AMOX | 4.04 ± 0.18 a,C | 7.9 ± 0.03 a,B | 8.41 ± 0.25 b,A |
-
Different lowercase letters in the same row and different uppercase letters in the same column indicate a significant difference. Two-way ANOVA and Bonferroni’s post-hoc multiple comparisons (α=5%).
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