Antimicrobial and immunomodulatory in vitro profile of double antibiotic paste

Abstract

Aim To evaluate the antimicrobial and immunomodulatory activity of double antibiotic paste (DAP) in an in vitro infection model. Methodology The minimum inhibitory and bactericidal concentrations (MIC and MBC) and the antibiofilm activities (TTC assay) of DAP and its components (ciprofloxacin and metronidazole) were evaluated against Staphylococcus aureus and Enterococcus faecalis compared with triple antibiotic paste (TAP). The cellular viability of RAW 264.7 macrophages (24 and 72 h) and L929 fibroblasts (48 and 72 h) was evaluated by MTT. Furthermore, the production of TNF‐α, IL‐12, IL‐6, IL‐1α, IL‐10 and NO (on RAW 264.7), besides IL‐6, TGF‐β and NO (on L929), stimulated with DAP in baseline and associated with heat‐killed microbial‐antigen conditions was measured by ELISA and Griess reaction. Data were analysed using the one‐way ANOVA test with Bonferroni's corrections. Results The MBC of pharmacopoeia DAP was similar to TAP for E. faecalis (0.25 μg. mL−1) and lower for S. aureus (DAP 1 μg. mL−1 and TAP 2 μg. mL−1; p < .001). Ciprofloxacin was the most effective antibiofilm drug from the pastes (35% of reduction for E. faecalis and S. aureus; p < .0001), and both pastes had a similar antibiofilm eradication against both biofilm species (29% and 35% for S. aureus and 76% and 85% for E. faecalis; p < .0001). DAP was cytotoxic against the tested cells. DAP significantly upregulated IL‐1α (p < .001), IL‐6 (p < .0001), TNF‐α (p < .01) and IL‐12 (p < .05; in the absence of antigens) and significantly reduced IL‐6 (p < .0001; in the presence of HK‐S. aureus) and IL‐10 (p < .05; in the presence of both antigens) on macrophages. Furthermore, DAP upregulated IL‐6 (p < .001) and NO (p < .05; in the absence of antigens), IL‐6 (p < .001; in the presence of HK‐S. aureus) and reduced NO (p < .001; in the presence of HK‐S. aureus). Conclusions Double antibiotic paste and TAP had similar antimicrobial activity against S. aureus and E. faecalis. DAP upregulated pro‐inflammatory cytokines mainly in the absence of antigens and had pro‐ and anti‐inflammatory activity in RAW 264.7 macrophages and L929 fibroblasts in the presence of antigens involved in pulp infections.

Antimicrobial potential of new diclofenac hydrogels for disinfection in regenerative endodontics: An in vitro and ex vivo study

“…Low concentrations of TAP and DAP reportedly have a similar capacity to reduce E. faecalis in vitro (Ferrer-Luque et al, 2021;Sabrah et al, 2015;Sousa et al, 2021). Whilst showing high clinical efficacy, TA has the disadvantage of inducing tooth discolouration due to minocycline (Ribeiro et al, 2020).…”

Association between host defense peptide IDR-1002 and ciprofloxacin: effects on human dental pulp cells

“…Among various peptides, IDR-1002 has demonstrated biotechnological potential for tissue engineering in endodontics [14]. A synergistic antimicrobial effect between the combination of IDR-1002 and cipro oxacin against E. faecalis and S. aureus was demonstrated [14]. This synergistic association presented interesting results not only in relation to the antimicrobial effect, but also interesting results relating to an immunomodulatory effect.…”

“…In addition, several studies have explored the immunomodulatory pro le of these pastes. Until now, researchers have shown that TAP and DAP have pro-in ammatory activity [13,14]. It is known that the immune response plays an essential role in the regeneration of pulp tissue in IPT [15].…”

“…Host defense peptides (HDPs) are biomolecules with antimicrobial capacity that also participate in the innate immune defense of several organisms [16]. The biotechnological potential of HDPs has been studied as a way of modulating the response to infections, pathologies, and tissue regeneration, in addition to having a broad spectrum against microorganism [14]. HDPs are promising agents in antibio lm and immunomodulatory strategies, factors of paramount importance in different areas of dentistry.…”

“…HDPs are promising agents in antibio lm and immunomodulatory strategies, factors of paramount importance in different areas of dentistry. Among various peptides, IDR-1002 has demonstrated biotechnological potential for tissue engineering in endodontics [14]. A synergistic antimicrobial effect between the combination of IDR-1002 and cipro oxacin against E. faecalis and S. aureus was demonstrated [14].…”