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The accumulated dental biofilm can be a source of oral bacteria that are aspirated into the lower respiratory tract causing ventilator-associated pneumonia in hospitalized patients. The aim of this study was to evaluate the synergistic antibiofilm action of the produced and phytochemically characterized extracts of Cinnamomum verum and Brazilian green propolis (BGP) hydroethanolic extracts against multidrug-resistant clinical strains of Acinetobacter baumannii and Pseudomonas aeruginosa, in addition to their biocompatibility on human keratinocyte cell lines (HaCaT). For this, High-performance liquid chromatography analysis of the plant extracts was performed; then the minimum inhibitory and minimum bactericidal concentrations of the extracts were determined; and antibiofilm activity was evaluated with MTT assay to prevent biofilm formation and to reduce the mature biofilms. The cytotoxicity of the extracts was verified using the MTT colorimetric test, evaluating the cellular enzymatic activity. The data were analyzed with one-way ANOVA and Tukey’s tests as well as Kruskal–Wallis and Dunn’s tests, considering a significance level of 5%. It was possible to identify the cinnamic aldehyde in C. verum and p-coumaric, caffeic, and caffeoylquinic acids as well as flavonoids such as kaempferol and kaempferide and Artepillin-C in BGP. The combined extracts were effective in preventing biofilm formation and reducing the mature biofilms of A. baumannii and P. aeruginosa. Moreover, both extracts were biocompatible in different concentrations. Therefore, C. verum and BGP hydroethanolic extracts have bactericidal and antibiofilm action against multidrug resistant strains of A. baumannii and P. aeruginosa. In addition, the combined extracts were capable of expressively inhibiting the formation of A. baumannii and P. aeruginosa biofilms (prophylactic effect) acting similarly to 0.12% chlorhexidine gluconate.

Green propolis may represent a promising therapeutic alternative against dental anaerobic pathogens because of its antimicrobial action. This study aimed to evaluate the antimicrobial and antibiofilm actions of Brazilian green propolis aqueous extract (BGP-AqExt) against dental anaerobic bacteria. The minimum inhibitory concentration (MIC) and minimum microbicide concentration (MMC) of the extract were determined against the standard strains (ATCC) of Fusobacterium nucleatum, Parvimonas micra, Prevotella intermedia, Porphyromonas gingivalis and Porphyromonas endodontalis. BGP-AqExt was chemically characterized by high-performance liquid chromatography with diode-array detection (HPLC-DAD) analysis. Antibiofilm action was measured by MTT and crystal violet tests. The data were statistically analyzed by ANOVA and Tukey (5%) tests. The extract had antimicrobial action against all tested anaerobic bacteria, with an MIC value of 55 mg/mL for all bacteria, an MMC of 27.5 mg/mL for F. nucleatum and P. micra and 55 mg/mL for P. intermedia. Chemically, BGP-AqExt is composed of quercetin, gallic acid, caffeic and p-coumaric acid, drupani, kaempferol and Artepillin C. Significant reductions in biomass and metabolic action of biofilms were found after BGP-AqExt application. Therefore, BGP-AqExt has an antimicrobial and antibiofilm effect against dental anaerobic bacteria.

This study was performed to evaluate the biocompatibility and antifungal effect of Rosmarinus officinalis against Candida albicans in Galleria mellonella model. Five different concentrations of R. officinalis glycolic extract (50; 25; 12.5 e 6.25 mg/mL) were used to evaluate its biocompatibility in G. mellonella model, in which the nystatin suspension (100; 50; 25; 12.5 e 6.25%) was used as a control group. The antifungal action of R. officinalis glycolic extract was evaluated on C. albicans for 72, 48 and 12 h at two different phases: (1) using the extract as therapeutic agent; and (2) using the extract as prophylactic agent. PBS was used as a negative control group. G. mellonella survival curves were plotted using the Kaplan–Meier method and statistical analysis was performed using the log-rank test (Mantel–Cox) and the significance level was set at (α ≤ 0.05). There was no significant difference among the groups in which all were biocompatible except of a significant death rate of 26.6% with nystatin 100%. In phase 1, it was found that after 7 days, there was no statistically significant difference among the prophylactic treatment groups. In phase 2, the groups of R. officinalis 6.25 mg/mL for 72 h and R. officinalis of 12.5 mg/mL for 24 h promoted the survival rate of the larvae in comparison with the control group with a significant difference (p = 0.017) and (p = 0.032) respectively. Therefore, R. officinalis extract is biocompatible in different concentrations and can be used as a prophylactic agent against fungal infection.

Enterococcus faecalis and Enterococcus faecium are frequently associated with persistent endodontic infections due to their ability to form biofilms and resist conventional treatments. The aim of this study was to evaluate the antibacterial and antibiofilm effects of Lavandula dentata L. essential oil at 100% against the standard and clinical strains of Enterococcus spp. The minimum bactericidal concentration (MBC) of L. dentata essential oil for each bacterial strain was determined. The antibiofilm effect was evaluated by an MTT assay. Data were statistically analyzed by the ANOVA and Tukey test, with a significance level of 5%. The MBC value was 32% (292.8 mg/mL) against all the tested strains. L. dentata significantly reduced E. faecalis and E. faecium biofilms at 16–64% (146.4–585.6 mg/mL) concentrations, with effects comparable to 2% chlorhexidine (CHX) (p ≥ 0.05). Biofilm reduction was strain-dependent at 30 min but showed broader efficacy after 24 h, particularly against E. faecium clinical strains (p ≤ 0.001). L. dentata demonstrated effective antimicrobial activity against planktonic and biofilm forms of E. faecalis and E. faecium as it reduced biofilm formation at a concentration of 16% and 64% (146.4 and 585.6 mg/mL), with results comparable to 2% CHX after 30 min and 24 h. L. dentata EO may serve as a potential alternative or adjunctive antimicrobial agent in endodontic treatment. However, the study’s limitations included the in vitro design and lack of cytotoxicity assessment.

Staphylococcus aureus and Acinetobacter baumannii are opportunistic pathogens, and both are involved in different oral infections. This work aimed to analyze the phytochemical composition of Myrciaria cauliflora hydroethanolic extract and to evaluate its antimicrobial and antibiofilm action against Staphylococcus aureus (ATCC 6538) and Acinetobacter baumannii (ATCC 19606; multi-resistant clinical strains 58004, 50098, 566006, and H557). Myrciaria cauliflora hydroethanolic extract was prepared, and the content of soluble solids, flavonoids, and phenols was quantified. High-performance liquid chromatography (HPLC) was performed later. The minimum inhibitory concentration was determined using the broth microdilution method according to the Clinical and Laboratory Standards Institute, standard M7-A6, and subsequently, its minimum bactericidal concentration was determined. Then, the most effective concentrations were analyzed against biofilms. Statistical analysis was performed using the ANOVA method with Tukey’s test. The soluble solids content in the prepared hydroethanolic extract of M. cauliflora was 2.22%. Additionally, the total flavonoid content, measured using the quercetin standard curve, was 0.040 mg/mL. Furthermore, the total phenol content, determined using the gallic acid standard curve, was 0.729 mg/mL. HPLC analysis presented peaks of gallic acid (11.80 m), p-coumaric acid (12.09 m), cinnamic acid derivative (19.02 m), and ellagic acid (29.83 m). The extract demonstrated antimicrobial and antibiofilm action against all tested strains. However, the most effective antibacterial concentration against all the tested bacteria was 5.55 mg/mL. Therefore, these chemical components justify that M. cauliflora hydroethanolic extract is effective in reducing biofilm formation in S. aureus (standard strain) and A. baumannii (standard and clinical strains).

The World Health Organization (WHO) has prioritized developing new drugs against specific bacteria and fungi, such as Enterobacteriaceae and Candida spp. While Pfaffia paniculata is commonly called the “cure-everything”, its scientifically proven benefits are limited to anti-inflammatory and antioxidant actions. Therefore, this study aims to determine the spectrum of antimicrobial activity of Pfaffia paniculata and assess its cytotoxicity. Thus, broth microdilution test was conducted according to the CLSI M7-A9 and M27-A3 reference methods. After screening, microbial species with minimum inhibitory concentration (MIC) values were selected for biofilm tests. These tests evaluated biomass using the crystal violet (CV) test, metabolic activity using the MTT assay, and structural analysis via Scanning Electron Microscopy (SEM). Cytotoxicity was evaluated in human gingival fibroblasts (FMM-1). There were reductions of 29.4 and 42.7% in CV and MTT assays for Candida spp. biofilm. S. mutans and P. aeruginosa biofilms showed a decrease of 15.7 and 28.6%, respectively. Cell viability tests indicated 55.1, 56.9, and 65.5% of viability after contact with 1.93, 0.96, and 0.48 mg/mL of the extract, respectively. The P. paniculata extract showed antimicrobial action, displayed MIC values, and antibiofilm action on P. aeruginosa, S. mutans, and C. albicans. The cytotoxicity on the FMM-1 cell line was dose-dependent. Therefore, P. paniculata extract holds significant potential for developing new drugs.

ABSTRACT Cryptococcus neoformans is a yeast that mainly affects immunocompromised individuals and causes meningoencephalitis depending on the immune status of the host. The present study aimed to validate the efficacy of selective serotonin reuptake inhibitors, fluoxetine hydrochloride (FLH) and paroxetine hydrochloride (PAH), alone and in combination with amphotericin B (AmB) against C. neoformans. Susceptibility tests were conducted using the broth microdilution method and synergistic effects of combining FLH and PAH with AmB were analyzed using the checkerboard assay. Effects of minimum inhibitory concentration (MIC) and synergistic concentration were evaluated in biofilms by quantifying the biomass, measuring the viability by counting the colony-forming units (CFU/mL) and examining the size of the induced capsules. Cryptococcus neoformans was susceptible to FLH and PAH and the synergistic effect of FLH and PAH in combination with AmB reduced the MIC of AmB by up to 8-fold. The isolated substances and combination with AmB were able to reduce biofilm biomass and biofilm viability. In addition, FLH and PAH alone or in combination with AmB significantly decreased the size of the yeast capsules. Collectively, our results indicate the use of FLH and PAH as a promising prototype for the development of anti-cryptococcal drugs. Serotonin reuptake inhibitors, fluoxetine and paroxetine, are a promising prototype for the development of anti-cryptococcal drugs (The graphical abstract was designed using BioRender).

Objective To evaluate the chemical composition and effects of Artemisia vulgaris (AV) hydroalcoholic extract (HEAV) on breast cancer cells (MCF-7 and SKBR-3), chronic myeloid leukemia (K562) and NIH/3T3 fibroblasts. Methods Phytochemical analysis of HEAV was done by high-performance liquid chromatography-mass (HPLC) spectrometry. Viability and cell death studies were performed using trypan blue and Annexin/FITC-7AAD, respectively. Ferrostatin-1 (Fer-1) and necrostatin-1 (Nec-1) were used to assess the mode of HEAV-induced cell death and acetoxymethylester (BAPTA-AM) was used to verify the involvement of cytosolic calcium in this event. Cytosolic calcium measurements were made using Fura-2-AM. Results HEAV decreased the viability of MCF-7, SKBR-3 and K562 cells ( P <0.05). The viability of HEAV-treated K562 cells was reduced compared to HEAV-exposed fibroblasts ( P <0.05). Treatment of K562 cells with HEAV induced cell death primarily by late apoptosis and necrosis in assays using annexin V-FITC/7-AAD ( P <0.05). The use of Nec-1 and Fer-1 increased the viability of K562 cells treated with HEAV relative to cells exposed to HEAV alone ( P <0.01). HEAV-induced Ca 2+ release mainly from lysosomes in K562 cells ( P <0.01). Furthermore, BAPTA-AM, an intracellular Ca 2+ chelator, decreased the number of non-viable cells treated with HEAV ( P <0.05). Conclusions HEAV is cytotoxic and activates several modalities of cell death, which are partially dependent on lysosomal release of Ca 2+ . These effects may be related to artemisinin and caffeoylquinic acids, the main compounds identified in HEAV.

Enterococcus spp. are a common culprit behind the failure of endodontic treatments, primarily due to their notorious resistance to antimicrobial agents. Considering this challenge, this study was conducted to assess the antimicrobial efficacy of a unique blend of hydroalcoholic extracts sourced from Salvia officinalis and Glycyrrhiza glabra against biofilms formed by Enterococcus faecalis and Enterococcus faecium. The chemical composition of these plant extracts was rigorously characterized, with primary compound quantification achieved through high-performance liquid chromatography (HPLC-DAD) analysis. Additionally, this study determined the minimal bactericidal concentrations of these extracts and evaluated their potential to combat biofilms by quantifying colony-forming units per milliliter (CFU/mL). The findings reveal that the simultaneous application of both extracts yielded additive and synergistic effects against E. faecalis and E. faecium, including both ATCC and clinical strains. Impressively, after a 24 h exposure, these extract combinations demonstrated efficacy comparable to that of a 0.12% chlorhexidine solution, establishing a statistically significant difference from the negative control group. Consequently, the concurrent use of these extracts emerges as a promising alternative antimicrobial strategy for addressing Enterococcus spp. in endodontic treatments, holding substantial potential for clinical applications in this context.

The use of invertebrates for in vivo studies in microbiology is well established in the scientific community. Larvae of Galleria mellonella are a widely used model for studying pathogenesis, the efficacy of new antimicrobial compounds, and immune responses. The immune system of G. mellonella larvae is structurally and functionally similar to the innate immune response of mammals, which makes this model suitable for such studies. In this review, cellular responses (hemocytes activity: phagocytosis, nodulation, and encapsulation) and humoral responses (reactions or soluble molecules released in the hemolymph as antimicrobial peptides, melanization, clotting, free radical production, and primary immunization) are discussed, highlighting the use of G. mellonella as a model of immune response to different human pathogenic microorganisms.