Enhancement of Antibiofilm Activity of Ciprofloxacin against Staphylococcus aureus by Administration of Antimicrobial Peptides

Staphylococcus aureus can develop resistance by mutation, transfection or biofilm formation. Resistance was induced in S. aureus by growth in sub-inhibitory concentrations of ciprofloxacin for 30 days. The ability of the antimicrobials to disrupt biofilms was determined using crystal violet and live/dead staining. Effects on the cell membranes of biofilm cells were evaluated by measuring release of dyes and ATP, and nucleic acids. None of the strains developed resistance to AMPs while only S. aureus ATCC 25923 developed resistance (128 times) to ciprofloxacin after 30 passages. Only peptides reduced biofilms of ciprofloxacin-resistant cells. The antibiofilm effect of melimine with ciprofloxacin was more (27%) than with melimine alone at 1X MIC (p < 0.001). Similarly, at 1X MIC the combination of Mel4 and ciprofloxacin produced more (48%) biofilm disruption than Mel4 alone (p < 0.001). Combinations of either of the peptides with ciprofloxacin at 2X MIC released ≥ 66 nM ATP, more than either peptide alone (p ≤ 0.005). At 2X MIC, only melimine in combination with ciprofloxacin released DNA/RNA which was three times more than that released by melimine alone (p = 0.043). These results suggest the potential use of melimine and Mel4 with conventional antibiotics for the treatment of S. aureus biofilms.

Aerosolized Hypertonic Saline Hinders Biofilm Formation to Enhance Antibiotic Susceptibility of Multidrug-Resistant Acinetobacter baumannii

Limited therapeutic options are available for multidrug-resistant Acinetobacter baumannii (MDR-AB), and the development of effective treatments is urgently needed. The efficacy of four aerosolized antibiotics (gentamicin, amikacin, imipenem, and meropenem) on three different MDR-AB strains was evaluated using hypertonic saline (HS, 7 g/100 mL) as the aerosol carrier. HS aerosol effectively hindered biofilm formation by specific MDR-AB strains. It could also interrupt the swarming dynamics of MDR-AB and the production of extracellular polymeric substances, which are essential for biofilm progression. Biofilms protect the microorganisms from antibiotics. The use of HS aerosol as a carrier resulted in a decreased tolerance to gentamicin and amikacin in the biofilm-rich MDR-AB. Moreover, we tested the aerosol characteristics of antibiotics mixed with HS and saline, and results showed that HS enhanced the inhaled delivery dose with a smaller particle size distribution of the four antibiotics. Our findings demonstrate the potential of using “old” antibiotics with our “new” aerosol carrier, and potentiate an alternative therapeutic strategy to eliminate MDR-AB infections from a biofilm-disruption perspective.

Photodynamic Therapy in Endodontics: A Helpful Tool to Combat Antibiotic Resistance? A Literature Review

Background: Antibiotic resistance has become a growing global problem where overprescription is a contributing factor for its development. In the endodontics field, complementary treatments, such as antimicrobial photodynamic therapy (aPDT), have been described to eliminate residual bacteria from the root canal space and reduce complications. The aim of this review is to describe the literature evidence up to now regarding the advantages, efficiency, and clinical outcomes of this therapy in endodontics as a possible tool to combat antibiotic resistance. Methods: A review of the literature from 2010 to 2021 was carried out using the PubMed and Web of Science databases. Two steps were taken: First, articles were compiled through the terms and MeSH terms “Photochesdmotherapy” and “endodontics.” Then, a second search was conducted using “photodynamic therapy” and “antibiotic resistance” or “drug resistance, microbial.” Results: A total of 51 articles were included for evaluation: 27 laboratory studies, 14 reviews, and 10 clinical studies. Laboratory studies show that aPDT achieves significant bacterial elimination, even against antibiotic-resistant species, and is also effective in biofilm disruption. Clinical studies suggest that aPDT can be considered a promising technique to reduce bacterial complications, and reviews about the issue confirm its advantages. Conclusion: The benefits of aPDT in reducing complications due to its antibacterial effects means a possible decrease in systemic antibiotic prescription in endodontics. In addition, it could be an alternative to local intracanal antibiotic therapy, avoiding the appearance of possible antibiotic resistance, as no bacterial resistance with aPDT has been described to date.

Enhancement of Antibiofilm Activity of Ciprofloxacin against Staphylococcus aureus by Administration of Antimicrobial Peptides

Staphylococcus aureus can develop resistance by mutation, tranfection or biofilm formation. Resistance was induced in S. aureus by growth in sub-inhibitory concentrations of ciprofloxacin for 30 days. The ability of the antimicrobials to disrupt biofilms was determined using crystal violet and live/dead staining. Effects on the cell membranes of biofilm cells was evaluated by measuring release of dyes and ATP and nucleic acids. S. aureus did not develop resistance to the AMPs but resistance increased to ciprofloxacin by 128 times after 30 passages. Only peptides reduced biofilms of ciprofloxacin resistant cells. The antibiofilm effect of melimine with ciprofloxacin was more (27%) than with melimine alone at 1X MIC (p &lt; 0.001). Similarly, at 1X MIC the combination of Mel4 and ciprofloxacin produced more (48%) biofilm disruption than Mel4 alone (p &lt; 0.001). Combinations of either of the peptides with ciprofloxacin at 2X MIC released  66 nM ATP, more than either peptide alone (p  0.005). At 2X MIC, only melimine in combination with ciprofloxacin released DNA/RNA which was 3 times more than released by melimine alone (p = 0.043). These results suggest the potential use of melimine and Mel4 with conventional antibiotics for the treatments of S. aureus biofilms.

Improving Phage-Biofilm In Vitro Experimentation

Bacteriophages or phages, the viruses of bacteria, are abundant components of most ecosystems, including those where bacteria predominantly occupy biofilm niches. Understanding the phage impact on bacterial biofilms therefore can be crucial toward understanding both phage and bacterial ecology. Here, we take a critical look at the study of bacteriophage interactions with bacterial biofilms as carried out in vitro, since these studies serve as bases of our ecological and therapeutic understanding of phage impacts on biofilms. We suggest that phage-biofilm in vitro experiments often may be improved in terms of both design and interpretation. Specific issues discussed include (a) not distinguishing control of new biofilm growth from removal of existing biofilm, (b) inadequate descriptions of phage titers, (c) artificially small overlying fluid volumes, (d) limited explorations of treatment dosing and duration, (e) only end-point rather than kinetic analyses, (f) importance of distinguishing phage enzymatic from phage bacteriolytic anti-biofilm activities, (g) limitations of biofilm biomass determinations, (h) free-phage interference with viable-count determinations, and (i) importance of experimental conditions. Toward bettering understanding of the ecology of bacteriophage-biofilm interactions, and of phage-mediated biofilm disruption, we discuss here these various issues as well as provide tips toward improving experiments and their reporting.

Mechanical biofilm disruption causes microbial and immunological shifts in periodontitis patients

Periodontitis is characterized by subgingival biofilm dysbiosis, inflammation and tissue destruction. Current treatment involves mechanical biofilm disruption known as non-surgical periodontal therapy (NSPT). This study sought to characterise the impact of treatment on microbial diversity and overall community, and the parallel impact on host inflammation in the oral cavity. Fourty-two periodontitis patients were included in this study, with periodontal clinical parameters, subgingival plaque and saliva samples collected at baseline and 90 days after treatment. Salivary cytokines were quantified, and subgingival plaque was analysed using 16S rRNA sequencing. After treatment, there were marked health-associated alterations in microbial composition and diversity, including differential abundance of 42 genera and 61 species. These changes were accompanied by substantial clinical improvement (pockets ≥ 5 mm, 27.50% to 9.00%, p < 0.001) and a decrease in salivary IL-1β (p < 0.001)—a putative marker of periodontal inflammation. Despite significant reductions in disease associated anaerobes, several genera (Fusobacterium, Prevotella, Tanenerella, Treponema) remained present and formed a distinct subnetwork associated with residual disease. Collectively, this study shows that current periodontal treatment results in partial restoration of a healthy microbial ecosystem, but features of biofilm dysbiosis and host inflammation remain in some patients, which were surprisingly independent of clinical response.

Effects of Octenidine on the Formation and Disruption of Dental Biofilms: An Exploratory In Situ Study in Healthy Subjects

Dental biofilms are highly structured, complex multispecies communities that, if left untreated, lead to severe oral complications such as caries and periodontal diseases. Therefore, antibiofilm agents are often recommended for both preventive and therapeutic measures. However, biofilm management can be challenging due to the low sensitivity of biofilms to antimicrobial treatments. Octenidine dihydrochloride (OCT) is a highly effective antibacterial agent. Because the OCT antibiofilm efficacy has not been studied in situ, this exploratory crossover study aimed to evaluate the effects of OCT mouth rinsing on biofilm formation and on the disruption of mature biofilms. Moreover, a comparison to the gold-standard chlorhexidine (CHX) was conducted. The biofilms were formed intraorally by 5 healthy volunteers on enamel specimens fixed to acrylic splints. For biofilm formation analysis, OCT, CHX, or water rinses were applied for 30 s every 12 h. The samples evaluation took place at 24-and 48-h time points. For biofilm disruption analysis, sample assessment was performed before and directly after the first OCT or CHX rinse on 48-h mature biofilms. A second rinse was carried out 12 h later. The last assessment was applied to 72-h mature biofilms. The biofilms were analyzed by fluorescence microscopy and transmission electron microscopy. The results showed OCT significantly reducing biofilm formation and bacterial vitality in situ. Simultaneously, the biofilm thickness was strongly decreased. Moreover, a single application of OCT to a 48-h mature biofilm induced substantial biofilm disruption. In addition, the efficacy of OCT compared favorably to CHX. These findings show that OCT rinses prevent biofilm formation and disrupt preexisting mature biofilms formed by healthy subjects. This work suggests that OCT might be used for dental biofilm management as a part of the medical treatment of oral diseases. Future studies with a larger subject heterogeneity and number are needed to confirm the observed OCT effects.