Prevention Of Bacterial Biofilm Formation Research Articles

The Antibacterial Properties of a Silver Multilayer Coating for the Prevention of Bacterial Biofilm Formation on Orthopedic Implants—An In Vitro Study

The prevention of biofilm formation on orthopedic implants is essential, as biofilms are the main challenge in the effective treatment of periprosthetic joint infection (PJI). A silver multilayer (SML) coating was developed to prevent biofilm formation on the implant surface. Previous studies have already demonstrated its antibacterial properties without cytotoxic effects. However, the coating has not been previously tested when applied to common titanium surfaces used in total joint arthroplasty implants. These surfaces often have increased roughness and porosity in the case of cementless implants, which can alter the antibacterial effect of the coating. In this study, we assessed the antibacterial and anti-biofilm properties of the SML coating on corundum-blasted and plasma-sprayed microporous-coated titanium alloy surfaces, using S. aureus, S. epidermidis, and E. coli. An antibacterial activity test following the principles of ISO 22196, ASTM E2180-18, and JIS Z 2801 standards was performed, as well as a biofilm proliferation assay investigating bacterial adhesion and biofilm formation. The SML coating exhibited strong antibacterial effects for all bacterial strains. After 24 h biofilm culture, a >4-log reduction in CFU was induced by the SML coating for S. epidermidis and E. coli on the corundum-blasted and plasma-sprayed microporous-coated titanium surfaces, respectively, when compared to the uncoated surfaces. The coating showed bactericidal properties against Gram-positive bacteria on the corundum-blasted discs. The SML coating on two common titanium surfaces demonstrates significant potential as an effective strategy in combating PJI across a wide range of orthopedic implants.

Prevention of bacterial biofilm formation on orthodontic brackets by non-crosslinked chitosan coating

During orthodontic treatment, the patients are susceptible to dental caries as a result of the bacterial adhesion and biofilm formation around the orthodontic brackets. Prevention of the caries-related biofilm formation is of significance for maintaining both aesthetics and health of the teeth. Herein, the brackets were functionalized with antibacterial activity via coating a layer of non-crosslinked chitosan (CS). We firstly demonstrated the ability of free CS scaffolds (not coated on brackets) to inhibit the formation of Streptococcus mutans biofilms (inhibition rate 94.3 % for CS-0.3 mg) and to eradicate the mature biofilms (biofilm loss rate 99.8 % for CS-1.2 mg). Further, the inhibition of S. mutans biofilm formation on brackets by CS coating was investigated for the first time. As a result, the CS-coated brackets (Br-CS) kept the great biofilm inhibition capacity of free CS scaffolds. In detail, the Br-CS, prepared by immersing brackets in CS solutions (containing 1.0, 2.5, 5.0 and 10 mg/mL CS) and freeze-drying, showed the biofilm inhibition rate of 48.5 %, 88.6 %, 96.4 % and 99.6 %, respectively. In conclusion, coating orthodontic brackets with the non-crosslinked CS is a potential approach for inhibiting biofilm formation and protecting patients from dental caries.

A multicenter, double-blind, placebo-controlled parallel study to evaluate the role of Yinhua Miyanling tablets in the prevention of bacterial biofilm formation on ureteral stents: a randomised trial.

Yinhua Miyanling tablet (YMT) not only has the functions of clearing away heat and toxin, dredging drenching and diuresis, but also has antibacterial activity. The formation of bacterial biofilm in ureteral stent and its related infection have plagued the clinic. Antibacterial traditional Chinese medicine is a potential method. This multicenter, randomized, double-blind, placebo-controlled study was designed to enroll patients who underwent ureteroscopic lithotripsy associated with indwelling ureteral stents at six centers between March 2019 and June 2020. The eligible patients were randomly assigned to the experimental group to take YMT 2 g qid orally or the control group to take dummy YMT 2 g qid orally from the first day after the operation according to a random number table. The unused drugs were recalled 14±3 days after the operation and record the body temperature. Relevant laboratory tests (urinalysis and urine culture) were performed before extubation. The ureteral stent was removed. The specimen was collected for scanning electron microscopy (SEM). Biofilm formation, USSQ scores, postoperative infectious complications, stone formation, and adverse drug reactions were compared between the two groups. Of the 211 patients enrolled, 165 were included in the per-protocol set (PPS), including 86 in the control group and 79 in the experimental group. No significant difference was found between the two groups in baseline parameters (P>0.05). The prevalence of biofilm formation in the control group (47%) was significantly higher than that in the experimental group (22.7%, P=0.001). There was no significant difference in total USSQ score and domain score between the two groups (all P>0.05). There were more patients with symptomatic urinary tract infection (UTI) in the control group (12.9%) than in the experimental group (2.6%, P=0.017). The incidence of other complications did not show a significant difference between the two groups (all P>0.05). The incidence of stone formation on the ureteral stent surface and adverse drug reactions did not show a significant difference between the two groups (all P>0.05). YMT is helpful to reduce the formation of bacterial biofilms on ureteral stents and the incidence of symptomatic UTIs related to ureteral stenting after surgery for ureteral calculi. Chinese Clinical Trial Registry ChiCTR2000041399.

Multiple Drug-Induced Stress Responses Inhibit Formation of Escherichia coli Biofilms.

In most ecosystems, bacteria exist primarily as structured surface-associated biofilms that can be highly tolerant to antibiotics and thus represent an important health issue. Here, we explored drug repurposing as a strategy to identify new antibiofilm compounds, screening over 1,000 compounds from the Prestwick Chemical Library of approved drugs for specific activities that prevent biofilm formation by Escherichia coli Most growth-inhibiting compounds, which include known antibacterial but also antiviral and other drugs, also reduced biofilm formation. However, we also identified several drugs that were biofilm inhibitory at doses where only a weak effect or no effect on planktonic growth could be observed. The activities of the most specific antibiofilm compounds were further characterized using gene expression analysis, proteomics, and microscopy. We observed that most of these drugs acted by repressing genes responsible for the production of curli, a major component of the E. coli biofilm matrix. This repression apparently occurred through the induction of several different stress responses, including DNA and cell wall damage, and homeostasis of divalent cations, demonstrating that biofilm formation can be inhibited through a variety of molecular mechanisms. One tested drug, tyloxapol, did not affect curli expression or cell growth but instead inhibited biofilm formation by suppressing bacterial attachment to the surface.IMPORTANCE The prevention of bacterial biofilm formation is one of the major current challenges in microbiology. Here, by systematically screening a large number of approved drugs for their ability to suppress biofilm formation by Escherichia coli, we identified a number of prospective antibiofilm compounds. We further demonstrated different mechanisms of action for individual compounds, from induction of replicative stress to disbalance of cation homeostasis to inhibition of bacterial attachment to the surface. Our work demonstrates the potential of drug repurposing for the prevention of bacterial biofilm formation and suggests that also for other bacteria, the activity spectrum of antibiofilm compounds is likely to be broad.

Articles of Significant Interest in This Issue

The clinical importance of bacterial biofilms has become increasingly appreciated, creating demand to identify new antibiofilm drugs. Teteneva et al. (e01113-20) used the drug repurposing strategy, screening the library of approved drugs to identify compounds that specifically suppress the formation of Escherichia coli biofilms. Proposed mechanisms of action of compounds range from induction of different stress responses to inhibition of surface attachment. This work demonstrates the potential of drug repurposing for the prevention of bacterial biofilm formation.

ε-Polylysine Nanoconjugates: Value-Added Antimicrobials for Drug-Resistant Bacteria.

Antimicrobial resistance poses a serious threat to human health and is evidently not restricted to any one part of the globe. Over the last few decades, no new antibiotics have been discovered, and many antibiotics currently available are failing against several critical pathogenic strains due to emerging drug resistance. We have designed a strategy to combat deadly drug-resistant bacteria by using nanocargos that consist of gold nanoparticles (AuNPs) conjugated to ε-polylysine (PLL) and octadecanethiol (C18) either alone or in combination. These nanocargos when tested against reference strains of carbapenem-resistant Acinetobacter baumannii (CRAB) and methicillin-resistant Staphylococcus aureus (MRSA) showed 15-20-fold higher antibacterial activity compared to free PLL. The minimum inhibitory concentration (MIC) of the nanoconjugates was found to lie between 8 and 15 μg/mL for both these bacteria, and they were also found to be nonhemolytic and nontoxic to mammalian cells. The mechanistic evaluation of antibacterial action showed alternate pathways of uptake for free PLL and the nanoconjugates. Further, the nanocargos were successfully used and found to be superior to free PLL in preventing biofilm formation in MRSA and CRAB. The PLL nanoconjugates may find applications in prevention of bacterial biofilm formation on surfaces such as surgical instruments and indwelling devices like stents, catheters, cannulas, orthopedic implants, and pacemakers.

Antimicrobial Activity and Prevention of Bacterial Biofilm Formation of Silver and Zinc Oxide Nanoparticle-Containing Polyester Surfaces at Various Concentrations for Use.

Food contact surfaces are primary sources of bacterial contamination in food industry processes. With the objective of preventing bacterial adhesion and biofilm formation on surfaces, this study evaluated the antimicrobial activity of silver (Ag-NPs) and zinc oxide (ZnO-NPs) nanoparticle-containing polyester surfaces (concentration range from 400 ppm to 850 ppm) using two kinds of bacteria, Gram-positive (Staphylococcus aureus) and Gram-negative (Escherichia coli), and the prevention of bacterial biofilm formation using the pathogen Listeria monocytogenes. The results of antimicrobial efficacy (reductions ≥ 2 log CFU/cm2) showed that at a concentration of 850 ppm, ZnO-NPs were effective against only E. coli (2.07 log CFU/cm2). However, a concentration of 400 ppm of Ag-NPs was effective against E. coli (4.90 log CFU/cm2) and S. aureus (3.84 log CFU/cm2). Furthermore, a combined concentration of 850 ppm Ag-NPs and 400 ppm ZnO-NPs showed high antimicrobial efficacy against E. coli (5.80 log CFU/cm2) and S. aureus (4.11 log CFU/cm2). The results also showed a high correlation between concentration levels and the bacterial activity of Ag–ZnO-NPs (R2 = 0.97 for S. aureus, and R2 = 0.99 for E. coli). They also showed that unlike individual action, the joint action of Ag-NPs and ZnO-NPs has high antimicrobial efficacy for both types of microorganisms. Moreover, Ag-NPs prevent the biofilm formation of L. monocytogenes in humid conditions of growth at concentrations of 500 ppm. Additional studies under different conditions are needed to test the durability of nanoparticle containing polyester surfaces with antimicrobial properties to optimize their use.

Prevention of Bacterial Biofilm Formation on Soft Contact Lenses Using Natural Compounds

PurposeIn eye care field, contact lenses (CL) have a great impact on improving vision, but their use can be limited by ocular infection. CL- associated infections can be reduced by good attention to CL storage case practice. CL-care solutions should be able to control microbial growth on CL.The aim of the study was to evaluate and compare the efficacy of CL-care solutions (found in Egyptian market) with some natural compounds in removal and inhibition of bacterial biofilm formed on soft CL.Clinical isolates were recovered from patients having conjunctivitis from Benha University Hospital and identified microbiologically. Quantification of biofilm was done using microtiter plate assay. Three multipurpose CL-care solutions were examined for their ability to remove and inhibit biofilm. Also four natural extracts having antibacterial activity and are safe on eye were tested for their anti-biofilm activity.ResultsThe major bacterial isolates from eye infections were Pseudomonas aeruginosa (36%) and Staphylococcus spp. (37.8%). Only 33.3% of isolates showed ability to produce weak to moderate biofilm. The tested multi-purpose CL-care solutions showed moderate ability to remove preformed biofilm. Among the tested natural compounds, Calendula officinalis and Buddleja salviifolia extracts showed an excellent efficacy in inhibition of biofilm and also removal of preformed biofilm.ConclusionThis study demonstrated that isolates from infected eye and CL-cases showed weak to moderate biofilm formation. Calendula officinalis and Buddleja salviifolia extracts showed excellent effect on inhibition and removal of biofilm, these extracts could be added into CL-care solutions which could markedly reduce eye-infections during CL-wear.

A small-molecule norspermidine and norspermidine-hosting polyelectrolyte coatings inhibit biofilm formation by multi-species wastewater culture.

Norspermidine is a potent and non-bactericidal small-molecule inhibitor of biofilm growth. In this study, impacts of norspermidine on biofilm control and existing biofilm dispersal by a mixed culture from wastewater treatment systems were investigated. A surface-mediated releasing approach for prevention of bacterial biofilm formation was established via encapsulating norspermidine into polyelectrolyte multilayer coatings. Results showed that the presence of norspermidine (500-1000μM) in medium remarkably prevented biofilm formation. Norspermidine was also effective in disassembling pre-formed biofilms. Norspermidine-containing multilayer coatings were successfully fabricated on glass slides via layer-by-layer deposition in polyethylenimine (PEI) and poly(acrylic acid) (PAA) solution. This coating exhibited a high anti-biofilm property against a mixed culture and three pure strains (Bacillus subtilis, Pseudomonas aeruginosa, and Escherichia coli). The loading amount and space distribution of norspermidine in the multilayer coating were key factors influencing its anti-biofilm efficacy. The polymer coating with norspermidine loaded in each bilayer (each-layer-type) exhibited better anti-biofilm efficacy than the bottom-type and the top-type coating, which showed a stable biofilm inhibition rate of about 60% even after 5-day leaching in aqueous solution. Norspermidine could retard bacterial adhesion and destruct biofilm matrix by reducing exopolysaccharides and extracellular DNA (eDNA) associated with bacteria instead of growth inhibition. Norspermidine and the norspermidine-hosting coatings in this study offer a great potential for the control of biofilms in the settings of water purification and wastewater treatment systems, which shows the advantage of broad spectrum and less risk of evolved bacterial resistance compared to conventional microbicidal agents (e.g., antibiotics).

New tryptophanase inhibitors: Towards prevention of bacterial biofilm formation

Tryptophanase (tryptophan indole-lyase, Tnase, EC 4.1.99.1), a bacterial enzyme with no counterpart in eukaryotic cells, produces from L-tryptophan pyruvate, ammonia and indole. It was recently suggested that indole signaling plays an important role in the stable maintenance of multicopy plasmids. In addition, Tnase was shown to be capable of binding Rcd, a short RNA molecule involved in resolution of plasmid multimers. Binding of Rcd increases the affinity of Tnase for tryptophan, and it was proposed that indole is involved in bacteria multiplication and biofilm formation. Biofilm-associated bacteria may cause serious infections, and biofilm contamination of equipment and food, may result in expensive consequences. Thus, optimal and specific factors that interact with Tnase can be used as a tool to study the role of this multifunctional enzyme as well as antibacterial agents that may affect biofilm formation. Most known quasi-substrates inhibit Tnase at the mM range. In the present work, the mode of Tnase inhibition by the following compounds and the corresponding Ki values were: S-phenylbenzoquinone-L-tryptophan, uncompetitively, 101 μM; α-amino-2-(9,10-anthraquinone)-propanoic acid, noncompetitively, 174 μM; L-tryptophane-ethylester, competitively, 52 μM; N-acetyl-L-tryptophan, noncompetitively, 48 μM. S-phenylbenzoquinone-L-tryptophan and α-amino-2-(9,10-anthraquinone)-propanoic acid were newly synthesized.

Prevention of biofilm formation by polymer modification

Bacterial biofilm formation on synthetic polymers plays an important role in industry and in modern medicine, leading, for example, to difficult-to-treat infections caused by colonized foreign bodies. Prevention of biofilm formation is a necessary step in the successful prophylaxis of such infections. One approach is to inhibit bacterial adherence by polymer surface modification. We have investigated polymer modification by glow discharge treatment in order to study the influence of the modified surface on bacterial adherence. Surface roughness, surface charge density and contact angles of the modified polymers were determined and related to the adherence of Staphylococcus epidermidis KH6. Although no influence of surface roughness and charge density on bacterial adherence was noticed, a correlation between the free enthalpy of adhesion (estimated from contact angle measurements) and adherence was observed. There seems to exist a certain minimum bacterial adherence, independent of the nature of the polymer surface. Modified polymers with negative surface charge allow for bacterial adherence close to the adherence minimum. These polymers could be improved further by the ionic bonding of silver ions to the surface. Such antimicrobial polymers are able to prevent bacterial colonization, which is a prerequisite for biofilm formation. It is suggested that modification of polymers and subsequent surface coupling of antimicrobials might be an effective approach for the prevention of bacterial biofilm formation.