Recent advances and future challenges in the use of nanoparticles for the dispersal of infectious biofilms

Increasing occurrence of intrinsically antimicrobial-resistant,human pathogens and the protective biofilm-mode in which they grow,dictates a need for the alternative control of infectious biofilms.Biofilm bacteria utilize dispersal mechanisms to detach parts of a biofilm as part of the biofilm life-cycle during times of nutrient scarcity or overpopulation.We here identify recent advances and future challenges in the development of dispersants as a new infection-control strategy.Deoxyribonuclease(DNase)and other extracellular enzymes can disrupt the extracellular matrix of a biofilm to cause dispersal.Also,a variety of small molecules,reactive oxygen species,nitric oxide releasing compounds,peptides and molecules regulating signaling pathways in biofilms have been described as dispersants.On their own,dispersants do not inhibit bacterial growth or kill bacterial pathogens.Both natural,as well as artificial dispersants,are unstable and hydrophobic which necessitate their encapsulation in smart nanocarriers,like pH-responsive micelles,liposomes or hydrogels.Depending on their composition,nanoparticles can also possess intrinsic dispersant properties.Bacteria dispersed from an infectious biofilm end up in the blood circulation where they are cleared by host immune cells.However,this sudden increase in bacte-rial concentration can also cause sepsis.Simultaneous antibiotic loading of nanoparticles with dispersant properties or combined administration of dispersants and antibiotics can counter this threat.Importantly,biofilm remaining after dispersant administration appears more susceptible to existing antibiotics.Being part of the natural biofilm life-cycle,no signs of"dispersant-resistance"have been observed.Dispersants are therewith promising for the control of infectious biofilms.     

Artificial Channels in an Infectious Biofilm Created by Magnetic Nanoparticles Enhanced Bacterial Killing by Antibiotics

The poor penetrability of many biofilms contributes to the recalcitrance of infectious biofilms to antimicrobial treatment. Here, a new application for the use of magnetic nanoparticles in nanomedicine to create artificial channels in infectious biofilms to enhance antimicrobial penetration and bacterial killing is proposed. Staphylococcus aureus biofilms are exposed to magnetic‐iron‐oxide nanoparticles (MIONPs), while magnetically forcing MIONP movement through the biofilm. Confocal laser scanning microscopy demonstrates artificial channel digging perpendicular to the substratum surface. Artificial channel digging significantly (4–6‐fold) enhances biofilm penetration and bacterial killing efficacy by gentamicin in two S. aureus strains with and without the ability to produce extracellular polymeric substances. Herewith, this work provides a simple, new, and easy way to enhance the eradication of infectious biofilms using MIONPs combined with clinically applied antibiotic therapies.     

Thermo-responsive copolymer coated MnFe2O4 magnetic nanoparticles for hyperthermia therapy and controlled drug delivery

This study is about multifunctional magnetic nanoparticles surface-modified with bilayer oleic acid, and coated with a thermo-responsive copolymer poly(N-isopropylacrylamide-co-acrylamide) by emulsion polymerization, for controlled drug delivery and magnetic hyperthermia applications. Nanoparticles were loaded with anticancer drug doxorubicin into the copolymer chains at 25 °C. Composite nanoparticles (hydrated) of average diameter 45 nm were of core–shell structure having magnetic core of about 18 nm and shell was composed of organic compounds and water. Magnetic core was superparamagnetic lacking coercive force and remanance due to the pseudo-single domain nanostructure. Lower critical solution temperature (LCST) of the thermo-responsive copolymer was observed to be around 39 °C. Below this temperature, copolymer was hydrophilic, hydrated and swelled. But above LCST, copolymer became hydrophobic, dehydrated and shrank in volume. UV visible spectrophotometer was used to investigate the drug loading and releasing profile at different temperatures as well as under magnetic heating. There was almost absence of drug release at around 37 °C (normal body temperature). Drug was released at temperatures above LCST, which is significant for controlled drug delivery. Magnetic heat-generation was studied by exposing the magnetic fluid to alternating magnetic field of 7.2 kA m⁻¹ having frequency 70 kHz. A simple magnetic capturing system (simulating a blood vessel) was used to analyze the capturing of magnetic nanoparticles under various applied fields for drug targeting purpose.     

Mechanical robustness of Pseudomonas aeruginosa biofilms

Biofilms grow on various surfaces and in many different environments, a phenomenon that constitutes major problems in industry and medicine. Despite their importance little is known about the viscoelastic properties of biofilms and how these depend on the chemical microenvironment. Here, we find that the mechanical properties of Pseudomonas aeruginosa (P.a.) biofilms are highly robust towards chemical perturbations. Specifically, we observe that P.a. biofilms are able to fully regain their initial stiffness after yielding is enforced, even in the presence of chemicals. Moreover, only trivalent ions and citric acid significantly affect the biofilm elasticity, the first of which also alter the texture of the material. Finally, our results indicate that biofilm mechanics and bacteria viability inside the biofilm are not necessarily linked which suggests that targeting bacteria alone might not be sufficient for biofilm removal strategies.     

Recent Nanotechnologies to Overcome the Bacterial Biofilm Matrix Barriers

Bacterial biofilm-related infectious diseases severely influence human health. Under typical situations, pathogens can colonize inert or biological surfaces and form biofilms. Biofilms are functional aggregates that coat bacteria with extracellular polymeric substances (EPS). The main reason for the failure of biofilm infection treatment is the low permeability and enrichment of therapeutic agents within the biofilm, which results from the particular features of biofilm matrix barriers such as negatively charged biofilm components and highly viscous compact EPS structures. Hence, developing novel therapeutic strategies with enhanced biofilm penetrability is crucial. Herein, the current progress of nanotechnology methods to improve therapeutic agents’ penetrability against biofilm matrix, such as regulating material morphology and surface properties, utilizing the physical penetration of nano/micromotors or microneedle patches, and equipping nanoparticles with EPS degradation enzymes or signal molecules, is first summarized. Finally, the challenges, perspectives, and future implementations of engineered delivery systems to manage biofilm infections are presented in detail.     

In vitro evaluation of antimicrobial and antibiofilm potentials of silver nanoparticles biosynthesised by Streptomyces griseorubens

This study was performed to determine the antimicrobial and antibiofilm activities of silver nanoparticles (AgNPs) biosynthesised using Streptomyces griseorubens AU2 isolated from soil. The antimicrobial activity of the AgNPs was determined by agar well diffusion, disc diffusion and broth microdilution methods. Diameters of the zone of inhibition results clearly displayed that the microbially biosynthesised AgNPs have potent antimicrobial activity against Candida albicans, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. The minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) of the nanoparticles that had been determined by broth microdilution method were found to be 20 and 50 µg/ml for C. albicans, B. subtilis and S. aureus; 10 and 20 µg/ml for E. coli and P. aeruginosa, respectively. For determining the effect of AgNPs on biofilm formation under in vitro conditions, MIC and subMICs were studied on P. aeruginosa and S. aureus biofilms by using microplate biofilm assay. Treatment of the AgNPs resulted in a decrease in the biofilm formation of S. aureus and P. aeruginosa as 26.52 and 25.50%, respectively. As a result of this study, it can be suggested that actinobacterially synthesised AgNPs have an effective potential to be used for pharmaceutical applications against multi‐resistant microorganisms.Inspec keywords: silver, nanoparticles, nanomedicine, antibacterial activity, biomedical materials, microorganismsOther keywords: antimicrobial potentials, antibiofilm potentials, silver nanoparticles, antimicrobial activity, antibiofilm activity, Streptomyces griseorubens AU2, disc diffusion, microdilution method, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, nanoparticle minimum inhibitory concentration, nanoparticle minimum lethal concentration, biofilm formation, in vitro conditions, microplate biofilm assay, pharmaceutical applications, multiresistant microorganisms, Ag     

Towards a versatile platform based on magnetic nanoparticles for in vivo applications

Magnetic nanoparticles have attracted wide attention because of their usefulness as contrast agents for magnetic resonance imaging (MRI) or colloidal mediators for cancer magnetic hyperthermia. This paper examines these in vivo applications through an understanding of the problems involved and the current and future possibilities for resolving them. A special emphasis is made on magnetic nanoparticle requirements from a physical viewpoint, the factors affecting their biodistribution and the solutions envisaged for enhancing their half-life in the blood compartment and targeting tumour cells. Then, our synthesis strategies are presented and focused on covalent platforms based on maghemite and dextran and capable to be tailorderivatized by surface molecular chemistry. The opportunity of taking advantage of temperature-dependence of magnetic properties of some complex oxides for controlling the in vivo temperature is also discussed.     

Nanomaterial-based strategies in antimicrobial applications: Progress and perspectives

The dramatic increase of microbial resistances against conventional available antibiotics is a huge challenge to the effective treatment of infectious disease and thus becoming a daunting global threat of major concern, which necessitates the development of innovative therapeutics. Nanomaterial-based antimicrobial strategies have emerged as novel and promising tools to combat lethal bacteria and recalcitrant biofilm, featuring the abilities to evade existing drug resistance-related mechanisms. In this review, recent advances in “state-of-the-art” nanosystems which acting either as inherent therapeutics or nanocarriers for the precise delivery of antibiotics, are comprehensively summarized. Those nanosystems can effectively accumulate at the infectious sites, achieve multifunctional synergistic antibacterial efficacy, and provide controlled release of antibiotics in response to endogenous or exogenous stimulus (e.g., low pH, enzymes, or illumination). Especially, the nanoplatform that integrated with photothermal/photodynamic therapy (PTT/PDT) can enhance the bacterial destruction and biofilm penetration or ablation. In addition, nanoparticle-based approaches with enzymatically promoting bacterial killing, anti-virulence, and other mechanisms were also involved. Overall, this review provides crucial insights into the recent progress and remaining limitations of various antimicrobial nanotherapeutic strategies, and enlightens the further developments in this field simultaneously, which eventually benefiting public health.

     

Living cells of Staphylococcus aureus immobilized onto the capillary surface in electrochromatography: a tool for screening of biofilms

Microorganisms attach to nonliving surfaces in many natural, industrial, and medical environments, enveloped within extracellular polymeric substances. The result is a biofilm. Biofilms are reported to exist in 65-80% of bacterial infections refractory to host defenses and antibiotics therapy and are regarded as a central problem in present-day medical microbiology. Understanding of the parameters governing the interaction of antimicrobials with biofilms is thus of great interest in any attempt to increase biocide efficacy. In this work, study was made of the feasibility of using open tubular capillary electrochromatography (CEC) in bacterial biofilm studies with living cells. Staphylococcus aureus was selected as model bacterium. First, S. aureus was shown, under various conditions, to form a biofilm on the inner wall of a fused-silica capillary coated with poly(L-lysine). Optimal conditions for biofilm formation, such as bacterial concentration, growing time, and the stability of the ensemble, were preliminarily defined with conventional 96-microtiter well plates. Continuous flushing of the capillary with fresh cells meant that no growth medium was needed. The presence of biofilm in the capillary was confirmed by atomic force microscopy. Interactions between S. aureus biofilms and different antibiomicrobial agents were studied by capillary electrochromatography. The effect of five antibiotics (penicillin G, oxacillin, fusidic acid, rifampicin, vancomycin) on biofilms was examined in terms of retention factors and reduced mobilities of the antibiotics. The antibiotic susceptibility profile for S. aureus is similar as the result of minimal inhibitory concentrations registered on the 96-microtiter well plates for both planktonic and biofilm cells. The results show, for the first time, that bacterial biofilms can be studied by CEC. The technique allows highly efficient and easy characterization of interactions between S. aureus biofilms and potentially active antimicrobial compounds under different conditions. Reagent and cell consumption are minimal.     

Ferrofluids and magnetically guided superparamagnetic particles in flows: a review of simulations and modeling

Ferrofluids are typically suspensions of magnetite nanoparticles, and behave as a homogeneous continuum. The ability of the ferrofluid to respond to an external magnetic field in a controllable manner has made it emerge as a smart material in a variety of applications, such as seals, lubricants, electronics cooling, shock absorbers and adaptive optics. Magnetic nanoparticle suspensions have also gained attraction recently in a range of biomedical applications, such as cell separation, hyperthermia, MRI, drug targeting and cancer diagnosis. In this review, we provide an introduction to mathematical modeling of three problems: motion of superparamagnetic nanoparticles in magnetic drug targeting, the motion of a ferrofluid drop consisting of chemically bound nanoparticles without a carrier fluid, and the breakage of a thin film of a ferrofluid.     

磁性纳米颗粒介导的电磁神经治疗的最新进展

目的 介绍磁性纳米颗粒的性质和生物医学应用,以及通过磁性纳米颗粒介导的电磁神经刺激治疗的最新进展.为今后优化刺激参数、提高磁神经刺激效率提供参考.方法 总结近年来国内外对磁性纳米颗粒的研究进展,并重点分析基于磁性纳米颗粒的神经磁刺激方法及效果.结果 磁性纳米颗粒具有成像、靶向给药、磁热疗等生物医学应用,以磁性纳米颗粒为基础进行神经磁刺激的类型可分为磁热刺激、磁电刺激及磁机械力刺激三种.这种刺激方式安全、高效且精准性高,能够改善传统磁刺激方式的缺陷.结论 磁性纳米颗粒性质独特,是近年来研究最多、发展速度最快的纳米材料之一.利用磁性纳米颗粒介导的神经磁刺激具有广阔的应用前景.     

Experimental and mathematical modelling of magnetically labelled mesenchymal stromal cell delivery

A key challenge for stem cell therapies is the delivery of therapeutic cells to the repair site. Magnetic targeting has been proposed as a platform for defining clinical sites of delivery more effectively. In this paper, we use a combined in vitro experimental and mathematical modelling approach to explore the magnetic targeting of mesenchymal stromal cells (MSCs) labelled with magnetic nanoparticles using an external magnet. This study aims to (i) demonstrate the potential of magnetic tagging for MSC delivery, (ii) examine the effect of red blood cells (RBCs) on MSC capture efficacy and (iii) highlight how mathematical models can provide both insight into mechanics of therapy and predictions about cell targeting in vivo. In vitro MSCs are cultured with magnetic nanoparticles and circulated with RBCs over an external magnet. Cell capture efficacy is measured for varying magnetic field strengths and RBC percentages. We use a 2D continuum mathematical model to represent the flow of magnetically tagged MSCs with RBCs. Numerical simulations demonstrate qualitative agreement with experimental results showing better capture with stronger magnetic fields and lower levels of RBCs. We additionally exploit the mathematical model to make hypotheses about the role of extravasation and identify future in vitro experiments to quantify this effect.     

Control of bacterial biofilm growth on surfaces by nanostructural mechanics and geometry

Surface-associated communities of bacteria, called biofilms, pervade natural and anthropogenic environments. Mature biofilms are resistant to a wide range of antimicrobial treatments and therefore pose persistent pathogenic threats. The use of surface chemistry to inhibit biofilm growth has been found to only transiently affect initial attachment. In this work, we investigate the tunable effects of physical surface properties, including high-aspect-ratio (HAR) surface nanostructure arrays recently reported to induce long-range spontaneous spatial patterning of bacteria on the surface. The functional parameters and length scale regimes that control such artificial patterning for the rod-shaped pathogenic species Pseudomonas aeruginosa are elucidated through a combinatorial approach. We further report a crossover regime of biofilm growth on a HAR nanostructured surface versus the nanostructure effective stiffness. When the 'softness' of the hair-like nanoarray is increased beyond a threshold value, biofilm growth is inhibited as compared to a flat control surface. This result is consistent with the mechanoselective adhesion of bacteria to surfaces. Therefore by combining nanoarray-induced bacterial patterning and modulating the effective stiffness of the nanoarray--thus mimicking an extremely compliant flat surface--bacterial mechanoselective adhesion can be exploited to control and inhibit biofilm growth.     

Dynamic Sessile‐Droplet Habitats for Controllable Cultivation of Bacterial Biofilm

Bacterial biofilms play essential roles in biogeochemical cycling, degradation of environmental pollutants, infection diseases, and maintenance of host health. The lack of quantitative methods for growing and characterizing biofilms remains a major challenge in understanding biofilm development. In this study, a dynamic sessile‐droplet habitat is introduced, a simple method which cultivates biofilms on micropatterns with diameters of tens to hundreds of micrometers in a microfluidic channel. Nanoliter plugs are utilized, spaced by immiscible carrier oil to initiate and support the growth of an array of biofilms, anchored on and spatially confined to the micropatterns arranged on the bottom surface of the microchannel, while planktonic or dispersal cells are flushed away by shear force of aqueous plugs. The performance of the aforementioned method of cultivating biofilms is demonstrated by Pseudomonas aeruginosa PAO1 and its derived mutants, and quantitative antimicrobial susceptibility testing of PAO1 biofilms. This method could significantly eliminate corner effects, avoid microchannel clogging, and constrain the growth of biofilms for long‐term observations. The controllable sessile droplet‐based biofilm cultivation presented in this study should shed light on more quantitative and long‐term studies of biofilms, and open new avenues for investigation of biofilm attachment, growth, expansion, and eradication.     

Bacterial inactivation using silver-coated magnetic nanoparticles as functional antimicrobial agents

The ability for silver nanoparticles to function as an antibacterial agent while being separable from the target fluids is important for bacterial inactivation in biological fluids. This report describes the analysis of the antimicrobial activities of silver-coated magnetic nanoparticles synthesized by wet chemical methods. The bacterial inactivation of several types of bacteria was analyzed, including Gram-positive bacteria ( Staphylococcus aureus and Bacillus cereus ) and Gram-negative bacteria ( Pseudomonas aeruginosa , Enterobacter cloacae , and Escherichia coli ). The results have demonstrated the viability of the silver-coated magnetic nanoparticles for achieving effective bacterial inactivation efficiency comparable to and better than that of silver nanoparticles conventionally used. The bacteria inactivation efficiency of our silver-coated MnZn ferrite (MZF@Ag) nanoparticles was also determined for blood platelets samples, demonstrating the potential of utilization in inactivating bacterial growth in platelets prior to transfusion to ensure blood product safety, which also has important implications for enabling the capability of effective separation, delivery, and targeting of the antibacterial agents.     

Composite polymeric magnetic nanoparticles for co-delivery of hydrophobic and hydrophilic anticancer drugs and MRI imaging for cancer therapy

Exercising complementary roles of polymer-coated magnetic nanoparticles for precise drug delivery and image contrast agents has attracted significant attention in biomedical applications. The objective of this study was to prepare and characterize magnetic nanoparticles embedded in polylactide-co-glycolide matrixes (PLGA-MNPs) as a dual drug delivery and imaging system capable of encapsulating both hydrophilic and hydrophobic drugs. PLGA-MNPs were capable of encapsulating both hydrophobic and hydrophilic drugs in a 2:1 ratio. Biocompatibility, cellular uptake, cytotoxicity, membrane potential, and apoptosis were carried out in two different cancer cell lines (MCF-7 and PANC-1). The molecular basis of induction of apoptosis was validated by Western blotting analysis. For targeted delivery of drugs, targeting ligand such as Herceptin was used, and such a conjugated system demonstrated enhanced cellular uptake and an augmented synergistic effect in an in vitro system when compared with native drugs. Magnetic resonance imaging was carried out both in vitro and in vivo to assess the efficacy of PLGA-MNPs as contrast agents. PLGA-MNPs showed a better contrast effect than commercial contrast agents due to higher T(2) relaxivity with a blood circulation half-life ～ 47 min in the rat model. Thus, our results demonstrated the dual usable purpose of formulated PLGA-MNPs toward either, in therapeutics by delivering different hydrophobic or hydrophilic drugs individually or in combination and imaging for cancer therapeutics in the near future.     

Stable water-soluble iron oxide nanoparticles using Tiron

Success in biological and nanomaterial applications that rely on magnetic iron oxide nanoparticles (IONPs) often depends on monodispersity, size, and aqueous stability of the synthesized particles. Here we report a simple and efficient strategy to prepare monodisperse, ultrasmall, water dispersible superparamagnetic IONPs. Monodisperse IONPs are initially synthesized in organic solvents using oleic acid as a dispersant. The subsequent ligand exchange of oleic acid for dopamine and Tiron (4,5-dihydroxy-1,3-benzenedisulfonic acid disodium salt) allows for superior colloidal stability in aqueous media. Zeta potential measurements confirm the stability of the nanoparticles upon redispersal in water or biologically relevant buffers. The synthesized particles also preserve their general shape, size, and crystallinity after ligand exchange as evidenced by TEM and SAED measurements. Magnetic properties are also maintained after the ligand exchange as verified by magnetometry and magnetic force microscopy (MFM). An analysis of potential issues regarding this and other prior ligand exchanges is also highlighted, which may aid others in future investigations.     

Detection of C-reactive protein based on immunoassay using antibody-conjugated magnetic nanoparticles

We report a detection method for C-reactive protein (CRP) based on competitive immunoassay using magnetic nanoparticles under magnetic fields. Functional magnetic nanoparticles were prepared and conjugated with anti-CRP for immunoassay. Magnetic nanoparticles labeled with anti-CRP were flowed through a separation channel to form depositions for selective capture of CRP under magnetic fields. Free CRP and a fixed number of CRP-labeled particles were used to compete for a limited number of anti-CRP binding sites on the magnetic nanoparticles. The deposited percentages of CRP-labeled particles at various concentrations of free CRP were determined and used as a reference plot. The determination of CRP in the unknown sample was deduced from the reference plot using the deposited percentages. The running time was less than 10 min. The CRP concentration of serum sample was linearly over the range of 1.2-310 microg/mL for deposited percentages of CRP-labeled particles. The detection limit of this method was 0.12 microg/mL which was approximately 8-fold lower than the typical clinical cutoff concentration (1 microug/mL). This method can provide a fast, simple, and sensitive way for protein detection based on competitive immunoassay using magnetic nanoparticles under magnetic fields.     

In vitro study on infectious ureteral encrustation resistance of Cu-bearing stainless steel

Cu-bearing stainless steel has been found to have obvious inhibition performance against encrustation in vitro. This study was aiming to further investigate the inhibitory effect of a Cu-bearing stainless steel(316 L-Cu SS) on the infectious encrustation based on its antimicrobial activity. The encrustation in presence of bacteria, antibacterial performance, urease production and Ca and Mg precipitation were examined by scanning electron microscopy, antibacterial assay, enzyme-linked immunosorbent assay and inductively coupled plasma-mass spectrometry, respectively. It was found that 316 L-Cu SS could inhibit the formation of bacterial biofilm due to the release of Cu~(2+) ions and then decrease the urease amount splitting by bacteria, which produced a neutral environment with pH around 7. However, more encrustations coupled with bacterial biofilms on the surface of comparison stainless steel(316 L SS) with an alkaline environment were recorded. It can thus be seen that the 316 L-Cu SS highlights prominent superiority against encrustation in the presence of microorganisms.     

Insights into Cronobacter sakazakii Biofilm Formation and Control Strategies in the Food Industry

Cronobacter sakazakii (C. sakazakii) is a foodborne opportunistic pathogen that can cause life-threatening invasive diseases, such as necrotizing enterocolitis, meningitis, and sepsis in infants. The potential risk of C. sakazakii contamination of powdered infant formula (PIF) is an issue that has attracted considerable attention from manufacturers, regulators, and consumers. C. sakazakii biofilms on the surfaces of equipment and in diverse food-production environments constitute a mode of cell growth that protects the pathogen from hostile environments, and are an important source of persistent contamination of food products. Bacterial biofilms are difficult to remove due to their resistant properties. Conventional cleaning and sterilizing procedures may be insufficient for biofilm control, and may lead to further biofilm development and dispersal. Consequently, novel control strategies are being developed, such as nanotechnology-based delivery systems, interspecies interactions, antimicrobial molecules of microbial origin, natural extracts, and phages. This review focuses on describing the mechanisms underlying the biofilm formation and behavior of C. sakazakii and discussing potential control strategies.