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

doi:10.1016/j.jmst.2021.02.007

Abstract

Abstract:

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 bacterial 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.

Key words: Dispersant, Enzymes, EPS, eDNA, Antibiotic resistance, Dispersal mechanism, Nanoparticles

Shuang Tian, Henny C. van der Mei, Yijin Ren, Henk J. Busscher, Linqi Shi. Recent advances and future challenges in the use of nanoparticles for the dispersal of infectious biofilms[J]. J. Mater. Sci. Technol., 2021, 84: 208-218.

Figures/Tables 9

Fig. 1. Sequential steps and consequences of biofilm dispersal. (a) Undisturbed biofilm, in which bacteria are glued together by their self-produced EPS matrix. (b) Dispersant action, continuing from the outside of the biofilm, disrupting the EPS matrix and leading to less dense regions in a biofilm. (c) Ongoing dispersant action into deeper layers and detachment of biofilm inhabitants. (d) Biofilm remaining after dispersal, with lower bacterial density than undisturbed biofilm in which antimicrobials can more readily penetrate.

Fig. 2. eDNA and DNase and their roles in biofilm dispersal. (a) eDNA acting as a biofilm glue. (b) Disruption of EPS by DNase I coating attacking the eDNA component of the EPS matrix to prevent bacterial adhesion to a substratum surface. (c) Average thickness of P. aeruginosa and S. aureus biofilms on DNase I coated surfaces. Panels (a?c) are reprinted with permission [30]. Copyright 2013, Wiley-VCH. (d) Pre-exposure of existing S. aureus biofilms to DNase rendered staphylococcal susceptibility to killing by different antimicrobials. Reprinted with permission [35]. Copyright 2011, Springer Nature.

Fig. 3. Enzyme inactivation and artificial enzymes. (a) Factors controlling inactivation of biofilm dispersing enzymes. (b) Preparation scheme of a DNase-mimetic artificial enzyme by confining multiple multinuclear AuNPs covered with Ce complexes on the surface of Fe3O4/SiO2 nanoparticles. (c) Effects of exposing an existing S. aureus biofilm grown for different periods to natural DNase I and DNase-mimetic artificial enzyme (DMAE) on biofilm mass. (d) Same as panel (c), now for biofilm thickness. Panels (b-d) are reprinted with permission [50]. Copyright 2016, Wiley-VCH.

Fig. 4. Biofilm disruption and dispersal by ROS. (a) Different types of ROS. (b) EPS biofilm matrix components oxidized by ROS. (c) EPS degradation within S. mutans biofilms and glucan breakdown by a combination of catalytic nanoparticles (CAT-NP) and H2O2 yielding ROS generation. Reprinted with permission [54]. Copyright 2016, Elsevier. (d) Dispersal of existing P. aeruginosa, E. coli, S. marcescens and L. monocytogenes biofilms by ROS generating iron oxide (Fe3O4) nanoparticles at different concentrations in suspension. Reprinted with permission [55]. Copyright 2018, Frontiers Media S.A.

Table 1 Summary of uncoated and surface-coated nanoparticles as dispersant and dispersant loaded nanoparticles for use against biofilms of different bacterial strains.

Nanoparticles with intrinsic dispersant properties
Type of nanoparticle	Mechanism		Bacterial strain	Refs.
Citrate-capped gold nanospheres	Induce structural changes in the biofilm, leading to dispersal		Legionella pneumophila	[82]
Citrate-coated platinum nanoparticles	Reduce biovolume to induce biofilm dispersal		L. pneumophila	[83]
PEG-coated gold nanoparticles	Reduce biovolume to induce biofilm dispersal		L. pneumophila	[83]
PEG-coated iron oxide nanoparticles	Reduce biovolume to induce biofilm dispersal		L. pneumophila	[83]
Iron oxide nanoparticles	Decrease intracellular c-di-GMP levels to disperse biofilms		P. aeruginosa	[84]
Iron oxide nanoparticles	Generate ROS by interaction with bacteria to disperse biofilms		S. marcescens, E. coli, P. aeruginosa and L. monocytogenes	[55]
Ferumoxytol nanoparticles	EPS matrix degradation by generating free radicals from H₂O₂		S. mutans	[85]
Graphene quantum dots	Disrupt amyloid fibrils to disperse biofilms		S. aureus	[86]
2,3-dimethylmaleic-anhydride modified carbon-dots	Reduce volumetric-bacterial-density to disperse non-EPS-producing biofilms		S. epidermidis	[87]
Cationic dextran-block copolymer nanoparticles	Interpose in between bacteria and the biofilm matrix to cause dispersal		S. aureus, vancomycin-resistant Enterococci, and Enterococcus faecalis	[24]
Cationic, poly(2-(dimethylamino)ethyl methacrylate) based micelles	Penetrate biofilms to disrupt the EPS matrix		E. coli, P. aeruginosa, B. subtilis and S. aureus	[88]
Cationic polymer micelles bearing silver ions	Interact with acidic moieties of EPS to solubilize the matrix		P. aeruginosa	[89]
Zwitterionic, mixed-shell polymeric micelles	Interact with major EPS components to disperse biofilms		S. aureus	[23]
DNase-mimetic artificial enzymes based on multinuclear metal complexes	Cleave eDNA to disperse biofilms		S. aureus	[50]
A silver-binding peptide fused to Dispersin B	Hydrolyze PNAG in the matrix to disperse biofilms		S. epidermidis	[90]
Dispersant-loaded nanoparticle carriers
Type of nanoparticle	Nanoparticle loading	Mechanism	Bacterial strain	Refs.
Nanostructured lipid carriers	DNase	Degrade eDNA to disrupt the biofilm matrix	P. aeruginosa and S. aureus	[91]
Nanocapsules	DNase	Degrade eDNA to disperse biofilms	S. aureus	[92]
Chitosan hydrogel	Dispersin B	Dispersin B degrades PNAG to detach biofilm	S. aureus, S. epidermidis and Aggregatibacter actinomycetemcomitans	[93]
Alginate	CaO₂ and hemin-loading graphene	Convert H₂O into ROS to degrade matrix components in biofilms	S. aureus	[51]
Graphene-mesoporous silica nanosheets	Ascorbic acid and ferromagnetic nanoparticles	Catalyze ascorbic acid forming ^-OH to disperse biofilms	S. aureus and E. coli	[52]
Hybrid micelles	D-tyrosine	Affect amyloid fibers to disperse biofilms	P. aeruginosa	[94]
Chitin-based nanocomposite containing D-amino acids and iron oxide nanoparticles	D-tyrosine, D-tryptophan, and D-phenylalanine	D-amino acids disrupt biofilms	S. aureus	[95]
Polymeric nanoparticle	NO	Sustained release of NO to disperse biofilms	P. aeruginosa	[96]
Polydopamine-coated iron oxide nanoparticles	NO	Localized NO release to disperse biofilms	P. aeruginosa	[97]
Polymer/gold hybrid nanoparticles	NO	NO triggered dispersal of biofilms	P. aeruginosa	[98]
Polyethylenimine/ diazeniumdiolate -doped PLGA nanoparticles	NO	NO triggered dispersal of MRSA biofilms in vitro and in vivo	methicillin-resistant S. aureus	[99]
Micelles	NO	Visible light triggered release of NO to disperse biofilms	P. aeruginosa	[100]
Liposomes	Biosurfactants isolated from Lactobacillus gasseri	Biosurfactants dispersed biofilms	S. aureus	[101]
Dispersant-coated nanoparticles
Type of nanoparticle	Nanoparticle coating	Mechanism	Bacterial strain	Refs.
Poly(L-lysine) coated PLGA nanoparticles	DNase	Disperse biofilm by degrading eDNA	P. aeruginosa	[102]
Chitosan nanoparticles	DNase	Disperse biofilm by degrading eDNA	P. aeruginosa	[103]
Gold nanoparticles	DNase	Disperse biofilm by degrading eDNA	S. aureus, S. epidermidis, E. coli, and P. aeruginosa	[104]
Silica nanobeads	Proteinase K	Degrade proteins in biofilm matrix	P. fluorescens	[105]
Gold nanoparticles	Proteinase K	Degrade proteins in biofilm matrix	P. fluorescens	[106]
Silver nanoparticles	D-cysteine	“Disperse-then-kill”	E. coli, P. aeruginosa and S. aureus	[107]

Table 1 Summary of uncoated and surface-coated nanoparticles as dispersant and dispersant loaded nanoparticles for use against biofilms of different bacterial strains.

Nanoparticles with intrinsic dispersant properties
Type of nanoparticle	Mechanism		Bacterial strain	Refs.
Citrate-capped gold nanospheres	Induce structural changes in the biofilm, leading to dispersal		Legionella pneumophila	[82]
Citrate-coated platinum nanoparticles	Reduce biovolume to induce biofilm dispersal		L. pneumophila	[83]
PEG-coated gold nanoparticles	Reduce biovolume to induce biofilm dispersal		L. pneumophila	[83]
PEG-coated iron oxide nanoparticles	Reduce biovolume to induce biofilm dispersal		L. pneumophila	[83]
Iron oxide nanoparticles	Decrease intracellular c-di-GMP levels to disperse biofilms		P. aeruginosa	[84]
Iron oxide nanoparticles	Generate ROS by interaction with bacteria to disperse biofilms		S. marcescens, E. coli, P. aeruginosa and L. monocytogenes	[55]
Ferumoxytol nanoparticles	EPS matrix degradation by generating free radicals from H₂O₂		S. mutans	[85]
Graphene quantum dots	Disrupt amyloid fibrils to disperse biofilms		S. aureus	[86]
2,3-dimethylmaleic-anhydride modified carbon-dots	Reduce volumetric-bacterial-density to disperse non-EPS-producing biofilms		S. epidermidis	[87]
Cationic dextran-block copolymer nanoparticles	Interpose in between bacteria and the biofilm matrix to cause dispersal		S. aureus, vancomycin-resistant Enterococci, and Enterococcus faecalis	[24]
Cationic, poly(2-(dimethylamino)ethyl methacrylate) based micelles	Penetrate biofilms to disrupt the EPS matrix		E. coli, P. aeruginosa, B. subtilis and S. aureus	[88]
Cationic polymer micelles bearing silver ions	Interact with acidic moieties of EPS to solubilize the matrix		P. aeruginosa	[89]
Zwitterionic, mixed-shell polymeric micelles	Interact with major EPS components to disperse biofilms		S. aureus	[23]
DNase-mimetic artificial enzymes based on multinuclear metal complexes	Cleave eDNA to disperse biofilms		S. aureus	[50]
A silver-binding peptide fused to Dispersin B	Hydrolyze PNAG in the matrix to disperse biofilms		S. epidermidis	[90]
Dispersant-loaded nanoparticle carriers
Type of nanoparticle	Nanoparticle loading	Mechanism	Bacterial strain	Refs.
Nanostructured lipid carriers	DNase	Degrade eDNA to disrupt the biofilm matrix	P. aeruginosa and S. aureus	[91]
Nanocapsules	DNase	Degrade eDNA to disperse biofilms	S. aureus	[92]
Chitosan hydrogel	Dispersin B	Dispersin B degrades PNAG to detach biofilm	S. aureus, S. epidermidis and Aggregatibacter actinomycetemcomitans	[93]
Alginate	CaO₂ and hemin-loading graphene	Convert H₂O into ROS to degrade matrix components in biofilms	S. aureus	[51]
Graphene-mesoporous silica nanosheets	Ascorbic acid and ferromagnetic nanoparticles	Catalyze ascorbic acid forming ^-OH to disperse biofilms	S. aureus and E. coli	[52]
Hybrid micelles	D-tyrosine	Affect amyloid fibers to disperse biofilms	P. aeruginosa	[94]
Chitin-based nanocomposite containing D-amino acids and iron oxide nanoparticles	D-tyrosine, D-tryptophan, and D-phenylalanine	D-amino acids disrupt biofilms	S. aureus	[95]
Polymeric nanoparticle	NO	Sustained release of NO to disperse biofilms	P. aeruginosa	[96]
Polydopamine-coated iron oxide nanoparticles	NO	Localized NO release to disperse biofilms	P. aeruginosa	[97]
Polymer/gold hybrid nanoparticles	NO	NO triggered dispersal of biofilms	P. aeruginosa	[98]
Polyethylenimine/ diazeniumdiolate -doped PLGA nanoparticles	NO	NO triggered dispersal of MRSA biofilms in vitro and in vivo	methicillin-resistant S. aureus	[99]
Micelles	NO	Visible light triggered release of NO to disperse biofilms	P. aeruginosa	[100]
Liposomes	Biosurfactants isolated from Lactobacillus gasseri	Biosurfactants dispersed biofilms	S. aureus	[101]
Dispersant-coated nanoparticles
Type of nanoparticle	Nanoparticle coating	Mechanism	Bacterial strain	Refs.
Poly(L-lysine) coated PLGA nanoparticles	DNase	Disperse biofilm by degrading eDNA	P. aeruginosa	[102]
Chitosan nanoparticles	DNase	Disperse biofilm by degrading eDNA	P. aeruginosa	[103]
Gold nanoparticles	DNase	Disperse biofilm by degrading eDNA	S. aureus, S. epidermidis, E. coli, and P. aeruginosa	[104]
Silica nanobeads	Proteinase K	Degrade proteins in biofilm matrix	P. fluorescens	[105]
Gold nanoparticles	Proteinase K	Degrade proteins in biofilm matrix	P. fluorescens	[106]
Silver nanoparticles	D-cysteine	“Disperse-then-kill”	E. coli, P. aeruginosa and S. aureus	[107]

Fig. 5. Examples of the use of nanoparticles for biofilm dispersal. (a) Scanning electron micrographs of graphene quantum dots (GQD)-mediated staphylococcal biofilm dispersal, showing patches that are entirely devoid of biofilm after GQD exposure and remaining biofilms arranged in a pattern that presumably reflects drying artifacts due to low cohesivity of the EPS matrix after exposure to GQD. Reprinted with permission [86]. Copyright 2019, American Chemical Society. (b) Zwitterionic micelles dispersed existing S. aureus biofilms in vitro (green-fluorescence: bacteria, red-fluorescence: EPS). Reprinted with permission [23]. Copyright 2020, Science Publishing Group. (c) Confocal laser scanning microscopy images of methicillin-resistant S. aureus (MRSA) biofilms exposed to cationic dextran-block copolymeric nanoparticles, showing biofilm dispersal over time (Green fluorescence: bacteria, red fluorescence: DA95B5 dispersant). Reprinted with permission [24]. Copyright 2018, American Chemical Society. (d) Volumetric bacterial densities of S. epidermidis biofilms after 4 h exposure to buffer or suspensions with 125 μg mL-1 carbon-dots without 2,3-dimethylmaleic-anhydride (DMMA) (C-dots) or CDMMA-dots (CDMMA-dots) at pH 7.4 and pH 5.0. Reprinted with permission [87]. Copyright 2020, Elsevier.

Fig. 6. Protective encapsulation of dispersants in nanoparticle carriers. (a) Advantages of the use of dispersant loaded nanocarriers above non-encapsulated dispersants. (b) Sustained NO release from polymer/gold hybrid nanoparticles in a phosphate buffer (pH 6.8). Reprinted with permission [98]. Copyright 2014, Royal Society of Chemistry. (c) Relative activity of free recombinant Dispersin B (DspB) and DspB loaded in chitosan nanoparticles as a function of temperature. Activity was assessed from the ability of DspB to enzymatically release of p-nitrophenolate from 4-nitrophenyl-N-acetyl β-D-glucosaminide. (d) Relative activity upon storage at 37 ℃ of free Dispersin B and Dispersin B loaded in chitosan nanoparticles as a function of time. (c and d) Reprinted with permission [93]. Copyright 2015, Elsevier.

Fig. 7. Spreading of infection in a murine model after dispersal of a subcutaneous, bioluminescent P. aeruginosa biofilm using glycoside hydrolase (GH) as a dispersant in absence of simultaneously administered antibiotics. Infection is visualized using bioluminescent imaging. Reprinted with permission [112]. Copyright 2018, Springer Nature.

Fig. 8. Biofilm dispersants enhance efficacy of existing antibiotics. (a) Breaking the biofilm barrier by dispersant action makes remaining biofilms more penetrable to antibiotics. (b) Survival of S. aureus in biofilms pre-exposed DNase loaded nanocapsules (nDNase) and subsequently exposed to ciprofloxacin (CPFX). For control, similarly encapsulated bovine serum albumin (nBSA) was employed for pre-exposure. Reprinted with permission [92]. Copyright 2020, Royal Society of Chemistry. (c) Log10 CFU (colony forming units) of S. epidermidis in biofilms after pre-exposure to buffer or carbon dots with or without DMMA (C-dots and CDMMA-dots, respectively) at different pH followed by 72 h growth in medium supplemented with vancomycin at different multiple concentrations of its minimal bactericidal concentration (MBC). Reprinted with permission [87]. Copyright 2020, Elsevier. (d) Effect of gentamicin-NO (GEN-NO) nanoparticles on P. aeruginosa biofilm viability after combined release of NO and gentamicin. Reprinted with permission [96]. Copyright 2016, Royal Society of Chemistry.

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