Enzyme-Linked Lipid Nanocarriers for Coping Pseudomonal Pulmonary Infection. Would Nanocarriers Complement Biofilm Disruption or Pave Its Road?

Overview

Journal Int J Nanomedicine

Publisher Dove Medical Press

Specialty Biotechnology

Date 2024 May 6

PMID 38708178

Authors

Noha Nafee

Dina M Gaber

Alaa Abouelfetouh

Mustafa Alseqely

Martin Empting

Marc Schneider

Affiliations

Soon will be listed here.

Abstract

Introduction: Cystic fibrosis (CF) is associated with pulmonary infections persistent to antibiotics.

Methods: To eradicate biofilms, solid lipid nanoparticles (SLNs) loaded with quorum-sensing-inhibitor (QSI, disrupting bacterial crosstalk), coated with chitosan (CS, improving internalization) and immobilized with alginate lyase (AL, destroying alginate biofilms) were developed.

Results: SLNs (140-205 nm) showed prolonged release of QSI with no sign of acute toxicity to A549 and Calu-3 cells. The CS coating improved uptake, whereas immobilized-AL ensured >1.5-fold higher uptake and doubled SLN diffusion across the artificial biofilm sputum model. Respirable microparticles comprising SLNs in carbohydrate matrix elicited aerodynamic diameters MMAD (3.54, 2.48 µm) and fine-particle-fraction FPF (65, 48%) for anionic and cationic SLNs, respectively. The antimicrobial and/or antibiofilm activity of SLNs was explored in reference mucoid/nonmucoid strains as well as clinical isolates. The full growth inhibition of planktonic bacteria was dependent on SLN type, concentration, growth medium, and strain. OD measurements and live/dead staining proved that anionic SLNs efficiently ceased biofilm formation and eradicated established biofilms, whereas cationic SLNs unexpectedly promoted biofilm progression. AL immobilization increased biofilm vulnerability; instead, CS coating increased biofilm formation confirmed by 3D-time lapse confocal imaging. Incubation of SLNs with mature biofilms of isolates increased biofilm density by an average of 1.5-fold. CLSM further confirmed the binding and uptake of the labeled SLNs in biofilms. Considerable uptake of CS-coated SLNs in non-mucoid strains could be observed presumably due to interaction of chitosan with LPS glycolipids in the outer cell membrane of .

Conclusion: The biofilm-destructive potential of QSI/SLNs/AL inhalation is promising for site-specific biofilm-targeted interventional CF therapy. Nevertheless, the intrinsic/extrinsic fundamentals of nanocarrier-biofilm interactions require further investigation.

References

Suk J, Lai S, Wang Y, Ensign L, Zeitlin P, Boyle M . The penetration of fresh undiluted sputum expectorated by cystic fibrosis patients by non-adhesive polymer nanoparticles. Biomaterials. 2009; 30(13):2591-7. PMC: 2661768. DOI: 10.1016/j.biomaterials.2008.12.076. View

Alipour M, Suntres Z, Omri A . Importance of DNase and alginate lyase for enhancing free and liposome encapsulated aminoglycoside activity against Pseudomonas aeruginosa. J Antimicrob Chemother. 2009; 64(2):317-25. DOI: 10.1093/jac/dkp165. View

Nafee N, Husari A, Maurer C, Lu C, De Rossi C, Steinbach A . Antibiotic-free nanotherapeutics: ultra-small, mucus-penetrating solid lipid nanoparticles enhance the pulmonary delivery and anti-virulence efficacy of novel quorum sensing inhibitors. J Control Release. 2014; 192:131-40. DOI: 10.1016/j.jconrel.2014.06.055. View

Lamppa J, Griswold K . Alginate lyase exhibits catalysis-independent biofilm dispersion and antibiotic synergy. Antimicrob Agents Chemother. 2012; 57(1):137-45. PMC: 3535929. DOI: 10.1128/AAC.01789-12. View

Germoni L, Bremer P, Lamont I . The effect of alginate lyase on the gentamicin resistance of Pseudomonas aeruginosa in mucoid biofilms. J Appl Microbiol. 2016; 121(1):126-35. DOI: 10.1111/jam.13153. View

Schutz C, Ho D, Hamed M, Abdelsamie A, Rohrig T, Herr C . A New PqsR Inverse Agonist Potentiates Tobramycin Efficacy to Eradicate Pseudomonas aeruginosa Biofilms. Adv Sci (Weinh). 2021; 8(12):e2004369. PMC: 8224453. DOI: 10.1002/advs.202004369. View

Omri A, Beaulac C, Bouhajib M, Montplaisir S, Sharkawi M, Lagace J . Pulmonary retention of free and liposome-encapsulated tobramycin after intratracheal administration in uninfected rats and rats infected with Pseudomonas aeruginosa. Antimicrob Agents Chemother. 1994; 38(5):1090-5. PMC: 188155. DOI: 10.1128/AAC.38.5.1090. View

Drulis-Kawa Z, Gubernator J, Dorotkiewicz-Jach A, Doroszkiewicz W, Kozubek A . In vitro antimicrobial activity of liposomal meropenem against Pseudomonas aeruginosa strains. Int J Pharm. 2006; 315(1-2):59-66. DOI: 10.1016/j.ijpharm.2006.02.017. View

Nafee N, Taetz S, Schneider M, Schaefer U, Lehr C . Chitosan-coated PLGA nanoparticles for DNA/RNA delivery: effect of the formulation parameters on complexation and transfection of antisense oligonucleotides. Nanomedicine. 2007; 3(3):173-83. DOI: 10.1016/j.nano.2007.03.006. View

10.

Wan B, Zhu Y, Tao J, Zhu F, Chen J, Li L . Alginate Lyase Guided Silver Nanocomposites for Eradicating from Lungs. ACS Appl Mater Interfaces. 2020; 12(8):9050-9061. DOI: 10.1021/acsami.9b21815. View

11.

Lu C, Maurer C, Kirsch B, Steinbach A, Hartmann R . Overcoming the unexpected functional inversion of a PqsR antagonist in Pseudomonas aeruginosa: an in vivo potent antivirulence agent targeting pqs quorum sensing. Angew Chem Int Ed Engl. 2013; 53(4):1109-12. DOI: 10.1002/anie.201307547. View

12.

Yang M, Yu Y, Yang S, Shi X, Mou H, Li L . Expression and Characterization of a New PolyG-Specific Alginate Lyase From Marine Bacterium sp. Q7. Front Microbiol. 2018; 9:2894. PMC: 6281962. DOI: 10.3389/fmicb.2018.02894. View

13.

Gunday Tureli N, Torge A, Juntke J, Schwarz B, Schneider-Daum N, Tureli A . Ciprofloxacin-loaded PLGA nanoparticles against cystic fibrosis P. aeruginosa lung infections. Eur J Pharm Biopharm. 2017; 117:363-371. DOI: 10.1016/j.ejpb.2017.04.032. View

14.

Ungaro F, dAngelo I, Coletta C, dEmmanuele di Villa Bianca R, Sorrentino R, Perfetto B . Dry powders based on PLGA nanoparticles for pulmonary delivery of antibiotics: modulation of encapsulation efficiency, release rate and lung deposition pattern by hydrophilic polymers. J Control Release. 2011; 157(1):149-59. DOI: 10.1016/j.jconrel.2011.08.010. View

15.

Sana T, Lomas R, Gimenez M, Laubier A, Soscia C, Chauvet C . Differential Modulation of Quorum Sensing Signaling through QslA in Pseudomonas aeruginosa Strains PAO1 and PA14. J Bacteriol. 2019; 201(21). PMC: 6779463. DOI: 10.1128/JB.00362-19. View

16.

Schlafer S, Meyer R . Confocal microscopy imaging of the biofilm matrix. J Microbiol Methods. 2016; 138:50-59. DOI: 10.1016/j.mimet.2016.03.002. View

17.

Marangon C, Martins V, Ling M, Melo C, Plepis A, Meyer R . Combination of Rhamnolipid and Chitosan in Nanoparticles Boosts Their Antimicrobial Efficacy. ACS Appl Mater Interfaces. 2020; 12(5):5488-5499. DOI: 10.1021/acsami.9b19253. View

18.

Meers P, Neville M, Malinin V, Scotto A, Sardaryan G, Kurumunda R . Biofilm penetration, triggered release and in vivo activity of inhaled liposomal amikacin in chronic Pseudomonas aeruginosa lung infections. J Antimicrob Chemother. 2008; 61(4):859-68. DOI: 10.1093/jac/dkn059. View

19.

Liu Y, Ahator S, Wang H, Feng Q, Xu Y, Li C . Microevolution of the and Reinforces the Bias of Quorum Sensing System in Laboratory Strains of PAO1. Front Microbiol. 2022; 13:821895. PMC: 9041413. DOI: 10.3389/fmicb.2022.821895. View

20.

Schutz C, Hodzic A, Hamed M, Abdelsamie A, Kany A, Bauer M . Divergent synthesis and biological evaluation of 2-(trifluoromethyl)pyridines as virulence-attenuating inverse agonists targeting PqsR. Eur J Med Chem. 2021; 226:113797. DOI: 10.1016/j.ejmech.2021.113797. View