Liposomal Encapsulation of Two Novel Lytic Bacteriophages targeting Klebsiella pneumoniae: Preliminary Ex Vivo Evaluation of a Protective Delivery Platform for Pulmonary Infections

Theodor Bilharz Research Institute

Bibliographic Information

Authors: Ribera Guerrero B.; Samir S.; Vila M.M.D.C.; Balcão V.M.C.F.

Journal: Brazilian Archives of Biology and Technology

Publisher: Instituto de Tecnologia do Parana

Publication Date: 2026

Volume / Issue: Volume 69

Article No.: e26250853

ISSN: 15168913

DOI: 10.1590/1678-4324-2026250853

Scopus: View on Scopus

Document Type: Article

Access: All Open Access; Gold Open Access

Authors and Affiliations

Ribera Guerrero B., Universidade de Sorocaba, VBlab – Laboratório de Vírus Bacterianos, SP, Sorocaba, Brazil; Samir S., Theodor Bilharz Research Institute, Department of Biochemistry and Molecular Biology, Giza, Egypt; Vila M.M.D.C., Universidade de Sorocaba, VBlab – Laboratório de Vírus Bacterianos, SP, Sorocaba, Brazil; Balcão V.M.C.F., Universidade de Sorocaba, VBlab – Laboratório de Vírus Bacterianos, SP, Sorocaba, Brazil, Universidade de Aveiro, Departamento de Biologia e CESAM, Aveiro, Portugal

Abstract

Multidrug resistance, exacerbated by inappropriate antibiotic use and limited development of new antimicrobials, represents a critical global health challenge. Klebsiella pneumoniae, characterized by its protective capsule and strong colonization capacity, is a major cause of hospital-acquired pneumonia, emphasizing the urgency of alternative therapeutic strategies. Phage therapy has emerged as a promising option, and a novel “Trojan Horse” approach using liposome-encapsulated phages (LEP) may enhance treatment by enabling phage delivery while evading immune detection. This study investigated the encapsulation of a lytic bacteriophage cocktail within liposomes to target multidrug-resistant K. pneumoniae. LEP presented a negative Zeta potential similar to that of K. pneumoniae cells, suggesting potential electrostatic repulsion between LEP and bacterial surfaces. Antimicrobial susceptibility testing confirmed that the evaluated strain was sensitive only to amikacin, gentamicin, and tetracycline. Physicochemical characterization showed that LEP exhibited a 21% smaller diffusion coefficient than empty liposomes, consistent with the increased size of the loaded structures. In vitro, LEP demonstrated sustained phage release and effectively prevented bacterial regrowth after 9 hours of treatment. Ex vivo assays using artificially contaminated canine lung tissue revealed that LEP achieved a maximal bacterial reduction of 1.04 log CFU/mL after 12 hours, markedly outperforming free phages, which allowed bacterial resurgence after 9 hours. The limited phage diffusion within the solid lung tissue matrix likely reduced phage–bacterium interactions, indicating that higher multiplicity of infection may be required to enhance therapeutic efficacy in structured biological environments. © 2026 by the authors.

Keywords

Bacteriophage; Klebsiella pneumoniae; liposomes; lung infections

Citation Information

Scopus Citations: 0

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