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A Bacteriophage-Loaded Microparticle Laden Topical Gel for the Treatment of Multidrug-Resistant Biofilm-Mediated Burn Wound Infection

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Abstract

Klebsiella pneumoniae is regarded as one of the most profound bacteria isolated from the debilitating injuries caused by burn wounds. In addition, the multidrug resistance (MDR) and biofilm formation make treating burn patients with clinically available antibiotics difficult. Bacteriophage therapy has been proven an effective alternative against biofilm-mediated wound infections caused by MDR bacterial strains. In the current study, the bacteriophage (BPKPФ1) against MDR Klebsiella pneumoniae was isolated and loaded into the chitosan microparticles (CHMPs), which was later incorporated into the Sepineo P 600 to convert into a gel (BPKPФ1-CHMP-gel). BPKPФ1 was characterized for lytic profile, morphological class, and burst size, which revealed that the BPKPФ1 belongs to the family Siphoviridae. Moreover, BPKPФ1 exhibited a narrow host range with 128 PFU/host cell of burst size. The BPKPФ1-loaded CHMPs showed an average particle size of  1.96 ± 0.51 μm, zeta potential 32.16 ± 0.41 mV, and entrapment efficiency in the range of 82.44 ± 1.31%. Further, the in vitro antibacterial and antibiofilm effectiveness of BPKPФ1-CHMPs-gel were examined. The in vivo potential of the BPKPФ1-CHMPs-gel was assessed using a rat model with MDR Klebsiella pneumoniae infected burn wound, which exhibited improved wound contraction (89.22 ± 0.48%) in 28 days with reduced inflammation, in comparison with different controls. Data in hand suggest the potential of bacteriophage therapy to be developed as personalized therapy in case of difficult-to-treat bacterial infections.

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Abbreviations

MDR:

Multidrug resistance

AMR:

Antimicrobial resistance

BP:

Bacteriophage

K. pneumoniae :

Klebsiella pneumoniae

BPKPФ1:

Bacteriophage against K. pneumoniae

CHMPs:

Chitosan microparticles

BPKPФ1-CHMPs:

Bacteriophage chitosan microparticles

DLAO:

Double-layer agar overlay

TEM:

Transmission electron microscopy

SEM:

Scanning electron microscopy

SSD:

Silver sulfadiazine

USG/PA:

Ultrasound/photoacoustic

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Acknowledgements

Author D. Dehari would like to thank the Indian Institute of Technology (BHU) Varanasi for providing financial assistance in the form of a research support grant to conduct the study. SEM and Confocal microscopy were performed at IIT (BHU) Varanasi, for which the authors express their appreciation to the Central Instrument Facility Centre. The authors are also grateful to the Virus Research & Development Laboratory in the Department of Microbiology at the Institute of Medical Science at Banaras Hindu University in Varanasi for providing lab space for the study of Bateriophages as part of scheme 5066 and the use of an Optima XPN-100 Ultracentrifuge (supported by the Department of Science and Technology’s PURSE program). Department of Geology (BHU) is also acknowledged for the facility of scanning electron microscope used to examine biological samples. Also acknowledged is SATHI (Banaras Hindu University) for providing the Photoacoustic imaging platform.

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Authors and Affiliations

  1. Department of Pharmaceutical Engineering and Technology, Indian Institute of Technology (BHU), Varanasi, U.P., 221005, India

    Deepa Dehari, Aiswarya Chaudhuri, Dulla Naveen Kumar, Meraj Anjum, Dinesh Kumar & Ashish Kumar Agrawal

  2. Department of Microbiology, Institute of Medical Science, Banaras Hindu University, Varanasi, U.P., 221005, India

    Rajesh Kumar, Akshay Kumar & Gopal Nath

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  1. Deepa Dehari
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Contributions

Deepa Dehari: conceptualization, methodology, formal analysis and data curation, and writing; Aiswarya Chaudhuri, Dulla Naveen Kumar, and Meraj Anjum: formulation, data collection, participated in animal experiments; Rajesh Kumar and Akshay Kumar: bacterial strain collection, bacteriophage isolation, and data curation; Dinesh Kumar: validation, review, and editing, Gopal Nath: provided microbiology lab facility, conceptualization, supervised, writing—review and editing; and Ashish Kumar Agrawal: conceived and supervised the project, reviewing, editing, and approved the final manuscript. All authors have approved the final version of the manuscript.

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Correspondence to Ashish Kumar Agrawal.

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IAEC approved all animal experiments with approval no-IIT(BHU)/IAEC/2022/007 from the Department of Pharmaceutical Eng. & Tech., Indian Institute of Technology (BHU), Varanasi, U.P., India.

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Dehari, D., Chaudhuri, A., Kumar, D.N. et al. A Bacteriophage-Loaded Microparticle Laden Topical Gel for the Treatment of Multidrug-Resistant Biofilm-Mediated Burn Wound Infection. AAPS PharmSciTech 24, 165 (2023). https://doi.org/10.1208/s12249-023-02620-w

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