Substrate morphology modulates bacteriophage mobility and alters bactericidal potency

Abstract

Bacteriophages (phages) are regaining attention as alternatives to antibiotics in the antimicrobial resistance crisis. Although successful cases are widely reported, the influence of physical, abiotic substrate morphology on phage performance remains largely unexamined, despite morphology being an intrinsic topographical feature present at many infection and decontamination sites, such as granular chronic wound beds or porous bone-implant interfaces.

1. Introduction

An urgent call to address antimicrobial resistance (AMR) has been rising, driven primarily by the accelerating emergence of resistant pathogens, compounded by antibiotic discovery becoming increasingly slower. Phages, as a promising solution, possess several advantages , such as a long record of clinical history, a rich genetic reservoir generated during co-evolution with bacteria, and high specificity for specific strains.

Materials and Methods

Materials

Tofu (Tofu King, Birmingham, UK) was selected as texture model material. Staphylococcus aureus and its lytic phage K were obtained from NCTC. S. aureus selective nutrient medium, Mannitol Salt Agar, was purchased from OXOID. Tris(hydroxymethyl)aminomethane (Tris) buffer, ethanol, glutaraldehyde (25% v/v, GLA), and hexamethyldisilazane (HMDS) were purchased from Sigma-Aldrich

Engineered substrate morphology

Substrate morphology was engineered on fresh tofu blocks to obtain smooth, rough, and porous morphologies. Smooth substrates were prepared by trimming tofu with a sterile scalpel. Rough textures were produced by pressing the cut surfaces against coarse fabric. Porous structures were generated by freezing at −20 °C overnight .

Results

Phage and host

S. aureus and phage K were selected as the test model as shown in Phage K TEM image demonstrated typical myophage morphology, including a head around 50 nm and a contractile tail with around 250 nm length. S. aureus SEM images demonstrated typical grape-like clusters with 0.86 ± 0.06 µm size and rough surfaces with undefined nodes.

Substrate morphology effects on phage bactericidal potency

Phage potency was investigated via phage administration on flat, rough, and porous engineered substrates as shown. The morphologies remained consistent with the profiler information, in which continuous smooth surfaces were observed on flat substrates, while periodic fluctuations were observed on rough substrates, and spongy structures were shown on porous substrates

Conclusion

This study demonstrates that abiotic substrate morphology is a fundamental and previously overlooked factor affecting bacteriophage (phage) bactericidal potency during surface-associated decontamination, with implications for phage therapy where analogous local morphology at the infection site may similarly influence potency. 

Citation: Wang H, Awolusi OO, Jaganathan SK, Lyu J, Liu C, Tucker N, et al. (2026) Substrate morphology modulates bacteriophage mobility and alters bactericidal potency. PLoS One 21(9): e0356919. https://doi.org/10.1371/journal.pone.0356919

Editor: Geelsu Hwang, University of Pennsylvania, UNITED STATES OF AMERICA

Received: June 16, 2026; Accepted: August 10, 2026; Published: September 16, 2026

Copyright: © 2026 Wang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Data Availability: All relevant data supporting the findings of this study are included within the manuscript and the figure files submitted with this article.

Funding: This work was supported by Quality-related Research (QR) funding from the University of Lincoln to Dr Bukola Adenike Onarinde. No specific grant number was associated with this funding. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Competing interests: The authors have declared that no competing interests exist.