Abstract
Despite the hope that was raised with the implementation of antibiotics to the treatment of infections in medical practice, the initial enthusiasm has substantially faded due to increasing drug resistance in pathogenic microorganisms. Therefore, there is a need for novel analytical and diagnostic methods in order to extend our knowledge regarding the mode of action of the conventional and novel antimicrobial agents from a perspective of single microbial cells as well as their communities growing in infected sites, i.e., biofilms. In recent years, atomic force microscopy (AFM) has been mostly used to study different aspects of the pathophysiology of noninfectious conditions with attempts to characterize morphological and rheological properties of tissues, individual mammalian cells as well as their organelles and extracellular matrix, and cells’ mechanical changes upon exposure to different stimuli. At the same time, an ever-growing number of studies have demonstrated AFM as a valuable approach in studying microorganisms in regard to changes in their morphology and nanomechanical properties, e.g., stiffness in response to antimicrobial treatment or interaction with a substrate as well as the mechanisms behind their virulence. This review summarizes recent developments and the authors’ point of view on AFM-based evaluation of microorganisms’ response to applied antimicrobial treatment within a group of selected bacteria, fungi, and viruses. The AFM potential in development of modern diagnostic and therapeutic methods for combating of infections caused by drug-resistant bacterial strains is also discussed.
Highlights
Application of Atomic Force Microscopy (AFM) in the Field of MicrobiologyA set of specialized equipment and experimental methods that are currently used to develop new antibiotics and antiviral compounds includes, among others, such techniques as flow cytometry [1], spectroscopy [2], fluorometry [3], scanning electron microscopy (SEM) [4], and transmission electron microscopy (TEM) [5]
In this field of research, the most widespread application of atomic force microscopy includes (i) the detection of changes in microbes’ morphology or abnormalities in their structure upon antibiotic-induced killing, indirectly measuring the microbe susceptibility profile, (ii) analysis of the nanomechanical changes in the microbial cell envelope, e.g., stiffness, in order to understand mechanisms associated with drug resistance as well as (iii) the investigation of colonization and adhesion mechanisms of microbial cells, which is crucial for the biofilm-forming ability of most pathogens [17,60]
Alam et al tested forces between S. aureus bacteria immobilized on the tipless cantilever and biomaterials such as titanium alloys (Ti-6AL-4V), hydroxyapatite (HA), stainless steel (SS), and ultra-high molecular weight poly ethylene (UHMWPE), i.e., materials commonly used in the production of bone implants [44]
Summary
A set of specialized equipment and experimental methods that are currently used to develop new antibiotics and antiviral compounds includes, among others, such techniques as flow cytometry [1], spectroscopy [2], fluorometry [3], scanning electron microscopy (SEM) [4], and transmission electron microscopy (TEM) [5]. The search for novel and innovative analytical and diagnostic methods that will facilitate characterization and elimination of those antibiotic-resistant bacteria is of great significance and is challenging at the same time In this field of research, the most widespread application of atomic force microscopy includes (i) the detection of changes in microbes’ morphology or abnormalities in their structure upon antibiotic-induced killing, indirectly measuring the microbe susceptibility profile, (ii) analysis of the nanomechanical changes in the microbial cell envelope, e.g., stiffness, in order to understand mechanisms associated with drug resistance as well as (iii) the investigation of colonization and adhesion mechanisms of microbial cells, which is crucial for the biofilm-forming ability of most pathogens [17,60]. Analysis of Nanotopography of Pathogens as an Approach to Elucidate Viability of Microbes and Antibiotics’ Mechanism of Action
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