Abstract
Ureteral stenting is a common surgical procedure, which is associated with a high morbidity and economic burden, but the knowledge on the link between biofilms on these stents, morbidity, and the impact of the involved microbiota is still limited. This is partially due to a lack of methods that allow for a controlled extraction of the biofilms from stents. Development of an appropriate in vitro model to assess prevention of biofilm formation by antimicrobial coatings and biomaterials requires a profound understanding of the biofilm composition, including the involved microbiota. This work describes an analytical pipeline for the extraction of native biofilms from ureteral stents for both cultivation-dependent and -independent analysis, involving a novel mechanical abrasion method of passing stent samples through a tapered pinhole. The efficiency of this novel method was evaluated by quantifying the removed biofilm mass, numbers of cultivable bacteria, calcium content, and microscopic stent analysis after biofilm removal using 30 clinical stent samples. Furthermore, the extraction of in vitro formed Escherichia coli biofilms was evaluated by universal 16S quantitative PCR, a cultivation-independent method to demonstrate efficient biofilm removal by the new approach. The novel method enables effective contamination-free extraction of the biofilms formed on ureteral stents and their subsequent quantification, and it represents a useful tool for comprehensive examinations of biofilms on ureteral stents.
Highlights
The insertion of medical devices into the urinary tract, such as ureteral stents and urinary catheters, frequently causes irritations and urinary tract infections, which are the most common nosocomial infection (Warren, 2001). This is related to the encrustation with urine components that crystallize on the surface of biomaterials, and to the formation of microbial biofilms, complex communities of microorganisms embedded into a matrix of extracellular polymeric substances (EPS)
Biofilms from ureteral stents were effectively extracted by repetitively passing the ureteral stent through a tapered pinhole in a steel plate with a slightly smaller diameter than the stent (i.e., 1.9 and 2.0 mm, respectively)
To validate the pinhole method (PM) in a setting with realistic mechanical properties of the specimen, extraction of in vivo biofilms was performed from clinical ureteral stents
Summary
The insertion of medical devices into the urinary tract, such as ureteral stents and urinary catheters, frequently causes irritations and urinary tract infections, which are the most common nosocomial infection (Warren, 2001) This is related to the encrustation with urine components that crystallize on the surface of biomaterials, and to the formation of microbial biofilms, complex communities of microorganisms embedded into a matrix of EPS. We validated the method using in vivo clinical and in vitro model biofilms, and we demonstrated that the established biofilm extraction pipeline allows for efficient recovery of biofilm mass It avoids introduction of detectable DNA contamination, enabling cultivation-independent qPCR analysis. This work will facilitate quantitative microbiological, biochemical, and molecular analyses of biofilms from ureteral stents
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