Staphylococcus aureus Strain-Dependent Biofilm Formation in Bone-Like Environment (original) (raw)
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Human Osteoblast-Conditioned Media Can Influence Staphylococcus aureus Biofilm Formation
International Journal of Molecular Sciences
Osteoblasts are bone-forming and highly active cells participating in bone homeostasis. In the case of osteomyelitis and more specifically prosthetic joint infections (PJI) for which Staphylococcus aureus (S. aureus) is mainly involved, the interaction between osteoblasts and S. aureus results in impaired bone homeostasis. If, so far, most of the studies of osteoblasts and S. aureus interactions were focused on osteoblast response following direct interactions with co-culture and/or internalization models, less is known about the effect of osteoblast factors on S. aureus biofilm formation. In the present study, we investigated the effect of human osteoblast culture supernatant on methicillin sensitive S. aureus (MSSA) SH1000 and methicillin resistant S. aureus (MRSA) USA300. Firstly, Saos-2 cell line was incubated with either medium containing TNF-α to mimic the inflammatory periprosthetic environment or with regular medium. Biofilm biomass was slightly increased for both strains in...
Frontiers in Microbiology
Prosthesis and joint infections are an important threat in public health, especially due to the development of bacterial biofilms and their high resistance to antimicrobials. Biofilmassociated infections increase mortality and morbidity rates as well as hospitalization costs. Prevention is the best strategy for this serious issue, so there is an urgent need to understand the signals that could induce irreversible bacterial adhesion on a prosthesis. In this context, we investigated the influence of the bone environment on surface adhesion by a methicillin-susceptible Staphylococcus aureus strain. Using static and dynamic biofilm models, we tested various bone environment factors and showed that the presence of Mg 2+ , lack of oxygen, and starvation each increased bacterial adhesion. It was observed that human osteoblast-like cell culture supernatants, which contain secreted components that would be found in the bone environment, increased bacterial adhesion capacity by 2-fold (p = 0.015) compared to the medium control. Moreover, supernatants from osteoblast-like cells stimulated with TNF-α to mimic inflammatory conditions increased bacterial adhesion by almost 5-fold (p = 0.003) without impacting on the overall biomass. Interestingly, the effect of osteoblast-like cell supernatants on bacterial adhesion could be counteracted by the activity of synthetic antibiofilm peptides. Overall, the results of this study demonstrate that factors within the bone environment and products of osteoblast-like cells directly influence S. aureus adhesion and could contribute to biofilm initiation on bone and/or prosthetics implants.
Study of Staphylococcus aureus Biofilm Ability to Destroy Bone Matrix in Absences of Host Immunity
2020
Purpose: Osteomyelitis is an infectious bone process leading to bone necrosis and destruction. Published reports on pathogen biofilm thus far have focused on indirect bone resorption mediated by host cells and factors secondary to immune system activation. However, direct bone resorption due to biofilm pathogen has not been adequately studied yet. This study aims to investigate the effect of biofilm pathogen in ex-vivo human bones in the absence of the host immune response using Raman spectroscopy and Scanning electron microscopy. Methods: Bone samples collected from patients who underwent knee replacement surgeries were inoculated with Staphylococcus aureus bacteria. Bacterial direct effects on the bone quality were then examined, at various time intervals, using Raman spectroscopy and scanning electron microscopy. Results: Raman spectroscopy and scanning electron demonstrated the destruction of bone structure and drop in bone quality. Conclusion: This experiment shows the direct effect of bacteria on bone during osteomyelitis in addition to the recognised destruction caused by the host immune system. Introduction: Osteomyelitis (OM) is bone inflammatory process secondary to an infectious organism leading to bone necrosis and destruction. Bone is a sterile organ resistant to bacterial colonisation. However, events like trauma, surgery, or hematogenous spread of virulent pathogen may disturb structural integrity and results in infection (1). Several confounding factors may influence the inflammatory process of OM leading to diagnostic difficulties and treatment delays. Due to the marked variability in osteomyelitis presentation and treatment, many researchers tried to investigate disease pathogenesis and introduce a controlled approach to active management. However, the primary focus of most of the published reports was a collection of infection-related parameters: haematological, radiological, histology or bacterial culture (2). Some reports focused on the process of bone destruction due to immune system activation. However, there is limited emphasis on biofilm formation and pathogen-induced bone destruction (3). Most causative pathogens can adhere to and grow on the surface of bone, joints and prosthesis causing bone resorptions. The pathophysiology of bacterium-induced bone resorption has not been identified. However, reports have suggested that pathogens such as Staphylococcus aureus express an array of cell surface and soluble molecules (e.g. acids and proteases) that have the potential to promote bone destruction at the infection site (4). These molecules can either induce immune system activation or direct bone resorption due to their osteolytic properties. Published reports on pathogen biofilm thus far have focused on indirect bone resorption mediated by host cells and factors secondary to immune system activation (3, 5, 6). However, direct bone resorption due to biofilm pathogen excretions has not been adequately studied yet. Therefore, we planned to investigate the effect of biofilm pathogen in ex-vivo human bones in the absence of the .
BMC Musculoskeletal Disorders, 2013
Background: Osteomyelitis is a severe and often debilitating disease characterized by inflammatory destruction of bone. Despite treatment, chronic infection often develops which is associated with increased rates of treatment failure, delayed osseous-union, and extremity amputation. Within affected bone, bacteria exist as biofilms, however the impact of biofilms on osteoblasts during disease are unknown. Herein, we evaluated the effect of S. aureus biofilms on osteoblast viability, osteogenic potential, and the expression of the pro-osteoclast factor, receptor activator of NF-kB ligand (RANK-L). Methods: Osteoblasts were exposed to biofilm conditioned media (BCM) from clinical wound isolates of Staphylococcus aureus under normal growth and osteogenic conditions to assess cellular viability and osteoblast differentiation, respectively. Cell viability was evaluated using a live/dead assay and by quantifying total cellular DNA at days 0, 1, 3, 5, and 7. Apoptosis following treatment with BCM was measured by flow-cytometry using the annexin V-FITC/PI apoptosis kit. Osteogenic differentiation was assessed by measuring alkaline phosphatase activity and intracellular accumulation of calcium and osteocalcin for up to 21 days following exposure to BCM. Expression of genes involved in osteogenic differentiation and osteoclast regulation, were also evaluated by quantitative real-time PCR. Results: BCM from clinical strains of S. aureus reduced osteoblast viability which was accompanied by an increase in apoptosis. Osteogenic differentiation was significantly inhibited following treatment with BCM as indicated by decreased alkaline phosphatase activity, decreased intracellular accumulation of calcium and inorganic phosphate, as well as reduced expression of transcription factors and genes involved in bone mineralization in viable cells. Importantly, exposure of osteoblasts to BCM resulted in up-regulated expression of RANK-L and increase in the RANK-L/ OPG ratio compared to the untreated controls. Conclusions: Together these studies suggest that soluble factors produced by S. aureus biofilms may contribute to bone loss during chronic osteomyelitis simultaneously by: (1) reducing osteoblast viability and osteogenic potential thereby limiting new bone growth and (2) promoting bone resorption through increased expression of RANK-L by osteoblasts. To our knowledge these are the first studies to demonstrate the impact of staphylococcal biofilms on osteoblast function, and provide an enhanced understanding of the pathogenic role of staphylococcal biofilms during osteomyelitis.
Environmental factors modulate biofilm formation by Staphylococcus aureus
Science Progress, 2020
Biofilm formation on indwelling medical devices represents an exclusive evasion mechanism for many pathogenic bacteria to establish chronic infections. Staphylococcus aureus is one of the major bacterial pathogens that are able to induce both animal and human infections. The continued emergence of multiple drug-resistant S. aureus, especially methicillin-resistant S. aureus, is problematic due to limited treatment options. Biofilm formation by S. aureus complicates the treatment of methicillin-resistant S. aureus infections. Therefore, elucidating the mechanisms of biofilm formation in this pathogen is important for the development of alternative therapeutic strategies. Various environmental and genetic factors contribute to biofilm formation. In this review, we address the environmental factors and discuss how they affect biofilm formation by S. aureus.
PLOS ONE
Biofilm formation is a critical virulence factor responsible for treatment failure and chronicity in orthopaedic device-related infections (ODIs) caused by Staphylococcus aureus. Clonal lineages differ in terms of their biofilm forming capacities. The aim of this study was to investigate the correlation between the clonal complex (CC) affiliation and biofilm phenotype of 30 clinical S. aureus isolates responsible of ODI based on i) early biofilm formation using BioFilm Ring Test® and mature biofilm formation using crystal violet assays, ii) biofilm composition using DNase and proteinase K activity, and iii) prevention of biofilm formation by cloxacillin, teicoplanin and vancomycin using Antibiofilmogram® (biofilm minimal inhibitory concentration-bMIC). In terms of early biofilm formation, the CC30 strains were significantly slower than the CC5, CC15 and CC45 strains. CC45 strains produced significantly more mature biofilm than other group of strains did. The formation of biofilms was highly dependent on the presence of extracellular DNA in the CC5, CC15 and CC30 strains whereas it was mostly dependent on the presence of proteins in CC45. Finally, the CC30 group highlighted higher proportion of susceptible (bMIC < breakpoints of EUCAST guidelines) for cloxacillin, teicoplanin and vancomycin compared to the other CCs. These results demonstrate that the biofilm phenotype of clinical S. aureus isolates from ODIs is strongly related to their respective CC affiliation.
International Journal of Molecular Sciences
The biofilm-associated infections of bones are life-threatening diseases, requiring application of dedicated antibiotics in order to counteract the tissue damage and spread of microorganisms. The in vitro analyses on biofilm formation and susceptibility to antibiotics are frequently carried out using methods that do not reflect conditions at the site of infection. To evaluate the influence of nutrient accessibility on Staphylococcus aureus biofilm development in vitro, a cohesive set of analyses in three different compositional media was performed. Next, the efficacy of four antibiotics used in bone infection treatment, including gentamycin, ciprofloxacin, levofloxacin, and vancomycin, against staphylococcal biofilm, was also assessed. The results show a significant reduction in the ability of biofilm to grow in a medium containing elements occurring in the serum, which also translated into the diversified changes in the efficacy of used antibiotics, compared to the setting in which...
Antibiotics
The need for bone and joint prostheses is currently growing due to population aging, leading to an increase in prosthetic joint infection cases. Biofilms represent an adaptive and quite common bacterial response to several stress factors which confer an important protection to bacteria. Biofilm formation starts with bacterial adhesion on a surface, such as an orthopedic prosthesis, further reinforced by matrix synthesis. The biofilm formation and structure depend on the immediate environment of the bacteria. In the case of infection, the periprosthetic joint environment represents a particular interface between bacteria, host cells, and the implant, favoring biofilm initiation and maturation. Treating such an infection represents a huge challenge because of the biofilm-specific high tolerance to antibiotics and its ability to evade the immune system. It is crucial to understand these mechanisms in order to find new and adapted strategies to prevent and eradicate implant-associated i...
Anin vitrobiofilm model ofStaphylococcus aureusinfection of bone
Letters in Applied Microbiology, 2019
Significance and Impact of the Study: The majority of studies of antibiotic efficacy in the treatment of chronic osteomyelitis are carried out in animals. We developed an in vitro model of Staphylococcus aureus infection of bone to evaluate the ability of antibiotics to eradicate mature biofilms on surfaces analogous to necrotic bone. The results demonstrated the difficulties which occur in osteomyelitis treatment, with only very high concentrations of antibiotic able to penetrate the bone sufficiently to reduce bacterial survival whilst still failing to eradicate biofilms. This model could be of use in initial screening of novel compounds intended for use in the treatment of osteomyelitis.
PloS one, 2017
Bone infections are a significant public health burden associated with morbidity and mortality in patients. Microbial biofilm pathogens are the causative agents in chronic osteomyelitis. Research on the pathogenesis of osteomyelitis has focused on indirect bone destruction by host immune cells and cytokines secondary to microbial insult. Direct bone resorption by biofilm pathogens has not yet been seriously considered. In this study, common osteomyelitis pathogens (Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, and Streptococcus mutans) were grown as biofilms in multiple in vitro and ex vivo experiments to analyze quantitative and qualitative aspects of bone destruction during infection. Pathogens were grown as single or mixed species biofilms on the following substrates: hydroxyapatite, rat jawbone, or polystyrene wells, and in various media. Biofilm growth was evaluated by scanning electron microscopy and pH levels were monitored over time. Histomorphologic and q...