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Synthetic synovial fluid drives biofilm and virulence expression in Staphylococcus aureus

ABSTRACT: Staphylococcus aureus is a leading cause of implant-associated infections, including periprosthetic joint infections (PJI). Its high virulence and ability to form biofilms make these infections particularly difficult to treat. Traditional in vitro studies rely on conventional growth media that lack key components of the infectious microenvironment, limiting their physiological relevance. To this end, a synthetic synovial fluid (SSF2) model was developed incorporating essential host compounds such as amino acids, hyaluronic acid, albumin and fibrinogen. However, how accurately this model reflects S. aureus behavior during PJI remains unclear. To investigate this, RNA sequencing was performed on a PJI isolate of S. aureus grown in SSF2 and Luria-Bertani (LB) medium, followed by differential gene expression analysis. Functional classification using Clusters of Orthologous Groups (COG) and gene ontology (GO) enrichment was applied, and known and putative virulence and resistance genes were evaluated. To assess how closely SSF2 mimics in vivo conditions, transcriptomic profiles were compared with previously published in vivo gene expression data for the same strain. Approximately half of the protein-coding genes were differentially expressed in SSF2 compared to LB (56.8%). Consistent with transcriptomic findings, growth in SSF2 resulted in reduced growth rates and enhanced biofilm formation. Furthermore, SSF2 induced a large fraction of virulence-associated genes (69.5%), including those encoding toxins, iron acquisition systems, adhesins and efflux pumps. These findings are in line with previously published in vivo data of the same strain. These results highlight the potential of the SSF2 medium to approximate in vivo conditions encountered by S. aureus during PJI. Nonetheless, more comprehensive in vivo transcriptomic datasets are needed to fully validate this model.

ORGANISM(S): Staphylococcus aureus

PROVIDER: GSE312058 | GEO | 2026/02/23

REPOSITORIES: GEO

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Project description:Periprosthetic joint infection (PJI), a devastating complication of total joint replacement, is of incompletely understood pathogenesis and may sometimes be challenging to clinically distinguish from other causes of arthroplasty failure. We characterized human gene expression in 93 specimens derived from surfaces of resected arthroplasties, comparing transcriptomes of subjects with infection- versus non-infection-associated arthroplasty failure. Differential gene expression analysis confirmed 28 previously reported potential biomarkers of PJI, including bactericidal/permeability increasing protein (BPI), cathelicidin antimicrobial peptide (CAMP), C-C-motif chemokine ligand 3 (CCL3), 4(CCL4), and C-X-C-motif chemokine ligand 2 (CXCL2), colony stimulating factor 2 receptor beta (CSF2RB), colony stimulating factor 3 (CSF3), alpha-defensin (DEFA4), Fc fragment of IgG receptor 1B (CD64B), intercellular adhesion molecule 1 (ICAM1), interferon gamma (IFNG), interleukin 13 receptor subunit alpha 2 (IL13RA2), interleukin 17D (IL17D), interleukin 1 (IL1A, IL1B, IL1RN), interleukin 2 receptors (IL2RA, IL2RG), interleukin 5 receptor (IL5RA), interleukin 6 (IL6), interleukin 8 (IL8), lipopolysaccharide binding protein (LBP), lipocalin (LCN2), lactate dehydrogenase C (LDHC), lactotransferrin (LTF), matrix metallopeptidase 3 (MMP3), peptidase inhibitor 3 (PI3), and vascular endothelial growth factor A (VEGFA), and identified three novel molecules of potential diagnostic use for detection of PJI, namely C-C-motif chemokine ligand CCL20, coagulation factor VII (F7), and B cell receptor FCRL4. Comparative analysis of infections caused by staphylococci versus bacteria other than staphylococci and Staphylococcus aureus versus Staphylococcus epidermidis showed elevated expression of interleukin 13 (IL13), IL17D, and MMP3 in staphylococcal infections, and of IL1B, IL8, and platelet factor PF4V1 in S. aureus compared to S. epidermidis infections. Pathway analysis of over-represented genes suggested activation of host immune response and cellular maintenance and repair functions in response to invasion of infectious agents. The data presented provides new potential targets for diagnosis of PJI and for differentiation of PJI caused by different infectious agents.

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Synthetic synovial fluid drives biofilm and virulence expression in Staphylococcus aureus

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