Journal
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Volume 23, Issue 15, Pages -Publisher
MDPI
DOI: 10.3390/ijms23158308
Keywords
Pseudomonas aeruginosa; chlorhexidine; cross-resistance; biofilms; Psl; cell aggregate; wound
Funding
- Chulalongkorn University through Fundamental Fund 65 [CUFRB65_hea [33] _040_30_21]
- National Research Council of Thailand [NRCT-N41A640076]
- NSRF via the Program Management Unit for Human Resources & Institutional Development, Research, and Innovation [811/2563, B16F640175, B05F640144]
- Second Century Fund (C2F) for Postdoctoral Fellowship, Chulalongkorn University
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In this study, it was found that Pseudomonas aeruginosa adapted to Chlorhexidine exhibited differences in morphology, metabolism, and biofilm characteristics compared to the parent strain. Moreover, the Chlorhexidine-adapted strain developed resistance to Chlorhexidine and colistin through the overexpression of psl genes. Additionally, it was observed that Chlorhexidine-induced activation of bacteria resulted in lower inflammatory reactions compared to the parent strain.
Because Pseudomonas aeruginosa is frequently in contact with Chlorhexidine (a regular antiseptic), bacterial adaptations are possible. In comparison with the parent strain, the Chlorhexidine-adapted strain formed smaller colonies with metabolic downregulation (proteomic analysis) with the cross-resistance against colistin (an antibiotic for several antibiotic-resistant bacteria), partly through the modification of L-Ara4N in the lipopolysaccharide at the outer membrane. Chlorhexidine-adapted strain formed dense liquid-solid interface biofilms with enhanced cell aggregation partly due to the Chlorhexidine-induced overexpression of psl (exopolysaccharide-encoded gene) through the LadS/GacSA pathway (c-di-GMP-independence) in 12 h biofilms and maintained the aggregation with SiaD-mediated c-di-GMP dependence in 24 h biofilms as evaluated by polymerase chain reaction (PCR). The addition of Ca2+ in the Chlorhexidine-adapted strain facilitated several Psl-associated genes, indicating an impact of Ca2+ in Psl production. The activation by Chlorhexidine-treated sessile bacteria demonstrated a lower expression of IL-6 and IL-8 on fibroblasts and macrophages than the activation by the parent strain, indicating the less inflammatory reactions from Chlorhexidine-exposed bacteria. However, the 14-day severity of the wounds in mouse caused by Chlorhexidine-treated bacteria versus the parent strain was similar, as indicated by wound diameters and bacterial burdens. In conclusion, Chlorhexidine induced psl over-expression and colistin cross-resistance that might be clinically important.
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