Abstract
Staphylococcus epidermidis, is a common microflora of human body that can cause opportunistic infections associated with indwelling devices. It is resistant to multiple antibiotics necessitating the need for naturally occurring antibacterial agents. Malaysian propolis, a natural product obtained from beehives exhibits antimicrobial and antibiofilm properties. Chitosan-propolis nanoparticles (CPNP) were prepared using Malaysian propolis and tested for their effect against S. epidermidis. The cationic nanoparticles depicted a zeta potential of +40 and increased the net electric charge (zeta potential) of S. epidermidis from -17 to -11 mV in a concentration-dependent manner whereas, ethanol (Eth) and ethyl acetate (EA) extracts of propolis further decreased the zeta potential from -17 to -20 mV. Confocal laser scanning microscopy (CLSM) depicted that CPNP effectively disrupted biofilm formation by S. epidermidis and decreased viability to ~25% compared to Eth and EA with viability of ~60–70%. CPNP was more effective in reducing the viability of both planktonic as well as biofilm bacteria compared to Eth and EA. At 100 μg/mL concentration, CPNP decreased the survival of biofilm bacteria by ~70% compared to Eth or EA extracts which decreased viability by only 40%-50%. The morphology of bacterial biofilm examined by scanning electron microscopy depicted partial disruption of biofilm by Eth and EA extracts and significant disruption by CPNP reducing bacterial number in the biofilm by ~90%. Real time quantitative PCR analysis of gene expression in treated bacteria showed that genes involved in intercellular adhesion such as IcaABCD, embp and other related genes were significantly downregulated by CPNP. In addition to having a direct inhibitory effect on the survival of S. epidermidis, CPNP showed synergism with the antibiotics rifampicin, ciprofloxacin, vancomycin and doxycycline suggestive of effective treatment regimens. This would help decrease antibiotic treatment dose by at least 4-fold in combination therapies thereby opening up ways of tackling antibiotic resistance in bacteria.
Highlights
Staphylococcus epidermidis survives on the skin as normal flora under the epithelium and is recognized as an opportunistic pathogen commonly encountered in hospital-acquired infections without critical implications
The bacteria attach to solid surfaces forming biofilms; this process is characterized by distinct phases–initiation of establishment and colonization leading to infectious stage, primary reversible adhesion developing into secondary irreversible adhesion, and biofilm formation [1]
The regulatory gene sarA was only downregulated with Chitosan-propolis nanoparticles (CPNP) treatment but not with propolis Eth treatment. These results show that treatment with CPNP is more effective than propolis extracts and all the treatments downregulate the genes involved in biofilm formation of S. epidermidis, causing inhibition of the same
Summary
Staphylococcus epidermidis survives on the skin as normal flora under the epithelium and is recognized as an opportunistic pathogen commonly encountered in hospital-acquired infections without critical implications. S. epidermidis can adhere to both biotic and abiotic surfaces of indwelling or implanted medical devices or tissues and form biofilms, leading to treatment failure and relapse of infections [2]. It is implicated in cardiac prosthetic valve infections causing endocarditis, which might lead to intra-cardiac abscesses and mortality. S. epidermidis exhibits resistance to β-lactam antibiotics, which are commonly used in clinical settings [3]. Its antimicrobial resistance is mainly linked to its capability to colonize and produce biofilms
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