Vol.:(0123456789) 1 3
World Journal of Microbiology and Biotechnology (2020) 36:174
https://doi.org/10.1007/s11274-020-02948-6
ORIGINAL PAPER
Anti-Staphylococcal and cytotoxic activities of the short anti-microbial
peptide PVP
Hamed Memariani
1
· Mojtaba Memariani
1
· Reza Mahmoud Robati
1,2
· Soheila Nasiri
1,2
·
Fahimeh Abdollahimajd
1
· Zohre Baseri
3
· Hamideh Moravvej
1
Received: 23 June 2020 / Accepted: 8 October 2020
© Springer Nature B.V. 2020
Abstract
Over the past years, short anti-microbial peptides have drawn growing attention in the research and trade literature because
they are usually capable of killing a broad spectrum of pathogens by employing unique mechanisms of action. This study
aimed to evaluate the anti-bacterial efects of a previously designed peptide named PVP towards the clinical strains of methi-
cillin-resistant Staphylococcus aureus (MRSA) in vitro. Secondary structure, cytotoxicity, and membrane-permeabilizing
efects of the peptide were also assessed. PVP had a tendency to adopt alpha-helical conformation based upon structural
predictions and circular dichroism spectroscopy (in 50% trifuoroethanol). The peptide showed MIC values ranging from 1
to 16 µg/mL against 10 strains of MRSA. In contrast to ciprofoxacin and gentamicin, PVP at sub-lethal concentration (1 µg/
mL) did not provoke the development of peptide resistance after 14 serial passages. Remarkably, 1 h of exposure to 4 × MBC
of PVP (8 µg/mL) was sufcient for total bacterial clearance, whereas 4 × MBC of vancomycin (8 µg/mL) failed to totally
eradicate bacterial cells, even after 8 h. PVP showed negligible cytotoxicity against human dermal fbroblasts at concentra-
tions required to kill the MRSA strains. The results of fow cytometric analysis and fuorescence microscopy revealed that
PVP caused bacterial membrane permeabilization, eventually culminating in cell death. Owing to the potent anti-bacterial
activity, fast bactericidal kinetics, and negligible cytotoxicity, PVP has the potential to be used as a candidate antibiotic for
the topical treatment of MRSA infections.
Keywords Anti-microbial peptide · Methicillin-resistant Staphylococcus aureus · Bactericidal activity · Human dermal
fbroblasts · Membrane permeabilization
Abbreviations
AMPs Anti-microbial peptides
AO/EtBr Acridine orange/ethidium bromide
CD Circular dichroism
CFUs Colony forming units
DMEM Dulbecco’s Modifed Eagle’s Medium
HDFs Human dermal fbroblasts
MBC Minimum bactericidal concentration
MDR Multidrug resistant
MHB Müeller-Hinton broth
MIC Minimum inhibitory concentration
MRSA Methicillin-resistant Staphylococcus aureus
PBS Phosphate-bufered saline
PCR Polymerase chain reaction
PI Propidium iodide
RP-HPLC Reversed-phase high performance liquid
chromatography
SI Selectivity index
SD Standard deviation
TFE Trifuoroethanol
TSB Tryptic soy broth
* Mojtaba Memariani
memaryani@gmail.com
* Hamideh Moravvej
hamidehmoravej@sbmu.ac.ir
1
Skin Research Center, Shahid Beheshti University
of Medical Sciences, Tehran, Iran
2
Department of Dermatology, Loghman Hakim Hospital,
Shahid Beheshti University of Medical Sciences, Tehran,
Iran
3
Department of Pathology and Laboratory Medicine, Shariati
Hospital, Tehran University of Medical Sciences, Tehran,
Iran