Assessment of antibacterial efficacy of a biocompatible nanoparticle PC@AgNPs against Staphylococcus aureus

doi:10.1016/j.micpath.2018.10.029

Microbial Pathogenesis

Volume 126, January 2019, Pages 27-39

https://doi.org/10.1016/j.micpath.2018.10.029 Get rights and content

Highlights

•
Bactericidal effect of PC@AgNPs on Staphylococcus aureus established by SEM, AFM, fatty acid profile and ion efflux data.
•
The nanoparticle also exhibits anti-biofilm property, evidenced by SEM, CLSM and dye staining method.
•
The anti-coagulase activity with drug Dabigatran indicated down regulation of Coa gene validating anti-bacterial activity.
•
Bio-compatibility of the nanoparticle was amply demonstrated at hemostatic level.

Abstract

A plant, Priva cordifolia mediated silver nanoparticle was prepared and characterized by UV-Vis spectroscopy, fourier-transform infrared spectroscopy (FT-IR), energy dispersive X-ray (EDX), scanning electron microscopy (SEM), transmission electron microscopy (TEM), dynamic light scattering (DLS) and x-ray diffraction (XRD) analysis. The minimum inhibitory concentration of the synthesized nanoparticle against Staphylococcus aureus was found to be 100 ± 0.80 μg/mL with 9.38 ± 0.04 mm zone of inhibition. The bactericidal activity was shown primarily due to membrane damage evident from SEM, atomic force microscopy (AFM), potassium efflux, cellular material leakage, and bio-electrochemical changes in electron transport chain data. It was also of interest to find PC@AgNPs interfering with biofilm formation by S.aureus, assessed qualitatively by SEM, confocal laser scanning microscopy (CLSM) and quantitatively by dye staining method. The bio-compatibility of PC@AgNPs was established by anti-coagulant, thrombolytic, partial thromboplastin time, thrombin-like activity and fibrinolytic activity that suggested its good maintenance of hemostatic conditions. PC@AgNPs also prevented the coagulation of rabbit plasma which as per the standard drug Dabigatran reaction was indicative of the down-regulation of virulence Coa gene expression.

Graphical abstract

Introduction

The need to find new ways and means to combat bacterial infections is keenly felt owing to alarming emergence of drug-resistant pathogenic bacteria [1]. The current focus is on metal nanoparticle and natural anti-bacterial agents. Inhibition of pathogenic bacteria, fungi, and yeast by nanoparticle preparations of silver, gold and titanium have been studied [2,3]. Metal nanoparticles target cellular components (DNA, RNA and ribosome's) and effectively alter cellular processes [4]. Lawrence et al. have reported the plant Priva cordifolia assisted synthesis of silver nanoparticles employing water extract of the plant's leaves [5]. However, they have not reported its antibacterial activity or other attributes of significance for its therapeutical potential. The present study thus became exigent to take forward the nanoparticle synthesis to examine in detail its potential for therapeutic applications. Priva cordifolia, though known a long time ago in traditional medicine as a wound-healing agent remains less studied.

S. aureus being a common pathogen was chosen as a model in the present study. It is among the normal flora colonizing in human nose, skin and gastrointestinal tract, but can turn into a perilous pathogen sometimes, to invade the bloodstream of humans to cause bacteremia, endocarditis more often as medical devices-associated (surgical tools, catheter etc.) infections on skin, soft tissues, pleuropulmonary and osteoarticular sites [6]. S.aureus secretes coagulase to activate the conversion of host prothrombin to the fibrin-fibrils, which protect the pathogen from phagocytosis in the immune system. Additionally, this also causes the diminution of drug activity on the pathogen [7]. S. aureus is also a biofilm forming pathogen and in this adaptation becomes formidable with multiple drug-resistances [8,9].

Section snippets

Chemicals, reagents and standard bacterial culture

Brain heart infusion broth(BHI), Rabbit plasma and Baird Parker agar (BHA) were purchased from Hi-Media (Mysore, India). Dabigatran (Pradaxa) was procured from the regional medical store. All other chemicals were purchased from Sigma & Merck, India. S.aureus (MTCC-96) was procured from Microbial Type Culture Collection (MTCC), Chandigarh, India.

Green synthesis of P.cordifolia silver nanoparticles (PC@AgNPs)

Priva cordifolia plants were collected from Chamundi hill terrain of Mysore, Karnataka, India. The plant was identified and authenticated [10]. The

Green synthesis and characterization of PC@AgNPs

The synthesized PC@AgNPs was confirmed by reduction of Ag⁺ ions and formation of a brown colored colloidal solution. The intensity of colloidal solution was recorded the λ max at 415 nm by UV–visible spectroscopy at the end of the reaction. The change of color in colloidal solution was indicative of the reduction of silver ions by the plant extract and act as a potent reducing and capping agent (Fig. 1A).

The FT-IR study was carried out for PC@AgNPs to find out the functional group involved in

Conclusion

The present study indicated the silver nanoparticle preparation, PC@AgNPs as having potent bactericidal activity compared to the crude plant extract, evident by bacterial membrane damage, cellular leakage and potassium efflux. The PC@AgNPs also showed capacity to prevent the bacterial biofilm formation, anticoagulant property and down-regulation of S.aureus Coa gene. In view of these attributes, PC@AgNPs could be regarded a worthy candidate to examine further its potential as an effective

Disclosure statement

The authors declare no competing financial interest. The authors alone are responsible for the content and writing of this article.

Conflicts of interest

The authors declare no competing interests.

Author contributions

Ananda, A.P., (APA) and Krishnamurthy, N.B., (KNB) designed the study, KNB supervised, review the manuscript and APA conducted all the experiments, prepared the manuscript. Manukumar, H.M., Nagendra, B.S., and Savitha, K.R. were given valuable suggestion and helped in the preparation of the manuscript.

Acknowledgements

The author A.P. Ananda, greatly acknowledges the A.R.Subramanya, CEO of the Ganesh consultancy and Analytical Services, Mysore for provided the facility to carry out this study. And also heartly thankful to Dr.Richard Joseph, former scientist (microbiology) CFTRI, Mysore for valuable inputs and suggestions on the manuscript.

References (44)

I.M. Hamouda
Current perspectives of nanoparticles in medical and dental biomaterials
J. Biomed. Res.
(2012)
A.S. Kwon et al.
Higher biofilm formation in multidrug-resistant clinical isolates of Staphylococcus aureus
Int. J. Antimicrob. Agents
(2008)
H.M. Manukumar et al.
Promising biocidal activity of thymol loaded chitosan silver nanoparticles (T-C@AgNPs) as anti-infective agents against perilous pathogens
Int. J. Biol. Macromol.
(2017)
P. Eaton et al.
Atomic force microscopy study of the antibacterial effects of chitosans on Escherichia coli and Staphylococcus aureus
Ultramicroscopy
(2008)
A.K. Chauhan et al.
Thymol disrupts the membrane integrity of Salmonella ser. typhimurium in vitro and recovers infected macrophages from oxidative stress in an ex vivo model
Res. Microbiol.
(2014)
M. Wang et al.
Mediated electrochemical method for the analysis of membrane damage effects of phenolic compounds to Staphylococcus aureus
J. Electroanal. Chem.
(2015)
R.I. Evans et al.
The effect of growth temperature on the phospholipid and fatty acyl compositions of non-proteolytic Clostridium botulinum
Int. J. Food Microbiol.
(1998)
F. Gambinossi et al.
Aggregation kinetics and colloidal stability of functionalized nanoparticles
Adv. Colloid Interface Sci.
(2015)
M.J. Kim et al.
Toxic effects of silver nanoparticles and nanowires on erythrocyte rheology
Food Chem. Toxicol.
(2014)
B.S. Harish et al.
Synthesis of fibrinolytic active silver nanoparticle using wheat bran xylan as a reducing and stabilizing agent
Carbohydr. Polym.
(2015)

K. Jyoti et al.

Characterization of silver nanoparticles synthesized using Urtica dioica Linn. leaves and their synergistic effects with antibiotics

J. Radiat. Res. Appl. Sci.

(2016)

E. Torlak et al.

Biofilm formation by Staphylococcus aureus isolates from a dental clinic in Konya, Turkey

J. Infect. Public Health

(2017)

W. Huang et al.

Chemical analysis and in vitro antimicrobial effects and mechanism of action of Trachyspermum copticum essential oil against Escherichia coli

Asian Pac. J. Trop. Med.

(2017)

D. Chudobova et al.

Oxidative stress in Staphylococcus aureus treated with silver (I) ions revealed by spectrometric and voltammetric assays

Int. J. Electrochem. Sci

(2013)

R.J. Fair et al.

Antibiotics and bacterial resistance in the 21st century

Perspect. Med. Chem.

(2014)

Linlin Wang et al.

The antimicrobial activity of nanoparticles: present situation and prospects for the future

Int. J. Nanomed.

(2017)

L. Shang et al.

Engineered nanoparticles interacting with cells: size matters

J. Nanobiotechnol.

(2014)

R.A.R. Lawrence et al.

Green synthesis and characterization of silver nanoparticles of leaf extracts of priva cordifolia (LF) druce

Int. J. Ecotoxicol. Ecobiol.

(2017)

Steven YC. Tong et al.

Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management

Clin. Microbiol. Rev.

(2015)

L. Thomer et al.

Pathogenesis of Staphylococcus aureus bloodstream infections

Annu. Rev. Pathol.

(2016)

T. Oyama et al.

Biofilm-forming methicillin-resistant Staphylococcus aureus survive in Kupffer cells and exhibit high virulence in mice

Toxins

(2016)

A.P. Ananda et al.

A taxonomical note on priva cordifolia (LF) druce an Indian ethno-medicinal plant

World J. Pharm. Pharmaceut. Sci.

(2016)

Cited by (30)

A trinuclear Zn (II) schiff base dicyanamide complex attenuates bacterial biofilm formation by ROS generation and membrane damage and exhibits anticancer activity
2024, Microbial Pathogenesis
A trinuclear Zn (II) complex, [(ZnL{N(CN)₂})₂Zn], termed complex 1 has been synthesized by the reaction of an aqueous solution of sodium dicyanamide to the methanolic solution of Zn (CH₃COO)₂, 2H₂O and corresponding Schiff base (H₂L) which is derived from 1:2 condensation of 1, 4 butane diamine with 3-ethoxy salicylaldehyde. Complex 1 is characterized by elemental analysis, IR, UV and Single X-ray diffraction study. Drug resistance is a growing global public health concern that has prompted researchers to look into advanced alternative treatment modalities. In this context, complex 1 has shown promising antibacterial and antibiofilm efficacy against gram-positive Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus strains. Complex 1 attenuated Staphylococcal biofilm formation by reducing several virulence factors including the formation of extracellular polysaccharide matrix, slime, haemolysin, staphyloxanthin, auto-aggregation, cell surface hydrophobicity, and motility. Notably, complex 1 mechanistically potentiated Reactive Oxygen Species (ROS) generation within the bacterial cells, leading to the damage of bacterial cell membrane followed by DNA leakage and thereby impeding the growth of Staphylococcus aureus. Furthermore, complex 1 significantly exhibited anticancer activity by reducing the growth of prostate adenocarcinoma cells. It obstructed the migration of cancer cells by potentiating apoptosis and arresting the cell cycle at the G2/M phase. In summary, complex 1 could act as a potent candidate for the generation of novel antibacterial, antibiofilm as well as anticancer treatment regimens for the management of drug-resistant biofilm-mediated Staphylococcus aureus infection and lethal prostate malignancy.
Exploring the biological application of Penicillium fimorum-derived silver nanoparticles: In vitro physicochemical, antifungal, biofilm inhibitory, antioxidant, anticoagulant, and thrombolytic performance
2023, Heliyon
This study showed the anti-candida, biofilm inhibitory, antioxidant, anticoagulant, and thrombolytic properties of biogenic silver nanoparticles (AgNPs) fabricated by using the supernatant of Penicillium fimorum (GenBank accession number OQ568180) isolated from soil. The biogenic AgNPs were characterized by using different analytical techniques. A sharp surface plasmon resonance (SPR) peak of the colloidal AgNPs at 429.5 nm in the UV–vis spectrum confirmed the fabrication of nanosized silver particles. The broth microdilution assay confirmed the anti-candida properties of AgNPs with a minimum inhibitory concentration (MIC) of 4 μg mL⁻¹. In the next step, the protein and DNA leakage assays as well as reactive oxygen species (ROS) assay were performed to evaluate the possible anti-candida mechanisms of AgNPs representing an increase in the total protein and DNA of supernatant along with a climb-up in ROS levels in AgNPs-treated samples. Flow cytometry also confirmed a dose-dependent cell death in the AgNPs-treated samples. Further studies also confirmed the biofilm inhibitory performance of AgNPs against Candia albicans. The AgNPs at the concentrations of MIC and 4*MIC inhibited 79.68 ± 14.38% and 83.57 ± 3.41% of biofilm formation in C. albicans, respectively. Moreover, this study showed that the intrinsic pathway may play a significant role in the anticoagulant properties of AgNPs. In addition, the AgNPs at the concentration of 500 μg mL⁻¹, represented 49.27%, and 73.96 ± 2.59% thrombolytic and DPPH radical scavenging potential, respectively. Promising biological performance of AgNPs suggests these nanomaterials as a good candidate for biomedical and pharmaceutical applications.
Design, synthesis, and in-silico studies of pyrazolylpyridine analogues: A futuristic antibacterial contender against coagulase positive superbug-MRSA
2022, Journal of Molecular Structure
Citation Excerpt :
For all β-lactams, PBP2a has a low affinity and the active PBP2a site is, indeed, capable of blocking the binding of β-lactams while enabling cross-linking to progress at the same time. All β-lactams are ineffective against PBP2a expressing bacterial strains, including penicillin's, cephalosporins, and carbapenems [22]. Targeting the active site and allosteric sites of PBP2a by synthesized compound 4e results the inactivation of cell wall synthesis and helps the β-lactam drugs to inhibit the cellwall synthesis (Fig. 11a and 11c) which validated by cellular material (DNA) leakage and potassium efflux of MRSA (Fig. 3).
In the coagulation cascade during host infection by MRSA, staphylococci are blocked within a fibrin clot mesh, and very hard to clear these pathogens by host tissues and also enable the pathogen to cause lethal sepsis. To overcome this complication, this work aimed at synthesis of pyrazolylpyridine analogs as potential anti-superbug agents and using a simple synthetic protocol. The synthesized pyrazolylpyridine analogs were characterized using spectroscopic techniques like FTIR, NMR, and mass spectroscopy. All synthesized pyrazolylpyridine analogs were evaluated for the in-vitro antibacterial activity. The antibacterial activity was further confirmed with cellular content leakage and measurement of potassium efflux. The anti-biofilm activity was studied qualitatively and quantitatively with crystal violet dye technique. The compounds binding ability with 1NU7 and 1MWT proteins were tested using molecular docking and further analogs were exposed to dynamic simulations. The synthesized compounds were also inspected for their ADMET analysis. It was observed that 4(a g) series showed a potential antibacterial activity compared to 3(a g) analogs to standard drug vancomycin. The MIC value of 4e was found to 30±0.4 µg/mL and also showed an excellent anti-biofilm activity. The revealed data was strongly correlated with molecular docking studies on staphylocoagulase enzyme (PDB: 1NU7) and Penicillin-binding protein 2a (PBP2a) (PDB: 1MWT) with a good docking score, and glide score with high binding energy. By dynamic simulation it was obtained that 4d exhibit stable interaction with staph-coagulase enzyme up to 20 ns with PBP2a protein, and compound 4e exhibited stable interaction at both active and allosteric site. The potent pyrazolylpyridine was designed and synthesized to treat infectious agent MRSA. Among the analogs synthesized, compound 4e showed antibacterial activity validated by insilico studies. In this study, experimental results show significant anti-coagulase, anti-biofilm and antimicrobial activity against MRSA by synthesised compound 4e. This leads to the compound 4e can serve as a potent drug against MRSA in upcoming days.
Antibacterial activity and mechanism of slightly acidic electrolyzed water against Shewanella putrefaciens and Staphylococcus saprophytic
2022, Biochemical and Biophysical Research Communications
The purpose of this study was to investigate the antimicrobial effect and mechanism of slightly acidic electrolyzed water (SAEW) against Shewanella putrefaciens (S. putrefaciens) and Staphylococcus saprophytic (S. saprophyticus). The results showed that SAEW exhibited strong antimicrobial activity against tested bacteria, which was positively correlated to the available chlorine concentration (ACC) of SAEW. The mortality rate of S. putrefaciens and S. saprophyticus was up to 96% and 85%, respectively, when the ACC of SAEW was 60.0 mg/L. The results of scanning electron microscopy (SEM) showed that the cell morphology and structure were destroyed by SAEW. Besides, the results of confocal laser scanning microscopy (CLSM), leakage of DNA and protein provided evidence that SAEW induced membrane permeabilization in cells. Compared with the control, the intracellular reactive oxygen species (ROS) generated by SAEW was strengthened significantly with the increase of ACC, and the cells were injured to death accordingly.
Design, synthesis, molecular docking and DFT computational insight on the structure of Piperazine sulfynol derivatives as a new antibacterial contender against superbugs MRSA
2022, Journal of Molecular Structure
In this investigation, a series of piperazine sulfynol derivatives 5(a-l) were synthesized and amply characterized by various spectral techniques viz, LC-MS, ¹H NMR, ¹³C NMR, and FT-IR. The energy and other quantum chemical computations of all the piperazine sulfynol derivatives were achieved using Density functional theory (DFT), Absorption, Distribution, Metabolism and Excretion (ADME), Blood Brain Barrier (BBB) and their Prediction of Activity Spectra of Computational Screening (PASS) for their possible approaches for biological applications were evaluated. The synthesized compounds were checked for the physicochemical properties such as clogP, clogS, drug-likeness, total surface area, polar surface area, H-acceptor, and H-donor parameters. The considerable change in the design with sulfonyl moiety on the piperazine core motivates to find out new antibacterial contender of the resultant molecules against superbugs-MRSA. All 5(a-l) moieties were tested for antibacterial potency against MRSA. The 5e moiety shows a MIC value of 35±0.41 µg/mL and 9.90±0.03 ZOI in mm compared to drug bacitracin 10 µg/disk (10.21±0.04) and streptomycin 10 µg/disk (14.10±0.06) ZOI in mm. The action of biocidal properties against MRSA was confirmed by an in silico docking study on the protein 1FZP and 3SRW of MRSA. The revealed data strongly recommended 5e having a good docking score, binding energy, and glide energy with high binding affinity. The antibacterial activity was validated by SEM, cellular leakage, potassium efflux and inhibitory effect on electron transport chain. The behavior of the 5e analog in haemostatic condition was well established and cytotoxicity was evaluated against L6 cell lines. The revealed data showed that 5e analog is a potential antibacterial contender against MRSA and it can be used as a futuristic drug in the eradication of MRSA infections.
Designing bi-functional silver delafossite bridged graphene oxide interfaces: Insights into synthesis, characterization, photocatalysis and bactericidal efficiency
2021, Chemical Engineering Journal
Citation Excerpt :
The bacterial strain was cultured in prescribed broth as per the revival procedure provided by MTCC. The disc diffusion method was adopted to observe the bactericidal activity was AgFeO2, GO and AgFeO2/GO catalysts following the procedure reported [45]. The bacterial cultures were prepared from the overnight culture, then 1 × 108 CFU/mL (0.5 McFarland) of cells was inoculated onto nutrient agar, further, the sterile disc (6 mm) was loaded with 5 µl of different serially diluted test samples (100, 200, 300, 400, and 500 μg/mL).
The bi-functional nanoscale catalysts are of growing attention to address environmental problems. The present work successfully fabricated the hexagonal silver delafossite oxide bridged graphene oxide ultrathin nanosheets interfaces (AgFeO₂/GO) to target photodegradation of an antibiotic and inactivation of model waterborne pathogens for the first time. The AgFeO₂/GO composite was prepared via facile hydrothermal treatment followed by ultrasonic agitation by varying the GO loading. The AgFeO₂/GO composite and their parent constituents were verified by advanced analytical, microscopic, and spectroscopic techniques like FESEM, EDX, HR-TEM, DLS, XRD, FTIR, XPS, UV–Vis, and PL. The AgFeO₂/GO3 composite showed excellent photocatalytic activity towards mineralization of a broad-spectrum antibiotic, lomefloxacin (LMF) under the irradiation of visible light (30 W, LED). A maximum of ~88 % of LMF (20 mg/L) was degraded within 75 min of catalysis. The superoxide and hydroxyl radicals were predominant in LMF breakdown. In addition, AgFeO₂/GO3 was proved effective up to three consecutive trials retaining 78.85 % efficiency. On the other hand, AgFeO₂/GO3 heterojunctions demonstrated fair bactericidal activity against pathogens, Escherichia coli and Staphylococcus aureus. Disc diffusion method of antimicrobial assay confirmed the activity of AgFeO₂/GO3 on par with standard drugs. Therefore, in both the functionalities, the GO incorporation along with AgFeO₂ standpoints enhanced photoactivity and catalytic inactivation making it a sought-after, bi-functional, and efficient environmental catalyst.

View all citing articles on Scopus

View full text

Assessment of antibacterial efficacy of a biocompatible nanoparticle PC@AgNPs against Staphylococcus aureus

Highlights

Abstract

Graphical abstract

Introduction

Section snippets

Chemicals, reagents and standard bacterial culture

Green synthesis of P.cordifolia silver nanoparticles (PC@AgNPs)

Green synthesis and characterization of PC@AgNPs

Conclusion

Disclosure statement

Conflicts of interest

Author contributions

Acknowledgements

J. Biomed. Res.

Int. J. Antimicrob. Agents

Int. J. Biol. Macromol.

Ultramicroscopy

Res. Microbiol.

J. Electroanal. Chem.

Int. J. Food Microbiol.

Adv. Colloid Interface Sci.

Food Chem. Toxicol.

Carbohydr. Polym.

J. Radiat. Res. Appl. Sci.

J. Infect. Public Health

Asian Pac. J. Trop. Med.

Int. J. Electrochem. Sci

Antibiotics and bacterial resistance in the 21st century

Perspect. Med. Chem.

The antimicrobial activity of nanoparticles: present situation and prospects for the future

Int. J. Nanomed.

Engineered nanoparticles interacting with cells: size matters

J. Nanobiotechnol.

Green synthesis and characterization of silver nanoparticles of leaf extracts of priva cordifolia (LF) druce

Int. J. Ecotoxicol. Ecobiol.

Staphylococcus aureus infections: epidemiology, pathophysiology, clinical manifestations, and management

Clin. Microbiol. Rev.

Pathogenesis of Staphylococcus aureus bloodstream infections

Annu. Rev. Pathol.

Biofilm-forming methicillin-resistant Staphylococcus aureus survive in Kupffer cells and exhibit high virulence in mice

Toxins

A taxonomical note on priva cordifolia (LF) druce an Indian ethno-medicinal plant

World J. Pharm. Pharmaceut. Sci.