Startseite Bactericidal and cytotoxic properties of green synthesized nanosilver using Rosmarinus officinalis leaves
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Bactericidal and cytotoxic properties of green synthesized nanosilver using Rosmarinus officinalis leaves

  • Maha Daghestani , Sarah A. Al Rashed , Wadha Bukhari , Badryah Al-Ojayan , Eiman M. Ibrahim , Asma M. Al-Qahtani , Nada M. Merghani , Rasha Ramadan und Ramesa Shafi Bhat EMAIL logo
Veröffentlicht/Copyright: 3. Mai 2020
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Green Processing and Synthesis
Aus der Zeitschrift Green Processing and Synthesis Band 9 Heft 1

Abstract

Green synthesized nanoparticles from plant extracts are being used in various biomedical applications, particularly because of their bactericidal and cytotoxic activities. In this study, silver nanoparticles (AgNPs) were successfully synthesized from the Rosmarinus officinalis aqueous leaf extract. Different spectroscopic and microscopic analyses such as ultraviolet-visible (UV-vis) spectroscopy, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy, and energy-dispersive X-ray spectroscopy were performed to verify the biosynthesized AgNPs in our sample. The formation of nanosilver particles was preliminarily confirmed by UV-vis spectroscopy at 400 nm. The presence of carboxylic or amide groups was confirmed by FTIR, for the reduction of the silver ion. Transmission electron microscopy confirmed a particle size of 12–22 nm. The prepared AgNPs showed good antibacterial activity against human pathogens and good cytotoxic activity against the human breast cancer cell line (MDA MB 231). The nanoparticles prepared from R. officinalis can be used for various biomedical applications.

Keywords: Rosmarinus officinalis; green synthesis; nanosilver

List of abbreviations

AgNPs

Silver nanoparticles

DLS

Dynamic light scattering

EDX

energy-dispersive X-ray spectrum

FTIR

Fourier transform infrared spectroscopy

PDI

Polydispersity index

SPR

Surface plasmon resonance

TEM

Transmission electron microscopy

UV-vis

Ultraviolet-visible absorption

1 Introduction

Silver has been used by mankind for a long time because of its low toxicity and good biocidal properties. It is frequently used in water-purifying systems in hospitals, for healing injuries, and in the field of nanoparticle synthesis [1]. Some bacteria are resistant to silver and are able to accumulate this metal up to 25% of their dry weight biomass in their cells [2], which makes them suitable for extracting silver from ores [3]. Silver nanoparticles (AgNPs) are used as an effective nano-drug against many diseases [4]. Also, they are more efficient in killing silver-resistant bacteria due to their highly developed surface area [5], whereas highly reactive silver ions (Ag+ ions) form insoluble precipitates so are less efficient in killing bacteria [6].

Nanobiotechnology is constantly exploring the synthesis of metal nanoparticles in a nontoxic and ecofriendly manner [7]. The main requirements for the synthesis of a metal nanoparticle are the solvent medium, a reducing agent, and a nontoxic stabilizer of nanoparticles [8]. Plants are rich in active compounds and, thus, can be used as a good source of reducing agents for synthesizing nanoparticles [9]; this has prompted a new field of green synthesis [10]. Almost all parts of the plants are rich in active biomolecules such as amino acids, alkaloids, proteins, polysaccharides, phenolics, terpenoids, and flavones, which can exhibit good reducing and capping properties useful in synthesizing nanoparticles [11]. Additionally, hydroxyl and carboxyl groups present in phenols and flavonoids wrap the nanoparticles and act as capping agents [12].

Rosmarinus officinalis, commonly known as rosemary, contains many different bioactive compounds, and it is used as a flavoring spice in cooking for many years [13]. It is one of the most important commercial crops in Saudi Arabia [14]. Its essential oil has been reported to have analgesic, antispasmodic, antibacterial, and hypnotic effects [15]. It is also used in the treatment of depression and stress-related disorders [16]. Same plants grown under different climatic conditions show different active components [17,18]. Some environmental factors like type of soil and temperature can alter the plant structure and its phytochemical activity, especially the bioactive compounds present in the leaves [19]. Some studies even reported a direct relationship between vitamins and phenolic compounds of plant and soil mineral content of the area where the plant is grown [20]. Natural compounds of R. officinalis leaves change significantly if grown under differential environmental conditions [21]. It was of great interest to synthesize AgNPs by the reduction of aqueous Ag+ ions with the R. officinalis leaf extract collected from Riyadh, Saudi Arabia, and synthesis of AgNPs from R. officinalis collected from deserts has not been reported. The green synthesized AgNPs were characterized using various microscopic and analytical techniques, including ultraviolet-visible absorption (UV-vis) spectroscopy, Fourier transform infrared spectroscopy, and transmission electron microscopy (TEM). The bactericidal and cytotoxic properties of the synthesized nanoparticles were also analyzed.

2 Materials and methods

2.1 Leaf extract

Fresh green leaves of rosemary cultivated in the Riyadh region of Saudi Arabia were used for this study. Ten grams of healthy leaves were washed, chopped, and boiled in 0.1 L of double-distilled water for 30 min. After cooling to room temperature, the broth was filtered and stored at 4°C.

2.2 Biosynthesis of AgNPs

The freshly prepared leaf extract was mixed with a silver nitrate solution at a final concentration of 5 mM at room temperature. The reduction of silver ions to AgNPs was indicated by a color change from yellow to brown and recorded by scanning through a UV-vis spectrophotometer at different intervals for 3 h.

2.3 Characterization of the prepared nanoparticles

The as-prepared nanoparticles were analyzed by observing the color change of the reaction mixture. The absorbance of the reaction mixture was measured over the range of 200–700 nm every 30 min. The average size of the as-prepared nanoparticles was determined by carrying out the zeta potential analysis. The hydrodynamic size and zeta potential of the prepared AgNPs were measured at the scattering angle of 173° with a red laser at the wavelength of 633 nm. The average size of the synthesized particles was measured with the help of a TEM. Infrared spectroscopy was used to determine the nature of bio-reducing functional groups.

2.4 Antimicrobial activity

Bacterial strains, namely Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis, Salmonella typhi, Staphylococcus aureus, Staphylococcus epidermidis, and Streptococcus pneumoniae, were provided by King Khalid Hospital, Riyadh, Saudi Arabia. The antibacterial activity of the synthesized AgNPs against all the mentioned bacteria was determined by the disk diffusion method.

2.5 Screening of cytotoxic activity of the prepared nanoparticles

R. officinalis nanoparticles were screened for cytotoxicity on the breast cancer cell line MDA-231 by the MTT assay. Different concentrations of the synthesized nanoparticles (3.125, 6.25, 12.5, 25, 50, and 100 µL) were used to treat the cancerous cells. The inhibition percentage was calculated.

3 Results

UV-vis spectroscopy was used as a primary method to confirm the synthesis of nanoparticles due to their interaction with specific wavelengths. The synthesis of nanoparticles from silver ions in the presence of the rosemary plant extract was initially confirmed by a color change from yellow to brown. The surface plasmon resonance (SPR) peak at 400 nm confirmed the presence of nanoparticles in the extract (Figure 1). Free electrons in the nanoparticles resonate at certain wavelengths, resulting in SPR. A size reduction of the particles can lower the SPR peak, thus relating the absorption to the particle size [22].

Figure 1 UV-vis spectral analysis of the biosynthesized AgNPs prepared with aqueous R. officinalis leaf extract.
Figure 1

UV-vis spectral analysis of the biosynthesized AgNPs prepared with aqueous R. officinalis leaf extract.

Recorded infrared (IR) spectra point toward the active compounds of the R. officinalis extract, which facilitate the green synthesis of nanoparticles. Nanoparticles are synthesized in three phases, starting with the activation phase, where the plant metabolites with reducing capacities help to split the metal ion from the salt. Next is the growth phase, where metal ions combine to form nanoparticles, and finally, the termination phase, where capping around the synthesized particle is done with the help of the metabolites present in the plant extract [23]. The IR spectrum of Figure 2 shows an intense peak at 1636.75 cm−1, which is recognized as C═O and C═C bond stretches and identified as carboxylic or amide groups, and a peak at 3304.40 cm−1, which represents the hydroxyl group and suggests the presence of phenols, flavonoids, and alcohols in the plant extract.

Figure 2 IR spectra of the biosynthesized AgNPs prepared with aqueous R. officinalis leaf extract.
Figure 2

IR spectra of the biosynthesized AgNPs prepared with aqueous R. officinalis leaf extract.

In this study, we used dynamic light scattering (DLS) to determine the particle size distribution of the synthesized AgNPs (Figure 3). The Z-average mean of the synthesized AgNPs was 66.79 nm, with a polydispersity index (PDI) of 0.177. The PDI is best measured with DLS. Our results with respect to nanoparticles size are acceptable, as a PDI value of more than 0.7 indicates a broad size distribution [24]. Plant extracts are good reducing agents for the synthesis of metal nanoparticles. The interface of the particles mainly depends on the zeta potential since a larger zeta potential results in a particle abundance, thus providing stability to the particles (Figure 4) [25]. The energy-dispersive X-ray spectrum (EDX) of the synthesized nanoparticles exhibited strong signals in the oxygen, potassium, and chlorine regions (Figure 5). The presence of other elements such as carbon, calcium, sodium, and magnesium was also observed in the spectrum. A TEM image was used to find the average size of the prepared nanoparticles, which was in the range of 12–22 nm (Figure 6). TEM is typically an ultrathin sample image showing the 2D structure with the help of an absorption beam passing through the sample [26]. Nowadays, AgNPs are highly important in detergent and disinfectant manufacturing industries because of the resistance level of microbes to chemical fungicides and disinfectants [27]. We examined the antibacterial efficiency of AgNPs prepared from R. officinalis against some pathogenic bacteria. The varying degree of inhibition by both the Gram-positive and Gram-negative bacteria is shown in Table 1.

Figure 3 Average size of the biosynthesized AgNPs prepared with aqueous R. officinalis leaf extract.
Figure 3

Average size of the biosynthesized AgNPs prepared with aqueous R. officinalis leaf extract.

Figure 4 Zeta potential distribution of the biosynthesized AgNPs prepared with aqueous R. officinalis leaf extract.
Figure 4

Zeta potential distribution of the biosynthesized AgNPs prepared with aqueous R. officinalis leaf extract.

Figure 5 EDX elemental analysis of the biosynthesized AgNPs prepared with aqueous R. officinalis leaf extract.
Figure 5

EDX elemental analysis of the biosynthesized AgNPs prepared with aqueous R. officinalis leaf extract.

Figure 6 TEM micrograph of the biosynthesized AgNPs prepared with aqueous R. officinalis leaf extract (particle size shown in range from 12 to 22 nm).
Figure 6

TEM micrograph of the biosynthesized AgNPs prepared with aqueous R. officinalis leaf extract (particle size shown in range from 12 to 22 nm).

Table 1

Antibacterial activity of the biosynthesized AgNPs prepared with aqueous R. officinalis leaf extract against eight bacterial species tested by disc diffusion assay

StrainsZone of inhibition (mm)
Gram positiveS. aureus11 ± 0.30
S. epidermidis8 ± 0.13
S. pneumonia10 ± 0.25
E. faecalis10 ± 0.50
B. subtilis11 ± 0.20
Gram negativeE. coli13 ± 0.10
S. typhi14 ± 0.31
K. pneumoniae9 ± 0.11

4 Discussion

The antibacterial efficacy of a particle depends on its small size and round shape. Our results agree with many previous studies reporting high antimicrobial activities of round-shaped nanoparticles with a size of less than 10 nm [28]. It has been suggested that the antibacterial properties of the plant-derived AgNPs could be due to either the particles’ ability to permeate the cell and interact with the genetic material deoxyribonucleic acid (DNA) and other important constituents, leading to cell death, or their adherence to the negatively charged cell surface that alters the chemical and physical properties of the cell wall and cell membrane [29]. In this study, the green synthesized AgNPs from R. officinalis were evaluated at various concentrations for the cytotoxic effects on the breast cancer cell lines (MB 231) in vitro. As observed in Figure 7, both plant extracts and nanoparticles decrease the viability of cancer cell lines (MB 231), but the cytotoxicity of AgNPs was higher than the cytotoxicity of the plant extract. The cytotoxic effect of nanoparticles on cell viability has a major role in antitumor activity, thereby reducing disease progression [30]. The cytotoxic effects of silver result from the active physiochemical interaction of silver atoms with functional groups of intracellular proteins, as well as with the nitrogen bases and phosphate groups in DNA [31].

Figure 7 Cytotoxicity of aqueous R. officinalis leaf extract and its AgNPs on breast cancer cell lines (MDA MB 231) following 24 h exposure.
Figure 7

Cytotoxicity of aqueous R. officinalis leaf extract and its AgNPs on breast cancer cell lines (MDA MB 231) following 24 h exposure.

Acknowledgments

This research project was supported by a grant from the Research Centre for Female Scientific and Medical Colleges at King Saud University. The authors thank the Deanship of Scientific Research and RSSU at King Saud University for their technical support.

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Received: 2019-11-12
Revised: 2020-01-15
Accepted: 2020-02-04
Published Online: 2020-05-03

© 2020 Maha Daghestani et al., published by De Gruyter

This work is licensed under the Creative Commons Attribution 4.0 International License.

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https://doi.org/10.1515/gps-2020-0025
Schlagwörter für diesen Artikel
Rosmarinus officinalis; green synthesis; nanosilver
Creative Commons
BY 4.0

Artikel in diesem Heft

  1. Obituary for Prof. Dr. Jun-ichi Yoshida
  2. Regular Articles
  3. Optimization of microwave-assisted manganese leaching from electrolyte manganese residue
  4. Crustacean shell bio-refining to chitin by natural deep eutectic solvents
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  24. Potential of adsorbents from agricultural wastes as alternative fillers in mixed matrix membrane for gas separation: A review
  25. Bactericidal and cytotoxic properties of green synthesized nanosilver using Rosmarinus officinalis leaves
  26. Synthesis of biomass-supported CuNi zero-valent nanoparticles through wetness co-impregnation method for the removal of carcinogenic dyes and nitroarene
  27. Synthesis of 2,2′-dibenzoylaminodiphenyl disulfide based on Aspen Plus simulation and the development of green synthesis processes
  28. Catalytic performance of the biosynthesized AgNps from Bistorta amplexicaule: antifungal, bactericidal, and reduction of carcinogenic 4-nitrophenol
  29. Optical and antimicrobial properties of silver nanoparticles synthesized via green route using honey
  30. Adsorption of l-α-glycerophosphocholine on ion-exchange resin: Equilibrium, kinetic, and thermodynamic studies
  31. Microwave-assisted green synthesis of silver nanoparticles using dried extracts of Chlorella vulgaris and antibacterial activity studies
  32. Preparation of graphene oxide/chitosan complex and its adsorption properties for heavy metal ions
  33. Green synthesis of metal and metal oxide nanoparticles from plant leaf extracts and their applications: A review
  34. Synthesis, characterization, and electrochemical properties of carbon nanotubes used as cathode materials for Al–air batteries from a renewable source of water hyacinth
  35. Optimization of medium–low-grade phosphorus rock carbothermal reduction process by response surface methodology
  36. The study of rod-shaped TiO2 composite material in the protection of stone cultural relics
  37. Eco-friendly synthesis of AuNPs for cutaneous wound-healing applications in nursing care after surgery
  38. Green approach in fabrication of photocatalytic, antimicrobial, and antioxidant zinc oxide nanoparticles – hydrothermal synthesis using clove hydroalcoholic extract and optimization of the process
  39. Green synthesis: Proposed mechanism and factors influencing the synthesis of platinum nanoparticles
  40. Green synthesis of 3-(1-naphthyl), 4-methyl-3-(1-naphthyl) coumarins and 3-phenylcoumarins using dual-frequency ultrasonication
  41. Optimization for removal efficiency of fluoride using La(iii)–Al(iii)-activated carbon modified by chemical route
  42. In vitro biological activity of Hydroclathrus clathratus and its use as an extracellular bioreductant for silver nanoparticle formation
  43. Evaluation of saponin-rich/poor leaf extract-mediated silver nanoparticles and their antifungal capacity
  44. Propylene carbonate synthesis from propylene oxide and CO2 over Ga-Silicate-1 catalyst
  45. Environmentally benevolent synthesis and characterization of silver nanoparticles using Olea ferruginea Royle for antibacterial and antioxidant activities
  46. Eco-synthesis and characterization of titanium nanoparticles: Testing its cytotoxicity and antibacterial effects
  47. A novel biofabrication of gold nanoparticles using Erythrina senegalensis leaf extract and their ameliorative effect on mycoplasmal pneumonia for treating lung infection in nursing care
  48. Phytosynthesis of selenium nanoparticles using the costus extract for bactericidal application against foodborne pathogens
  49. Temperature effects on electrospun chitosan nanofibers
  50. An electrochemical method to investigate the effects of compound composition on gold dissolution in thiosulfate solution
  51. Trillium govanianum Wall. Ex. Royle rhizomes extract-medicated silver nanoparticles and their antimicrobial activity
  52. In vitro bactericidal, antidiabetic, cytotoxic, anticoagulant, and hemolytic effect of green-synthesized silver nanoparticles using Allium sativum clove extract incubated at various temperatures
  53. The green synthesis of N-hydroxyethyl-substituted 1,2,3,4-tetrahydroquinolines with acidic ionic liquid as catalyst
  54. Effect of KMnO4 on catalytic combustion performance of semi-coke
  55. Removal of Congo red and malachite green from aqueous solution using heterogeneous Ag/ZnCo-ZIF catalyst in the presence of hydrogen peroxide
  56. Nucleotide-based green synthesis of lanthanide coordination polymers for tunable white-light emission
  57. Determination of life cycle GHG emission factor for paper products of Vietnam
  58. Parabolic trough solar collectors: A general overview of technology, industrial applications, energy market, modeling, and standards
  59. Structural characteristics of plant cell wall elucidated by solution-state 2D NMR spectroscopy with an optimized procedure
  60. Sustainable utilization of a converter slagging agent prepared by converter precipitator dust and oxide scale
  61. Efficacy of chitosan silver nanoparticles from shrimp-shell wastes against major mosquito vectors of public health importance
  62. Effectiveness of six different methods in green synthesis of selenium nanoparticles using propolis extract: Screening and characterization
  63. Characterizations and analysis of the antioxidant, antimicrobial, and dye reduction ability of green synthesized silver nanoparticles
  64. Foliar applications of bio-fabricated selenium nanoparticles to improve the growth of wheat plants under drought stress
  65. Green synthesis of silver nanoparticles from Valeriana jatamansi shoots extract and its antimicrobial activity
  66. Characterization and biological activities of synthesized zinc oxide nanoparticles using the extract of Acantholimon serotinum
  67. Effect of calcination temperature on rare earth tailing catalysts for catalytic methane combustion
  68. Enhanced diuretic action of furosemide by complexation with β-cyclodextrin in the presence of sodium lauryl sulfate
  69. Development of chitosan/agar-silver nanoparticles-coated paper for antibacterial application
  70. Preparation, characterization, and catalytic performance of Pd–Ni/AC bimetallic nano-catalysts
  71. Acid red G dye removal from aqueous solutions by porous ceramsite produced from solid wastes: Batch and fixed-bed studies
  72. Review Articles
  73. Recent advances in the catalytic applications of GO/rGO for green organic synthesis
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Heruntergeladen am 12.10.2025 von https://www.degruyterbrill.com/document/doi/10.1515/gps-2020-0025/html?lang=de
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