Anti-bacterial and Anti-cancer Activities of Silver and Gold Nanoparticles Synthesized using Sasa borealis and Agrimonia pilosa
- Alternative Title
- 조릿대와 짚신나물 추출물을 이용하여 합성한 은과 금 나노입자의 항균 및 항암 활성
- Abstract
- Nanobiotechnology is emerging as a cutting-edge technology involving many academic disciplines like biology, chemistry, physics, material science and medicines. The word “nano” is derived from the Greek word nanos, which means “dwarf”. The size of a nanoparticle (NPs) is one billionth of a meter (10-9m), and materials with nanodimensions (1-100 nm) have significant activity compared to the same material in bulk form. Novel strategies are needed to improve efficiency of biomedicines. Presumptions are directed towards Nanobiotechnology-based applications in microbiology and cell biology. We have developed a green and eco-friendly route for synthesis of silver-silver chloride (Ag-AgCl) and gold (Au) NPs using medicinal plants (Sasa borealis, and Agrimonia pilosa). The visual observation indicates color change of reaction solution is the first sign of NPs formation, and confirmed by UV-visible spectroscopy. The shape, size and morphology of NPs were observed by transmission electron microscopy and field emission-scanning electron microscopy. The elemental composition and crystalline nature of synthesized NPs were analyzed by energy dispersive X-ray spectroscopy and X-ray diffraction spectroscopy, respectively. The synthesized NPs stability was determined by zeta potential study. The biomolecules present in plant extract and their interaction in NPs synthesis were studied by Fourier transform-infrared spectroscopy and gas chromatography-mass spectrometry.
S. borealis leaves extract-mediated Ag-AgCl NPs were spherical, oval shape with particle size around 10-30 nm and -12.95 mV zeta potential value indicates NPs covered with negative charged material and which is responsible to prevent aggregation of NPs by electrostatic repulsion. The synthesized Ag-AgCl NPs were tested for antibacterial activity against Gram-positive (Listeria monocytogenes, Staphylococcus aureus, Staphylococcus saprophyticus) and Gram-negative bacteria (Escherichia coli, Pseudomonas putida). Among tested bacteria, Gram-positive bacteria (S. saprophyticus) show more sensitivity, and Gram-negative (P. putida) shows less sensitivity to the synthesized Ag-AgCl NPs. The cytotoxic activity was observed on gastric adenocarcinoma AGS cells. The concentration dependent cytotoxicity was observed and IC50 at 10 µg ml-1 recorded; same concentration was applied to check nuclear morphological observation by DAPI staining. A fragmentation and condensation is the characteristic of apoptosis which were observed in AGS cell.
Likewise, S. borealis extract-mediated Au NPs were observed a spherical, oval shape with about 10-30 nm particle size. The toxic effect against embryonic kidney HEK293 cells and cytotoxic activity against gastric adenocarcinoma AGS cells were perform; upto 300 µg ml-1 of Au NPs were nontoxic to HEK293 cells and concentration dependent cell viability was observed on AGS cells. In DAPI staining, fragmentation and concensation of nuclear material were observed in treated AGS cells with IC50 concentration (120 µg ml-1).
In next studies, A. pilosa aerial part extract was used to synthesize Ag-AgCl NPs from reduction of silver nitrate; small size of particles around 10-20 nm with a spherical and oval shapes were observed. The zeta potential value (-3.84 mV) indicates synthesized Ag-AgCl NPs carried negative charges. The antibacterial activity of synthesized Ag-AgCl NPs indicates, Gram-positive bacteria were more sensitive compared to Gram-negative bacteria. An FE-SEM study of Ag-AgCl NPs with and without treated bacterial samples indicates the possible mechanism behind bacterial growth inhibition is changes in cell wall morphology.
Overall for the first time, these findings showed that S. borealis leaves extract-mediate Ag-AgCl NPs, Au NPs and A. pilosa aerial parts extract-mediated Ag-AgCl NPs have antibacterial and anticancer activity. This study will provide significant information for finding molecular mechanism behind bacterial and cancer cells death.
- Issued Date
- 2017
- Awarded Date
- 2017. 8
- Type
- Dissertation
- Keyword
- Agrimonia pilosa Anti-bacterial Anti-cancer Gold nanoparticles Nanotechnology Sasa borealis Silver nanoparticles
- Publisher
- Pukyong National University
- Affiliation
- 부경대학교 대학원
- Department
- 대학원 미생물학과
- Advisor
- Gun-Do Kim
- Table Of Contents
- 1. General introduction 1
1.1. Sasa borealis and Agrimonia pilosa 2
1.2. Nanotechnology 4
1.2.1. Chemical and physical method for nanoperticles (NPs) synthesis 5
1.2.2. Biological method for NPs synthesis 6
1.3. Mechanism of NPs formation 8
1.4. Characterization of NPs 10
1.5. Application of silver (Ag) and gold (Au) NPs 13
1.5.1. Antibacterial approach of Ag NPs 13
1.5.2. Anticancer approach of Ag NPs 16
1.5.3. Anticancer approach of Au NPs 17
1.6. The purpose of this research 20
1.7. References 21
PART I. Silver and gold nanoparticles synthesis using Sasa borealis 33
2. Sasa borealis leaf extract-mediated green synthesis of silver-silver chloride nanoparticles and their antibacterial and anticancer activities 34
2.1. Abstract 34
2.2. Introduction 35
2.3. Materials and methods 37
2.3.1. Materials 37
2.3.2. Extraction of Sasa borealis leaves 37
2.3.3. Biosynthesis of Ag-AgCl NPs 37
2.3.4. Characterization of Ag-AgCl NPs 40
2.3.5. Antibacterial activity 40
2.3.6. Anticancer activity 41
2.4. Results and discussion 44
2.4.1. Effect of pH on Ag-Ag-Cl NPs synthesis 44
2.4.2. Effect of S. borealis leaf extract contents 46
2.4.3. Effect of AgNO3 cocentration on Ag-AgCl NP synthesis 48
2.4.4. Effect of reaction temperature on Ag-AgCl NPs synthesis 50
2.4.5. Effect of reaction incubation time 52
2.4.6. Energy dispersive X-ray spectroscopy and transmission electron microscopic analysis 54
2.4.7. X-ray diffraction analysis and Zeta potential analysis 56
2.4.8. Fourier transform infrared spectroscopy 58
2.4.9. Antibacterial activity 60
2.4.10. Anticancer activity 63
2.5. Conclusion 65
2.6. References 66
3. Anticancer activity of Sasa borealis leaf extract-mediated gold nanoparticles 70
3.1. Abstract 70
3.2. Introduction 71
3.3. Materials and methods 73
3.3.1. Materials 73
3.3.2. Sasa borealis leaf extraction 73
3.3.3. Green synthesis of gold nanoparticles (Au NPs) 73
3.3.4. Characterization studies 75
3.3.5. Anticancer activity 75
3.3.6. In vitro cell viability assay 75
3.3.7. Crystal violet staining 76
3.3.8. Immunofluorescence staining 76
3.4. Results and discussion 77
3.4.1. UV-visible and Energy dispersive X-ray spectroscopy analysis of synthesized Au NPs 77
3.4.2. Size, shape and crystalline nature of synthesized Au NPs 79
3.4.3. S. borealis biomolecules interaction in Au NPs synthesis 81
3.4.4. Phytocomposition of S. borealis leaf extract 83
3.4.5. Anticancer activity 86
3.4.6. Immunofluoroscence staining 88
3.5. Conclusion 90
3.6. References 91
PART II. Silver nanoparticles synthesis using Agrimonia pilosa 94
4. Antibacterial efficiency of silver-silver chloride nanoparticles synthesized using Agrimonia pilosa against pathogenic bacteria 95
4.1. Abstract 95
4.2. Introduction 96
4.3. Materials and methods 98
4.3.1. Materials 98
4.3.2. Preparation of extract from aerial part of A. pilosa 98
4.3.3. Facile synthesis of Ag-AgCl NPs 98
4.3.4. Characterization of Ag-AgCl NPs 98
4.3.5. Antibacterial activity of Ag-AgCl NPs 99
4.3.6. Detection of MIC and MBC 100
4.3.7. Study of bacterial morphology 100
4.4. Results and discussion 101
4.4.1. UV-visible spectroscopy 101
4.4.2. Elemental composition and crystalline nature of Ag-AgCl NPs 103
4.4.3. Ag-AgCl NPs size, morphology and zeta potential analysis 105
4.4.4. Interaction of biomolecules in Ag-AgCl NPs formation 107
4.4.5. Phytocompositions of A. pilosa 109
4.4.6. Antibacterial activity of Ag-AgCl NPs 112
4.4.7. Bacterial cell morphology 115
4.5. Conclusion 117
4.6. References 118
국문요약 121
Acknowledgements 124
- Degree
- Doctor
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