A review on green synthesis of silver nanoparticles and their applications

Figures & data

Table 1. Flow chart describing the techniques for synthesis of Ag-NPs.

Synthesis of nanoparticles
Bottom-up approach		Top-down approach
Green methods	Chemical methods	Physical methods
• Using bacteria• Using fungi• Using plant and their extracts• Using yeast• Using enzymes and biomolecules• Using microorganism	• Chemical reduction• Sonochemical• Microemulsion• Photochemical• Electrochemical• Pyrolysis• Microwave• Solvothermal• Coprecipition	• Pulsed laser ablation• Evaporation–condensation• Arc discharge• Spray Pyrolysis• Ball milling• Vapour and gas phase• Pulse wire discharge• Lithography
Non-toxic	Toxic

Figure 1. Schematic diagram for synthesis of Ag-NPs by using bacteria.

Table 2. Green synthesis of Ag-NPs using bacteria.

Bacteria	Precursor	Intracellular/extracellular	Size	Morphology	References
Cyanobacteria Spirulina platensis and actinobacteria Streptomyces spp. 211A	AgNO3	Extracellular	7–15	–	(Tsibakhashvil et al. 2010)
Aeromonas sp. THG-FG1.2	AgNO3		8–16	Face Centered Cubic (FCC), spherical	(Singh et al. 2016b)
Serratia nematodiphila	AgNO3	Extracellular	10–31	Crystalline and spherical	(Malarkodi et al. 2013)
Pseudomonas putida NCIM 2650	AgNO3	Extracellular	70	Spherical	(Thamilselvi and Radha 2013)
Escherichia coli DH5α	AgNO3	Extracellular	10–100	Spherical	(Ghorbani 2013)
Lactobacillus strains	AgNO3	–	15–500	Cluster triangular, hexagonal and crystalline	(Nair and Pradeep 2002)
Lactobacillus casei subsp. casei	AgNO3	–	25–50	Spherical	(Korbekandi et al. 2012)
Nocardiopsis sp.MBRC-1	AgNO3	–	45 ± 0.15	Spherical	(Manivasagan et al. 2013)
Pseudomonas stutzeri AG259	AgNO3	–	35–46 and upto 200	Crystalline, hexagonal and equilateral triangle	(Haefeli et al. 1984)
Shewanella oneidensis	AgNO3	Extracellular	4 ± 1.5	Spherical	(Suresh et al. 2010)
Penicillium glabrum (MTCC 1985)	AgNO3	Extracellular	26–32	Spherical	(Nanda and Majeed 2014)
Bacillus strain CS 11	AgNO3	Extracellular	45 ± 0.15	FCC, spherical	(Das et al. 2014)
Actinobacteria	AgNO3	Intracellular	5–50	Spherical	(Suman et al., 2014)
Bacillus sp.	AgNO3	Intracellular	5–15	Crystalline	(Pugazhenthiran et al. 2009)
Vibrio alginolyticus	AgNO3	Extra and Intracellular	50–100	Crystalline and spherical	(Rajeshkumar et al. 2013)
Marine Ochrobactrum sp.	AgNO3	Intracellular	38–85	Spherical	(Thomas et al. 2014)
Exiguobacterium mexicanum	Ag NO3	Extracellular	5–40	–	(Padman et al. 2014)
Endosymbiotic Bacterium	AgNO3	Extracellular	10–60	Cubic, spherical, hexagonal, crystalline, oval	(Syed et al. 2016)
Bacillus subtilis (MTCC441)	AgNO3	Extracellular	10–100	Spherical	(Venkatesan et al. 2013)
Lactobacillus spp.	AgNO3	Extra and intracellular	2–20	Spherical	(Ranganath et al. 2012)
Novosphingobium sp.HG-C3	AgNO3	Extracellular	8–25	Spherical and crystalline	(Du et al. 2016)
Kinneretia THG-SQ14	AgNO3	Extracellular	15–20	Mono-disperse, FCC, spherical	(Singh et al. 2016a)
Bacillus methylotrophicus		Extracellular	10–30	Spherical	(Wang et al. 2015)
S1–S10	AgNO3	Intra/Extracellular	25–30	–	(Nanda et al. 2011)
Rhodococcus spp.	AgNO3	Intracellular	5–50	Spherical	(Otari et al. 2015)

Figure 2. Schematic diagram for synthesis of Ag-NPs by using fungi.

Table 3. Green synthesis of Ag-NPs using fungi.

Fungi	Precursor	Intracellular/Extracellular	Size (nm)	Morphology	References
Humicola sp.	AgNO3	Extracellular	5–25	Spherical	(Syed et al. 2013)
Penicillium citrinum	AgNO3	Extracellular	109	Uniform spherical	(Honary et al. 2013)
Filamentous fungus Penicillium sp.	AgNO3	–	58.35 ± 17.88	–	(Hemath Naveen et al. 2010)
Endophytic fungus Penicillium sp.	AgNO3	Extracellular	25 and 30	Spherical	(Singh et al. 2014)
Endophytic bryophilous	AgNO3	Extracellular	25–50	Ellipsoidal	(Raudabaugh et al. 2013)
Aspergillus niger, Fusarium oxysporum and Alternaria solani	AgNO3	–	20	Spherical	(Ghazwani 2015)
Schizophyllum commune	AgNO3	Intracellular & Extracellular	51–93	Spherical	(Arun et al. 2014)
Trichoderma reesei	AgNO3	Extracellular	5–50	Random	(Vahabi et al. 2011)
Fusarium semitectum	AgNO3	Extracellular	1–50	Ellipsoid and polydispersed spherical	(Shelar and Chavan 2014)
Aspergillus niger	AgNO3	–	1–20	Polydispersed spherical	(Sagar and Ashok 2012)
Trichoderma harzianum	AgNO3	–	34.77	Ellipsoid and spherical	(Shelar and Chavan 2015)
Fusarium acuminatum	AgNO3	Extracellular	13	Spherical	(Ingle et al. 2008)
Penicillium fellutanum	AgNO3	Extracellular	5–25	Spherical	(Kathiresan et al. 2009)
Endophytic fungus Aspergillus clavatus	AgNO3	Extracellular	10–25	Hexagonal and polydispersed spherical	(Verma et al. 2010)
Pestalotia sp.	AgNO3	Extracellular	12.40	Polydispersed and spherical	(Raheman et al. 2011)
Alternaria alternata	AgNO3	Extracellular	32.5	Polydispersed and spherical	(Gajbhiye et al. 2009)
Fusarium semitectum	AgNO3	Extracellular	10–60	Crystalline and spherical	(Basavaraja et al. 2008)
Phoma glomerata	AgNO3	Extracellular	60–80	Spherical	(Birla et al. 2009)
Trichoderma viride	AgNO3	Extracellular	5–40	Spherical	(Fayaz et al. 2010)

Table 4. Green synthesis of Ag-NPs using plant and plant extracts.

Plants/plant extracts	Precursor	Reducing and capping agent	Size (nm)	Structure/shape	References
Helicteres isora	AgNO3	Root extract	30–40	Crystalline and spherical	(Bhakya et al. 2015)
C. olitorus Linn and I.batatas Lam	AgNO3	Leaves extract	37.9 & 67.3	Crystalline	(Meva et al. 2016)
Rheum palmatum	AgNO3	Root extract	121 ± 2	Cubic, spherical and hexagonal	(Arokiyaraj et al. 2016)
Excoecaria agallocha	AgNO3	Leaves extract	20	Crystalline, hexagonal and spherical	(Bhuvaneswari et al. 2016)
Banana	AgNO3	Peel extract	23.7	Crystalline and spherical	(Ibrahim 2015)
Eucalyptus globulus	AgNO3	Leaves extract	1.9–4.3 and 5–25	Spherical	(Ali et al. 2015)
Oranges and pineapples	AgNO3	Fruit extract	Depends upon fruit extract	Depends upon fruit extract	(Hyllested et al. 2015)
Solanum nigrum andclitoria ternatea	AgNO3	Leaves extract	23	Spherical	(Jassal et al. 2016)
Cranberry	AgNO3	Powder extract	2.8 ± 2.1, 1.4 ± 0.8, 8.6 ± 2.5	Spherical	(Ashour et al. 2015)
Phyllanthus niruri	AgNO3	Leaves extract	30–60	Crystalline FCC and spherical	(Suresh et al. 2015)
Bryophyllum and Asiatic Pennywort	AgNO3	Leaves extract	18–21	Crystalline FCC and spherical	(Saikia et al. 2015)
Zingiber officinale rhizome	AgNO3	Broth extract	3.1	Crystalline and spherical	(Shalaby et al. 2015)
Tagetes erecta	AgNO3	Flower broth	10–90	Irregular, hexagonal and spherical	(Padalia et al. 2015)
Orange	AgNO3	Peel extract	91	Spherical	(Awad et al. 2014)
Piper betle	AgNO3	Leaves extract	–	–	(Prabha et al. 2014)
Justica adhatoda	AgNO3	Leaves extract	11–20	Smooth and spherical	(Kudle et al. 2014)
Eucalyptus leucoxylon	AgNO3	Leaves extract	50	FCC and spherical	(Rahimi-Nasrabadi et al. 2014)
Oak	AgNO3	Fruit extract	40	Cubic and spherical	(Heydari and Rashidipour 2015)
Chrysanthemum indicum L	AgNO3	Flower extract	37.71–71.99	Crystalline FCC and spherical	(Arokiyaraj et al. 2014)
Asclepias curassavica	AgNO3	Leaves extract	75–95	Spherical	(G.S.O.S.N. 2014)
Dextran	AgNO3	Dextran solution	50–70	Spherical	(Hussain et al. 2014)
Alstonia scholaris	AgNO3	Bark extract	50	Spherical	(Shetty et al. 2014)
Piper nigrum	AgNO3	Leaves extract	7–50 and 9–30	Crystalline and spherical	(Paulkumar et al. 2014)
Achillea bieberstennii	AgNO3	Flower extract	12 ± 2	Hexagonal, pentagonal and spherical	(Baharara et al. 2014)
Ocimum tenuiforum, Azadirachta indica and Musa balbisiana	AgNO3	Leaves extract	Up to 200	Cubical, spherical and triangular	(Banerjee et al. 2014)
Vitex negundo	AgNO3	Leaves extract	60	Spherical	(Kathireswari et al. 2014)
Pithophora oedogonia	AgNO3	Algal extract	34.03	Hexagonal and cubical	(Sinha et al. 2015)
Sida acuta and Artemisia annua	AgNO3	Leaves extract	–	–	(Johnsona et al. 2014)
Datura metel	AgNO3	Leaves extract	5–50	Spherical	(Kumar Ojha et al. 2013)
Partheniumhysterophorus L.	AgNO3	Leaves extract	20–50	Roughly spherical	(Thombre et al. 2013)
Rumex hymenosepalus	AgNO3	Root extract and plant extract	2–40	Hexagonal and FCC	(Rodríguez-León et al. 2013)
Tecoma stans	AgNO3	Leaves extract	15	FCC and spherical	(Arunkumar et al. 2013)
Lens culinaris	AgNO3	Seed exudates	13	Crystalline and spherical	(Shams et al. 2013)
Citrus sinensis, Solanumtricobatum, Cuminiasiatica and Syzygium cumini	AgNO3	Plant powders	41, 42. 52 and 53	Crystalline and irregular	(Logeswari et al. 2013)
Dalbergia sissoo	AgNO3	Leaves extract	5-55	Crystalline and spherical	(Singh et al. 2012)
Nelumbo nucifera	AgNO3	Leaves extract	45	Decahedral, truncated triangles, spherical and triangle	(Santhoshkumar et al. 2011)
Macrotyloma uniflorum	AgNO3	Seed extract	12	Anisotropic, FCC and nearly spherical	(Vidhu et al. 2011)
Desmodium triflorum	AgNO3	Plant Broth	10	Spherical	(Ahmad et al. 2011)
Melia azedarach	AgNO3	Seed extract	–	–	(Shams et al. 2014)
Calliandra haematocephala	AgNO3	Leaves extract	70	Spherical and FCC	(Raja et al. 2015)
Trilobata	AgNO3	Leaves extract	–	–	(Billacura and Mimbesa 2015)
Pongam pinnata L. Pierre	AgNO3	Leaves biomass	38	Spherical	(Raut et al. 2010)
Achyranthes aspera L.	AgNO3	Leaves extract	1.0 ± 18.3	Spherical	(Gude et al. 2013)
Nerium oleander	AgNO3	Leaves extract	380–420	–	(Subbaiya et al. 2014)
Premna serratifolia L.	AgNO3	Leaves extract	22.97	Cubic	(Paul et al. 2015)
Catharanthus roseus	AgNO3	Root extract	35–55	Crystalline FCC and spherical	(Rajagopal et al. 2015)
Calotropis procera	AgNO3	Latex serum extract	12.33	Spherical	(Mohamed et al. 2014)
Aloe vera	AgNO3	Leaves extract	70	Spherical, rectangular, cubical and triangular	(Medda et al. 2015)
Allium cepa	AgNO3	Extract	33.6	Spherical	(Saxena et al. 2010)
Garlic	AgNO3	Extract	4–6	–	(Von White et al. 2012)
Prunus armeniaca	AgNO3	Fruit extract	Less than 20	Crystalline and spherical	(Dauthal and Mukhopadhyay 2013)
Peach gum	AgNO3	Peach gum powder	23.56 ± 7.87	FCC	(Yang et al. 2015)
Psidium guajava	AgNO3	Leaves extract	26 ± 5	Crystalline and spherical	(Raghunandan et al. 2011)
Solanum lycopersicum	AgNO3	Fruit extract	10	Spherical	(Umadevi et al. 2013)
Solanum tuberosum	AgNO3	Potato infusion	10–12	Crystalline, FCC and spherical	(Roy et al. 2015)
Juglans Regia L.	AgNO3	Leaves extract	10–50	Quasi spherical	(Korbekandi et al. 2013)
Citrus limon	AgNO3	Limon extract	Less than 50	Spheroidal and spherical	(Prathna et al. 2011)
Chenopodium album	AgNO3	Leaves extract	10–30	Quasi spherical	(Dwivedi and Gopal 2010)
Honey	AgNO3	Honey solution	4	Crystalline and spherical	(Philip 2010b)
Mentha piperita	AgNO3	Leaves extract	5–30	Spherical	(Parashar et al. 2009)
Henna	AgNO3	Leaves extract	50	Spherical	(Daniel et al. 2013)
Hibiscusrosa sinensis	AgNO3	Extract	14	Prism or spherical	(Philip 2010a)
Tea	AgNO3	Leaves extract	20	Prism or spherical	(Sun et al. 2014)
Albizia lebbeck	AgNO3	Plant extract	–	Roughly spherical	(Parvathy et al. 2014)
Syzygium cumini	AgNO3	Bark extract	20–60	Spherical	(Prasad and Swamy 2013)
Tribulus terrestris	AgNO3	Fruit extract	16–28	Spherical	(Gopinath et al. 2012)
Ziziphora tenuior	AgNO3	Leaves extract	8–40	Spherical	(Sadeghi and Gholamhoseinpoor 2015)
Melia dubia	AgNO3	Leaves extract	35	Spherical	(Kathiravan et al. 2014)
Thevetia peruviana	AgNO3	Latex extract	10–30	Spherical	(Rupiasih et al. 2015)
Vitex negundo	AgNO3	Leaves extract	5 and 10–30	FCC and spherical	(Zargar et al. 2011)
Brassica rapa	AgNO3	Leaves extract	16.4	–	(Narayanan and Park 2014)
Vitis vinifera	AgNO3	Fruit extract	30–40	–	(Gnanajobitha et al. 2013)
Musa paradisiacal	AgNO3	Peel extract	20	–	(Bankar et al. 2010)
Eclipta prostrate	AgNO3	Leaves extract	35–60	Hexagons, triangles and pentagons	(Rajakumar and Rahuman 2011)
Cuminum cyminum	AgNO3	Seeds powder	12	Smooth surface and spherical	(Kudle et al. 2012)
Securinega Leucopyrus	AgNO3	Leaves and fruit extract	11–20	Spherical, oval and smooth	(Donda et al. 2013)
Lantana camara	AgNO3	Fruit extract	12.55–12.99	Spherical	(Sivakumar et al. 2012)
Pistacia atlantica	AgNO3	Seeds extract	10–50	Spherical	(Sadeghi et al. 2015)
Carica papaya	AgNO3	Fruit extract	15	Hexagonal and cubic	(Jain et al. 2009)
Ficus carica	AgNO3	Leaves extract	13	–	(Geetha et al. 2014)
Memecylon edule	AgNO3	Leaves extract	20–50	Hexagonal, circular, triangular	(Elavazhagan and Arunachalam 2011)
Swietenia mahogany	AgNO3	Leaves extract	50	–	(Mondal et al. 2011)
Trachyspermum ammi	AgNO3	Seed extracts	87, 99.8	–	(Vijayaraghavan et al. 2012)
Olea europaea	AgNO3	Seed extracts	34	Crystalline	(Sadeghi 2014)
Prunus japonica (Rosaceae)	AgNO3	Leaves extract	26	Hexagonal, spherical, crystalline and irregular	(Saravanakumar et al. 2016)
Terminalia arjuna	AgNO3	Bark extract	2–100	Spherical	(Ahmed et al. 2016a)
Catharanthus roseus	AgNO3	Leaves extract	20	Crystalline, FCC and spherical	(Al-Shmgani et al. 2016)
Turbinaria ornata (Turner)	AgNO3	Seaweed aqueous extract	20–32	Spherical and crystalline	(Deepak et al. 2016)
Althaea officinalis radix	AgNO3	Hydroalcoholic extract		Polydispersed and spherical	(Korbekandi et al. 2016)
Quercus brantii	AgNO3	Leaves hydroalcoholic extract	6	Polydispersed and spherical	(Korbekandi et al. 2015)

Figure 3. Schematic diagram for synthesis of Ag-NPs by using plan/plant extracts.

Table 5. Potential applications of Ag-NPs.

Medical applications	• Dental cements	(Magalhães et al. 2012)
	• Healing of burns and wound care	(Chaloupka et al. 2010, Huang et al. 2007)
	• Skin therapy and crucial in cosmetics	(Kokura et al. 2010, Silver 2003)
	• Reconstructive orthopedic surgery and bone cements	(Caplan 2005, Shi et al. 2006)
	• Medical devices and plastic catheters	(Roe et al. 2008, Monteiro et al. 2009)
	• Target drug delivery	(Ahamed et al. 2010)
	• Medical imaging, biolabeling and detection	(Loo et al. 2005, Houk et al. 2009) (Alivisatos 2004)
	• Cancer therapy	(Boca et al. 2011)
	• Coating of hospital textiles such as face mask surgical	(Lu et al. 2008, Van Duyne et al. 2003)
	• gowns, coating of breath mask patent and ultrasonic detection	(Tran and Le 2013, Morley et al. 2007)
	• Coating of implants for joint replacement	(Alt et al. 2004, Zheng et al. 2010,)
	• Orthodontic and orthopedic fixations and implants	(Belt et al. 2001, Liu et al. 2012)
Environmental applications	• Air disinfection	(Yoon et al. 2008)
	• Drinking water purification	(Zhang 2013)
	• Biological wastewater and ground water disinfection	(Sheng and Liu 2011, Mpenyana-Monyatsi et al. 2012)
	• Surface disinfection	(Kumar et al. 2008)
	• Wastewater treatment	(Benn and Westerhoff 2008)
Consumer goods and personal care products	• Textiles, paints and sunscreen cosmetics. Toys and humidifiers, filters, kitchen utensilstoothpaste, washers systems, shampoo, Bedding, deodorants, and fabrics.	(Ahamed et al. 2010, Baker et al. 2005)
Food	• Nutraceutical, food safety and food packaging	(Ramos et al. 2014, Morones et al. 2005, Duncan 2011)
	• Biosensors for food analysis	(Pérez-López and Merkoçi 2011)
Electronic components and transportation	• Sensors, electrodes and integrated circuits	(Kotthaus et al. 1997, Hau et al. 2009, Fukuda et al. 2012)
	• Nano wires and high energy batteries.	(Jana et al. 2001, Jasinski et al. 1967)
Antibacterial activity	• Ag-NPs exhibits inhibitory activity against Gram positive, Gram negative, Pseudomonasaeruginoss, Bacillus cereus, Escherichia hermanii, Bacillus subtilis, Klebsiella pneumonia,agricultural plant pathogens, and antibiotic resistant bacteria.	(Prabha et al. 2014, Paulkumar et al. 2014, Sinha et al. 2015, Bhakya et al. 2015, Meva et al. 2016, Shalaby et al. 2015, Kumar Ojha et al. 2013, Saxena et al. 2012, Govarthanan et al. 2016)
Antimicrobial activity	• Ag-NPs have antimicrobial activity against S. epidermidis, P. fluorescens, Klebsiella Pneumonia, Pseudomonas aeruginosa, Pseudomonous sp., Gram positive, Gram negative, Salmonella typhi, R. arrhizus, and Proteus vulgaris, and multidrug resistant bacteria.	(Govarthanan et al. 2016, Hussain et al. 2014, Kumar Ojha et al. 2013, Logeswari et al. 2013, Shalaby et al. 2015, Shetty et al. 2014, Singh et al. 2016b)
Antiviral activity	• Ag-NPs exhibits antiviral activity against human immunodeficiency virus-1, Influenza virus,(HIV-1), Herpes simplex virus, HSV-2, Monkey pox virus, hepatitis B, HSV-1, and Respiratory syncytial virus.	(Ahmed et al. 2015, Baram-Pinto et al. 2009, Elechiguerra et al. 2005, Lara et al. 2010, Lu et al. 2008, Rogers et al. 2008, Sun et al. 2008)
Antifungal activity	• Ag-NPs have fungicidal and antifungal activity against Candida albicans, Candida krusei, Candida species, Candida glabrata, Trichophyton mentagrophytes, and Candida tropicalis.	(Coker et al. 2011, Gajbhiye et al. 2009, Kim et al. 2008, 2009, Manjumeen et al. 2014)

Table 6. In-vitro toxicity effects of Ag-NPs.

Particles size (nm)	Dose	Concentration	Exposure time	Cell/organisms	Major outcomes	References
13.2 ± 4.72	0.5–8.0μg Ag/mL	0.5–8.0 μg Ag/mL	4–24 hours	Suspension cells (U937) and adhesive cells (HeLa)	• Cell death is dose dependent. • Ag-NPs exhibits significant toxicity toward the U937 and HeLa tumor cells.• Cytotoxicity was determined by change in cell viability.	(Kaba and Egorova 2015)
29.57	7.74 mg/l and1.16 mg/l	2.5–5 mg/l	48–72 hours	HaCat and HeLa cell lines	• Apoptotic cell population was significantly increase by increasing Concentration of Ag-NPs in HeLa and HaCat cell lines. • It is studied that Adenosine diphosphate (ADP):Adenosine triphosphate (ATP) ratios were increased than their associated controls.• At high concentration, huge variations inadenylate kinase (AK) activity was occurred.• Upon exposure of Ag-NPs HeLa cells exhibits higher sensitivity as compare to HeCaT cells.	(Mukherjee et al. 2012)
1.98–64.9	1.73 × 10^4/cm^3, 1.27 × 10^5/cm^3 1.32 × 10^6particles/cm^3	p < 0.1	4 hours	Sprague–Dawley rat	• Higher concentration of Ag-NPs decreased the cell viability and mitochondrial function as rising lactate dehydrogenase (LDH) leakage. • Due to exposure of Ag-Nps no significant organ weight was changed.• Doses of Ag-Nps caused no significant health effect.	(Ji et al. 2007)
10, 40, and 75	5–50 μg/mL	5–50 μg/mL	4 and 24 hours	In the lung cells of human	• Viability of human lung cells is effected byexposure of Ag-NPs. • Intracellular metal release due to the difference in intracellular localization and cellular uptake are explained by the difference in ecotoxicity.	(Gliga et al. 2014)
15 and 100	10–50μg/ml	10–50 μg/ml	24 hours	BRL 3A rat liver cells	• Ag-NPs indicate depletion of reduced glutathione (GSH), reduction of mitochondrial membrane potential and generation of reactive oxygenspecies (ROS). • At higher concentration of Ag-NPs membrane was damaged.	(Hussain et al. 2005)
61.2 ± 33.9	25–75 mg l^−1	25–75 mg l^−1	24 hours	Kidney cells of mammalian	• Ag-NPs induced genotoxicity in porcine kidney (PK15) cells.• Deoxyribonucleic acid (DNA) damage like DNA–DNA/DNA–protein cross links oxidatively, single and double strand becomes an cause of alkaline damages upon the exposure of Ag-Nps. • Dose dependent DNA damage.	(Milić et al. 2015)
22.18	1.91 × 107particles/cm^3	1.91 × 10^7	6 hours/day for 14 days	Male C57BL/6 mice	• Potential immunotoxicity and neurotoxicity	(Lee et al. 2010)
254 ± 27	10.7 μg/mL	0.0345–345 μg/mL	24 hours	In the range of Rainbow Trout cell Lines (RTG, RTL-W1, and RTH-149) also with primary Hepatocytes	• Ag-NPs exhibits toxic effect on fresh waterenvironments. • Specific effects of Ag-NPs were indicated by using lysosomal damage as an indicator.	(Connolly et al. 2015)
15	8.75 μg/ml or21.3 μg/ml	10 μg/ml and above	48 hours	Dependent Toxicity stem cell line Reduced (C 18-4) of Mouse spermatogonial	• Increased membrane leakage and reducedmitochondrial function.	(Braydich-Stolle et al. 2005)
18	1.73 × 104/cm^3–1.27 × 105/cm^3 1.32 × 106 particles/cm^3	1.323 × 10^6 particles/cm^3, 61 μg/m^3	6 hours/day during 90 days	Sprague–Dawley rat	• Enhanced liver inflammation and bile ducthyperplasia. Reduced the alveolar inflammation and tidal volume.	(Sung et al. 2008)
20	278 ng/cm^2	6μg/mL	24 hours	Lung cells	• No significant induction of proinflammatorymediators and cytotoxicity was observed by applying realistic dose of Ag-NPs.	(Herzog et al. 2013)

Table 7. In-vivo toxicity effects of Ag-NPs.

Particles size (nm)	Dose	Concentration	Exposure time	Cell/organisms	Major outcomes	References
2.5–150	0.7 × 10^6, 1.4 × 10^6 and 2.9 × 10^6	32 × 10^6 particles/cm^3, 61 μg/m^3	90 days	Bone marrow of male and female rat	• No significant organ weight change and bone marrow cytotoxicity was indicated in male and female rats. • Dose dependent MNPECs number was increased in male rat and no detection was found treatment related to enhanced number of MNPCEs in female rats	(Kim et al. 2011)
100	2.8 μg/ml	20–4000 μg/ml	7 days	Rat ear and BALB/c3T3 cell lines	• In the permeability of biological barrierssignificant changes was occurred.	(Zou et al. 2014)
35 and ≤100	0.625, 1.25, 2.5, 5, 7.5, 10, 15, 20, and 25 mg/L	50 μg/ml	24–72 hours	Fathead minnow	• In embryos morphological abnormalities and high mortality are mostly edema	(Scown et al. 2010)
˜180	0, 50, 100, 250, 500, and 1000 mg/kg/day	5.50, 3.56, 2.87, 2.10, 11.28 μg/g wet weight	2 weeks	Rat hepatic cytochrome P450(CYP)	• To inhibit CYP enzymes activities.Loss of drug efficacy for patients due to unwanted changes in plasma drug levels.	(Kulthong et al. 2012)
81	25–84 μg L^−1	>72 μg L^−1	48 hours	Zebrafish (Danio rerio)	• At higher concentration all fishes were dead.	(Bilberg et al. 2012)
10	50 and 100 μg/ml	p < 0.05	2 times/day for 10 minutes during 5 days	Fruit fly (Drosophila melanogaster)	• Apoptosis in the larvae, heat shock protein 70, induction of oxidative stress and DNA damage markers.	(Siddique et al. 2009)
20, 25, 35, 50, and 80	0.34–1.7 μgml^−1	1.7–0.000544 μg/mL	14 days	Porcine skin	• After 14 days intercellular and intra consecutive cellular epidermal days edema.	(Samberg et al. 2010)
18–19	0.63 × 10^6 particles/cm^3, 49 mg/m^3, 1.43 × 10^6 particles/cm^3, 133 mg/m^3 and 3.03 × 10^6 particles/cm^3, 515 mg/m^3	1.32 × 10^6 particles/cm^3, 61 μg/m^3	For 13 weeks 6 hours/day, 5 days/week.	Lungs and liver in Sprague–Dawley rats.	• Dose dependent bile-duct hyperplasia was increased in the liver of female and male rat.• In the lungs of female and male macrophage accumulation, chronic alveolar inflammation was reported.	(Sung et al. 2009)
243.8 ± 176.7	121.1 ± 7.6 pg/mL, 47.4 ± 1.8 pg/mL and 32.2 ± 2.8 pg/mL, 8.2 ± 1.9 pg/mL, and 2.2 ± 0.4 pg/mL	125 μg/kg, 250 μg/kg, and 500 μg/kg	7–28 days	Male mice	• Adverse effects of Ag-NPs on mice and due to disturbance in homeostasis of body may cause many inflammatory diseases such as mesothelioma, pneumo coniosis, and emphysema.	(Park et al. 2011)
25	100, 500, and 1000 mg/kg	100, 500, and 1000 mg/kg	24 hours	Adult-male C57BL/6N mice	• Generate neurotoxicity by producing free radical induced oxidative stress and also by changing gene expression generate neurotoxicity and apoptosis.	(Rahman et al. 2009)

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