Spectrofluorometric method for detection glycogen using chemically gold nanoparticles: Cyanobacteria as biological model.

There is a need for simple spectrofluorimetric method for detection of glycogen molecule based on binding to nanogold. Here we propose such a quantification method for glycogen using cyanobacteria as a biological model. Biologically, two strains of cyanobacteria were selected based on their previously tested nanogold biosynthetic abilities. Chemically, spherical gold nanoparticles were prepared and tested for binding to the glycogen molecule. Experimental analyses were conducted to determine the morphological and optical properties of the Au-glycogen hydrocolloids. Results: The plasmon band of biosynthesized AuNPs-glycogen was centered at 520-540 nm with size diameter was 41.7 ± 0.2 nm. The vibrational bands of glycogen were observed at 1,000 to 1,200 cm-1. The Au3+/Au0 redox coupling cycle was observed. The luminescence of AuNPs showed more stability by the addition of gradual concentrations of glycogen molecules. The detection (LOD) and quantitation limits (LOQ) were observed to be 0.89 and 2.95 µmol L-1 respectively (R2 = 0.99). The good chemical stability of this colloidal system and the glycogen molecule studied via density functional theory (DFT). The HOMO level of glycogen unit was closed near to LUMO level of Au3+. Conclusion: The associations formed between the gold nanoparticles and glycogen resulted in good chemical stability. This indicates that the quantification method proposed can be stably applied.

Multidrug-Resistant Bacterial Pathogens and Public Health: The Antimicrobial Effect of Cyanobacterial-Biosynthesized Silver Nanoparticles.

BACKGROUND: Cyanobacteria are considered as green nano-factories. Manipulation of the size of biogenic silver nanoparticles is needed to produce particles that suit the different applications such as the use as antibacterial agents. The present study attempts to manipulate the size of biosynthesized silver nanoparticles produced by cyanobacteria and to test the different-sized nanoparticles against pathogenic clinical bacteria. METHODS: Cyanothece-like. coccoid unicellular cyanobacterium was tested for its ability to biosynthesize nanosilver particles of different sizes. A stock solution of silver nitrate was prepared from which three different concentrations were added to cyanobacterial culture. UV-visible spectroscopy and FTIR were conducted to characterize the silver nanoparticles produced in the cell free filtrate. Dynamic Light Scattering (DLS) was performed to determine the size of the nanoparticles produced at each concentration. The antimicrobial bioassays were conducted on broad host methicillin-resistant Staphylococcus aureus (MRSA), and Streptococcus sp., was conducted to detect the nanoparticle size that was most efficient as an antimicrobial agent. RESULTS: The UV-Visible spectra showed excellent congruence of the plasmon peak characteristic of nanosilver at 450 nm for all three different concentrations, varying peak heights were recorded according to the concentration used. The FTIR of the three solutions revealed the absence of characteristic functional groups in the solution. All three concentrations showed spectra at 1636 and 2050-2290 nm indicating uniformity of composition. Moreover, DLS analysis revealed that the silver nanoparticles produced with lowest concentration of precursor AgNO3 had smallest size followed by those resulting from the higher precursor concentration. The nanoparticles resulting from highest concentration of precursor AgNO3 were the biggest in size and tending to agglomerate when their size was above 100 nm. The three types of differently-sized silver nanoparticles were used against two bacterial pathogenic strains with broad host range; MRSA-(Methicillin-resistant Staphylococcus aureus) and Streptococcus sp. The three types of nanoparticles showed antimicrobial effects with the smallest nanoparticles being the most efficient in inhibiting bacterial growth. DISCUSSION: Nanosilver particles biosynthesized by Cyanothece-like cyanobacterium can serve as antibacterial agent against pathogens including multi-drug resistant strains. The most appropriate nanoparticle size for efficient antimicrobial activity had to be identified. Hence, size-manipulation experiment was conducted to find the most effective size of nanosilver particles. This size manipulation was achieved by controlling the amount of starting precursor. Excessive precursor material resulted in the agglomeration of the silver nanoparticles to a size greater than 100 nm. Thereby decreasing their ability to penetrate into the inner vicinity of microbial cells and consequently decreasing their antibacterial potency. CONCLUSION: Antibacterial nanosilver particles can be biosynthesized and their size manipulated by green synthesis. The use of biogenic nanosilver particles as small as possible is recommended to obtain effective antibacterial agents.

Silver Nanoparticle Production by the Cyanobacterium Cyanothece sp.: De Novo Manipulation of Nano-Biosynthesis by Phytohormones.

BACKGROUND: Numerous cyanobacteria have the potential to reduce metallic ions to form pure metal nanoparticles in a green biosynthesis process. AIM: To investigate the production capacity of silver nanoparticles by the cyanobacterium Cyanothece sp. and to examine the effect of five different phytohormones, indole acetic acid, kinetin; gibberellic acid; abscisic acid; and methyl jasmonate, on this capacity. METHODS: The cyanobacterial strain was grown for 60 days and the harvested cyanobacterium biomass was incubated with 0.1 mM of AgNO3. Percentage conversion of Ag+ to Ag0 was calculated to indicate the AgNPs' production capacity. Different concentrations of the five phytohormones were added to cultures and the AgNP production was monitored throughout different time intervals. RESULTS: Cyanothece sp. biosynthesized spherical AgNPs (diameter range 70 to 140 nm, average diameter 84.37 nm). The addition of indole acetic acid and kinetin provoked the maximum conversion (87.29% and 55.16%, respectively) of Ag+ to Ag0, exceeding or slightly below that of the control (56%). Gibberellic and abscisic acids failed to elevate the Ag+ to Ag0 conversion rate (45.23% and 47.95%, respectively) above that of the control. Methyl jasmonate increased the Ag+ to Ag0 conversion rate to 90.29%, although nearly all the cyanobacterial cultures died at the end. CONCLUSION: Phytohormones could be used to induce or inhibit the green production of AgNPs with the cyanobacterium Cyanothece sp. This novel manipulation technique may have several applications in agriculture or biomedicine.

Investigating the Production of Antimicrobial Nanoparticles by Chlorella vulgaris and the Link to Its Loss of Viability.

Chlorella vulgaris from Al-Ahsa, KSA was proved to be an active silver and gold nanoparticle producer. Nanogold and nanosilver particles were characterized using UV-visible spectroscopy, Fourier-transform infrared spectroscopy, and scanning electronmicroscopy. Both nanoparticles were used in the antimicrobial bioassay. The two nanoparticles showed antibacterial activities, with the silver nanoparticles being the most effective. To investigate the argumentative nature of their biosynthesis (i.e., whether it is a biotic or abiotic process), we isolated total ribonucleic acid RNA as an indicator of vitality. RNA was completely absent in samples taken after one week of incubation with silver nitrate and even after one or two days. However, successful extraction was only achievable in samples taken after incubation for one and four hours with silver nitrate. Most importantly, the gel image showed recognizable shearing of the nucleic acid after 4 h as compared to the control. An assumption can be drawn that the synthesis of nanoparticles may start biotically by the action of enzyme(s) and abiotically by action of reducing entities. Nonetheless, with prolonged incubation, excessive nanoparticle accumulation can be deadly. Hence, their synthesis continues abiotically. From the RNA banding profile, we suggest that nanosilver production starts both biotically and abiotically in the first few hours of incubation and then continues abiotically. Nanosilver particles proved to have more of an antimicrobial impact than nanogold and hence are recommended for different applications as antibacterial agents.

Green Synthesis of Silver Nanoparticles by the Cyanobacteria Synechocystis sp.: Characterization, Antimicrobial and Diabetic Wound-Healing Actions.

Green nanotechnology is now accepted as an environmentally friendly and cost-effective advance with various biomedical applications. The cyanobacterium Synechocystis sp. is a unicellular spherical cyanobacterium with photo- and hetero-trophic capabilities. This study investigates the ability of this cyanobacterial species to produce silver nanoparticles (AgNPs) and the wound-healing properties of the produced nanoparticles in diabetic animals. METHODS: UV-visible and FT-IR spectroscopy and and electron microscopy techniques investigated AgNPs' producibility by Synechocystis sp. when supplemented with silver ion source. The produced AgNPs were evaluated for their antimicrobial, anti-oxidative, anti-inflammatory, and diabetic wound healing along with their angiogenesis potential. RESULTS: The cyanobacterium biosynthesized spherical AgNPs with a diameter range of 10 to 35 nm. The produced AgNPs exhibited wound-healing properties verified with increased contraction percentage, tensile strength and hydroxyproline level in incision diabetic wounded animals. AgNPs treatment decreased epithelialization period, amplified the wound closure percentage, and elevated collagen, hydroxyproline and hexosamine contents, which improved angiogenesis factors' contents (HIF-1α, TGF-ß1 and VEGF) in excision wound models. AgNPs intensified catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) activities, and glutathione (GSH) and nitric oxide content and reduced malondialdehyde (MDA) level. IL-1ß, IL-6, TNF-α, and NF-κB (the inflammatory mediators) were decreased with AgNPs' topical application. CONCLUSION: Biosynthesized AgNPs via Synechocystis sp. exhibited antimicrobial, anti-oxidative, anti-inflammatory, and angiogenesis promoting effects in diabetic wounded animals.

Cyanobacteria as Nanogold Factories II: Chemical Reactivity and anti-Myocardial Infraction Properties of Customized Gold Nanoparticles Biosynthesized by Cyanothece sp.

Cyanothece sp., a coccoid, unicellular, nitrogen-fixing and hydrogen-producing cyanobacterium, has been used in this study to biosynthesize customized gold nanoparticles under certain chemical conditions. The produced gold nanoparticles had a characteristic absorption band at 525-535 nm. Two types of gold nanoparticle, the purple and blue, were formed according to the chemical environment in which the cyanobacterium was grown. Dynamic light scattering was implemented to estimate the size of the purple and blue nanoparticles, which ranged from 80 ± 30 nm and 129 ± 40 nm in diameter, respectively. The highest scattering of laser light was recorded for the blue gold nanoparticles, which was possibly due to their larger size and higher concentration. The appearance of anodic and cathodic peaks in cyclic voltammetric scans of the blue gold nanoparticles reflected the oxidation into gold oxide, followed by the subsequent reduction into the nano metal state. The two produced forms of gold nanoparticles were used to treat isoproterenol-induced myocardial infarction in experimental rats. Both forms of nanoparticles ameliorated myocardial infarction injury, with a slight difference in their curative activity with the purple being more effective. Mechanisms that might explain the curative effect of these nanoparticles on the myocardial infarction were proposed. The morphological, physiological, and biochemical attributes of the Cyanothece sp. cyanobacterium were fundamental for the successful production of "tailored" nanoparticles, and complemented the chemical conditions for the differential biosynthesis process. The present research represents a novel approach to manipulate cyanobacterial cells towards the production of different-sized gold nanoparticles whose curative impacts vary accordingly. This is the first report on that type of manipulated gold nanoparticles biosynthesis which will hopefully open doors for further investigations and biotechnological applications.

Cyanobacteria as Nanogold Factories: Chemical and Anti-Myocardial Infarction Properties of Gold Nanoparticles Synthesized by Lyngbya majuscula.

To the best of our knowledge, cyanobacterial strains from the Arabian Gulf have never been investigated with respect to their potential for nanoparticle production. Lyngbya majuscula was isolated from the AlOqair area, Al-Ahsa Government, Eastern Province, Kingdom of Saudi Arabia. The cyanobacterium was initially incubated with 1500 mg/mL of HAuCl4 for two days. The blue-green strain turned purple, which indicated the intracellular formation of gold nanoparticles. Prolonged incubation for over two months triggered the extracellular production of nanogold particles. UV-visible spectroscopy measurements indicated the presence of a resonance plasmon band at ~535 nm, whereas electron microscopy scanning indicated the presence of gold nanoparticles with an average diameter of 41.7 ± 0.2 nm. The antioxidant and anti-myocardial infarction activities of the cyanobacterial extract, the gold nanoparticle solution, and a combination of both were investigated in animal models. Isoproterenol (100 mg/kg, SC (sub cutaneous)) was injected into experimental rats for three days to induce a state of myocardial infarction; then the animals were given cyanobacterial extract (200 mg/kg/day, IP (intra peritoneal)), gold nanoparticles (200 mg/kg/day, IP), ora mixture of both for 14 days. Cardiac biomarkers, electrocardiogram (ECG), blood pressure, and antioxidant enzymes were determined as indicators of myocardial infarction. The results showed that isoproterenol elevates ST and QT segments and increases heart rate and serum activities of creatine phosphokinase (CPK), creatine kinase-myocardial bound (CP-MB), and cardiac troponin T (cTnT). It also reduces heart tissue content of glutathione peroxidase (GRx) and superoxide dismutase (SOD), and the arterial pressure indices of systolic arterial pressure (SAP), diastolic arterial pressure (DAP), and mean arterial pressure (MAP). Gold nanoparticles alone or in combination with cyanobacterial extract produced an inhibitory effect on isoproterenol-induced changes in serum cardiac injury markers, ECG, arterial pressure indices, and antioxidant capabilities of the heart.