Development of a bacterial cellulose-gelatin composite as a suitable scaffold for cardiac tissue engineering.

PURPOSE: Cardiac tissue engineering is suggested as a promising approach to overcome problems associated with impaired myocardium. This is the first study to investigate the use of BC and gelatin for cardiomyocyte adhesion and growth. METHODS: Bacterial cellulose (BC) membranes were produced by Komagataeibacter xylinus and coated or mixed with gelatin to make gelatin-coated BC (BCG) or gelatin-mixed BC (mBCG) scaffolds, respectively. BC based-scaffolds were characterized via SEM, FTIR, XRD, and AFM. Neonatal rat-ventricular cardiomyocytes (nr-vCMCs) were cultured on the scaffolds to check the capability of the composites for cardiomyocyte attachment, growth and expansion. RESULTS: The average nanofibrils diameter in all scaffolds was suitable (~ 30-65 nm) for nr-vCMCs culture. Pore diameter (≥ 10 µm), surface roughness (~ 182 nm), elastic modulus (0.075 ± 0.015 MPa) in mBCG were in accordance with cardiomyocyte requirements, so that mBCG could better support attachment of nr-vCMCs with high concentration of gelatin, and appropriate surface roughness. Also, it could better support growth and expansion of nr-vCMCs due to submicron scale of nanofibrils and proper elasticity (~ 0.075 MPa). The viability of nr-vCMCs on BC and BCG scaffolds was very low even at day 2 of culture (~ ≤ 40%), but, mBCG could promote a metabolic active state of nr-vCMCs until day 7 (~ ≥ 50%). CONCLUSION: According to our results, mBCG scaffold was the most suitable composite for cardiomyocyte culture, regarding its physicochemical and cell characteristics. It is suggested that improvement in mBCG stability and cell attachment features may provide a convenient scaffold for cardiac tissue engineering.

Combinational antimicrobial activity of biogenic TiO2 NP/ZnO NPs nanoantibiotics and amoxicillin-clavulanic acid against MDR-pathogens.

The development of effective strategies against multidrug-resistant (MDR) pathogens is an urgent need in modern medicine. Nanoantibiotics (nABs) offer a new hope in countering the surge of MDR-pathogens. The aim of the current study was to evaluate the antibacterial activity of two attractive nABs, TiO2 NPs and ZnO NPs, and their performance in improving the antimicrobial activity of defined antibiotics (amoxicillin-clavulanic acid, amox-clav) against MDR-pathogens. The nABs were synthesized using a green method. The physicochemical characteristics of the synthesized nanoparticles were determined using standard methods. The results showed the formation of pure anatase TiO2 NPs and hexagonal ZnO NPs with an average particle size of 38.65 nm and 57.87 nm, respectively. The values of zeta potential indicated the high stability of the samples. At 8 mg/mL, both nABs exhibited 100 % antioxidant activity, while ZnO showed significantly higher activity at lower concentrations. The antibiofilm assay showed that both nABs could inhibit the formation of biofilms of Acinetobacter baumannii 80 and Escherichia coli 27G (MDR-isolates). However, ZnO NPs showed superior antibiofilm activity (100 %) against E. coli 27G. The MIC values were determined to be 8 (1), 2 (2), and 4 (4) mg/mL for amox-clav, TiO2 NPs, and ZnO NPs against A. baumannii 80 (E. coli 27G), respectively. The results showed that both nABs had synergistically enhanced antibacterial performance in combination with amox-clav. Specifically, an 8-fold reduction in MIC values of antibiotics was observed when they were combined with nABs. These findings highlight the potential of TiO2 NPs and ZnO NPs as effective nanoantibiotics against MDR-pathogens. The synergistic effect observed when combining nABs with antibiotics suggests a promising approach for combating antibiotic resistance. Further research and development in this area could lead to the development of more effective treatment strategies against MDR infections.

Plant elicitation and TiO2 nanoparticles application as an effective strategy for improving the growth, biochemical properties, and essential oil of peppermint.

Mentha piperita L., which is an abundant source of essential oils (EO) and phenolic acids, is well known for its medicinal significance. The present research aimed to evaluate the impact of various concentrations of methyl jasmonate (MeJA; 0, 0.1, and 0.5 mM), titanium dioxide nanoparticles (TiO2 NPs; 0 and 150 mg L-1), and salicylic acid (SA; 0, 0.1, and 1 mM) on growth, EOs, and phenolic compounds of M. piperita L. The results demonstrated that the simultaneous application of SA (0.1 mM) and TiO2 NPs (150 mg L-1) enhanced shoot dry weight, the shoot length, and membrane stability index of peppermint by 56.17, 19.52, and 36%, respectively, compared to control. Moreover, phenolic content (76%), caffeic acid content (78%), rosmarinic acid content (87%), 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging ability (78%), and catalase (155%), ascorbate peroxidase activities (95%) were further improved by simultaneously applying MeJA (0.1 mM) and TiO2 NPs (150 mg L-1) compared to control. The highest menthol production (44.51%) was obtained with exogenous application of MeJA (0.1 mM) with 150 mg L-1 TiO2 NPs. The findings of the current study presented an ideal combination of TiO2 NPs with plant growth regulators for promoting antioxidant activities and increasing major components of EO in peppermint plants.

Antibacterial and antibiofilm efficacy of Ag NPs, Ni NPs and Al2O3 NPs singly and in combination against multidrug-resistant Klebsiella pneumoniae isolates.

BACKGROUND: Although traditional antibiotic therapy provided an effective approach to combat pathogenic bacteria, the long-term and widespread use of antibiotic results in the evolution of multidrug-resistant bacteria. Recent progress in nanotechnology offers an alternative opportunity to discover and develop novel antibacterial agents. METHODS: A total of 51 K. pneumoniae strains were collected from several specimens of hospitalized patients and identified by two parallel methods (biochemical tests and Vitek-2 system). The antibiotic sensitivity of isolates was evaluated by disk diffusion antibiogram and Vitek-2 system. The biofilms formation ability of antibiotic-resistant strains was examined by microtiter plate and tube methods based on crystal violet staining. The molecular technique was used to determine key genes responsible for biofilms formation of clinical isolates. The antibacterial and antibiofilm activities of Ag NPs, Ni NPs, Al2O3 NPs singly (NPs) and in combination (cNPs) were investigated against selected strains using standard methods. Moreover, the cytotoxicity of NPs was evaluated on mouse neural crest-derived (Neuro-2A) cell line. RESULTS: The results of bacterial studies revealed that more than 80 % of the isolates were resistant to commonly used antibiotics and about 95 % of them were able to form biofilms. Moreover, the presence of fimA and mrkA genes were determined in all biofilm-producing strains. The results of antibacterial and antibiofilm activities of NPs and cNPs demonstrated the lower MIC and MBEC values for Al2O3 NPs singly as well as for Ag/Ni cNPs and Ag/Al2O3 cNPs in combination, respectively. Overall, the inhibitory effects of cNPs were superior to NPs against all strains. Furthermore, the results of the checkerboard assays showed that Ag NPs act synergistically with two other NPs against multidrug-resistant Klebsiella pneumoniae (MDR-K. pneumoniae) isolates. The in vitro cytotoxicity assay revealed no significant toxicity of NPs against Neuro-2A cells. CONCLUSION: In the present study, the combination of Ag NPs, Ni NPs, and Al2O3 NPs were used against MDR-K. pneumoniae strains and antibacterial and antibiofilm activities were observed for Ag/Ni cNPs and Ag/Al2O3 cNPs.

Cell-free extract assisted synthesis of ZnO nanoparticles using aquatic bacterial strains: Biological activities and toxicological evaluation.

The introduction of novel bacterial strains and the development of microbial approaches for nanoparticles biosynthesis could minimize the negative environmental impact and eliminate the concern and challenges of the available approaches. In this study, a biological method based on microbial cell-free extract was used for biosynthesis of ZnO NPs using two new aquatic bacteria, Marinobacter sp. 2C8 and Vibrio sp. VLA. The synthesized ZnO NPs were characterized by UV-Visible spectroscopy, Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), transmission electron microscopy (TEM), atomic force microscope (AFM), dynamic light scattering (DLS) and zeta potential. The UV-Visible absorption peak was found to be at 266 and 250 nm for ZnO-2C8 NPs and ZnO-VLA NPs, respectively. FTIR study suggested that the hydroxyl, amine, and carboxyl groups of bacterial proteins are mainly responsible for stabilizing the biosynthesized ZnO NPs. The formation of hexagonal wurtzite structure of ZnO NPs was confirmed by the XRD pattern. The morphology of the nanoparticles was found to be spherical with the average particle size of about 10.23 ± 2.48 nm and 20.26 ± 4.44 nm for ZnO-2C8 NPs and ZnO-VLA NPs, respectively. The values of zeta potential indicate the high stability of the biosynthesized ZnO NP. Zeta potential values indicated the high stability of the biosynthesized ZnO NP and were obtained -20.54 ± 7.15 and -23.87 ± 2.29 mV for ZnO-2C8 NPs and ZnO-VLA NPs, respectively. The biosynthesized ZnO NPs had antibacterial activity against Gram-negative and Gram-positive strains and possessed excellent antibiofilm activity with the maximum inhibition of about 96.55% at 250 µg/mL. The DPPH activity of ZnO-2C8 NPs and ZnO-VLA NPs were found 88.9% and 85.7% for 2500 µg/mL concentration, respectively. The toxicity test revealed the biocompatibility of the biosynthesized ZnO NPs. The results suggested that this approach is a very good route for synthesizing ZnO NPs with potential applications in biotechnology.

Application of a marine luminescent Vibrio sp. B4L for biosynthesis of silver nanoparticles with unique characteristics, biochemical properties, antibacterial and antibiofilm activities.

Biosynthesis of silver nanoparticles (AgNPs) by marine bacteria especially luminescent Vibrio species is least investigated. In this study, AgNPs were first synthesized by the culture supernatant of a luminescent bacterium (Vibrio sp. B4L) and then, the prepared samples were characterized employing several techniques. The antibacterial activity of the AgNPs was investigated against Escherichia coli and Staphylococcus aureus using disk diffusion agar and broth microdilution methods. The growth curve, Reactive Oxygen Species (ROS) formation, and Lactate Dehydrogenase (LDH) activity of the samples were measured along with Field Emission Scanning Electron Microscopy (FESEM) observation and inhibition of biofilm formation. Dynamic light scattering (DLS) analysis showed that the average particle size of the synthesized AgNPs was in the range of about 32.67-107.18 nm and the polydispersity index (PDI) of 0.1120 indicated the formation of monodispersed particles. The average zeta potential of AgNPs obtained -36.15 mV, showing the high stability of biosynthetic nanoparticles. Antibacterial studies indicated that not only the AgNPs had antibacterial activity but also increased the antibacterial properties of tetracycline when used in combination. ROS production was enhanced in a dose-dependent manner. A high difference in LDH activities was found between AgNPs treated cells and the control group. FESEM images revealed membrane disruption and lysis in AgNPs treated cells. The formation of E. coli biofilm was 100% inhibited at 62.5 µg/ml showing that our bacteriogenic AgNPs can be a potential alternative remedies for controlling antibiotic-resistant pathogens.

Highly efficient solar photocatalytic degradation of a textile dye by TiO2/graphene quantum dots nanocomposite.

Herein, two sunlight responsive photocatalysts including TiO2 nanoparticles (NPs) and TiO2/graphene quantum dots (GQDs) nanocomposite for degrading a textile dye, Reactive Black 5 (RB5), were prepared. The results showed that 100% of 50 ppm RB5 could be degraded by TiO2 NPs and TiO2/GQDs within 60 and 30 min sunlight irradiation, respectively. Hence, much better photocatalytic activity in degradation of RB5 was achieved by TiO2/GQDs under sunlight irradiation compared with pure TiO2 NPs due to its lower band gap (2.13 eV) and electron/hole recombination rate. The photocatalytic degradation mechanism of RB5 by TiO2 NPs was elucidated by adding some scavengers to the solution. The main reactive species contributing to RB5 degradation were surface hydroxyl radicals. The first-order solar degradation rate constant of RB5 for TiO2/GQDs is greater than that of TiO2 NPs under sunlight illumination.

Microbial cell lysate supernatant (CLS) alteration impact on platinum nanoparticles fabrication, characterization, antioxidant and antibacterial activity.

Microbial mediated biological synthesis of nanoparticles is of enormous interest to modern nanotechnology due to its simplicity and eco-friendliness. In the present study, a novel green method for the synthesis of platinum nanoparticles (PtNPs) has been developed using bio-derived product-cell lysate supernatant (CLS) from various microorganisms including Gram-negative bacteria: Pseudomonas kunmingensis ADR19, Psychrobacter faecalis FZC6, Vibrio fischeri NRRL B-11177, Gram-positive bacteria: Jeotgalicoccus coquinae ZC15, Sporosarcina psychrophila KC19, Kocuria rosea MN23, genetically engineered bacterium: Pseudomonas putida KT2440 and yeast: Rhodotorula mucilaginosa CCV1. The biogenic PtNPs were characterized by UV-visible spectroscopy, X-ray diffraction (XRD), Fourier transform infrared (FTIR), transmission electron microscopy (TEM), high resolution transmission electron microscopy (HRTEM), and atomic force microscopy (AFM). The UV-visible spectra showed a red shift in the absorbance of H2PtCl6.6H2O from 260 nm to 330 nm for all prepared PtNPs. The XRD patterns of the samples indicated the formation of high purity of the cubic phase. The FTIR spectra and EDS profiles of the samples demonstrated the existence of proteins on fabricated and stabilized PtNPs. The TEM and AFM images analysis showed the synthesis of smallest PtNPs by a bacterium strain (FZC6) and yeast while genetically engineered bacteria produced the largest NPs. Also, the HRTEM analysis showed the high crystallinity of PtNPs and the interplanar spacing of 0.2 nm, corresponds to the (1 1 1) of plane of PtNPs. The results of zeta potential indicated the high stability of PtNPs in neutral pH. Moreover, the suitability of PtNPs antioxidant and antibacterial activity was correlated to the size and zeta potential of microbe used for NPs biosynthesis. In conclusion, it was found that the type of microorganisms can have influences on PtNPs characteristics and properties as Gram-negatives produced smaller PtNPs while more negatively charged NPs were obtained by Gram-positives. These findings could facilitate the selection of appropriate green approaches for more effective biotechnological production of PtNPs.

Development and evaluation of a novel beneficent antimicrobial bioscaffold based on animal waste-fish swim bladder (FSB) doped with silver nanoparticles.

Treated fish wastes have found many applications in industry and medicine. Besides, nowadays low-cost scaffold with antimicrobial activity which can accelerates the process of wound healing is very demanding. In this study fish swim bladder (FSB), taken from Rutilus frisii, which is a disposable waste was doped with silver nanoparticles (AgNPs) and evaluated as antimicrobial wound dressing. The scanning electron microscopy (SEM) micrographs showed the presence of AgNPs on the scaffold. Histological observation confirmed cells and muscle removal from FSB and collagen preservation. There was significant antibacterial activity even in 50 ppm AgNPs concentration against pathogenic bacteria, swelling ratio was rather low, and cytotoxic assay revealed that the AgNPs-FSB scaffold had no toxic effect on human foreskin fibroblast (HFF) cells. Interestingly, despite the porous structure, the AgNPs-FSB scaffold was found to be a suitable barrier to microbial penetration even after 72 h. Further study showed the gradual release of AgNPs during 24 h. In conclusion, biofabricated FSB prepared in this study have appropriate characteristics notably encompassing a high quantity of collagen and broad-spectrum antimicrobial activity. Also, its porous structure made it suitable as a 3-D structure for the growth of cells and adding other antimicrobial nano-sized materials.

Comparative evaluation of silver nanoparticles biosynthesis by two cold-tolerant Streptomyces strains and their biological activities.

The present study reflected on high-priority biological activities of novel silver nanoparticles (AgNPs) synthesized via two cold-tolerant strains; namely, Streptomyces sp.OSIP1 and Streptomyces sp.OSNP14. These AgNPs were synthesized through a green method using culture supernatant of bacteria at 20 °C and characterized by several instrumental techniques. The TEM results revealed that the NPs obtained from OSIP1 were smaller (8 nm, average) than those taken from OSIP14 (15 nm, average). Both AgNPs-OSP1 and AgNPs-OSNP14 also posed the strongest growth inhibitory effect against several pathogenic bacteria alone and especially in combination with antibiotics. Smaller NPs especially at 3.9-31.25 µg/ml concentrations were assumed more effective biofilm inhibitors of Pseudomonas aeruginosa. Cytotoxic activity of both AgNPs (at 25 and 50 µg/mL concentrations) on mouse colorectal carcinoma cells (CT26) were then studied using methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay. The findings demonstrated that smaller AgNPs at a 50 µg/mL concentration had 7% more cytotoxicity effects. In conclusion; although AgNPs produced by diverse strains of cold-adapted Streptomyces had close characteristics and biological activities, they showed some multifarious properties.

Exploring the potentials of halophilic prokaryotes from a solar saltern for synthesizing nanoparticles: The case of silver and selenium.

Halophiles are the organisms that thrive in extreme high salt environments. Despite the extensive studies on their biotechnological potentials, the ability of halophilic prokaryotes for the synthesis of nanoparticles has remained understudied. In this study, the archaeal and bacterial halophiles from a solar saltern were investigated for the intracellular/extracellular synthesis of silver and selenium nanoparticles. Silver nanoparticles were produced by the archaeal Haloferax sp. (AgNP-A, intracellular) and the bacterial Halomonas sp. (AgNP-B, extracellular), while the intracellular selenium nanoparticles were produced by the archaeal Halogeometricum sp. (SeNP-A) and the bacterial Bacillus sp. (SeNP-B). The nanoparticles were characterized by various techniques including UV-Vis spectroscopy, XRD, DLS, ICP-OES, Zeta potentials, FTIR, EDX, SEM, and TEM. The average particle size of AgNP-A and AgNP-B was 26.34 nm and 22 nm based on TEM analysis. Also, the characteristic Bragg peaks of face-centered cubic with crystallite domain sizes of 13.01 nm and 6.13 nm were observed in XRD analysis, respectively. Crystallographic characterization of SeNP-A and SeNP-B strains showed a hexagonal crystallite structure with domain sizes of 30.63 nm and 29.48 nm and average sizes of 111.6 nm and 141.6 nm according to TEM analysis, respectively. The polydispersity index of AgNP-A, AgNP-B, SeNP-A, and SeNP-B was determined as 0.26, 0.28, 0.27, and 0.36 and revealed high uniformity of the nanoparticles. All of the synthesized nanoparticles were stable and their zeta potentials were calculated as (mV): -33.12, -35.9, -31.2, and -29.34 for AgNP-A, AgNP-B, SeNP-A, and SeNP-B, respectively. The nanoparticles showed the antibacterial activity against various bacterial pathogens. The results of this study suggested that the (extremely) halophilic prokaryotes have great potentials for the green synthesis of nanoparticles.

Use of a genetically engineered E. coli overexpressing ß-glucuronidase accompanied by glycyrrhizic acid, a natural and anti-inflammatory agent, for directed treatment of colon carcinoma in a mouse model.

Bacteria-directed enzyme prodrug therapy (BDEPT), is an emerging alternative directed and tumor-specific approach. The basis of this method is the conversion of a non-toxic prodrug by a bacterial enzyme to a toxic drug within the tumor-microenvironment (TME). In the present study, the therapeutic efficacy of BDEPT was investigated based on the ability of Escherichia coli DH5α-lux/ßG in activation of glycyrrhizic acid (GL), a natural and non-toxic compound purified from licorice, to glycyrrhetinic acid (GA) only in TME. To do so, the anti-bacterial effects of GL on bacteria and the cytotoxic effects of the produced GA on survival rate of CT26 mouse colon carcinoma cells were evaluated. The IC50 values of the produced GA and cisplatin were determined as 210 µM and 100 µM, respectively. Comparing these values to GL treatment (1305 µM) indicates that bacteria could have efficiently activated GL to GA to inhibit the growth of tumor cells. Afterward, the anti-cancer effects of bacteria used in combination with GL was investigated in a mouse model of colon carcinoma. Results were indicative of targeted homing and even proliferation of luminescent bacteria in TME. Moreover, combined treatment greatly inhibited tumor growth. Histopathological analysis of dissected tissues also demonstrated increased apoptosis rate in tumor cells after combined treatment and interestingly, showed no obvious damage to the spleen and liver of treated mice. Accordingly, this BDEPT approach could be considered as an effective alternative tumor-specific therapy utilizing prodrug-activating enzymes expressing from tumor-targeting bacteria to allow the development of new tumor-specific pharmacotherapy protocols.

Multifunctional CuO nanoparticles with cytotoxic effects on KYSE30 esophageal cancer cells, antimicrobial and heavy metal sensing activities.

In this work, fluorescent copper oxide nanoparticles (CuO NPs) were green synthesized using viable cells, cell lysate supernatant (CLS) and protein extracts of luminescent Vibrio sp. VLC. Biogenic CuO NPs were then characterized by XRD, FTIR, UV/Vis spectroscopy, TEM, DLS, and PL spectroscopy. Results showed that CLS method was more efficient for CuO NPs production, therefore CuO NPs synthesized by this method from copper sulfate (CuO NPs-1) and/or copper nitrate (CuO NPs-2) were used for further studies. The crystallite size of polydispersed CuO NPs-1 and CuO NPs-2 were about 8.83 and 8.77 nm, respectively indicating their suitability for biological applications. Antibacterial activity of CuO NPs was determined using broth microdilution, well diffusion agar, and time-kill curves methods. Both CuO NP-1 and CuO NP-2 inhibited bacterial growth at the minimum inhibitory concentration (MIC) of 625 mg/L except St. mutants (MIC = 1250 mg/L). Emission of fluorescent light from the surface of NPs was increased when exposed to Cd2+, As2+ and Hg2+ ions but decreased by Pb2+ ions. Results showed that CuO NP-1 had anticancer properties against KYSE30 esophageal cancer cell line (IC50 = 13.96 mg/L) while no higher cytotoxic effects were observed on Human Dermal Fibroblasts (HDF) (IC50 = 48.88 mg/L).

Optimization of complete RB-5 azo dye decolorization using novel cold-adapted and mesophilic bacterial consortia.

Azo dyes are an important group of recalcitrant xenobiotics, which are difficult to degrade and deteriorate in cold environments. In this study, two microbial consortia consisting of cold-adapted and mesophilic bacteria were developed for effective decolorization of Reactive Black-5 azo dye. These bacteria were isolated from textile wastewater and soil of a cold region. Identification of bacterial isolates using 16s rRNA gene analysis revealed that they belong to genus Pseudoarthrobacter, Gordonia, Stenotrophomonas, and Sphingomonas. Decolorization assay was performed for every strain at dye concentrations of 25, 50 and 100 mg/L and the consortia PsGo consisting of mesophilic bacteria and StSp consisting of cold-adapted bacteria were constructed accordingly. Results showed that the consortia PsGo and StSp were able to decolorize 54 and 34 percent of RB-5 (50 mg/L) during 7 days. To improve the dye removal efficiency of the consortia, several parameters including temperature, pH, carbon and nitrogen sources were optimized. Over longer periods, StSp consortium managed to completely decolorize RB-5 (50 mg/L) at optimized conditions of 25-30 °C, pH 9, and using glucose and NH4H2PO4 as carbon and nitrogen source respectively, whereas PsGo consortium decolorized RB-5 (50 mg/mL) completely at 37 °C, pH 11, and with lactose and NH4H2PO4 used as carbon and nitrogen sources. Kinetic of reactions for StSp and PsGo consortia were found to be 0.05 and 0.13 day-1 respectively, but became 0.71 and 0.9 day-1 after optimization. In general, cold ecosystems are good sources for the isolation of novel bacterial strains with a potential application, especially when used as consortia, in environmental biotechnology such as decolorization of RB-5.

Selenium nanoparticle as a bright promising anti-nanobacterial agent.

The use of nanotechnology for nanobacteria (or calcifying nanoparticles) treatment is a new creative approach. Use of selenium nanoparticles (SeNPs) as anti-nanobacterial agents might be considered as a bright promising approach due to their critical role in the inhibition of crystal growth and aggregation of calcium oxalate. Hence, in this study, we investigated the probable outcome of SeNPs inhibitory effects on growth of nanobacteria. Fragments of thirty urinary tract stones were chemically analyzed by X-ray diffraction (XRD) and urinary stones Kits for calcifying nanoparticles presence. Then powder of stone fragments were resuspended in Dulbecco's modified Eagle's medium (DMEM), sterilized by filtration and cultured in presence of 1, 5, 30, 60, and 90 µmol/L SeNPs concentrations. Besides, calcifying nanoparticles growth in the culture without SeNPs was measured spectrophotometrically. Also, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) analyses were used, where calcifying nanoparticles formation occurred. Results showed that in the culture without SeNPs, the positive calcifying nanoparticles detection was 60% while after adding SeNPs at 90 µmol/L, not any calcifying nanoparticles were observed. Further confirmation came out when Energy-dispersive X-ray (EDX) analysis showed calcium and phosphate peaks in the culture medium without any SeNPs while in the culture containing 90 µm/L SeNPs a decrease in calcium and other minerals was obvious. Therefore, SeNPs clearly restricted the growth of nanobacteria due to their inhibitory effects on calcium oxalate deposition.

Multifunctional fluorescent titania nanoparticles: green preparation and applications as antibacterial and cancer theranostic agents.

Theranostic nanoparticles have attracted considerable attention in recently revolutionized medicine. Since the last decade, there has been a growing attempt to design various theranostic nanoparticles but difficulties still exist in the fabrication of their biocompatible one. Herein, fluorescent titania nanoparticles (FTN) were fabricated using a one-step green method. This FTN had ultra-high doxorubicin hydrochloride (DOX) loading capacity (encapsulation efficiency 95.50% and loading content 38.20%) that release the loaded drug in response to acidic pH. In vitro cytotoxicity experiments on human osteosarcoma (SaOs-2) and breast cancer (MCF-7) cell lines revealed superior anticancer efficacy (lowered the IC50 concentration by 3- and 5.5-fold for SaOs-2 and MCF-7 cells, respectively) and also better imaging for intracellular tracking of DOX/FTN relative to free DOX. Furthermore, the prepared nanoparticles showed efficient antibacterial activity against both gram-negative (Escherichia coli ATCC 25922) and gram-positive (Staphylococcus aureus ATCC 25923) bacteria. In this study, we have developed novel theranostic titania nanoparticles with inherent fluorescence property for cancer imaging and therapy.

Antibacterial activity of poly-l-arginine under different conditions.

BACKGROUND AND OBJECTIVES: Arginine-rich peptides are an important class of antimicrobial peptides (AMPs) that exert their antibacterial activity via a lytic mechanism. Although the antibacterial activity of arginine-rich peptides has been already evaluated, no reports have so far been evaluated the influence of reaction conditions on their antimicrobial potential. The aim of the present study was to investigate the influence of pH, temperature, and glycine on antibacterial activity of poly-l-arginine (PLA) with a molecular weight of 5-15 kDa against Escherichia coli O157:H7 and Staphylococcus aureus. MATERIALS AND METHODS: The percentage of growth inhibition of PLA against both bacteria was analyzed at various pH, temperatures and sub-inhibitory concentrations of glycine by two-fold broth microdilution method. RESULTS: The results showed that PLA had antibacterial activity against E. coli O157:H7 and S. aureus and the inhibitory effect increased with increasing PLA concentration. The antimicrobial activity of PLA against both microorganisms was higher in basic media than under acidic or neutral conditions. At 1/2 times the MIC, heat treatment intensified the toxicity of PLA against E. coli O157:H7 whereas the susceptibility to PLA seems to be temperature independent for S. aureus. The MICs of glycine against E. coli O157:H7 and S. aureus were 12.5 and 25 mg ml-1, respectively. The antibacterial activity of PLA against both microorganisms increased, as indicated by the increasing growth inhibition percentage of this peptide with increasing glycine concentration. CONCLUSION: The antibacterial activity of PLA against S. aureus and E. coli O157:H7 depends on pH and glycine concentration.

Genetically modified luminescent bacteria Ralostonia solanacerum, Pseudomonas syringae, Pseudomonas savastanoi, and wild type bacterium Vibrio fischeri in biosynthesis of gold nanoparticles from gold chloride trihydrate.

The effect of different genetically engineered bacteria, Pseudomonas syringae, Pseudomonas savastanoi, and Ralostonia solanacerum and also a natural marine bacterial species, Vibrio fischeri NRRL B-11177, is studied in producing gold nanoparticles. This is the first report about the biosynthesis of gold nanoparticles by natural and genetically engineered luminescent bacteria. These microorganisms reduced gold ions and produced fairly monodisperse nanoparticles. TEM analysis indicated that spherical nano gold particles in the different diameters and shapes were obtained at pH values of 6.64. In this biosynthesis protocol, the gold nanoparticles with desired shape and size can be prepared.

Evaluation of Antimicrobial and Healing Activities of Frog Skin on Guinea Pigs Wounds.

BACKGROUND: Frog skin secretions have potentials against a wide spectrum of bacteria. Also, frog skin compositions have healing properties. OBJECTIVES: The aim of this study was to investigate the antibacterial potentials along with healing properties of frog skin Rana ridibunda, a species which thoroughly lives in Iran marshes, as a biological dressing on wounds. MATERIALS AND METHODS: In this study, excisional wounds, dressed with frog skin, were compared between experimental and control groups of guinea pigs. In the experimental groups, wounds were dressed with the dermal (FS) and epidermal (RFS) sides of fresh frog R. ridibunda skin, while only usual cotton gauze covered the wounds of the control group. Furthermore, microbial samples were taken on different days (0, 3, 5, and 7 days post injury) to count the number of the colony-forming units. Additionally, the microbial penetration test was performed on frog skin and then the progression of wound closure was evaluated. RESULTS: In the microbial studies, the bacterial load considerably declined in the wounds treated with FS and RFS compared with the control wounds. On day 7 post injury, the numbers of the colony-forming units for the FS, RFS, and control groups were 6.75, 105, and 375, respectively. In the penetration test, fresh frog skin showed to be a bacterial resistant dressing. The results revealed that the rate of wound closure in the experimental groups significantly was accelerated in comparison with that in the control group. CONCLUSIONS: Our results demonstrated the antimicrobial properties of frog skin as a wound dressing, which has antimicrobial effects per se. This biological dressing shows promise as an effective biological wound dressing insofar as not only is it capable of resisting microbes and accelerating wound healing but also it is cost-effective and easy to use.

Optimization of biomass and biokinetic constant in Mazut biodegradation by indigenous bacteria BBRC10061.

Optimization based on appropriate parameters can be applied to improve a process. Mazut degradation as a critical issue in environment requires optimization to be efficiently done. To provide biodegradation conditions, experiments were designed on the least interactions among levels of parameters consisting of pH, Tween 80, glucose, phosphorous source, nitrogen source, and time. Kinetic constants and biomass were calculated based on 16 assays, designed using Taguchi method, which constructed various mazut biodegradation conditions. Kinetics of mazut degradation by newly isolated bacteria Enterobacter cloacae closely followed second order kinetic model. Results of the 16 experiments showed that biomass was in the range of 0.019 OD600 to 2.75 OD600, and biokinetic constant was in the range of 0.2 × 10(-5) L/ (mg day) to 10(-4) L/ (mg day). Optimal level for each parameter was obtained through data analysis. For optimal biomass equal to 2.75 OD600, optimal pH, Tween80, glucose, phosphorous source, and time were 8.3, 4 g/L, 4 g/L, 9 g/L, and 10 days, respectively. For biokinetic constant equal to 1.2 × 10(-4) L/ (mg day), optimal pH, Tween80, glucose, phosphorous source, and nitrogen source were 8.3, 1 g/L, 4 g/L, 1 g/L, and 9 g/L, respectively. The optimum levels for biomass and biokinetic constant were the same except the levels of the Tween 80, and phosphorous source. Consequently, mazut may be more degraded with adjusting the conditions on the optimum condition.