NIH 3T3 Mouse Embryonic Fibroblast Research Articles

Di-Sulfide Linked Polyethylenimine Coated Gold Nanoparticles as a Non-Viral Gene Delivery Agent in NIH-3T3 Mouse Embryonic Fibroblast.

Di-sulfide linked polyethylenimine coated gold nanoparticles (ssPEI-GNPs) of 20 nm size was prepared in order to deliver the genes to target site. DLS and TEM analysis demonstrated that the GNPs have average size of 13 nm in diameter. Upon coating the GNPs with ssPEI in the weight ratio of 1:3, the average hydrodynamic diameter of the ssPEI-GNPs was found to 19±1.14 nm and a zeta potential value 41±1.23 mV was observed. TEM analysis of ssPEI-GNPs demonstrated that the nanoparticles have spherical morphology. Thermogravemetric analysis of the prepared ssPEI-GNPs showed that the estimated composition of the ssPEI coated over the GNPs was approximately 5% (w/w). Gene expression capabilities of the nanoparticles were confirmed by fluorescent microscopy and luciferase assay, which demonstrated the transgene delivery capability of the ssPEI-GNPs. These results demonstrate that ssPEI-GNPs could be used as gene delivery agent.

Synthesis of Tolmetin Hydrazide-Hydrazones and Discovery of a Potent Apoptosis Inducer in Colon Cancer Cells.

Tolmetin hydrazide and a novel series of tolmetin hydrazide-hydrazones 4a-l were synthesized in this study. The structures of the new compounds were determined by spectral (FT-IR, (1)H NMR) methods. N'-[(2,6-Dichlorophenyl)methylidene]-2-[1-methyl-5-(4-methylbenzoyl)-1H-pyrrol-2-yl]acetohydrazide (4g) was evaluated in vitro using the MTT colorimetric method against the colon cancer cell lines HCT-116 (ATCC, CCL-247) and HT-29 (ATCC, HTB-38) to determine growth inhibition and cell viability at different doses. Compound 4g exhibited anti-cancer activity with an IC50 value of 76 μM against colon cancer line HT-29 (ATCC, HTB-38) and did not display cytotoxicity toward control NIH3T3 mouse embryonic fibroblast cells compared to tolmetin. In addition, this compound was evaluated for caspase-3, caspase-8, caspase-9, and annexin-V activation in the apoptotic pathway, which plays a key role in the treatment of cancer. We demonstrated that the anti-cancer activity of this compound was due to the activation of caspase-8 and caspase-9 involved in the apoptotic pathway. In addition, in this study, we investigated the catalytical effect of COX on the HT-29 cancer line, the apoptotic mechanism, and the moleculer binding of tolmetin and compound 4g on the COX enzyme active site.

Silk fibroin sponges with cell growth-promoting activity induced by genetically fused basic fibroblast growth factor.

Transgenic silkworm technology has enabled the biological properties of silk fibroin protein to be altered by fusion to recombinant bioactive proteins. However, few studies have reported the fabrication of genetically modified fibroin proteins into three-dimensional spongy structures to serve as scaffolds for tissue engineering. We generated a transgenic silkworm strain that produces fibroin fused to basic fibroblast growth factor (bFGF) and processed the fibroin into a spongy structure using a simple freeze/thaw method. NIH3T3 mouse embryonic fibroblasts grown on bFGF-fused fibroin sponges proliferated and spread out well, showing half the population doubling time of cells cultured on wild-type fibroin sponges. Furthermore, the number of primary rabbit articular chondrocytes growing on bFGF-fused fibroin sponges was around five-times higher than that of the wild-type control at 3-days post cell-seeding. As the physical properties of wild-type and bFGF-fused fibroin sponges were almost identical, it is suggested that bFGF fused to fibroin retained its biological activity, even after the bFGF-fused fibroin was fabricated into the spongy structure. The bFGF-fused fibroin sponge has the potential for widespread application in the field of tissue engineering, and the method of fabricating this structure could be applicable to other recombinant bioactive fibroin proteins.

Antibacterial and Antifungal Activity of ZnO Containing Glasses.

A new family of non-toxic biocides based on low melting point (1250°C) transparent glasses with high content of ZnO (15–40wt%) belonging to the miscibility region of the B2O3-SiO2-Na2O-ZnO system has been developed. These glasses have shown an excellent biocide activity (logarithmic reduction >3) against Gram- (E. coli), Gram+ (S. aureus) and yeast (C. krusei); they are chemically stable in different media (distilled water, sea-like water, LB and DMEN media) as well as biocompatible. The cytotoxicity was evaluated by the Neutral Red Uptake using NIH-3T3 (mouse embryonic fibroblast cells) and the cell viability was >80%. These new glasses can be considered in several and important applications in the field of inorganic non-toxic biocide agents such as medical implants, surgical equipment, protective apparels in hospitals, water purifications systems, food packaging, food storages or textiles.

Surface modifications of titanium implants by coating with bioactive and biocompatible poly (ε-caprolactone)/SiO2 hybrids synthesized via sol–gel

Class I organic/inorganic hybrid materials have been synthesized by sol–gel method from a multicomponent solution containing tetraethyl orthosilicate (TEOS), water, ethanol and nitric acid by adding different percentages of poly (ε-caprolactone) (PCL) dissolved in chloroform. Those hybrids have been used, in sol phase, to dip coat commercially pure titanium (CP Ti) grade 4 substrates with the aim of transferring to them the known biological properties of silica-based sol–gel materials. Particular attention has been directed to investigate the effect of PCL amount on both structure and coating performances.The chemical composition of the films was ascertained by attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) analysis. Scanning electron microscope (SEM) proved that polymer allows to make crack-free coatings.The biological performances of the un-coated and coated substrates were compared and evaluated by means of bioactivity test and WST-8 assay. SEM and energy dispersive X-ray (EDX) analyses have detected higher hydroxyapatite nucleation on the surface of the coated substrates after soaking in a fluid simulating the composition of the human blood plasma (SBF). Moreover, higher vitality of NIH 3T3 mouse embryonic fibroblast cells seeded on coated specimens was recorded. The results, thus, prove that coating application effectively improves the bioactivity and biocompatibility of CP Ti grade 4 substrates.

TiO2/PCL hybrid materials synthesized via sol-gel technique for biomedical applications.

The aim of the present work has been the synthesis of organic/inorganic hybrid materials based on titanium dioxide and poly(ε-caprolactone) (PCL) to be used in the biomedical field. Several materials have been synthesized using sol-gel methods by adding different amounts of polymer to the inorganic sol. The obtained gels have been characterized using Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscopy (SEM) and atomic force microscopy (AFM). The FT-IR data allowed us to hypothesize that the structure formed was that of an interpenetrating network, realized by hydrogen bonds between TiOH groups in the sol-gel intermediate species and carbonyl groups in the polymer repeating units. SEM and AFM analyses highlighted that the obtained materials were nanostructurated hybrids. To evaluate the biological properties of the hybrids, their bioactivity and cytotoxicity were investigated as a function of the PCL amount. The bioactivity of the synthesized systems was proven by the formation of a hydroxyapatite layer on the surface of samples soaked in a fluid simulating human blood plasma (SBF). MTT cytotoxicity tests and Trypan Blue dye exclusion tests were carried out exposing NIH-3T3 mouse embryonic fibroblasts for 24 and 48h to extracts from the investigated hybrid materials. The results showed that all the hybrids had a non-cytotoxic effect on target cells.

Designing and In-Vitro Characterization of Micelle Forming Amphiphilic PEGylated Rapamycin Nanocarriers for the Treatment of Gastric Cancer

The present study aims to develop and explore the use of PEGylated rapamycin (RP-MPEG) micelles for the treatment of gastric cancer. RP-MPEG was synthesized and characterized by using IR, H(1) NMR and C(13) NMR. RP-MPEG was prepared in the form of micelles and characterized by using field emission scanning electron microscopy, dynamic light scattering, zeta sizer, chromatographic analyses and photostability studies. The cytotoxicity studies of RP-MPEG micelles were conducted on specific CRL 1739 human gastric adenocarcinoma and CRL 1658 NIH-3T3 mouse embryonic fibroblast cell lines. RP-MPEG micelles showed the particle size distribution of 125±0.26 nm with narrow size distribution (polydispersity index 0.127±0.01). The surface charge of RP-MPEG micelles was found to be -12.3 mV showing enhanced anticancer activity against the CRL 1739 human gastric adenocarcinoma cell lines with an IC50 value of 1 mcg/ml.

Investigation of in vitro anticancer and DNA strap interactions in live cells using carboplatin type Cu(II) and Zn(II) metalloinsertors

A series of carboplatin type Cu(II) and Zn(II) metalloinsertors (1–8) having β-diketone analogues and biologically significant cyclobutane-1,1-dicarboxylic acid have been synthesized and characterized by spectral and analytical methods. The binding and cleavage propensity of these metalloinsertors on DNA and their cytotoxic effects in live cells have been explored. From the gel electrophoresis study, it is observed that the complexes 1–8 cleave pBR322 DNA via a hydrolytic mechanism induced by hydroxyl radical scavengers, DMSO and EtOH as the reactive oxygen species (ROS). In vivo antitumor efficacy has been studied on EAC tumor bearing mice which is assessed by mean survival time, effect on hematological parameters and solid tumor volume. The results strongly support that complex 1 shows potent antitumor effect against EAC and higher than the standard drug carboplatin. Moreover, the cytotoxicity of the complexes, screened on a panel of human cancerous cell lines viz., human cervical cancer cells (HeLa), human breast adenocarcinoma cells (MCF-7), human laryngeal epithelial carcinoma cells (HEp-2), human liver carcinoma cells (Hep G2) and non-cancerous NIH 3T3 mouse embryonic fibroblasts cell lines, reveals that complex 1 exhibits a better anticancer activity than other complexes and standards.

Biocompatibility of bio based calcium carbonate nanocrystals aragonite polymorph on NIH 3T3 fibroblast cell line.

Currently, there has been extensive research interest for inorganic nanocrystals such as calcium phosphate, iron oxide, silicone, carbon nanotube and layered double hydroxide as a drug delivery system especially in cancer therapy. However, toxicological screening of such particles is paramount importance before use as delivery carrier. In this study we examine the biocompatibility of CaCO3 nanocrystal on NIH 3T3 cell line. Transmission and field emission scanning electron microscopy (TEM and FESEM) were used for the characterisation of CaCO3 nanocrystals. Cytotoxicity and genotoxic effect of calcium carbonate nanocrystals in cultured mouse embryonic fibroblast NIH 3T3 cell line using various bioassays including MTT, and Neutral red/Trypan blue double-staining assays. LDH, BrdU and reactive oxygen species were used for toxicity analysis. Cellular morphology was examined by scanning electron microscopy (SEM) and confocal fluorescence microscope. The outcome of the analyses revealed a clear rod-shaped aragonite polymorph of calcium carbonate nanocrystal. The analysed cytotoxic and genotoxicity of CaCO3 nanocrystal on NIH 3T3 cells using different bioassays revealed no significance differences as compared to control. A slight decrease in cell viability was noticed when the cells were exposed to higher concentrations of 200 to 400 µg/ml, while increase in ROS generation and LDH released at 200 and 400 µg/ml was observed. The study has shown that CaCO3 nanocrystal is biocompatible and non toxic to NIH 3T3 fibroblast cells. The analysed results offer a promising potential of CaCO3 nanocrystal for the development of intracellular drugs, genes and other macromolecule delivery systems.

Functionally modified gelatin microspheres as a growth factor’s delivery system: development and characterization

One of the advances in biotechnology has been the development of the capability to produce large quantities of highly purified polypeptides and proteins. Unfortunately, the circulatory half-lives of many of these agents are short, usually of the order of minutes and the time required for a response in tissues is usually long compared to the half-life. Hence, there is always demand for polymeric systems which can deliver the proteins for prolonged period and also to protect the molecules from degradation. The present work was attempted to develop heparin-functionalized gelatin microspheres (HMS) to deliver heparin-binding growth factors particularly for wound-healing applications. The heparin conjugation was carried out using EDC/NHS coupling protocol. Heparin-binding EGF-like growth factor (HB-EGF) was loaded in HMS and its in vitro release behaviour in an environment with or without proteases was studied. The bioactivity of the HB-EGF released from the microspheres was assessed using NIH 3T3 mouse embryonic fibroblast culture. The extent of heparin modification was found to be 1.97 μmol/g of HMS and demonstrated significant protection against enzymatic degradation and sustained release of HB-EGF for more than 10 days. The bioactivity of HB-EGF released from the HMS was retained during the observed release period. The HMS was also found to be non-toxic as determined by calcein AM fluorescent staining. The overall study suggests that the HMS could be used as a growth factor’s delivery component in tissue engineering scaffolds particularly for wound-healing applications.

Biocompatibility improvement of titanium implants by coating with hybrid materials synthesized by sol-gel technique

Organic-inorganic hybrid materials based on zirconia and polyethylene glycol (PEG) have been synthesized via sol-gel method in the present study. Those materials, still in the sol phase, have been used to coat a titanium grade 4 (Ti-4) substrate to improve its biological properties. Dip-coating technique has been used to obtain thin films. PEG, a biocompatible polymer, used as the organic phase, has been incorporated with different percentages in an inorganic zirconium-based matrix. Those hybrids have been characterized by Fourier transform infrared spectroscopy (FTIR) to detect interactions between the two phases. The films have been examined using SEM to detect morphological changes with PEG percentages. The potential applications of the hybrid coatings in biomedical field have been evaluated by bioactivity and cytotoxicity tests. The coated titanium was immersed in simulated body fluid (SBF) for 21 days and the hydroxyapatite deposition on its surface was subsequently evaluated, as that feature can be used as an index of bone-bonding capability. SEM equipped with energy dispersive spectrometer (EDS) was used to examine hydroxyapatite formation. NIH 3T3 mouse embryonic fibroblast cells were seeded on specimens to evaluate cells-materials interactions and cell vitality was inspected using WST-8 Assay.

Genomic Organization and Identification of Promoter Regions for the BDNF Gene in the Pond Turtle Trachemys scripta elegans

Brain-derived neurotrophic factor (BDNF) is an important regulator of neuronal development and synaptic function. The BDNF gene undergoes significant activity-dependent regulation during learning. Here, we identified the BDNF promoter regions, transcription start sites, and potential regulatory sequences for BDNF exons I-III that may contribute to activity-dependent gene and protein expression in the pond turtle Trachemys scripta elegans (tBDNF). By using transfection of BDNF promoter/luciferase plasmid constructs into human neuroblastoma SHSY5Y cells and mouse embryonic fibroblast NIH3T3 cells, we identified the basal regulatory activity of promoter sequences located upstream of each tBDNF exon, designated as pBDNFI-III. Further, through chromatin immunoprecipitation (ChIP) assays, we detected CREB binding directly to exon I and exon III promoters, while BHLHB2, but not CREB, binds within the exon II promoter. Elucidation of the promoter regions and regulatory protein binding sites in the tBDNF gene is essential for understanding the regulatory mechanisms that control tBDNF gene expression.

Nanoscale engineering of low-fouling surfaces through polydopamine immobilisation of zwitterionic peptides

We report a versatile approach for the design of substrate-independent low-fouling surfaces via mussel-inspired immobilisation of zwitterionic peptides. Using mussel-inspired polydopamine (PDA) coatings, zwitterionic glutamic acid- and lysine-based peptides were immobilised on various substrates, including noble metals, metal oxides, polymers, and semiconductors. The variation of surface chemistry and surface wettability upon surface treatment was monitored with X-ray photoelectron spectroscopy (XPS) and water contact angle measurements. Following peptide immobilisation, the surfaces became more hydrophilic due to the strong surface hydration compared with PDA-coated surfaces. The peptide-functionalised surfaces showed resistance to human blood serum adsorption and also effectively prevented the adhesion of gram-negative and gram-positive bacteria (i.e., Escherichia coli and Staphylococcus epidermidis) and mammalian cells (i.e., NIH 3T3 mouse embryonic fibroblast cells). The versatility of mussel-inspired chemistry combined with the unique biological nature and tunability of peptides allows for the design of low-fouling surfaces, making this a promising coating technique for various applications.

Antitumoral Gold and Silver Complexes with Ferrocenyl-Amide Phosphines

The monosubstituted ferrocenyl-amide phosphine ligands PPh2CH2CH2NHCOFc and (PPh2CH2CH2)2NCOFc and the disubstituted ferrocenyl-amide diphosphine (PPh2CH2CH2NHCO)2Fc have been synthesized and used to prepare the gold chloride derivatives [AuCl(PPh2CH2CH2NHCOFc)], [Au2Cl2{(PPh2CH2CH2)2NCOFc}], and [Au2Cl2{(PPh2CH2CH2NHCO)2Fc}] or the silver species [Ag(OTf)(PPh2CH2CH2NHCOFc)2] and [Ag(OTf)(PPh3)(PPh2CH2CH2NHCOFc)]. In the gold complexes the chloro ligands can be easily substituted by several biologically relevant thiolates such as 2-mercaptonicotinic acid, 2-thiocytosine, 2-thiouracil, 2-mercaptopurine, and 2,3,4,6-tetra-6-acetyl-1-thiol-β-d-glucopyranosato, affording the gold phosphine thiolate derivatives. In addition, the gold phosphine thiolates of the closely related 1,1′-bis(diphenylphosphine)ferrocene ligand have been prepared in order to compare their biological activities. The antiproliferative activity of these compounds has been tested by the MTT viability assay in two murine cell lines, NIH-3T3 (mouse embryonic fibroblasts) and PC-12 (pheochromocytoma of the rat adrenal medulla), and two human cell lines, A-549 (adenocarcinomic human alveolar basal epithelial cells) and Hep-G2 (hepatocellular carcinoma). The amide-phosphine ligands are not active, whereas the chloro-gold derivatives have good antiproliferative activity in the murine cell lines and very low activity in the human cell lines. The silver complexes are less active than the gold derivatives. The gold thiolate complexes have moderate to very good cytotoxic activity for all of the ligands, showing excellent IC50 values for the thiolate complexes of the amide-phosphines PPh2CH2CH2NHCOFc and (PPh2CH2CH2N)2COFc and dppf.

The Influence of Laser-Induced 3-D Titania Nanofibrous Platforms on Cell Behavior

The current challenge in tissue engineering is to design a platform that can provide appropriate topography and suitable surface chemistry to encourage desired cellular activities and to guide 3-D tissue regeneration. Compared with traditional cell culture materials, 3-D nanofibrous platforms offer a superior environment for promoting cell functions by mimicking the architecture of extracellular matrix (ECM). In this study, we present a technique to engineer freestanding 3-D titania nanofibrous structures on titanium substrates using femtosecond laser processing. The crystallinity, surface adhesion, and surface energy of the synthesized nanostructures are discussed. The effects of synthesized nanoarchitectures on the proliferation, morphology, and viability of MC3T3-E1 mouse osteoblast-like cells and NIH 3T3 mouse embryonic fibroblasts are investigated. The nanofibrous structures show high surface energy and hydrophilicity. The results from in vitro studies reveal that the titania nanofibrous architectures possess excellent biocompatibility and significantly enhances proliferation of both cell lines compared to untreated titanium specimens. Study of the cell morphology shows dynamic cell migration and attachment on the titania nanofibrous architecture. The bioactivity and biocompatibility of the engineered 3-D nanostructures suggest noticeable perspective for developing bio-functionalized scaffolds and implantable materials in regenerative medicine and clinical tissue engineering.

BMP2-Induced Inflammation Can Be Suppressed by the Osteoinductive Growth Factor NELL-1

Bone-morphogenetic protein 2 (BMP2) is currently the only Food and Drug Administration-approved osteoinductive growth factor used in clinical settings for bone regeneration and repair. However, the use of BMP2 is encumbered by numerous clinical complications, including postoperative inflammation and life-threatening cervical swelling. Thus, methods to prevent BMP2-induced inflammation would have far-reaching clinical implications toward improving current BMP2-based methods for bone regeneration. For the first time, we investigate the potential role of the growth factor Nel-like molecule-1 (NELL-1) in inhibiting BMP2-induced inflammation. Adult rats underwent a femoral bone onlay procedure, treated with either BMP2 protein (4 mg/mL), NELL-1 protein (4 mg/mL), or both proteins combined. Animals were evaluated at 3, 7, and 14 days postoperatively by histology, histomorphometry, immunohistochemistry, and real-time PCR for markers of inflammation (TNFα, IL6). The relative levels of TNFα and IL6 in serum were also detected by ELISA. The mechanism for NELL-1's anti-inflammatory effect was further assessed through examining inflammatory markers and generation of reactive oxygen species (ROS) in the mouse embryonic fibroblast NIH3T3 cells. BMP2 significantly induced local inflammation, including an early and pronounced polymorphonuclear cell infiltration accompanied by increased expression of TNFα and IL6. Treatment with NELL-1 alone elicited no significant inflammatory response. However, NELL-1 significantly attenuated BMP2-induced inflammation by all markers and at all timepoints. These local findings were also confirmed using systemic serum inflammatory biomarkers (TNFα, IL6). In each case, NELL-1 fully reversed BMP2-induced systemic inflammation. Lastly, our findings were recapitulated in vitro, where NELL-1 suppressed BMP2 induced expression of inflammatory markers, as well as NF-κB transcriptional activity and generation of ROS. BMP2-induced inflammation is a serious public health concern with potentially life-threatening complications. In the present study, we observed that the growth factor, NELL-1, significantly attenuates or completely reverses BMP2-induced inflammation. The mechanisms of NELL-1's anti-inflammatory effect are only partially elucidated, and may include reduction of NF-κB transcriptional activity or ROS generation.

Pulsed ultrasound promotes melanoblast migration through upregulation of macrophage colony‐stimulating factor/focal adhesion kinase autocrine signaling and paracrine mechanisms

Repigmentation of vitiliginous lesions relies on the proliferation and migration of melanoblasts from hair follicles to the epidermis. Pulsed ultrasound has been demonstrated to have stimulatory effects on cell proliferation and migration and has been applied clinically to enhance tissue repair. To clarify the biologic effects and signaling mechanisms of pulsed ultrasound on melanoblast proliferation and migration, two melanoblast cell lines, the undifferentiated NCCmelb4 cells and the differentiated NCCmelan5 cells, were examined. We demonstrated that pulsed ultrasound increased cell migration in a dose-dependent manner without altering cell proliferation. Pulsed ultrasound enhanced autocrine secretion of macrophage colony-stimulating factor (M-CSF), which subsequently activated the focal adhesion kinase (FAK) pathway to promote melanoblast migration. Furthermore, conditioned medium from mouse embryonic fibroblasts NIH 3T3 and primary human keratinocytes treated with pulsed ultrasound could stimulate melanoblast migration through a paracrine effect. Our results provide a novel mechanism to promote migration of melanoblasts by pulsed ultrasound stimulation.

Trehalose Glycopolymers as Excipients for Protein Stabilization

Herein, the synthesis of four different trehalose glycopolymers and investigation of their ability to stabilize proteins to heat and lyophilization stress are described. The disaccharide, α,α-trehalose, was modified with a styrenyl acetal, methacrylate acetal, styrenyl ether, or methacrylate moiety resulting in four different monomers. These monomers were then separately polymerized using free radical polymerization with azobisisobutyronitrile (AIBN) as an initiator to synthesize the glycopolymers. Horseradish peroxidase and glucose oxidase were incubated at 70 and 50 °C, respectively, and β-galactosidase was lyophilized multiple times in the presence of various ratios of the polymers or trehalose. The protein activities were subsequently tested and found to be significantly higher when the polymers were present during the stress compared to no additive and to equivalent amounts of trehalose. Different molecular weights (10 kDa, 20 kDa, and 40 kDa) were tested, and all were equivalent in their stabilization ability. However, some subtle differences were observed regarding stabilization ability between the different polymer samples, depending on the stress. Small molecules such as benzyl ether trehalose were not better stabilizers than trehalose, and the trehalose monomer decreased protein activity, suggesting that hydrophobized trehalose was not sufficient and that the polymeric structure was required. In addition, cytotoxicity studies with NIH 3T3 mouse embryonic fibroblast cells, RAW 264.7 murine macrophages, human dermal fibroblasts (HDFs), and human umbilical vein endothelial cells (HUVECs) were conducted with polymer concentrations up to 8 mg/mL. The data showed that all four polymers were noncytotoxic for all tested concentrations. The results together suggest that trehalose glycopolymers are promising as additives to protect proteins from a variety of stressors.

Biological synthesis of silver nanoparticles using the fungus Humicola sp. and evaluation of their cytoxicity using normal and cancer cell lines

Nanoscience is a new born science of the modern era and taps into the potential of particles at nanoscale. Bulk materials reduced to nanoscale dimensions thus obtain unique properties such as electronic, optical, magnetic and chemical. As far as synthesis of nanoparticles is concerned, biological synthesis has recently sparked a great interest as compared to other available chemical and physical methods on account of its eco-friendliness and cost-effectiveness. Here we report, for the first time, the biosynthesis of silver nanoparticles by the thermophilic fungus Humicola sp. The fungus when reacted with Ag+ ions reduces the precursor solution and leads to the formation of extracellular nanoparticles as monitored by ultra violet visible spectroscopy (UV–Vis). The morphology of nanoparticles is found to be spherical with good dispersity as revealed by transmission electron microscopy (TEM). Cell viability assays were carried out to assess the cytotoxicity of silver nanoparticles on NIH3T3 mouse embryonic fibroblast cell line and MDA-MB-231 human breast carcinoma cell line.

Nanomagnetic Gene Transfection for Non-Viral Gene Delivery in NIH 3T3 Mouse Embryonic Fibroblasts.

The objective of this work was to examine the potential of oscillating nanomagnetic gene transfection systems (magnefect-nano™) for improving the transfection efficiency of NIH3T3 mouse embryonic fibroblasts (MEFs) in comparison to other non-viral transfection techniques—static magnetofection™ and the cationic lipid agent, Lipofectamine 2000™. Magnetic nanoparticles (MNPs) associated with the plasmid coding for green fluorescent protein (GFP) were used to transfect NIH3T3 cells. The magnefect-nano system was evaluated for transfection efficiency, and any potential associated effects on cell viability were investigated. MNPs associated with the plasmid coding for GFP were efficiently delivered into NIH3T3 cells, and the magnefect-nano system significantly enhanced overall transfection efficiency in comparison to lipid-mediated gene delivery. MNP dosage used in this work was not found to affect the cell viability and/or morphology of the cells. Non-viral transfection using MNPs and the magnefect-nano system can be used to transfect NIH3T3 cells and direct reporter gene delivery, highlighting the wide potential of nanomagnetic gene transfection in gene therapy.