Composite spheres made of bioengineered spider silk and iron oxide nanoparticles for theranostics applications

Bioengineered spider silk is a biomaterial that has exquisite mechanical properties, biocompatibility, and biodegradability. Iron oxide nanoparticles can be applied for the detection and analysis of biomolecules, target drug delivery, as MRI contrast agents and as therapeutic agents for hyperthermia-based cancer treatments. In this study, we investigated three bioengineered silks, MS1, MS2 and EMS2, and their potential to form a composite material with magnetic iron oxide nanoparticles (IONPs). The presence of IONPs did not impede the self-assembly properties of MS1, MS2, and EMS2 silks, and spheres formed. The EMS2 spheres had the highest content of IONPs, and the presence of magnetite IONPs in these carriers was confirmed by several methods such as SEM, EDXS, SQUID, MIP-OES and zeta potential measurement. The interaction of EMS2 and IONPs did not modify the superparamagnetic properties of the IONPs, but it influenced the secondary structure of the spheres. The composite particles exhibited a more than two-fold higher loading efficiency for doxorubicin than the plain EMS2 spheres. For both the EMS2 and EMS2/IONP spheres, the drug revealed a pH-dependent release profile with advantageous kinetics for carriers made of the composite material. The composite spheres can be potentially applied for a combined cancer treatment via hyperthermia and drug delivery

Silk-fibronectin protein alloy fibres support cell adhesion and viability as a high strength, matrix fibre analogue

Silk is a natural polymer with broad utility in biomedical applications because it exhibits general biocompatibility and high tensile material properties. While mechanical integrity is important for most biomaterial applications, proper function and integration also requires biomaterial incorporation into complex surrounding tissues for many physiologically relevant processes such as wound healing. In this study, we spin silk fibroin into a protein alloy fibre with whole fibronectin using wet spinning approaches in order to synergize their respective strength and cell interaction capabilities. Results demonstrate that silk fibroin alone is a poor adhesive surface for fibroblasts, endothelial cells, and vascular smooth muscle cells in the absence of serum. However, significantly improved cell attachment is observed to silk-fibronectin alloy fibres without serum present while not compromising the fibres' mechanical integrity. Additionally, cell viability is improved up to six fold on alloy fibres when serum is present while migration and spreading generally increase as well. These findings demonstrate the utility of composite protein alloys as inexpensive and effective means to create durable, biologically active biomaterials.T32 EB006359 - NIBIB NIH HH

Microrheological Studies of Regenerated Silk Fibroin Solution by Video Microscopy

We have carried out studies on the rheological properties of regenerated silk fibroin (RSF) solution using video microscopy. The degummed silk from the Bombyx mori silkworm was used to prepare RSF solution by dissolving it in calcium nitrate tetrahydrate-methanol solvent. Measurements were carried out by tracking the position of an embedded micron-sized polystyrene bead within the RSF solution through video imaging. The time dependent mean squared displacement (MSD) of the bead in solution and hence, the complex shear modulus of this solution was calculated from the bead's position information. An optical tweezer was used to transport and locate the bead at any desired site within the micro-volume of the sample, to facilitate the subsequent free-bead video analysis. We present here the results of rheological measurements of the silk polymer network in solution over a frequency range, whose upper limit is the frame capture rate of our camera, at full resolution. By examining the distribution of MSD of beads at different locations within the sample volume, we demonstrate that this probe technique enables us to detect local inhomogeneties at micrometer length scales, not detectable either by a rheometer or from diffusing wave spectroscopy.Comment: 5 page

Blueprint for a high-performance biomaterial: full-length spider dragline silk genes.

Spider dragline (major ampullate) silk outperforms virtually all other natural and manmade materials in terms of tensile strength and toughness. For this reason, the mass-production of artificial spider silks through transgenic technologies has been a major goal of biomimetics research. Although all known arthropod silk proteins are extremely large (>200 kiloDaltons), recombinant spider silks have been designed from short and incomplete cDNAs, the only available sequences. Here we describe the first full-length spider silk gene sequences and their flanking regions. These genes encode the MaSp1 and MaSp2 proteins that compose the black widow's high-performance dragline silk. Each gene includes a single enormous exon (>9000 base pairs) that translates into a highly repetitive polypeptide. Patterns of variation among sequence repeats at the amino acid and nucleotide levels indicate that the interaction of selection, intergenic recombination, and intragenic recombination governs the evolution of these highly unusual, modular proteins. Phylogenetic footprinting revealed putative regulatory elements in non-coding flanking sequences. Conservation of both upstream and downstream flanking sequences was especially striking between the two paralogous black widow major ampullate silk genes. Because these genes are co-expressed within the same silk gland, there may have been selection for similarity in regulatory regions. Our new data provide complete templates for synthesis of recombinant silk proteins that significantly improve the degree to which artificial silks mimic natural spider dragline fibers

Metrology for Bio Systems

The current paper addresses the advent of next generation bio system focussed Micro Nano Manufacturing Technologies (MNMT). These products and processes have placed significant new emphasis on specification and quality control systems, especially if these product and processes are to achieve economic scale up. Bio technology products and processes are a core element of MNMT and structured surfaces can be a key element in enabling bio system function. There examples of the application of such surfaces in bio systems for functions such as diverse as anti fouling and oseointegration. However a deficit exists in terms of metrology for bio structured surfaces and identifying suitable measurands and instrumentation remains a challenge for production engineers. Functional modelling would seem to point towards a better way of specifying metrology however for bio systems these are rare and often extensive function testing and clinical trials are used to inform the metrology selection. In the present paper the development of MNMT bio systems is discussed in the metrology context and several examples of developing metrology challenges. Four such bio related systems are discussed the solutions are outlined. The case studies cover traditional prosthetic implants, micro fluidic devices, cellular attachment and manufacture of cellular scaffolds

Second-order Nonlinear Optical Microscopy of Spider Silk

Asymmetric $\rm \beta$-sheet protein structures in spider silk should induce nonlinear optical interaction such as second harmonic generation (SHG) which is experimentally observed for a radial line and dragline spider silk by using an imaging femtosecond laser SHG microscope. By comparing different spider silks, we found that the SHG signal correlates with the existence of the protein $\rm \beta$-sheets. Measurements of the polarization dependence of SHG from the dragline indicated that the $\rm \beta$-sheet has a nonlinear response depending on the direction of the incident electric field. We propose a model of what orientation the $\rm \beta$-sheet takes in spider silk.Comment: 8 pages, 8 figures, 1 tabl

Graphene-based nanomaterials for tissue engineering in the dental field

The world of dentistry is approaching graphene-based nanomaterials as substitutes for tissue engineering. Apart from its exceptional mechanical strength, electrical conductivity and thermal stability, graphene and its derivatives can be functionalized with several bioactive molecules. They can also be incorporated into different scaffolds used in regenerative dentistry, generating nanocomposites with improved characteristics. This review presents the state of the art of graphene-based nanomaterial applications in the dental field. We first discuss the interactions between cells and graphene, summarizing the available in vitro and in vivo studies concerning graphene biocompatibility and cytotoxicity. We then highlight the role of graphene-based nanomaterials in stem cell control, in terms of adhesion, proliferation and differentiation. Particular attention will be given to stem cells of dental origin, such as those isolated from dental pulp, periodontal ligament or dental follicle. The review then discusses the interactions between graphene-based nanomaterials with cells of the immune system; we also focus on the antibacterial activity of graphene nanomaterials. In the last section, we offer our perspectives on the various opportunities facing the use of graphene and its derivatives in associations with titanium dental implants, membranes for bone regeneration, resins, cements and adhesives as well as for tooth-whitening procedure

Adsorption of olive leaf (Olea europaea L.) antioxidants on silk fibroin

The adsorption isotherms of oleuropein and rutin were evaluated at different temperatures, pH values, and solid/liquid ratios. The experimental data of adsorption isotherms were well fitted to a Langmuir model. The maximum adsorption capacities were determined as 108 mg of oleuropein/g of silk fibroin and 21 mg of rutin/g of silk fibroin. After adsorption of oleuropein and rutin, the antioxidant capacity of silk fibroin increased from 1.93 to 3.61 mmol of TEAC/g. Silk fibroin also gained antimicrobial activity against Staphylococcus aureus and Klebsiella pneumoniae after adsorption of olive leaf antioxidants. In a desorption process, 81% of rutin and 85% of oleuropein were removed from the adsorbent surface in 70% aqueous ethanol solution. Consequently, silk fibroin was found to be a promising biomaterial for the production of functional food or dietary supplements and for the purification of oleuropein and rutin from olive leaf extracts