Bio-inspired protein-based biomaterial

Protein functions are as diverse as protein structures. The tunability and biocompatibility of proteins make them attractive candidates for use as building blocks for biomaterials engineering. This strategy provides molecular-level material design, enabling straightforward and independent control over an array of biomaterial properties. A key challenge in our research is to unveil the mechanisms of formation of micro and nano-scale protein-based capsules-gels and shells, as well as to achieve their functionalization for uses in targeted delivery of bioactive materials. The aim to test protein microgels for their ability to act as drug carrier agents, and for the controlled release of different drug-like small molecules as well as the release of the component proteins. Advantages of these systems include compositional definition, control over topology and nanostructure, and the ability to combine multiple different functional components in a modular way. Please click Additional Files below to see the full abstract

The immobilization of polyethylene imine nano and microspheres on glass using high intensity ultrasound

The present article describes the creation and immobilization of Polyethylene imine (PEI) capsules on a glass surface. The synthesis and deposition were accomplished by short-time (3 min) one-step reaction. The preparation and immobilization of PEI spheres, was carried out using an environmental friendly method, the ultrasonic emulsification. The ultrasonic technique enables to control size and fulfillment of the internal part of the immobilized PEI spheres. Moreover, the ultrasonic emulsification method showed 100% efficiency in PEI spheres creation, which means no residues of aqueous PEI and oil solvents remained in the reaction flask after the nanosphere's creation. The immobilized PEI spheres have sizes varied from 50 to 500 nm. The PEI spheres were successfully filled either with organic solvent (hydrophobic) or with water (hydrophilic). This method provides us the perspective for future encapsulation of varies molecules which have hydrophobic or hydrophilic nature

Sonochemical coating of cotton and polyester fabrics with “Antibacterial” BSA and casein spheres

A novel antibacterial coating for cotton and polyester fabrics has been developed by using drug-loaded proteinaceous microspheres made of bovine serum albumin and casein proteins. The microbubbles were created and anchored onto the fabrics (see figure) in a one-step reaction that lasts 3 min. The sonochemically produced “antibacterial fabrics” have been characterized. The efficiency of the sonochemical process in converting the native proteins into microspheres, encapsulating the drug, and coating the fabric has also been studied.This research was carried out as part of the activities of the LIDWINE Consortium, Contract No. NMP2-CT-2006-026741. LIDWINE is an IP Project of the 6th European Customers (EC) Program

Nanoscale spatially resolved infrared spectra from single microdroplets

Droplet microfluidics has emerged as a powerful platform allowing a large number of individual reactions to be carried out in spatially distinct microcompartments. Due to their small size, however, the spectroscopic characterisation of species encapsulated in such systems remains challenging. In this paper, we demonstrate the acquisition of infrared spectra from single microdroplets containing aggregation-prone proteins. To this effect, droplets are generated in a microfluidic flow-focussing device and subsequently deposited in a square array onto a ZnSe prism using a micro stamp. After drying, the solutes present in the droplets are illuminated locally by an infrared laser through the prism, and their thermal expansion upon absorption of infrared radiation is measured with an atomic force microscopy tip, granting nanoscale resolution. Using this approach, we resolve structural differences in the amide bands of the spectra of monomeric and aggregated lysozyme from single microdroplets with picolitre volume.Comment: 5 pages, 3 Figure

Protein micro- and nano-capsules for biomedical applications

Micro- and nano-scale systems have emerged as important tools for developing clinically useful drug delivery systems. In this tutorial review, we discuss the exploitation of biomacromolecules for this purpose, focusing on proteins, polypeptides, nucleic acids and polysaccharides and mixtures thereof as potential building blocks for novel drug delivery systems. We focus on the mechanisms of formation of micro- and nano-scale protein-based capsules and shells, as well as on the functionalization of such structures for use in targeted delivery of bioactive materials. We summarise existing methods for protein-based capsule synthesis and functionalization and highlight future challenges and opportunities for delivery strategies based on biomacromolecules.U.S. is grateful to Professor Aharon Gedanken, Chemistry Department, Bar-Ilan University, Israel, for his support and supervision during her PhD research work. G.J.L.B. is a Royal Society University Research Fellow at the Department of Chemistry, University of Cambridge and an Investigador FCT at the Instituto de Medicina Molecular, Lisboa. We thank Nuno Azoia assistance with the preparation of the figures. The authors thank the European Union Seventh Framework Programme (FP7/20072013) under grant agreement NMP4-LA-2009-228827 NANOFOL

Gene silencing by siRNA nanoparticles synthesized via sonochemical method

The knowledge that small RNAs can affect gene expression has had a tremendous impact on basic and applied research, and gene silencing is currently one of the most promising new approaches for disease therapy. However, RNAs cannot easily penetrate cell membranes, therefore RNA delivery become one of the major challenges for gene silencing technology. In the current paper we discuss a general approach for converting siRNA molecules into a dense siRNA nanoparticles using environmentally friendly sonochemical method. The RNA nanoparticulation enhance its gene-silencing activity in vascular bovine endothelial as well as in cancer 293T/GFP-Puro cell lines without causing any toxic effect. We show that ultrasonic waves do not lead to RNA degradation or any changes in its chemical structure. Moreover, sonochemically produced siRNA nanoparticles have been shown to be resistant to a variety of environmental stresses including pH levels, enzymes and temperatures, hence solving problem of the short half-life of the RNA molecules. As the siRNA nanoparticles are biocompatibile and biodegradabile, and their RNA release properties may be controlled within limits, sonochemical formation of siRNA nanoparticles represent a new promising approach for generation of functional bionano materials.(undefined

Proteinaceous microspheres for targeted RNA delivery prepared by an ultrasonic emulsification method

In the present work we used sonochemically prepared proteinaceous BSA spheres as a novel RNA-delivery system. The preparation of RNA-loaded BSA spheres was accomplished using an environmental friendly method termed the “ultrasonic emulsification method”. It was demonstrated that ultrasonic waves do not cause the RNA chains to degrade and the RNA molecules remain untouched. The BSA–RNA complex was successfully introduced into mammalian (human) U2OS osteosarcoma cells and Trypanosoma brucei parasites. Using PVA coating of the RNA–BSA spheres we have achieved a significant increase in the number of microspheres penetrating mammalian cells. The mechanism of RNA encapsulation and the structure of the RNA–BSA complex are reported.Ulyana Shimanovich thanks Ministry of Science and Technology, Israel for the "Woman in Science" scholarship (3-8219)

Folic acid-tagged protein nanoemulsions loaded with CORM-2 enhance the survival of mice bearing subcutaneous A20 lymphoma tumors

Folic Acid (FA)-tagged protein nanoemulsions were found to be preferentially internalized on B-cell lymphoma cell line (A20 cell line), which, for the first time, are reported to express folate receptor (FR)-alpha. Carbon monoxide releasing molecule-2 (CORM-2) was incorporated in the oil phase of the initial formulation. FA-functionalized nanoemulsions loaded with CORM-2 exhibited a considerable antitumor effect and an increased survival of BALB/c mice bearing subcutaneous A20 lymphoma tumors. The developed nanoemulsions also demonstrated to be well tolerated by these immunocompetent mice. Thus, the results obtained in this study demonstrate that FA-tagged protein nanoemulsions can be successfully used in cancer therapy, with the important ability to delivery drugs intracellularly.SFRH/BD/81479/2011 and SFRH/BD/81269/2011 scholarships from Fundação para a Ciência e a Tecnologia (FCT). This work has received funding from the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement NMP4-LA-2009-228827 NANOFOL. This work was supported by FEDER through POFC-COMPETE and by Portuguese funds from FCT through the project PEst-OE/BIA/UI4050/2014

Releasing dye encapsulated in proteinaceous microspheres on conductive fabrics by electric current

The current paper reports on the relase properties of conductive fabrics coated with proteinaceous microspheres containing a dye. The release of the dye was achieved by passing an electric current through the fabric. The conductivity of the polyester fibers resulted from nanosilver (Ag NPs) coated on the surface of these fibers. Both types of coatings (nanosilver coating and the coating of the proteinaceous microspheres) were performed using high-intensity ultrasonic waves. Two different types of dyes, hydrophilic RBBR (Remazol Brilliant Blue R) and hydrophobic ORO (Oil Red O), were encapsulated inside the microspheres (attached to the surface of polyester) and then released by applying an electric current. The Proteinaceous Microsphere (PM)-coated conductive fabrics could be used in medicine for drug release. The encapsulated dye can be replaced with a drug that could be released from the surface of fabrics by applying a low voltage

Design of novel BSA/hyaluronic acid nanodispersions for transdermal pharma purposes

A novel transdermal hyaluronic acid (HA) conjugated with bovine serum albumin (BSA) was developed in the form of solid-in-oil (S/O) nanodispersion (129.7 nm mean diameter). Ex vivo skin penetration analysis by fluorescence and confocal observation of histological skin sections revealed the ability of BSA/HA nanodispersions to cross the stratum corneum and penetrate into the dermis. Furthermore, no significant toxicity was found in fibroblast and keratinocyte cells in vitro. These results proved the potential of the developed nanodispersion for transdermal delivery of hyaluronic acid constituting a high value to biopharmaceutical and cosmetics industries.We thank Matadouro Central Carnes de Entre Douro e Minho, Lda, for their support on pig samples. The histological studies were supported by the Department of Histology from Life and Health Sciences Research Institute (ICVS), University of Minho. The confocal 3D images were performed by Department of Chemistry and Kanbar Laboratory for Nanomaterials Bar-Ilan University Center for Advanced Materials and Nanotechnology. C.S. would like to thank Fundacao para a Ciencia e a Tecnologia for their funding (SFRH/BPD/46515/2008). T.M. would like to thank Fundacao para a Ciencia e a Tecnologia for their funding (SFRH/BPD/47555/2008)