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Natural and genetically engineered proteins for tissue engineering
Gomes, Sílvia C.; Leonor, I. B.; Mano, J. F.; Reis, R. L.; Kaplan, DavidTo overcome the limitations of traditionally used autografts, allografts and, to a lesser extent, synthetic materials, there is the need to develop a new generation of scaffolds with adequate mechanical and structural support, control of cell attachment, migration, proliferation and differentiation and with bio-resorbable features. This suite of properties would allow the body to heal itself at the same rate as...
Biological responses to spider silk - antibiotic fusion protein
Gomes, Sílvia C.; Gallego-Llamas, Jabier; Leonor, I. B.; Mano, J. F.; Reis, R. L.; Kaplan, DavidThe development of a new generation of multifunctional biomaterials is a continual goal for the field of materials science. The in vivo functional behaviour of a new fusion protein that combines the mechanical properties of spider silk with the antimicrobial properties of hepcidin was addressed in this study. This new chimeric protein, termed 6mer + hepcidin, fuses spider dragline consensus sequences (6mer) and...
Aligned silk-based 3-D architectures for contact guidance in tissue engineering
Oliveira, A. L.; Sun, L.; Kim, H. J.; Rice, W.; Kluge, J.; Reis, R. L.; Kaplan, DavidAn important challenge in the biomaterials field is to mimic the structure of functional tissues via cell and extracellular matrix (ECM) alignment and anisotropy. Toward this goal, silk-based scaffolds resembling bone lamellar structure were developed using a freeze-drying technique. The structure could be controlled directly by solute concentration and freezing parameters, resulting in lamellar scaffolds with ...
Development of silk-based scaffolds for tissue engineering of bone from human a...
Correia, Cristina; Bhumiratana, Sarindr; Yan, Leping; Oliveira, A. L.; Gimble, Jeffrey M.; Rockwood, Danielle; Kaplan, David; Sousa, R. A.; Reis, R. L.Silk fibroin is a potent alternative to other biodegradable biopolymers for bone tissue engineering (TE), because of its tunable architecture and mechanical properties, and its demonstrated ability to support bone formation both in vitro and in vivo. In this study, we investigated a range of silk scaffolds for bone TE using human adipose-derived stem cells (hASCs), an attractive cell source for engineering auto...
Spider silk-bone sialoprotein fusion proteins for bone tissue engineering
Gomes, Sílvia C.; Leonor, I. B.; Mano, J. F.; Reis, R. L.; Kaplan, DavidThe remarkable mechanical characteristics of the spider silk protein major ampullate spidroin protein suggest this polymer as a promising biomaterial to consider for the fabrication of scaffolds for bone regeneration. Herein, a new functionalized spider silk-bone sialoprotein fusion protein was designed, cloned, expressed, purified and the osteogenic activity studied. Bone sialoprotein (BSP) is a multidomain pr...
Antimicrobial functionalized genetically engineered spider silk
Gomes, Sílvia C.; Leonor, I. B.; Mano, J. F.; Reis, R. L.; Kaplan, DavidGenetically engineered fusion proteins offer potential as multifunctional biomaterials for medical use. Fusion or chimeric proteins can be formed using recombinant DNA technology by combining nucleotide sequences encoding different peptides or proteins that are otherwise not found together in nature. In the present study, three new fusion proteins were designed, cloned and expressed and assessed for function, b...
AFM study of morphology and mechanical properties of a chimeric 2 spider silk a...
Gomes, Sílvia C.; Numata, Keiji; Leonor, I. B.; Mano, J. F.; Reis, R. L.; Kaplan, DavidAtomic force microscopy (AFM) was used to assess a new chimeric protein consisting of a fusion protein of the consensus repeat for Nephila clavipes spider dragline protein and bone sialoprotein (6merþBSP). The elastic modulus of this protein in film form was assessed through force curves, and film surface roughness was also determined. The results showed a significant difference among the elastic modulus of the...
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