AFM study of morphology and mechanical properties of a chimeric 2 spider silk and bone sialoprotein protein for bone regeneration

Atomic 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 chimeric silk protein, 6merþBSP, and control films consisting of only the silk component (6mer). The behavior of the 6merþBSP and 6mer proteins in aqueous solution in the presence of calcium (Ca) ions was also assessed to determine interactions between the inorganic and organic components related to bone interactions, anchoring, and biomaterial network formation. The results demonstrated the formation of protein networks in the presence of Ca2þ ions, characteristics that may be important in the context of controlling materials assembly and properties related to bone formation with this new chimeric silk-BSP protein.Silvia Games thanks the Foundation for Science and Technology (FCT) for supporting her Ph.D. grant, SFRH/BD/28603/2006. This work was carried out under the scope of the European NoE EXPERTISSUES (NMP3-CT-2004-500283), the Chimera project (PTDC/EBB-EBI/109093/2008) funded by the FCT agency, the NIH (P41 EB002520) Tissue Engineering Resource Center, and the NIH (EB003210 and DE017207)

Solvent-Free Strategy Yields Size and Shape-Uniform Capsules

Capsules with a liquefied core were fabricated via the assembly of polymeric droplets induced by superamphiphobic surfaces. These highly repellent substrates exhibit distinct features such as (i) an easy and precise control over the particle size and shape, (ii) a high encapsulation efficiency, (iii) mild processing conditions, and (iv) the possibility to include any object in either a water or oil-based liquid core, which are not found on the current available strategies. As proof of concept, a photo-cross-linkable derivative of chitosan was used to produce the polymeric shell while a wealth variety of template cores were tested using a reversible cross-linking mechanism, interfacial gelation process or ice. Owing to the widespread application of polymeric capsules, the developed strategy is poised to usher the development of the next generation of materials not only for biomedical purposes but also for cosmetics, agriculture and electronics

Effect of Polyelectrolyte Multilayers Assembled on Ordered Nanostructures on Adhesion of Human Fibroblasts

Nanosphere lithography (NSL) and the layer-by-layer (LbL) technique are combined here for the first time to design a flexible system to achieve nanotopographical control of cell adhesion. NSL is used to generate regular patterns of tetrahedral gold nanodots of different size and distance. Besides the change in topography, LbL is used to generate a polyelectrolyte multilayer (PEM) system consisting of heparin (HEP) and poly­(ethylene imine) (PEI) on top of the gold dots. The localized formation of PEM on gold dots is achieved by prior passivation of the surrounding silicon or glass surface. Properties of PEM are changed by adjusting the pH value of HEP solution to either acidic or alkaline values. Studies with human dermal fibroblasts (HDF) reveal that cells spread to a higher extent on PEM formed at pH 5.0 in dependence on the structure dimension. Further, filopodia formation is highly increased in cells on nanostructures exhibiting HEP as a terminal layer. The new system offers a great potential to guide stem cell differentiation in the future owing to its high degree of chemical and topographical heterogeneity

Magnetically Multilayer Polysaccharide Membranes for Biomedical Applications

Self-standing nanocomposite films based on biopolymers and functional nanostructures have been widely used due to their potential applications as active elements in biomedical devices. The coupling between chitosan (CHI) and alginate (ALG) multilayered films and magnetic nanoparticles (MNPs) allowed to fabricate magnetic responsive freestanding membranes with a high structural control along the thickness, using the layer-by-layer (LbL) methodology. The mechanical characterization evidenced a trend for an increase of both Young modulus, and ultimate tensile strength with the inclusion of MNPs, or by cross-linking with genipin. Additionally, the multilayered membranes exhibited shape memory properties triggered by hydration. The in vitro biological performance studies showed that cells were more viable and adherent with higher proliferation rates when MNPs were included in the membranes. Our results suggested the potential of the developed magneto-active freestanding membranes for biomedical applications, such as in tissue engineering and biomedical applications

Magnetically Multilayer Polysaccharide Membranes for Biomedical Applications

Self-standing nanocomposite films based on biopolymers and functional nanostructures have been widely used due to their potential applications as active elements in biomedical devices. The coupling between chitosan (CHI) and alginate (ALG) multilayered films and magnetic nanoparticles (MNPs) allowed to fabricate magnetic responsive freestanding membranes with a high structural control along the thickness, using the layer-by-layer (LbL) methodology. The mechanical characterization evidenced a trend for an increase of both Young modulus, and ultimate tensile strength with the inclusion of MNPs, or by cross-linking with genipin. Additionally, the multilayered membranes exhibited shape memory properties triggered by hydration. The in vitro biological performance studies showed that cells were more viable and adherent with higher proliferation rates when MNPs were included in the membranes. Our results suggested the potential of the developed magneto-active freestanding membranes for biomedical applications, such as in tissue engineering and biomedical applications

Multilayered Hierarchical Capsules Providing Cell Adhesion Sites

Liquified capsules featuring (i) an external shell by layer-by-layer assembly of poly­(l-lysine), alginate, and chitosan, and encapsulating (ii) surface functionalized poly­(l-lactic acid) (PLLA) microparticles were developed. We hypothesize that, while the liquified environment enhances the diffusion of essential molecules for cell survival, microparticles dispersed in the liquified core of capsules provide the physical support required for cellular functions of anchorage-dependent cells. The influence of the incorporation of PLL on the regime growth, thickness, and stability was analyzed. Results show a more resistant and thicker film with an exponential build-up growth regime. Moreover, capsules ability to support cell survival was assessed. Capsules containing microparticles revealed an enhanced biological outcome in cell metabolic activity and proliferation, suggesting their potential to boost the development of innovative biomaterial designs for bioencapsulation systems and tissue engineering products

Multilayered Hierarchical Capsules Providing Cell Adhesion Sites

Liquified capsules featuring (i) an external shell by layer-by-layer assembly of poly­(l-lysine), alginate, and chitosan, and encapsulating (ii) surface functionalized poly­(l-lactic acid) (PLLA) microparticles were developed. We hypothesize that, while the liquified environment enhances the diffusion of essential molecules for cell survival, microparticles dispersed in the liquified core of capsules provide the physical support required for cellular functions of anchorage-dependent cells. The influence of the incorporation of PLL on the regime growth, thickness, and stability was analyzed. Results show a more resistant and thicker film with an exponential build-up growth regime. Moreover, capsules ability to support cell survival was assessed. Capsules containing microparticles revealed an enhanced biological outcome in cell metabolic activity and proliferation, suggesting their potential to boost the development of innovative biomaterial designs for bioencapsulation systems and tissue engineering products

Sequentially Moldable and Bondable Four-Dimensional Hydrogels Compatible with Cell Encapsulation

Hydrogels have captivated the attention of several research and industry segments, including bioengineering, tissue engineering, implantable/wearable sensors and actuators, bioactive agent delivery, food processing, and industrial processes optimization. A common limitation of these systems is their fixed shape. The concept of hydrogel moldability is often assigned to the injectability potential of liquid precursors, and this feature is often lost right after hydrogel formation. Hydrogel modulation is a recent trend that advocates the importance of designing materials with shape fitting ability targeting on-demand responses or defect filling purposes. Here, we present a compliant and cell encapsulation-compatible hydrogel prepared from unmodified natural origin polymers with the ability to undergo extreme sequential shape alterations with high recovery of its mechanical properties. Different fragments of these hydrogels could be bonded together in spatiotemporally controlled shape- and formulation-morphing structures. This material is prepared with affordable off-the-shelf polysaccharides of natural origin using a mild and safe processing strategy based solely on polyelectrolyte complexation followed by an innovative partial coacervate compaction and dehydration step. These unique hydrogels hold potential for multifield industrial and healthcare applications. In particular, they may find application as defect filling agents or highly compliant wound healing patches for cargo release and/or cell delivery for tissue regeneration and cell-based therapies

Sequentially Moldable and Bondable Four-Dimensional Hydrogels Compatible with Cell Encapsulation

Hydrogels have captivated the attention of several research and industry segments, including bioengineering, tissue engineering, implantable/wearable sensors and actuators, bioactive agent delivery, food processing, and industrial processes optimization. A common limitation of these systems is their fixed shape. The concept of hydrogel moldability is often assigned to the injectability potential of liquid precursors, and this feature is often lost right after hydrogel formation. Hydrogel modulation is a recent trend that advocates the importance of designing materials with shape fitting ability targeting on-demand responses or defect filling purposes. Here, we present a compliant and cell encapsulation-compatible hydrogel prepared from unmodified natural origin polymers with the ability to undergo extreme sequential shape alterations with high recovery of its mechanical properties. Different fragments of these hydrogels could be bonded together in spatiotemporally controlled shape- and formulation-morphing structures. This material is prepared with affordable off-the-shelf polysaccharides of natural origin using a mild and safe processing strategy based solely on polyelectrolyte complexation followed by an innovative partial coacervate compaction and dehydration step. These unique hydrogels hold potential for multifield industrial and healthcare applications. In particular, they may find application as defect filling agents or highly compliant wound healing patches for cargo release and/or cell delivery for tissue regeneration and cell-based therapies

Singularity of quantitative research: from collecting information to producing results

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