Bacteria metabolic adaptation to oxidative stress: the case of silica.

Although the presence of silica in many living organisms offers advanced properties including cell protection, the different in vitro attempts to build living materials in pure silica never favoured the cells viability. Thus, little attention has been paid to host-guest interactions to modify the expected biologic response. Here we report the physiological changes undergone by Escherichia coli K-12 in silica from colloidal solution to gel confinement. We show that the physiological alterations in growing cultures are not triggered by the initial oxidative Reactive Oxygen Species (ROS) response. Silica promotes the induction of alternative metabolic pathways along with an increase of growth suggesting the existence of rpoS polymorphisms. Since the functionality of hybrid materials depends on the specific biologic responses of their guests, such cell physiological adaptation opens perspectives in the design of bioactive devices attracting for a large field of sciences.

A multiscale study of bacterial proliferation modes within novel E. coli@Si(HIPE) hybrid macrocellular living foams.

For the first time the study at various length scales of E. coli proliferation modes within Si(HIPE) inorganic macrocellular foams is proposed. Both qualitatively and semi-quantitatively, bacterial proliferation within the foam is not homogeneous and is directly linked to the random distribution of Si(HIPE) macroscopic cells. When inoculated in a nutrient loaded Si(HIPE), the bacterial growth is enhanced within the Si(HIPE) matrices compared to the surrounding LB media. The bacterial growth kinetics tends to be faster and the concentration at saturation is roughly 100% times higher. In the case of a Si(HIPE) host free of nutrients, bacterial motion occurs as an infiltration wave; the peak of this propagation wave moves at a constant speed of 88 µm h-1, while bacterial concentrations within the Si(HIPE) host reach levels far above the ones reached in the presence of nutrients, suggesting a real synergetic relationship between the bacterial colony guests and the Si(HIPE) host. When a nutrient reservoir is present at the opposite position from which bacteria were inoculated, bacterial proliferation is associated with a coalescence process between the growing colonies that were rapidly established within the first hours. When the Si(HIPE) host was fully colonized we found out a specific distance between adjacent colonies of 5 and 15 µm in good correspondence with the repartition of the wall to wall distances of the Si(HIPE) macroscopic cells, meaning that bacterial repartition once colonization occurs is optimum. These results show that Si(HIPE) foams are outstanding candidates for strengthened bacterial proliferation without motion restriction imposed by conventional silica gels.

Silica@proton-alginate microreactors: a versatile platform for cell encapsulation.

As an alternative approach to the well known Ca(ii)-alginate encapsulation process within silica hydrogels, proton-driven alginate gelation was investigated in order to establish its capacity as a culture carrier, both isolated and embedded in an inorganic matrix. Control over the velocity of the proton-gelation front allows the formation of a hydrogel shell while the core remains liquid, allowing bacteria and microalgae to survive the strongly acidic encapsulation process. Once inside the inorganic host, synthesized by a sol-gel process, the capsules spontaneously redissolve without the aid of external complexing agents. The entrapped cells survive the two-step process to a significant extent; culture's growth restores the initial cell count in less than two weeks. Biosynthesis of Au nanoparticles mediated by the entrapped microalgae illustrates the preservation of the biosynthetic abilities supported by this platform.

In vitro studies and preliminary in vivo evaluation of silicified concentrated collagen hydrogels.

Hybrid and nanocomposite silica-collagen materials derived from concentrated collagen hydrogels were evaluated in vitro and in vivo to establish their potentialities for biological dressings. Silicification significantly improved the mechanical and thermal stability of the collagen network within the hybrid systems. Nanocomposites were found to favor the metabolic activity of immobilized human dermal fibroblasts while decreasing the hydrogel contraction. Cell adhesion experiments suggested that in vitro cell behavior was dictated by mechanical properties and surface structure of the scaffold. First-to-date in vivo implantation of bulk hydrogels in subcutaneous sites of rats was performed over the vascular inflammatory period. These materials were colonized and vascularized without inducing strong inflammatory response. These data raise reasonable hope for the future application of silica-collagen biomaterials as biological dressings.

Morphological evolution of (NH4)(0.5)V2O5·mH2O fibers into belts, triangles, and rings.

In this contribution, single-crystalline (NH(4))(0.5)V(2)O(5)·mH(2)O xerogels made of belts, rings, triangles, and ovals have been synthesized using a surfactant-free hydrothermal method. The analytical techniques of scanning electron microscopy (SEM), transmission electron microscopy (TEM), powder X-ray diffraction (PXRD), thermogravimetric analysis (TGA), energy dispersive X-ray spectroscopy (EDX), Fourier transform infrared (FTIR), high-resolution TEM (HRTEM), and selected area electron diffraction (SAED) have been used to characterize the morphology, composition, and structure of the as-prepared products. On the basis of SEM and TEM observations, we suggested that the as-prepared (NH(4))(0.5)V(2)O(5)·mH(2)O rings, triangles, and ovals have been formed by connecting two ends of the vanadium oxide sheet made of edge and corner sharing VO(5) square pyramids. The as-prepared (NH(4))(0.5)V(2)O(5)·mH(2)O nanobelts are up to several hundreds of micrometers long, 402-551 nm wide, and 235-305 nm thick. The thickness and width of the rings are respectively ∼454 nm and ∼1 µm. Triangles with three unequal sides having a thickness of ∼143 nm and a width of ∼1 µm were also formed. The crystalline orthorhombic phase of shcherbianite V(2)O(5) was obtained on calcination of (NH(4))(0.5)V(2)O(5)·mH(2)O at 350 °C for 2 h. The SEM image of this V(2)O(5) product retains the parent morphology of the preheated compound. A possible reaction mechanism and the growth process involved in the formation of belts/rings/triangles and ovallike microstructures are discussed.

Micro-algal biosensors.

Fighting against water pollution requires the ability to detect pollutants for example herbicides or heavy metals. Micro-algae that live in marine and fresh water offer a versatile solution for the construction of novel biosensors. These photosynthetic microorganisms are very sensitive to changes in their environment, enabling the detection of traces of pollutants. Three groups of micro-algae are described in this paper: chlorophyta, cyanobacteria, and diatoms.

From diatoms to silica-based biohybrids.

This critical review shows that diatoms can be a source of inspiration for the synthesis of advanced nanostructured biohybrids. These single cell microalgae are living inside a porous silica shell called 'frustule'. Mimicking this model, silica-based biohybrids have been produced via the so-called sol-gel process. Biomolecules such as proteins, enzymes or antibodies can be trapped within a silica matrix leading to hybrid biosensors and bioreactors. Whole cells remain viable and retain their metabolic activity leading to the formation of living biohybrids that offer new possibilities in the field of biotechnology and nanomedicine. Diatom frustules exhibit an incredible variety of sophisticated shapes; they can be used as 3D hierarchically structured materials for the realization of sensors, photonic devices or microfluidics. They can also be a model for the bio-templated synthesis of nanostructured materials. Diatom nanotechnology is becoming a new field of research where biologists and materials scientists are working together! (125 references).

Silica-collagen bionanocomposites as three-dimensional scaffolds for fibroblast immobilization.

Silica-collagen bionanocomposite hydrogels were obtained by addition of silica nanoparticles to a protein suspension followed by neutralization. Electron microscopy studies indicated that larger silica nanoparticles (80 nm) do not interact strongly with collagen, whereas smaller ones (12 nm) form rosaries along the protein fibers. However, the composite network structurally evolved with time due to the contraction of the cells and the dissolution of the silica nanoparticles. When compared to classical collagen hydrogels, these bionanocomposite materials showed lower surface contraction in the short term (1 week) and higher viability of entrapped cells in the long term (3 weeks). A low level of gelatinase MMP2 enzyme expression was also found after this period. Several proteins involved in the catabolic and anabolic activity of the cells could also be observed by immunodetection techniques. All these data suggest that the bionanocomposite matrices constitute a suitable environment for fibroblast adhesion, proliferation and biological activity and therefore constitute an original three-dimensional environment for in vitro cell culture and in vivo applications, in particular as biological dressings.

Hydrothermal Synthesis of Nanostructured Vanadium Oxides.

A wide range of vanadium oxides have been obtained via the hydrothermal treatment of aqueous V(V) solutions. They exhibit a large variety of nanostructures ranging from molecular clusters to 1D and 2D layered compounds. Nanotubes are obtained via a self-rolling process while amazing morphologies such as nano-spheres, nano-flowers and even nano-urchins are formed via the self-assembling of nano-particles. This paper provides some correlation between the molecular structure of precursors in the solution and the nanostructure of the solid phases obtained by hydrothermal treatment.

Photosynthetic microorganism-mediated synthesis of akaganeite (beta-FeOOH) nanorods.

Common Anabaena and Calothrix cyanobacteria and Klebsormidium green algae are shown to form intracellularly akaganeite beta-FeOOH nanorods of well-controlled size and unusual morphology at room temperature. X-ray diffraction (XRD), transmission electron microscopy (TEM), and scanning electron microscopy X-ray energy dispersive spectrometry (SEM-EDS) analyses are used to investigate particle structure, size, and morphology. A mechanism involving iron-siderophore complex formation is proposed and compared with iron biomineralization in magnetotactic bacteria.

Influence of decavanadate clusters on the rheological properties of gelatin.

The influence of polyoxovanadate clusters ([H(2)V(10)O(28)](4-)) on the thermo-reversible gelation of porcine skin gelatin solution (type A, M w approximately 40 000 g.mol (-1), pH = 3.4 << isoelectric point (IEP) approximately 8) has been investigated as a function of temperature and vanadate concentration by combining rheology and microcalorimetry. This work shows that the rheological properties of the system depend on electrostatic interactions between [H(2)V(10)O(28)](4-) and positively charged gelatin chains. In a first stage, we describe the renaturation of the gelatin triple helices in the presence of decavanadate clusters. We reveal that, when gelatin chains are in coil conformation (30 degrees C < T < 50 degrees C), the inorganic clusters act as physical cross-linkers that govern the visco-elastic properties of the mixture with an exponential dependence of the (G', G'') modulus with the vanadate concentration. Below 30 degrees C, we show that gelatin triple helix nucleation is slightly favored by the presence of vanadate, but above a helix concentration of 0.012 g.cm (-3), G' is fully governed by the helix concentration. During the melting process, we reveal the non-fully reversible behavior of the vanadate/gelatin rheological properties and the stabilization of gelatin triple helices due to vanadate species until 50 degrees C. This non-reversible character has also been observed in the same experimental conditions with collagen/vanadate solutions. This is the first time that such a stabilization of triple helices has been reported in the case of gelatin hydrogels chemically cross-linked or not. We propose to analyze these results by considering that triple helix aggregates should persist because of decavanadate bridging, that the nucleation of an extended triple helix network may induce a strong modification of the vanadate cross-linker distribution in the system, or both, thus promoting the formation of thermally stable vanadate/gelatin micro-gels in the dangling end of the triple helices.

Biomimetic dual templating of silica by polysaccharide/protein assemblies.

The control of silica growth by living organisms such as diatoms is known to involve the templating effect of several biomolecules working concomitantly. However, until now, biomimetic studies involving model molecules have mainly been performed with single templates. We show here that the addition of two biopolymers, gelatin and alginic acid, to silicate solutions allows the formation of complex structures resulting from the combined templating effect of both components at different scales. Gelatin is able to activate silica formation resulting in hybrid aggregates at the nanoscale. Alginic acid does not interfere with silica condensation but is able to control silica morphology through the assembly of these gelatin-silica aggregates at the microscale. For all materials, calcination up to 700 degrees C degrades the polymer component of the hybrid material and opens macroporosity in the silica network. In parallel, the high thermal stability of gelatin allows a good preservation of initial silica nanoparticle size upon heating whereas a coarsening process is observed in the sole presence of alginate. These results correlate well with previous models of biosilicification and suggest that the use of multiple templates is a suitable approach to elaborate more complex silica architectures.

Contribution of multi-nuclear solid state NMR to the characterization of the Thalassiosira pseudonana diatom cell wall.

A major issue in the study of biosilicification processes is the harsh chemical conditions required for silica dissolution, which often lead to denaturation of the associated bio-organic matter. In order to demonstrate the potential of solid state NMR for investigating silicified materials of natural origin, this technique was applied to isotopically enriched Thalassiosira pseudonana diatom cells. (29)Si, (1)H,(31)P, (13)C and (15)N solid state NMR studies were performed on whole cells, SDS-extracted and H(2)O(2)-cleaned silica shells. Cross-polarization techniques were useful for identifying the presence of mobile and rigid molecules, allowing loosely bound and silica-entrapped species to be discriminated. Successive cleaning procedures efficiently eliminated weakly associated organic matter. The H(2)O(2)-cleaned silica shell still contained carbohydrates (mainly chitin) and proteins as well as lipids. This suggests that the role of lipids in diatom shell formation may have been underestimated so far, demonstrating the potential of solid state NMR for studying composite biomaterials.

First example of biopolymer-polyoxometalate complex coacervation in gelatin- mixtures.

The first example of complex coacervation between a biopolymer and polyoxometalate clusters is identified in the gelatin-decavanadate system.

Sol-gel encapsulation of cells is not limited to silica: long-term viability of bacteria in alumina matrices.

A colloidal route to aqueous alumina gels is described, allowing the long-term viability of encapsulated bacteria.

Aqueous silicates in biological sol-gel applications: new perspectives for old precursors.

Identification of silica sol-gel chemistry with silicon alkoxide hydrolysis and condensation processes is common in modern materials science. However, aqueous silicates exhibit several features indicating that they may be more suitable precursors for specific fields of research and applications related to biology and medicine. In this Account, we illustrate the potentialities of such aqueous precursors for biomimetic studies, bio-hybrid material design, and bioencapsulation routes. We emphasize that the natural relevance, the biocompatibility, and the low ecological impact of silicate chemistry may balance its lack of diversity, flexibility, and processability.

Cyanobacteria as bioreactors for the synthesis of Au, Ag, Pd, and Pt nanoparticles via an enzyme-mediated route.

Common Anabaena, Calothrix, and Leptolyngbya cyanobacteria are shown to form Au, Ag, Pd, and Pt nanoparticles of well-controlled size. These nanoparticles are synthesized intra-cellularly, and naturally released in the culture medium, where they are stabilized by algal polysaccharides, allowing their easy recovery. The size of the recovered particles as well as the reaction yield is shown to depend on the cyanobacteria genus. Investigations of nanoparticle formation indicate that the intracellular nitrogenase enzyme is responsible for the metal reduction but that the cellular environment is involved in the colloid growth process.

Potentialities of silica/alginate nanoparticles as hybrid magnetic carriers.

The possibility to associate traditional bio-organic capsules, such as polymer nanoparticles or liposomes, with silica has been recently demonstrated, opening the route to the design of novel nanocomposites that exhibit promising properties as drug carriers. In this context, we describe here the elaboration of silica/alginate nanoparticles incorporating magnetic iron oxide colloids and fluorescent carboxy-fluoroscein. These nanocomposites were characterized by electron microscopy, X-ray diffraction and magnetic measurements. The release of the fluorophore was investigated in vitro and was demonstrated to occur in 3T3 fibroblast cells. Further grafting of organic moieties on particle surface is also described. These data suggest that hybrid nanoparticles are flexible platforms for the developments of multi-functional bio-capsules.

Influence of poly-L-lysine on the biomimetic growth of silica tubes in confined media.

Pore channels of polycarbonate membranes were recently used as biomimetic models to study the effect of confinement on silicate condensation, leading to the formation of silica tubes exhibiting a core-shell structure. In this work, we preimmobilized poly-L-lysine on the membrane pores, leading to modification of the tube shell formation process and variation in core particle size. These data strengthen previous assumptions on the role of confinement on silica growth, i.e., interfacial interactions and perturbation of the diffusion coefficient. They also suggest that this approach is suitable to investigate in more detail the contribution of confinement effects on silica biomineralization.

Design of iron oxide/silica/alginate hybrid magnetic carriers (HYMAC).

A large number of natural and synthetic polymers have already been evaluated for the design of nanomaterials incorporating magnetic nanoparticles for biomedical applications. The possibility to use hybrid (bio)-organic/inorganic nano-carriers have been much less studied. Here we describe the design of Hybrid MAgnetic Carriers (HYMAC) consisting of alginate/silica nanocomposites incorporating magnetite nanoparticles, based on a spray-drying approach. Transmission electron microscopy and X-ray energy dispersive spectrometry confirm the successful incorporation of magnetic colloids within homogeneous hybrid capsules. X-ray diffraction data suggest that surface iron ions are partially desorbed by the spray-drying process, leading to the formation of lepidocrocite and of an iron silicate phase. Magnetic measurements show that the resulting nanocomposites exhibit a superparamagnetic behaviour with a blocking temperature close to 225 K. Comparison with un-silicified capsules indicate that the mineral phase enhances the thermal stability of the polymer network and do not modify of the amount of incorporated iron oxide nanoparticles. Moreover, evaluation of nanocomposite up-take by fibroblasts indicates their possible internalization. A selective intracellular alginate degradation is observed, suggesting that these HYMAC nanomaterials may exhibit interesting properties for the design of drug delivery devices.