The binding of CNA35 contrast agents to collagen fibrils.

CryoTEM demonstrates that a CNA35-bearing liposomal MRI contrast agent selectively binds to poorly assembled collagen type I as opposed to well-assembled collagen fibrils, whereas monomeric CNA35 binds to all forms of collagen. It is shown that upon conjugation to liposomes and micelles CNA35 loses its ability to dissociate ordered collagen fibrils and thereby to create its own binding sites.

The development of morphology and structure in hexagonal vaterite.

Inspired by the remarkable shapes and properties of CaCO(3) biominerals, many studies have investigated biomimetic routes aiming at synthetic equivalents with similar morphological and structural complexity. Control over the morphology of CaCO(3) crystals has been demonstrated, among other methods, by the use of additives that selectively allow the development of specific crystal faces, while inhibiting others. Both for biogenic and biomimetic CaCO(3), the crystalline state is often preceded by an amorphous precursor phase, but still limited information is available on the details of the amorphous-to-crystalline transition. By using a combination of cryoTEM techniques (bright field imaging, cryo-tomography, low dose electron diffraction and cryo-darkfield imaging), we show for the first time the details of this transition during the formation of hexagonal vaterite crystals grown in the presence of NH(4)(+) ions. The formation of hexagonal plate-like vaterite occurs via an amorphous precursor phase. This amorphous phase converts into the crystalline state through a solid state transformation in which order and morphology develop simultaneously. The mineral initially develops as polycrystalline vaterite which transforms into a single crystal directed by an NH(4)(+)-induced crystal plane that acts as a templating surface.

The initial stages of template-controlled CaCO3 formation revealed by cryo-TEM.

Biogenic calcium carbonate forms the inorganic component of seashells, otoliths, and many marine skeletons, and its formation is directed by an ordered template of macromolecules. Classical nucleation theory considers crystal formation to occur from a critical nucleus formed by the assembly of ions from solution. Using cryotransmission electron microscopy, we found that template-directed calcium carbonate formation starts with the formation of prenucleation clusters. Their aggregation leads to the nucleation of amorphous nanoparticles in solution. These nanoparticles assemble at the template and, after reaching a critical size, develop dynamic crystalline domains, one of which is selectively stabilized by the template. Our findings have implications for template-directed mineral formation in biological as well as in synthetic systems.

Kinetics of avidin-induced clearance of biotinylated bimodal liposomes for improved MR molecular imaging.

Dual labeled liposomes, carrying both paramagnetic and fluorescent lipids, were recently proposed as potent contrast agents for MR molecular imaging. These nanoparticles are coated with poly(ethylene glycol) (PEG) to increase their blood circulation half-life, which should allow extensive accumulation at the targeted site. To eliminate nonspecific blood pool signal from the MR images, the circulating liposomes should ideally be cleared from the circulation when sufficient target-specific contrast enhancement is obtained. To that aim, we designed an avidin chase that allowed controlled and rapid clearance of paramagnetic biotinylated liposomes from the blood circulation in C57BL/6 mice. Avidin-induced alterations in blood clearance kinetics and tissue distribution were studied quantitatively by determination of the Gd content in blood and tissue samples ex vivo. Intrinsic liposomal blood clearance showed bi-exponential behavior with half-lives t(1/2alpha) = 2.1 +/- 1.1 and t(1/2beta) = 15.1 +/- 5.4 hours, respectively. In contrast, the contrast agent was cleared from the blood by the avidin infusion to <1% of the initial dose within 4 hours. Avidin-induced liposomal blood clearance was also demonstrated in vivo by dynamic T(1)-weighted MRI. The ability to rapidly clear circulating contrast agents opens up exciting possibilities to study targeting kinetics, to increase the specificity of molecular MRI and to optimize nanoparticulate contrast agent formulations.

Noncovalent triblock copolymers based on a coiled-coil peptide motif.

The formation of a noncovalent triblock copolymer based on a coiled-coil peptide motif is demonstrated in solution. A specific peptide pair (E and K) able to assemble into heterocoiled coils was chosen as the middle block of the polymer and conjugated to poly(ethylene glycol) (PEG) and polystyrene (PS) as the outer blocks. Mixing equimolar amounts of the polymer-peptide block copolymers PS-E and K-PEG resulted in the formation of coiled-coil complexes between the peptides and subsequently in the formation of the amphiphilic triblock copolymer PS-E/K-PEG. Aqueous self-assembly of the separate peptides (E and K), the block copolymers (PS-E and K-PEG), and equimolar mixtures thereof was studied by circular dichroism, dynamic light scattering, and cryogenic transmission electron microscopy. It was found that the noncovalent PS-E/K-PEG copolymer assembled into rodlike micelles, while in all other cases, spherical micelles were observed. Temperature-dependent studies revealed the reversible nature of the coiled-coil complex and the influence of this on the morphology of the aggregate. A possible mechanism for these transitions based on the interfacial free energy and the free energy of the hydrophobic blocks is discussed. The self-assembly of the polymer-peptide conjugates is compared to that of polystyrene-b-poly(ethylene glycol), emphasizing the importance of the coiled-coil peptide block in determining micellar structure and dynamic behavior.