M. Soledad Fernández

Improving the brain delivery of gold nanoparticles by conjugation with an amphipathic peptide

BACKGROUND & AIMS Gold nanoparticles (GNPs) have promising applications for drug delivery as well as for the diagnosis and treatment of several pathologies, such as those related to the CNS. However, GNPs are retained in a number of organs, such as the liver and spleen. Owing to their negative charge and/or processes of opsonization, GNPs are retained by the reticuloendothelial system, thereby decreasing their delivery to the brain. It is therefore crucial to modify the nanoparticle surface in order to increase its lipophilicity and reduce its negative charge, thus achieving enhanced delivery to the brain. RESULTS In this article, we have shown that conjugation of 12 nm GNPs with the amphipathic peptide CLPFFD increases the in vivo penetration of these particles to the rat brain. The C(GNP)-LPFFD conjugates showed a smaller negative charge and a greater hydrophobic character than citrate-capped GNPs of the same size. We administered intraperitoneal injections of citrate GNPs and C(GNP)-LPFFD in rats, and determined the gold content in the tissues by neutron activation. Compared with citrate GNPs, the C(GNP)-LPFFD conjugate improved the delivery to the brain, increasing the concentration of gold by fourfold, while simultaneously reducing its retention by the spleen 1 and 2 h after injection. At 24 h, the conjugate was partially cleared from the brain, and mainly accumulated in the liver. The C(GNP)-LPFFD did not alter the integrity of the blood-brain barrier, and had no effect on cell viability.

Nervous system of the snail Helix aspersa

SummaryThe fine structure of vascular channels and amebocytes associated with the sheath of the infraesophageal ganglion of Helix aspersa, is described. The extracellular stroma of the sheath, together with the hemocoel and blood vessels, forms an interconnected system of pathways which appears to be involved in the transport of metabolites, amebocytes, hemocyanin and experimentally introduced opaque tracers. The hemocoel, blood capillaries and precapillaries are lined by a discontinuous layer of single muscle cells whose luminal aspect is covered by a lamina of extracellular material named the vascular coat. This coat consists of a ground substance that forms a basement membrane and filamentous elements some of which are collagenous. Gaps in the blood vessel wall seem to provide the main routes for the movement of cells and large molecules to the hemocoel. Tracer experiments have given support to the idea that a diffusion barrier may be absent at the sheath-ganglion junction. Amebocytes have phagocytic properties; they appear associated in groups or scattered singly within the extracellular space of the sheath and the lumen of blood vessels. Single amebocytes have features of mobile cells and may function in the transport of hemocyanin as well as other proteins.

Embedded proteins and sacrificial bonds provide the strong adhesive properties of gastroliths

The adhesive properties of gastroliths from a freshwater crayfish (Cherax quadricarinatus) were quantified by colloidal probe atomic force microscopy (AFM) between heavily demineralized gastrolith microparticles and gastrolith substrates of different composition. Combined AFM and transmission electron microscopy studies demonstrated that the sequential detachment and large adhesion energies that characterise the adhesive behaviour of a native gastrolith substrate are dominated by sacrificial bonds between chitin fibres and between chitin fibres and CaCO(3). The sacrificial bonds were shown to be strongly related to the gastrolith proteins and when the majority of these proteins were removed by ethylenediaminetetraacetic acid (EDTA), the sequential detachment disappeared and the adhesive energy was reduced by more than two orders of magnitude.

CALCIUM CARBONATE GROWTH MODIFICATION BY CONSTITUENTS RELEASES FROM POROUS CELLULOSE FILTER MEMBRANES

Filtration of a suspension of calcium carbonate (CaCO 3 ) or other inorganic solutions are often part of the methodology for recovering crystals during biomimetic mineralization experiments performed by various procedures. However, the use of cellulose filter membranes (FM) may cause a problem in in vitro crystallization experiments, because constituents released from the filters into the filtrates can alter the morphology of CaCO 3 crystals. Therefore, it is possible to misinterpret data obtained when the effect of specific additives tested in the investigation of biomineralization mechanisms. Herein, we present essential information to avoid such misinterpretations of data obtained from mineralization experiments. CaCO 3 was precipitated at room temperature by the gas diffusion method in the presence of FM as support and particularly as filtrates of calcium chloride (CaCl 2 ) obtained from various commercial FM. The occurrence or absence of morphological modifications of the calcite and vaterite crystals obtained with different FM correlates well with the different elemental compositions of the solutions where crystals are grown because of the constituents released from the filters into the filtrate. X-ray photoelectron spectroscopy (XPS) data indicate significant differences in the filter elemental composition. We assume that the observed chronological changes in CaCO 3 crystal modification could be due to incorporation into the calcite lattice of constituents released from the FM, most likely monomers, oligomers, or short-chain polymers.

Effect of Carbonic Anhydrase Immobilized on Eggshell Membranes on Calcium Carbonate Crystallization In Vitro

The eggshell membranes (ESM) serve as the first interface with the inorganic phase during eggshell formation. During mineral growth, crystals nucleate on the outer side of the ESM at specialized sites called mammillae, mainly consisting of mammillan, a keratan sulfate proteoglycan together with the activity of carbonic anhydrase (CA).