Search

The membranolytic activity of silica particles toward red blood cells (RBCs) has been known for a long time and is sometimes associated with silica pathogenicity. However, the molecular mechanism and the reasons why hemolysis differs according to the silica form are still obscure. A panel of 15 crystalline (pure and commercial) and amorphous (pyrogenic, precipitated from aqueous solutions, vitreous) silica samples differing in size, origin, morphology, and surface chemical composition were selected and specifically prepared. Silica particles were grouped into six groups to compare their potential in disrupting RBC membranes so that one single property differed in each group, while other features were constant. Free radical production and crystallinity were not strict determinants of hemolytic activity. Particle curvature and morphology modulated the hemolytic effect, but silanols and siloxane bridges at the surface were the main actors. Hemolysis was unrelated to the overall concentration of silanols as fully rehydrated surfaces (such as those obtained from aqueous solution) were inert, and one pyrogenic silica also lost its membranolytic potential upon progressive dehydration. Overall results are consistent with a model whereby hemolysis is determined by a defined surface distribution of dissociated/undissociated silanols and siloxane groups strongly interacting with specific epitopes on the RBC membrane.

Amorphous silicas, opposite to crystalline polymorphs, have been regarded so far as nonpathogenic, but few studies have addressed the toxicity of the wide array of amorphous silica forms. With the advent of nanotoxicology, there has been a rising concern about the safety of silica nanoparticles to be used in nanomedicine. Here, we report a study on the toxicity of amorphous nanostructured silicas obtained with two different preparation procedures (pyrolysis vs. precipitation), the pyrogenic in two very different particle sizes, in order to assess the role of size and origin on surface properties and on the cell damage, oxidative stress, and inflammatory response elicited in murine alveolar macrophages. A quartz dust was employed as positive control and monodispersed silica spheres as negative control. Pyrogenic silicas were remarkably more active than the precipitated one as to cytotoxicity, reactive oxygen species production, lipid peroxidation, nitric oxide synthesis, and production of tumor necrosis factor-α, when compared both per mass and per unit surface. Between the two pyrogenic silicas, the larger one was the more active. Silanols density is the major difference in surface composition among the three silicas, being much larger than the precipitated one as indicated by joint calorimetric and infrared spectroscopy analysis. We assume here that full hydroxylation of a silica surface, with consequent stable coverage by water molecules, reduces/inhibits toxic behavior. The preparation route appears thus determinant in yielding potentially toxic materials, although the smallest size does not always correspond to an increased toxicity.

Siliceous sponges, one of the few animal groups involved in a biosilicification process, deposit hydrated silica in discrete skeletal elements called spicules. A multidisciplinary analysis of the structural features of the protein axial filaments inside the spicules of a number of marine sponges, belonging to two different classes (Demospongiae and Hexactinellida), is presented, together with a preliminary analysis of the biosilicification process. The study was carried out by a unique combination of techniques: fiber diffraction using synchrotron radiation, scanning electron microscopy (SEM), thermogravimetric analysis (TGA), differential scanning calorimetric (DSC), Fourier transform infrared spectroscopy (FTIR), and molecular modeling. From a phylogenetic point of view, the main result is the structural difference between the dimension and packing of the protein units in the spicule filaments of the Demospongiae and the Hexactinellida species. Models of the protein organization in the spicule axial filaments, consistent with the various experimental evidences, are given. The three different species of demosponges analyzed have similar general structural features, but they differ in the degree of order. The structural information on the spicule axial filaments can help shed some light on the still unknown molecular mechanisms controlling biosilicification.

The effect of grinding, heating, and etching was investigated on polymorphs of silicon dioxide exhibiting different biological responses. Diatomaceous earths were converted into cristobalite at 1000 degrees C. Dusts obtained by grinding crystalline minerals exhibited different micromorphology and a propensity to originate surface radicals which decrease in the sequence cristobalite --> quartz --> coesite --> stishovite. The production of surface radicals was suppressed by grinding in the presence of water. Thermal treatments selectively quenched the radicals and decreased surface hydrophilicity. Quartz treated with aluminum lactate exhibited higher surface acidity when compared with pure quartz, with a reduction in fibrogenicity. Etching by hydrofluoric acid smoothed the particles with loss of specific surface. Adsorption of water on three cristobalite dusts of different origin (ground mineral, ex-diatomite, heated quartz) indicated a loss in heated quartz (1300 degrees C) that was relatable to the corresponding reduction in fibrogenicity.

The hemolytic reaction to a dust is often used as a potential indicator of fibrogenicity of silicon dioxide polymorphs. However, occasionally the hemolytic response may not correlate with the observed fibrotic response in vivo. For example, amorphous silicas are very hemolytic but have little or no fibrogenic activity. In our study, heat treatment was used to alter alpha-cristobalite, a known fibrogenic dust, to a more hydrophobic surface. Comparisons were made between heated and unheated alpha-cristobalite for hemolytic activity in vitro and for lung response in vivo. Heat treatment resulted in decreased hemolytic response, but no change in the fibrotic response occurred in vivo. In addition, the heat treatment resulted in increased initial dust accumulation, reduced short-term clearance, and enhanced long-term clearance in vivo. Increased inflammatory cell recruitment was also observed in lungs of animals exposed to alpha-cristobalite. Thus, whereas heat-induced surface changes in alpha-cristobalite markedly altered the hemolytic activity of the particles, no changes were observed in the fibrotic response.

Reactive radicals at the surface of quartz or other SiO2 polymorphs have been studied by EPR in relation to their possible role in pathogenicity. All the examined dusts bear the characteristic radicals of silica ground in air: Si, SiO., SiO.2 (peroxyradical) and O2.- (superoxide ion), but some also show additional spectral lines belonging to other radical forms. Comparison of standard quartz dusts (DQ-12, Min-u-sil 5) with a natural quartz and with what obtained by grinding a very pure quartz crystal indicates that to a higher purity corresponds a higher radical population. Cristobalite and vitreous silica exhibit similar spectra, with larger proportion by respect to quartz, of partially reduced oxygen forms. The reactivity of the silica surface towards O2 and NaClO aqueous solutions are investigated by examining the modification in the EPR spectra induced by these treatments. A possible mechanism for fibrogenicity is proposed whereby, within the activated macrophage, a catalytic reaction occurs between surface functionalities and macrophage oxygen metabolites. This reaction would trigger the abnormal production of fibroblast stimulating factors, ending up with silicosis.