Mara Ghiazza

In search of the chemical basis of the hemolytic potential of silicas.

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.

Hematite Nanoparticles Larger than 90 nm Show No Sign of Toxicity in Terms of Lactate Dehydrogenase Release, Nitric Oxide Generation, Apoptosis, and Comet Assay in Murine Alveolar Macrophages and Human Lung Epithelial Cells

Three hematite samples were synthesized by precipitation from a FeCl₃ solution under controlled pH and temperature conditions in different morphology and dimensions: (i) microsized (average diameter 1.2 μm); (ii) submicrosized (250 nm); and (iii) nanosized (90 nm). To gain insight into reactions potentially occurring in vivo at the particle-lung interface following dust inhalation, several physicochemical features relevant to pathogenicity were measured (free radical generation in cell-free tests, metal release, and antioxidant depletion), and cellular toxicity assays on human lung epithelial cells (A549) and murine alveolar macrophages (MH-S) were carried out (LDH release, apoptosis detection, DNA damage, and nitric oxide synthesis). The decrease in particles size, from 1.2 μm to 90 nm, only caused a slight increase in structural defects (disorder of the hematite phase and the presence of surface ferrous ions) without enhancing surface reactivity or cellular responses in the concentration range between 20 and 100 μg cm⁻².

Crystalline silica incubated in ascorbic acid acquires a higher cytotoxic potential.

Quartz incubated in an aqueous solution of ascorbic acid is partially dissolved and the potential to generate hydroxyl radicals from hydrogen peroxide is enhanced. In order to investigate whether the surface activation triggered by the treatment with ascorbic acid would also involve an enhancement in cell toxicity, a murine macrophage cell line (RAW 264.7) was exposed to untreated and ascorbic acid-treated quartz. Ascorbic acid pretreated quartz was more toxic than untreated quartz and all cells died within 24 hours after exposure. Tetrandrine (a Chinese drug employed to retard or reverse fibrotic lesions of silicosis in humans) partially reduced cell toxicity generated by ascorbic acid pretreated quartz.

Inhibition of the ROS-mediated cytotoxicity and genotoxicity of nano-TiO2 toward human keratinocyte cells by iron doping

Nano-TiO2 powders are widely used in sunscreen lotions as UV filters in combination with other substances. The activation of TiO2 by UV rays leads to the release of reactive oxygen species (ROS, e.g., hydroxyl radicals and singlet oxygen) which are potentially harmful. For this reason the TiO2 particles are generally coated with inert materials (e.g., silica or alumina) that inhibit such reactivity. Alternatively, the release of ROS may be inhibited by introducing in the TiO2 lattice doping elements. In the present study we report a new modification consisting in a wet impregnation of TiO2 with iron salts followed by a thermal treatment that results in an inhibition of the surface reactivity. The insertion of iron ions also gradually reduces the ability of photo-activated TiO2 to cleave DNA and proteins. At the same time, a clear inhibition of cyto- and geno-toxicity toward human (HaCaT) keratinocytes was observed. The data presented herein suggest the insertion of Fe3+ ions at the surface of nano-TiO2 as a promising strategy to reduce the photo-induced toxicity of nano-TiO2 powders.

Carbon in Intimate Contact with Quartz Reduces the Biological Activity of Crystalline Silica Dusts

To evaluate the effect of carbonaceous materials on the pathogenic activity of quartz dusts, mixtures of carbon soot (1 and 10%) and quartz (Min-U-Sil) were prepared and then milled so to attain an intimate association of carbon and the quartz surface. Both cellular and cell-free tests show that carbon associated to quartz completely inhibits the typical free radical generation of quartz dusts (through Fenton activity and homolytic cleavage of a C-H bond) and suppresses the oxidative stress and inflammation induced by quartz alone on MH-S murine macrophage cells (lipid peroxidation, nitric oxide release, and tumor necrosis factor-α synthesis). The cytotoxic response to quartz is also largely reduced. An extremely pure quartz milled with 10% of soot showed inactivating effects on the adverse reactions to quartz similar to Min-U-Sil quartz. None of these effects takes place when the same experiments are carried out with mechanically mixed samples, which suggests that carbon acts not just as a radical quencher but because of its association to the quartz surface.