Silica nanoparticles as sources of silicic acid favoring wound healing in vitro.

There is good evidence that certain silicon-containing materials promote would healing and their common feature is the delivery of orthosilicic acid (Si(OH)4) either directly or following metabolism. In this respect, amorphous silica nanoparticles (NP), which dissolve in aqueous environments releasing up to 2mM orthosilicic acid, may be appropriate 'slow release' vehicles for bioactive silicon. Here we studied the impact of silica NP suspensions (primary particles∼10nm) in undersaturated conditions (below 2mM Si) with differing degrees of surface charge and dissolution rate on human dermal fibroblasts (CCD-25SK cells) viability, proliferation and migration in a cellular wound model. Silica was shown to be non-toxic for all forms and concentrations tested and whilst the anticipated stimulatory effect of orthosilicic acid was observed, the silica NPs also stimulated fibroblast proliferation and migration. In particular, the amine-functionalized particles promoted wound closure more rapidly than soluble orthosilicic acid alone. We suggest that this effect is related to easy cellular internalization of these particles followed by their intracellular dissolution releasing silicic acid at a faster rate than its direct uptake from the medium. Our findings indicate that amorphous silica-based NPs may favour the delivery and release of bioactive silicic acid to cells, promoting wound healing.

Nanometric emulsions encapsulating solid particles as alternative carriers for intracellular delivery.

AIM: Formulate nanometric oil droplets for encapsulating solid nanoparticles and assess their interactions with cells. MATERIALS & METHODS: Soybean oil droplets, stabilized by Pluronic F68 surfactant, incorporating hydrophobically modified fluorescent silica, nanoparticles were obtained. Cytotoxicity over time, internalization, subsequent intracellular localization and internalization pathways were assessed by microscopy (fluoresence and TEM) in vitro with HeLa cells. RESULTS: Oil droplets encapsulating solid nanoparticles are readily internalized by HeLa cells like free nanoparticles but the intracellular localization differs (nanoemulsions less colocalized with lysosomes) as well as internalization pathway is used (nanoemulsions partially internalized by nonendocytic transport). No cytotoxicity could be observed for either particles tested. CONCLUSION: Our results confirm that nanometric emulsions encapsulating solid nanoparticles can be used for alternative and multifunctional intracellular delivery.

Phagocytosis of immunoglobulin-coated emulsion droplets.

Phagocytosis by macrophages represents a fundamental process essential for both immunity and tissue homeostasis. The size of targets to be eliminated ranges from small particles as bacteria to large objects as cancerous or senescent cells. Most of our current quantitative knowledge on phagocytosis is based on the use of solid polymer microparticles as model targets that are well adapted to the study of phagocytosis mechanisms that do not involve any lateral mobility of the ligands, despite the relevance of this parameter in the immunological context. Herein we designed monodisperse, IgG-coated emulsion droplets that are efficiently and specifically internalized by macrophages through in-vitro FcγR-mediated phagocytosis. We show that, contrary to solid polymeric beads, droplet uptake is efficient even for low IgG densities, and is accompagnied by the clustering of the opsonins in the zone of contact with the macrophage during the adhesion step. Beyond the sole interest in the design of the material, our results suggest that lateral mobility of proteins at the interface of a target greatly enhances the phagocytic uptake.

Behaviour of silica nanoparticles in dermis-like cellularized collagen hydrogels.

A model of the fate of colloidal silica in the dermis was designed based on the diffusion of fluorescent silica nanoparticles through collagen hydrogels. The diffusion process was found to depend on particle size (10-200 nm) and surface charge, as well as on collagen concentration (1.5-5 mg mL-1). The presence of human dermal fibroblasts within the hydrogels also significantly impacted on the behaviour of the particles. In particular, the simultaneous monitoring of particulate and soluble forms of silica showed that both the hydrogel network and the cellular activity have a strong influence on the solubilization process of the silica particles, through a combination of surface sorption, uptake and intracellular dissolution. Interactions between silica and collagen in 3D environments also lower the cytotoxicity of 10 nm particles compared to traditional 2D cultures. The results emphasize the complexity of silica chemistry in living tissues and specifically indicate the need for further investigations of the in vivo behaviour of its soluble forms.

Introduction of disulfide bridges within silica nanoparticles to control their intra-cellular degradation.

Incorporation of disulfide bridges in the core structure of silica nanoparticles modifies their intracellular fate within dermal fibroblasts, especially influencing their degradation pathway.

Long-term fate of silica nanoparticles interacting with human dermal fibroblasts.

The long-term fate of fluorescent non-porous FITC-SiO(2) nanoparticles of various sizes (10-200 nm) and charge is studied in the presence of human dermal fibroblasts. Particle aggregates are formed in the culture medium and uptaken, at least partially, by macropinocytosis. The smallest particles have a strong impact on cell viability and genotoxic effects can be observed for negatively-charged colloids 10 nm in size. Largest particles do not impact on cellular activity and can be monitored in cellulo via fluorescence and transmission electron microscopy studies over two weeks. These observations reveal a significant decrease in the size of silica particles located in endocytic vesicles. The dissolution process is confirmed by monitoring the cell culture medium that contains both colloidal and soluble silica species. Such dissolution can be explained on the sole basis of silica solubility and has great implication for the use of non-porous silica particles as intra-cellular drug release systems.

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.