Subtle cytotoxicity and genotoxicity differences in superparamagnetic iron oxide nanoparticles coated with various functional groups.

Superparamagnetic iron oxide nanoparticles (SPIONs) have been widely utilized for the diagnosis and therapy of specific diseases, as magnetic resonance imaging (MRI) contrast agents and drug-delivery carriers, due to their easy transportation to targeted areas by an external magnetic field. For such biomedical applications, SPIONs must have multifunctional characteristics, including optimized size and modified surface. However, the biofunctionality and biocompatibility of SPIONs with various surface functional groups of different sizes have yet to be elucidated clearly. Therefore, it is important to carefully monitor the cytotoxicity and genotoxicity of SPIONs that are surfaced-modified with various functional groups of different sizes. In this study, we evaluated SPIONs with diameters of approximately 10 nm and 100~150 nm, containing different surface functional groups. SPIONs were covered with -O⁻ groups, so-called bare SPIONs. Following this, they were modified with three different functional groups--hydroxyl (-OH), carboxylic (-COOH), and amine (-NH2) groups--by coating their surfaces with tetraethyl orthosilicate (TEOS), (3-aminopropyl)trimethoxysilane (APTMS), TEOS-APTMS, or citrate, which imparted different surface charges and sizes to the particles. The effects of SPIONs coated with these functional groups on mitochondrial activity, intracellular accumulation of reactive oxygen species, membrane integrity, and DNA stability in L-929 fibroblasts were determined by water-soluble tetrazolium, 2',7'-dichlorodihydrofluorescein, lactate dehydrogenase, and comet assays, respectively. Our toxicological observations suggest that the functional groups and sizes of SPIONs are critical determinants of cellular responses, degrees of cytotoxicity and genotoxicity, and potential mechanisms of toxicity. Nanoparticles with various surface modifications and of different sizes induced slight, but possibly meaningful, changes in cell cytotoxicity and genotoxicity, which would be significantly valuable in further studies of bioconjugation and cell interaction for drug delivery, cell culture, and cancer-targeting applications.

Polyurethane foams electrophoretically coated with carbon nanotubes for tissue engineering scaffolds.

Carbon nanotubes (CNTs) were deposited on the surfaces of polyurethane (PUR) foams by electrophoretic deposition (EPD). The parameters of EPD were optimized in order to obtain homogeneous CNT coatings on PUR foams and adequate infiltration of the three-dimensional (3D) porous network. The microstructure of the composites was investigated by high-resolution scanning electron microscopy (HRSEM), revealing that optimal quality of the coatings was achieved by an EPD voltage of 20 V. The thermal properties of the CNT-coated specimens, determined by thermogravimetric analysis (TGA), were correlated to the foam microstructure. In vitro tests in concentrated simulated body fluid (1.5 SBF) were performed to study the influence of the presence of CNTs on the bioactivity of PUR-based scaffolds, assessed by the formation of calcium phosphate (CaP) compounds, e.g. hydroxyapatite (HA), on the foam surfaces. It was observed that CNTs accelerate the precipitation of CaP, which is thought to be due to the presence of more nucleation centres for crystal nucleation and growth, as compared with uncoated foams. Polyurethane foams with CNT coating have the potential to be used as bioactive scaffolds in bone tissue engineering due to their high interconnected porosity, bioactivity and nanostructured surface topography.

The use of murine embryonic stem cells, alginate encapsulation, and rotary microgravity bioreactor in bone tissue engineering.

The application of embryonic stem cells (ESCs) in bone tissue engineering and regenerative medicine requires the development of suitable bioprocesses that facilitate the integrated, reproducible, automatable production of clinically-relevant, scaleable, and integrated bioprocesses that generate sufficient cell numbers resulting in the formation of three-dimensional (3D) mineralised, bone tissue-like constructs. Previously, we have reported the enhanced differentiation of undifferentiated mESCs toward the osteogenic lineage in the absence of embryoid body formation. Herein, we present an efficient and integrated 3D bioprocess based on the encapsulation of undifferentiated mESCs within alginate hydrogels and culture in a rotary cell culture microgravity bioreactor. Specifically, for the first 3 days, encapsulated mESCs were cultured in 50% (v/v) HepG2 conditioned medium to generate a cell population with enhanced mesodermal differentiation capability followed by osteogenic differentiation using osteogenic media containing ascorbic acid, beta-glycerophosphate and dexamethasone. 3D mineralised constructs were generated that displayed the morphological, phenotypical, and molecular attributes of the osteogenic lineage, as well mechanical strength and mineralised calcium/phosphate deposition. Consequently, this bioprocess provides an efficient, automatable, scalable and functional culture system for application to bone tissue engineering in the context of macroscopic bone formation.