fumarates and thiazolyl-blue

As the use of superparamagnetic iron oxide nanoparticles (SPION) in biomedical applications increases (e.g. for targeting drug delivery and imaging), patients are likely to be exposed to products containing SPION. Despite their high biomedical importance, toxicity data for SPION are limited to date. The aim of this study is to investigate the cytotoxicity of SPION and its ability to change cell medium components. Bare and poly(ethylene glycol)-co-fumarate (PEGF)-coated SPION with narrow size distributions were synthesized. The particles were prepared by co-precipitation using ferric and ferrous salts with a molar Fe3+/Fe2+ ratio of 2. Dulbecco's modified Eagle's medium (DMEM) and primary mouse fibroblast (L929) cell lines were exposed to the SPION. Variation of cell medium components and cytotoxicity due to the interactions with nanoparticles were analyzed using ultraviolet and visible spectroscopy (UV/vis) and the 3-[4,5-dimethylthiazol-2yl]-2,5-diphenyltetrazolium bromide (MTT) assay methods, respectively. The toxicity amount has been traditionally identified by changes in pH and composition in cells and DMEM due to the tendency of SPION to adsorb proteins, vitamins, amino acids and ions. For in vitro toxicity assessments, a new surface passivation procedure is proposed which can yield more reliable quantitative results. It is shown that a more reliable way of identifying cytotoxicity for in vitro assessments is to use particles with saturated surfaces via interactions with DMEM before usage.

Topics: Adsorption; Animals; Cell Line; Cell Survival; Culture Media; Ferric Compounds; Fumarates; Hydrogen-Ion Concentration; Metal Nanoparticles; Mice; Microscopy, Electron, Transmission; Polyethylene Glycols; Spectrophotometry, Ultraviolet; Tetrazolium Salts; Thiazoles

The effect of temporary encapsulation of rat marrow stromal osteoblasts in crosslinked gelatin microparticles on cell viability and proliferation was investigated in this study for microparticles placed on a crosslinking poly(propylene fumarate) (PPF) composite over a 7 day time period. Encapsulated cells were seeded on crosslinking PPF composites at times up to 10 min following initiation of the crosslinking reaction, and also on fully crosslinked PPF composites and tissue culture polystyrene controls, with a cell seeding density of 5.3 x 10(4) cells/cm2. The crosslinked PPF composite exhibited an average gel point of 10.3 min and an average maximum crosslinking temperature of 47.5 degrees C. Cell viability and proliferation were assessed by DNA and 3H-thymidine assays and the results were compared with those for nonencapsulated cells. The results showed that the addition time of cells to a crosslinking PPF composite had a large effect on cell viability and proliferation for both encapsulated and nonencapsulated cells with more surviving cells added at later time points. Most importantly, the temporary encapsulation of cells significantly enhanced cell viability at earlier time points. The data indicate that the presence of gelatin microparticles does not affect the crosslinking of a PPF composite. They further suggest that the temporary encapsulation of cells in crosslinked gelatin microparticles may preserve the viability of cells contained in an actively crosslinking PPF composite used as an injectable polymeric scaffold serving also as a carrier for osteogenic cell populations.

Topics: Alkaline Phosphatase; Animals; Biocompatible Materials; Bone Substitutes; Cell Division; Cell Survival; Chromatography; Coloring Agents; Cross-Linking Reagents; DNA; Drug Compounding; Drug Delivery Systems; Fumarates; Gelatin; Gels; Injections; Osteoblasts; Osteocalcin; Phenotype; Polymers; Polypropylenes; Rats; Rats, Sprague-Dawley; Temperature; Tetrazolium Salts; Thiazoles; Thymidine; Time Factors; Tissue Engineering