The cytotoxicity of four catalysts commonly used for the synthesis of copolymers for biomedical use, such as segmented polyurethanes, was evaluated towards two types of cells, the first being the well-characterized cell line Swiss 3T3 mouse fibroblasts, the second the actual living system that faces any device in contact with blood, i.e. human endothelial cells (HEC). The catalysts were two tertiary aliphatic amines: TMBDA (tetramethylbutanediamine), and DABCO (1-4 diazo (2,2,2) octane); two alkyl tin compounds: DBTDL (dibutyl-tindilaurate), and SnOct (stannous octoate). Cytotoxicity tests were carried out by adding to the culture medium, after cell adhesion, different concentrations of each catalyst in dimethylsulphoxide, and keeping them in contact with the monolayer for 72 h. All the catalysts proved to be cytotoxic, although at different extent (in the order: DABCO < TMBDA < SnOct < DBTDL); their dose inhibiting 50% of cell growth (IC50) came out to be lower for 3T3 fibroblasts than for HEC, with the exception of DBTDL, which showed a similar toxicity for both the cell lines. As an example, the cytotoxicity of a polyurethane-amide, laboratory synthesized with DBTDL as catalysts, was checked with fibroblasts. By using both the method of the extract, and that of the direct contact (through a microporous membrane), a moderate to severe cell growth inhibition, related to the Sn content in the material, was observed.
A variety of natural polymers and proteins are considered to be 3D cell culture structures able to mimic the extracellular matrix (ECM) to promote bone tissue regeneration. Pectin, a natural polysaccharide extracted from the plant cell walls and having a chemical structure similar to alginate, provides interesting properties as artificial ECM. In this work, for the first time, pectin, modified with an RGD-containing oligopeptide or not, is used as an ECM alternative to immobilize cells for bone tissue regeneration. The viability, metabolic activity, morphology, and osteogenic differentiation of immobilized MC3T3-E1 preosteoblats demonstrate the potential of this polysaccharide to keep immobilized cells viable and differentiating. Preosteoblasts immobilized in both types of pectin microspheres maintained a constant viability up to 29 days and were able to differentiate. The grafting of the RGD peptide on pectin backbone induced improved cell adhesion and proliferation within the microspheres. Furthermore, not only did cells grow inside but also they were able to spread out from the microspheres and to organize themselves in 3D structures producing a mineralized extracellular matrix. These promising results suggest that pectin can be proposed as an injectable cell vehicle for bone tissue regeneration.
The relative chemical stability of two commercially available polyurethanes-Pellethane, currently used in biomedical devices, and Corethane, considered as a potential biomaterial-was investigated following aging protocols in hydrolytic and oxidative conditions (HOC, water, hydrogen peroxide, and nitric acid) and in physiological media (PHM, phosphate buffer, lipid dispersion, and bile from human donors). The chemical modifications induced on these polymers were characterized using differential scanning calorimetry (DSC), gel permeation chromatography (GPC), and Fourier transform infrared spectroscopy (FTIR). With the exception of nitric acid, all of the aging media promoted a mild hydrolytic reaction leading to a slight molecular weight loss in both polymers. When aged in water and hydrogen peroxide, Pellethane experienced structural modifications through microdomain phase separation along with an increase of the order within the soft-hard segment domains. The incubation of Pellethane in nitric acid also resulted in an important decrease of the melting temperature of its hard segments with chain scission mechanisms. Moreover, incubation in PHM led to an increase of the order within shorter hard-segment domains. FTIR data revealed the presence of aliphatic amide molecules used as additives on the Pellethane's surface. The incubation of Corethane under the same conditions promoted an almost uniform molecular reorganization through a phase separation between the hard and soft segments as well as an increase of the short-range order within the hard-segment domains. Incubation of this polymer in nitric acid also resulted in a chain scission process that was less pronounced than that measured for the Pellethane samples. Finally, lipid adsorption occurred on the Corethane sample incubated in bile for 120 days. Overall data indicate that polycarbonate urethane presents a greater chemical stability than does polyetherurethane.
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