ABSTRACT
OBJECTIVES: Transplantation of endothelial progenitor cells (EPCs) is a promising approach for revascularization of tissue. We have used a natural and biocompatible biopolymer, fibrin, to induce cell population growth, differentiation and functional activity of EPCs. MATERIALS AND METHODS: Peripheral blood mononuclear cells were cultured for 1 week to obtain early EPCs. Fibrin was characterized for stiffness and capability to sustain cell population expansion at different fibrinogen-thrombin ratios. Viability, differentiation and angiogenic properties of EPCs were evaluated and compared to those of EPCs grown on fibronectin. RESULTS: Fibrin had a nanometric fibrous structure forming a porous network. Fibrinogen concentration significantly influenced fibrin stiffness and cell growth: 9 mg/ml fibrinogen and 25 U/ml thrombin was the best ratio for enhanced cell viability. Moreover, cell viability was significantly higher on fibrin compared to being on fibronectin. Even though no significant difference was observed in expression of endothelial markers, culture on fibrin elicited marked induction of stem cell markers OCT 3/4 and NANOG. In vitro angiogenesis assay on Matrigel showed that EPCs grown on fibrin retain angiogenetic capability as EPCs grown on fibronectin, but significantly better release of cytokines involved in cell recruitment was produced by EPC grown on fibrin. CONCLUSION: Fibrin is a suitable matrix for EPC growth, differentiation and angiogenesis capability, suggesting that fibrin gel may be very useful for regenerative medicine.
Subject(s)
Cell Differentiation/physiology , Endothelial Cells/physiology , Fibrin/metabolism , Stem Cells/cytology , Biocompatible Materials/metabolism , Biomarkers/metabolism , Biomimetic Materials/metabolism , Cell Proliferation/drug effects , Cell Survival/drug effects , Cells, Cultured , Dose-Response Relationship, Drug , Endothelium/metabolism , Fibrin/ultrastructure , Fibrinogen/pharmacology , Fibronectins/metabolism , Homeodomain Proteins/biosynthesis , Humans , Nanog Homeobox Protein , Octamer Transcription Factor-3/biosynthesis , Porosity , Stem Cells/metabolism , Thrombin/pharmacologyABSTRACT
Boron nitride nanotubes (BNNTs) have generated considerable interest within the scientific community by virtue of their unique physical properties, which can be exploited in the biomedical field. In the present in vitro study, we investigated the interactions of poly-l-lysine-coated BNNTs with C2C12 cells, as a model of muscle cells, in terms of cytocompatibility and BNNT internalization. The latter was performed using both confocal and transmission electron microscopy. Finally, we investigated myoblast differentiation in the presence of BNNTs, evaluating the protein synthesis of differentiating cells, myotube formation, and expression of some constitutive myoblastic markers, such as MyoD and Cx43, by reverse transcription - polymerase chain reaction and Western blot analysis. We demonstrated that BNNTs are highly internalized by C2C12 cells, with neither adversely affecting C2C12 myoblast viability nor significantly interfering with myotube formation.
Subject(s)
Boron Compounds/administration & dosage , Boron Compounds/chemistry , Coated Materials, Biocompatible/administration & dosage , Coated Materials, Biocompatible/chemistry , Muscle Fibers, Skeletal/cytology , Muscle Fibers, Skeletal/drug effects , Nanotubes/chemistry , Polylysine/administration & dosage , Animals , Cell Differentiation/drug effects , Cell Line , Materials Testing , Muscle Fibers, Skeletal/chemistry , Nanotubes/ultrastructure , Polylysine/chemistry , RatsABSTRACT
Nanoscale structures and materials have been explored in many biological applications because of their extraordinary novel properties. Here we propose a study of cellular interactions with barium titanate nanoparticles, an interesting ceramic material that has received a lot of interest in the nanotechnology research, but without any attention about its biological potential. We introduced for the first time an efficient method for the preparation of stable aqueous dispersions of barium titanate nanoparticles, characterized with FIB, TEM and AFM imaging, light scattering, Z-potential and UV/vis analysis. Finally, we presented a systematic study of short-term cytotoxicity of the prepared dispersion based both on quantitative (metabolism, proliferation) and qualitative (apoptosis, viability, differentiation) assays.
