RESUMEN
BACKGROUND: Lonocyte-derived multipotential cells (MOMCs) include progenitors capable of differentiation into multiple cell lineages and thus represent an ideal autologous transplantable cell source for regenerative medicine. In this study, we cultured MOMCs, generated from mononuclear cells of peripheral blood, on the surface of nanocomposite thin films. METHODS: For this purpose, nanocomposite Poly(e-caprolactone) (PCL)-based thin films containing either 2.5 wt% silica nanotubes (SiO2ntbs) or strontium hydroxyapatite nanorods (SrHAnrds), were prepared using the spin-coating method. The induced differentiation capacity of MOMCs, towards bone and endothelium, was estimated using flow cytometry, real-time polymerase chain reaction, scanning electron microscopy and fluorescence microscopy after cells' genetic modification using the Sleeping Beauty Transposon System aiming their observation onto the scaffolds. Moreover, Wharton's Jelly Mesenchymal Stromal Cells were cultivated as a control cell line, while Human Umbilical Vein Endothelial Cells were used to strengthen and accelerate the differentiation procedure in semi-permeable culture systems. Finally, the cytotoxicity of the studied materials was checked with MTT assay. RESULTS: The highest differentiation capacity of MOMCs was observed on PCL/SiO2ntbs 2.5 wt% nanocomposite film, as they progressively lost their native markers and gained endothelial lineage, in both protein and transcriptional level. In addition, the presence of SrHAnrds in the PCL matrix triggered processes related to osteoblast bone formation. CONCLUSION: To conclude, the differentiation of MOMCs was selectively guided by incorporating SiO2ntbs or SrHAnrds into a polymeric matrix, for the first time.
Asunto(s)
Autoinjertos , Belleza , Línea Celular , Linaje de la Célula , Durapatita , Endotelio , Citometría de Flujo , Células Endoteliales de la Vena Umbilical Humana , Células Madre Mesenquimatosas , Métodos , Microscopía Electrónica de Rastreo , Microscopía Fluorescente , Nanocompuestos , Nanotubos , Osteoblastos , Osteogénesis , Polímeros , Reacción en Cadena en Tiempo Real de la Polimerasa , Medicina Regenerativa , Dióxido de Silicio , Estroncio , Gelatina de WhartonRESUMEN
Thermodynamically immiscible poly(lactic acid) (PLA) and poly(ε-caprolactone) (PCL) were blended and solution-cast by adding the 3% compatibilizer (tributyl citrate, TBC) of the PCL weight. In the PLA/PCL composition range of 99/1–95/5 wt%, mechanical properties of the PLA/PCL films with TBC were always superior to those of the films without TBC. The tensile strength of 42.9 ± 3.5 MPa and the elongation at break of 10.3 ± 2.7% were observed for the 93/7 PLA/PCL films without TBC, indicating that PCL addition is effective for strength and ductility. However, the tensile strength of 54.1 ± 3.4 MPa and the elongation at break of 8.8 ± 1.8% were found for the 95/5 PLA/PCL with TBC, indicating that the effect of co-addition of PCL and TBC on mechanical properties of the films is more pronounced. No cytotoxicity was observed for the PLA/PCL films regardless of TBC addition.