RESUMEN
In this study, azide and alkyne moieties were introduced to the structure of citric acid-modified hydroxyethyl cellulose (HEC) and then through a bioorthogonal click chemistry method: Strain-promoted azide-alkyne cycloaddition, a novel crosslinked HEC scaffold (click sample) was obtained. Chemical modifications and successful crosslinking of the samples were assessed with FTIR and 1H NMR spectroscopy. Lyophilized samples exhibited a porous interconnected microarchitecture with desirable features for commensurate cartilage tissue engineering applications. As the stability of scaffolds improved upon crosslinking, considerable water uptake and swelling degree of ~650% could still be measured for the click sample. Offering Young's modulus of ~10 MPa and tensile strength of ~0.43 MPa, the mechanical characteristics of click sample were comparable with those of normal cartilage tissue. Various in vitro biological assays, including MTT analysis, cellular attachment, histological staining with safranin O, and real-time PCR decisively approved significant biocompatibility, chondrogenic ability, and bioorthogonal features of click sample.
Asunto(s)
Materiales Biocompatibles/química , Cartílago/fisiología , Celulosa/análogos & derivados , Condrocitos/fisiología , Química Clic , Reactivos de Enlaces Cruzados/química , Ingeniería de Tejidos , Andamios del Tejido , Cartílago/metabolismo , Cartílago/ultraestructura , Adhesión Celular , Línea Celular , Supervivencia Celular , Celulosa/química , Condrocitos/metabolismo , Condrocitos/ultraestructura , Condrogénesis , Ácido Cítrico/química , Módulo de Elasticidad , Humanos , Porosidad , Resistencia a la TracciónRESUMEN
INTRODUCTION: In recent years, the extremely low frequency electromagnetic field (ELF-EMF) has attracted a great deal of scientific interest. The ELF-EMF signal is able to control ion transport across ion channels and therefore induce cell differentiation. AIM: The purpose of this study was to investigate the effect of ELF-EMF (50 Hz, 1 mT) on MAP2 and Nestin gene expression of dermal papilla mesenchymal cells (DPCs). METHODS: In order to examine the effect of chemical and electromagnetic factors on gene expression, 4 experimental groups, namely chemical (cell exposure to chemical signals), EMF (exposing cells to ELF-EMF), chemical-EMF (subjecting cells to chemical signals and ELF-EMF) and control (with no treatment) groups, were prepared, treated for 5 days, and studied. To assess the effect of extended test time on the expression of neural differentiation markers (Nestin and MAP2), an EMF group was prepared and treated for a period of 14 consecutive days. The beneficial role of EMF in inducing neural differentiation was shown by real-time PCR analysis. RESULTS: The higher expression of MAP2 after 14 days compared to that after 5 days and decrease of cell proliferation on days 5 to 20 were indicative of the positive effect of extending treatment time on neural differentiation by evaluation of gene expression in EMF group.