RESUMO
Surface initiated atom transfer radical polymerization (SI-ATRP) documented a simple but efficient technique to grow a dense polymer layer on any surface. Gold nanoparticles (AuNPs) give a broad surface to immobilize sulfhyryl group-containing initiators for SI-ATRP; in addition, AuNPs are the major nanoparticulate carriers for delivery of anti-cancer therapeutics, since they are biocompatible and bioinert. In this work, AuNPs with a disulfide initiator were polymerized with sulfoethyl methacrylate by SI-ATRP to decorate the particles with anionic corona, and branched polyethyeleneimine (PEI) and siRNA were sequentially layered onto the anionic corona of AuNP by electrostatic interaction. The in vitro anti-cancer effect confirmed that AuNP with anionic corona showed higher degrees of apoptosis as well as suppression of the oncogene expression in a siRNA dose-dependent manner. The in vivo study of tumor-bearing nude mice revealed that mice treated with c-Myc siRNA-incorporated AuNPs showed dramatically decreased tumor size in comparison to those with free siRNA for 4 weeks. Furthermore, histological examination and gene expression study revealed that the decorated AuNP significantly suppressed c-Myc expression. Thus, we envision that the layer-by-layer assembly on the anionic brushes can be potentially used to incorporate nucleic acids onto metallic particles with high transfection efficiency.
RESUMO
Cell sheet engineering has attracted great attention because thin layers of tissue can be easily transplanted to defect sites. Wound-dressing materials are required to support fast re-epithelization, both with keratinocytes and fibroblasts, to enhance the prognosis and therapeutic outcomes. We prepared self-assembled cell sheets composed of adipocyte-derived stem cells (ADSCs) and surface-engineered nanofibrils (NFs). NFs were surface-engineered with multilayers of gelatin so that the cell sheets could spontaneously assemble within 3 days in cell culture plates. Dorsal wounds transplanted with the cell sheets exhibited higher wound-healing rates when a high concentration of gelatin was immobilized on the surfaces of the NFs. Histochemical staining revealed that those with gelatin-immobilized NFs showed a higher expression of cytokeratin and collagen in the re-epithelized epidermis. Keratinocytic differentiation of the epidermis was molecularly evidenced by the higher expression of keratinocyte-specific genes.
Assuntos
Células-Tronco Mesenquimais , Nanofibras , Bandagens , Diferenciação Celular , GelatinaRESUMO
A free-standing, antibacterial hydrogel was fabricated using silver-nanoparticle-immobilized cellulose nanofibers (CNFs) and alginate. Surface hydroxyl groups of CNFs were oxidized to carboxylate groups using (2,2,6,6-tetramethylpiperidin-1-yl)oxidanyl (TCNF), followed by the treatment with silver nitrate solution for surface adsorption of silver ions. In situ reduction of silver ions to produce silver nanoparticles was performed for the silver-adsorbed CNFs. Electron microscopy, X-ray diffraction, and spectroscopic analysis revealed that higher amounts of silver nanoparticles were immobilized on the surface of TCNF than on the surface of native CNF. Silver-nanoparticle-immobilized TCNF was embedded in alginate gels and silver ions from the matrix were slowly released for 7 days. Silver-nanoparticle-loaded alginate gels showed comparable antibacterial activity to silver-ions-loaded alginate gels, although the former showed a significantly lower cytotoxicity against animal cells. Thus, the antibacterial gels can potentially be applied to various skin surfaces to prevent bacterial infection while minimizing skin damage.
RESUMO
Bilayered cell sheets were fabricated with coaxial hydro-nanofibrils for three-dimensional (3D) cultivation in a biomimetic environment. Polycaprolactone (PCL) was electrospun and hydrolyzed to release fragmented nanofibrils (NF) in an alkaline condition. Methacrylated gelatin (GelMA) was adsorbed and phototethered on the surface of the fibrils to prepare coaxial NF composed of hydrophilic shells and hydrophobic cores. GelMA layers on the NF were characterized by X-ray photoemission spectroscopy, Fourier-transform infrared spectroscopy, and thermogravimetric analysis. The GelMA showed higher decoration level on NF compared to that on native gelatin. GelMA-decorated NF significantly enhanced cell proliferation rate and phenotypic expression of human dermal fibroblasts when spontaneous formation of cell sheets was observed for 7 days. HaCaT cells were layered on top of the fibroblast sheets and further cultivated in air-water interfaces to prepare bilayered skin sheets. After 21 days of incubation, the top layers of the bilayered sheets showed higher expression of pan-keratin, and the dermal cells showed higher proliferation in the GelMA-decorated NF.