Thermo-Magnetic Properties of Fe3O4@Poly(N-Isopropylacrylamide) Core-Shell Nanoparticles and Their Cytotoxic Effects on HeLa and MDA-MB-231 Cell Lines.

In this study, we performed a systematic evaluation of the synthesis parameters of a multiresponsive core-shell nanoplatform (Fe3O4 nanoparticles coated by poly(N-isopropylacrylamide). These nanocomposites allow the possibility of controlling specific properties, such as particle size and morphology. The polymer coating was performed by in-situ free-radical polymerization of N-isopropylacrylamide on Fe3O4 nanoparticles to obtain a nanocomposite with a core-shell structure. We monitored the size and morphology of the nanocomposite by scanning transmission electron microscopy. Electrophoretic mobility determined the evolution of ζ-potentials during coating. We assessed the functionalization of the Fe3O4 nanoparticles by Fourier-transform infrared spectroscopy and ultraviolet-visible spectroscopy. The core-shell systems exhibited a collapsed structure when heated above the lower critical solution temperature, which was determined by dynamic light scattering. Squid based vibrating sample magnetometer tests were performed to verify superparamagnetic behavior at room temperature. The platform was approved as biocompatible for the HeLa and MDA-MB-231 cell lines by MTT assays, and it shows the desired properties for potential use in localized and controlled drug delivery.

Encapsulation and immobilization of ficin extract in electrospun polymeric nanofibers.

In this work, ficin extract (3.4.22.3, >1.0 u/mg) was immobilized for the first time in poly (vinyl alcohol) (PVA) electrospun nanofibers in order to preserve its tridimensional molecular structure. The analysis of the morphology of the electrospun nanofibers was carried out using scanning electron microscopy (SEM) showing a diameter in the range of 124-194 nm. The interaction of the ficin extract with the nanofibers structure was analyzed by infrared spectroscopy (FTIR) analysis. The immobilization step was achieved through crosslinking involving the exposure to glutaraldehyde (GA) vapor. The enzyme catalytic behavior was followed by ultraviolet spectroscopy (UV) using the Earlanger method using Nα-benzoyl-l-arginine-4-nitroanilide hydrochloride (BAPA) as substrate. The maximum catalytic activity was reached with 1 h of crosslinking, 20% of enzyme loading and pH 8. The immobilized ficin extract showed 92% of the enzyme activity of the crude ficin extract. The enzymatic activity of the immobilized ficin extract was conserved after a total of nine reuse cycles and maintained after being stored for 25 days. Finally, both the glass transition (Tg) and heat of fusion (Hf) were affected by number of enzyme molecules inside the polymeric nanofibers matrix according to the study of the thermal properties by differential scanning calorimetry (DSC).

Encapsulation and immobilization of papain in electrospun nanofibrous membranes of PVA cross-linked with glutaraldehyde vapor.

In this paper, papain enzyme (E.C. 3.4.22.2, 1.6 U/mg) was successfully immobilized in poly(vinyl alcohol) (PVA) nanofibers prepared by electrospinning. The morphology of the electrospun nanofibers was characterized by scanning electron microscopy (SEM) and the diameter distribution was in the range of 80 to 170 nm. The presence of the enzyme within the PVA nanofibers was confirmed by infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS) and energy dispersive X-ray spectroscopy (EDXS) analyses. The maximum catalytic activity was reached when the enzyme loading was 13%. The immobilization of papain in the nanofiber membrane was achieved by chemical crosslinking with a glutaraldehyde vapor treatment (GAvt). The catalytic activity of the immobilized papain was 88% with respect to the free enzyme. The crosslinking time by GAvt to immobilize the enzyme onto the nanofiber mat was 24h, and the enzyme retained its catalytic activity after six cycles. The crosslinked samples maintained 40% of their initial activity after being stored for 14 days. PVA electrospun nanofibers are excellent matrices for the immobilization of enzymes due to their high surface area and their nanoporous structure.