Immobilization of Pseudomonas cepacia lipase onto electrospun polyacrylonitrile fibers through physical adsorption and application to transesterification in nonaqueous solvent.

The lipase of Pseudomonas cepacia was immobilized onto electrospun polyacrylonitrile (PAN) fibers and used for the conversion of (S)-glycidol with vinyl n-butyrate to glycidyl n-butyrate in isooctane. The rate of reaction with the adsorbed lipase was 23-fold higher than the initial material. After 10 recyclings, the initial reaction rate was 80% of the original rate. This system of enzyme immobilization is therefore suitable for carrying out transesterification reactions in nonaqueous solvents.

Production of butyl-biodiesel using lipase physically-adsorbed onto electrospun polyacrylonitrile fibers.

Butyl-biodiesel production using electrospun polyacrylonitrile fibers with Pseudomonas cepacia lipase immobilized through physical adsorption was studied. About 80% conversion to butyl-biodiesel was achieved after 24h by suspending the catalyst at 2.4 mg/mL in a mixture of rapeseed oil and n-butanol at a molar ratio of 1:3, containing water at 8000 ppm at 40 degrees C. A further 24h of operation resulted in 94% conversion. The initial reaction rate detected for this process was 65-fold faster than those detected for Novozym 435 on a total catalyst mass basis. The immobilized lipase continued to work as a catalyst for 27 d, within a 15% reduction in conversion yield at the outlet of the reactor compared with the average value detected during the first 3d of operation in a continuous butyl-biodiesel production system.

Surface immobilization of poly(ethyleneimine) and plasmid DNA on electrospun poly(L-lactic acid) fibrous mats using a layer-by-layer approach for gene delivery.

The method of coating electrospun ultrafine poly(L-lactic acid) fibers with DNA, by building up polyelectrolyte layer(s) of poly(ethyleneimine) (PEI) and plasmid DNA using an electrostatic layer-by-layer deposition method, for gene delivery is presented. The pGL3 encoding luciferase was applied as plasmid DNA. The quantity of pGL3 immobilized on individual fibers increased with increasing pGL3 concentration in the immersion solution (0.017-0.870 mg/mL) and increasing bilayer number of PEI/pGL3 (single-triple). With the exception of one specimen prepared under the condition 0.870 mg/mL pGL3 solution and double PEI/pGL3 layers, the transfection efficiency of COS-7 cells, defined by the ratio of fluorescence intensity (resulting from the presence of luciferase) with respect to the quantity of cellular protein on the fibrous mat increased with increasing quantity of pGL3 on the fibers. In addition to the ease of controlling the quality of polyelectrolyte bilayer(s) by simply changing the concentrations of substances and number of immersing cycles, the features of the electrospun fibrous mat such as a very large surface-to-volume ratio and flexibility, could potentially be employed as a strategy for gene therapy combined with tissue engineering technology.

Transesterification by lipase entrapped in electrospun poly(vinyl alcohol) fibers and its application to a flow-through reactor.

We entrapped lipase in electrospun poly(vinyl alcohol) fibers of approximately 1 mum in diameter and evaluated the transesterification activity by converting (s)-glycidol to glycidyl n-butyrate with vinyl n-butyrate. The initial transesterification rate of the entrapped lipase was 5.2-fold faster than that of non-treated lipase. The fibrous membrane could be used as a component of a flow-through reactor for continuous transesterification.

Reinforcement of porous alginate scaffolds by incorporating electrospun fibres.

The mechanical properties of scaffolds play a vital role in transmitting input mechanical signals to the cells within them. We aimed to modify mechanical properties of porous scaffolds by incorporating electrospun fibres into their frameworks. Porous constructs containing electrospun silicate fibres were prepared from Na-alginate aqueous solutions suspending the silicate fibres with (ASF) or without amino groups (NASF) via an all-aqueous method based on a freeze-drying technique. The repulsion forces of constructs containing ASF towards compression increased as the fibre content increased. In contrast, constructs containing NASF showed no such increases in repulsion forces. Cells seeded onto constructs containing ASF exhibited suppressed growth, similar to cells seeded onto alginate scaffolds without fibres. In contrast, cells seeded onto scaffolds containing NASF showed about two-fold faster growth than cells seeded onto scaffolds containing ASF. The differences in the mechanical properties and cell growth profiles between the scaffolds containing ASF and NASF can be explained by the formation and non-formation of electrostatic bonds between the fibres and alginate, respectively. The results obtained in the present study demonstrate the feasibility of incorporating electrospun fibres for reinforcement of alginate scaffolds and enhancement of cell growth.

Prospective use of electrospun ultra-fine silicate fibers for bone tissue engineering.

Electrospinning involves the generation of a jet of viscous solution, and results in the formation of ultra-fine fibers. Silicate fibers prepared using this technology and via the sol-gel process were evaluated as scaffolds for bone tissue engineering. We found that human osteoblastic MG63 cells successfully adhered on individual silicate fibers, and proliferated on them. In an apatite-formation ability study, spherical particles covered the fibers after soaking in simulated body fluid for 7 days. Energy dispersive X-ray analysis revealed that Ca/P atomic ratio of the particles was similar to that of human bone. In addition, X-ray diffraction analysis revealed that crystalline structure of the particles agreed with that of apatite. These results suggest that electrospun silicate fibers are a potential candidate for scaffolding material in bone tissue engineering.