Jeroen Mattijs Bezemer

A novel method to obtain protein release from porous polymer scaffolds: emulsion coating.

To obtain the controlled release of proteins from macro-porous polymeric scaffolds, a novel emulsion-coating method has been developed. In this process, a water-in-oil emulsion, from an aqueous protein solution and a polymer solution, is forced through a prefabricated scaffold by applying a vacuum. After solvent evaporation, a polymer film, containing the protein, is then deposited on the porous scaffold surface. This paper reports the effect of processing parameters on the emulsion coating characteristics, scaffold structure, and protein release and stability. Poly(ether-ester) multiblock copolymers were chosen as the polymer matrix for both scaffolds and coating. Macro-porous scaffolds, with a porosity of 77 vol% and pores of approximately 500 microm were prepared by compression moulding/salt leaching. A micro-porous, homogeneous protein-loaded coating could be obtained on the scaffold surface. Due to the coating, the scaffold porosity was decreased, whereas the pore interconnection was increased. A model protein (lysozyme) could effectively be released in a controlled fashion from the scaffolds. Complete lysozyme release could be achieved within 3 days up to more than 2 months by adjusting the coated emulsion parameters. In addition, the coating process did not reduce the enzymatic activity. This new method appears to be promising for tissue engineering applications.

Control of vitamin B12 release from poly(ethylene glycol)/poly(butylene terephthalate) multiblock copolymers

The release of vitamin B12 (1355 Da) from matrices based on multiblock copolymers was studied. The copolymers were composed of hydrophilic poly(ethylene glycol)-terephthalate (PEGT) blocks and hydrophobic poly(butylene terephthalate) (PBT) blocks. Vitamin B12 loaded films were prepared by using a water-in-oil emulsion method. The copolymer properties, like permeability, could be varied by increasing the PEG-segment length from 300 up to 4,000 g/mol and by changing the wt% of PEGT. From permeation and release experiments. the diffusion coefficient of vitamin B12 through PEGT/PBT films of different compositions was determined. The diffusion coefficient of Vitamin B12 was strongly dependent on the composition of the copolymers. Although an increased wt% of PEGT (at a constant PEG-segment length) resulted in a higher diffusion coefficient, a major effect was observed at increasing PEG-segment length. By varying the copolymer composition, a complete release of vitamin B12 in 1 day up to a constant release for over 12 weeks was obtained. The release rate could be effectively tailored by blending copolymers with different PEG-segment lengths. The swelling and the crystallinity of the matrix could explain the effect of the matrix composition on the release behavior.

Poly(ethylene oxide)-modified carboxylated polystyrene latices - immobilization chemistry and protein adsorption

alpha,omega-Diamino poly(ethylene oxides) (PEOs) with different molecular weights (148, 1000, and 3400) were covalently immobilized onto carboxylated polystyrene latices. The immobilization of PEO was carried out with N-(3-dimethylaminopropyl)-N-ethylcarbodiimide (EDC) and N-hydroxysuccinimide (NHS) in aqueous media. The reaction conditions were optimized to obtain a maximal coupling of PEO. The degree of coupling was determined by the surface concentration of amino groups. The maximal surface concentrations of amino groups were close to what is expected for a complete coverage of the surface with PEO. Adsorption of albumin from a buffer solution onto PEO-containing surfaces was about 85% less than the albumin uptake by unmodified polystyrene latices. Protein adsorption from plasma dilutions was lower on surfaces containing PEO molecules with a higher molecular weight. The reduction of the protein uptake from plasma by surfaces containing PEO-3400 molecules was only 40% compared to the adsorption to unmodified surfaces. These results indicate that plasma proteins have a low affinity for surfaces modified with PEO. However the PEO modified surfaces are by no means protein resistant when exposed to plasma.