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Hydroxyapatite particles as drug carriers for proteins
Keishiro Tomoda1, Hidehiko Ariizumi, Takatomo Nakaji
1Faculty of Pharmaceutical Sciences, Tokyo University of Science, Noda, Chiba, Japan.
Recently, much attention has been paid to hydroxyapatite (HA) particles as protein drug carriers. HA is biological substance like tooth and bone, and HA has biodegradable and biocompatible property. Also HA has high affinity for versatile substances, such as proteins. The aim of this study was to prepare HA particles and to evaluate the effects of various experimental conditions on particles properties (i.e. shape and amount of protein adsorption to HA). HA particles were prepared by three methods including solvent diffusion methods, the method using sintering porous HA microgranules, and homogeneous precipitation method. Complicated spiky crystals were prepared by solvent diffusion method, whereas spherical amorphous agglomerates were prepared by sintering of porous HA granules. Spherical particles were obtained by homogeneous precipitation method and were well dispersed. The spherical particles were composed of minute spiky crystals. The temperature and time duration of the producing process and the amount of additives, such as urea and EDTANa(2), strongly affected the particle shape and size. When the production process is kept under low temperature, tiny crystals were not mutually sintered and also their aggregation was not deposited. Also, inhomogeneous crystal growth occurred when the reaction time was long. Addition of enough amount of EDTANa(2) to reaction solution made the small and spherical HA particles. Bovine serum albumin (BSA) or lysozyme hydrochloride (LSZ)-adsorbed HA particles were prepared. The affinity of HA particles and release profile of proteins from the particles were evaluated. X-ray powder diffraction measurements suggested that the surface area ratio of a plane against total surface area of HA crystal became smaller by decreasing EDTANa(2) concentration. The surface of plane a is positively charged but that of plane c is negatively charged. HA particle size decreased as EDTANa(2) concentration increased. The amounts of BSA and LSZ adsorbing onto HA particle surface were different, since BSA was negatively charged and LSZ was positively charged in the solution. Therefore BSA seems to adsorb onto plane c, whereas LSZ adsorb onto plane a. The differences of the ratio of surface a plane against the total surface area of HA particles and particles size influenced the amount of protein adsorption. The sustained release of BSA and LSZ from HA particles were observed for two weeks.
Recently, much attention has been paid to hydroxyapatite (HA) particles as protein drug carriers. HA is biological substance like tooth and bone, and HA has biodegradable and biocompatible property. Also HA has high affinity for versatile substances, such as proteins. The aim of this study was to prepare HA particles and to evaluate the effects of various experimental conditions on particles properties (i.e. shape and amount of protein adsorption to HA). HA particles were prepared by three methods including solvent diffusion methods, the method using sintering porous HA microgranules, and homogeneous precipitation method. Complicated spiky crystals were prepared by solvent diffusion method, whereas spherical amorphous agglomerates were prepared by sintering of porous HA granules. Spherical particles were obtained by homogeneous precipitation method and were well dispersed. The spherical particles were composed of minute spiky crystals. The temperature and time duration of the producing process and the amount of additives, such as urea and EDTANa(2), strongly affected the particle shape and size. When the production process is kept under low temperature, tiny crystals were not mutually sintered and also their aggregation was not deposited. Also, inhomogeneous crystal growth occurred when the reaction time was long. Addition of enough amount of EDTANa(2) to reaction solution made the small and spherical HA particles. Bovine serum albumin (BSA) or lysozyme hydrochloride (LSZ)-adsorbed HA particles were prepared. The affinity of HA particles and release profile of proteins from the particles were evaluated. X-ray powder diffraction measurements suggested that the surface area ratio of a plane against total surface area of HA crystal became smaller by decreasing EDTANa(2) concentration. The surface of plane a is positively charged but that of plane c is negatively charged. HA particle size decreased as EDTANa(2) concentration increased. The amounts of BSA and LSZ adsorbing onto HA particle surface were different, since BSA was negatively charged and LSZ was positively charged in the solution. Therefore BSA seems to adsorb onto plane c, whereas LSZ adsorb onto plane a. The differences of the ratio of surface a plane against the total surface area of HA particles and particles size influenced the amount of protein adsorption. The sustained release of BSA and LSZ from HA particles were observed for two weeks.
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