Hideaki Nagahama

Preparation and characterization of novel β-chitin-hydroxyapatite composite membranes for tissue engineering applications

Beta-chitin is a biopolymer principally found in shells of squid pen. It has the properties of biodegradability, biocompatibility, chemical inertness, wound healing, antibacterial and anti-inflammatory activities. Hydroxyapatite (HAp) is a natural inorganic component of bone and teeth and has osteoconductive property. In this work, beta-chitin-HAp composite membranes were prepared by alternate soaking of beta-chitin membranes in CaCl2 (pH 7.4) and Na2HPO4 solutions for 2 h in each solution. After 1, 3 and 5 cycles of immersion, beta-chitin membranes were characterized using the SEM, FT-IR, EDS and XRD analyses. The results showed the presence of apatite layer on surface of beta-chitin membranes, and the amounts of size and deposition of apatite layers were increased with increasing number of immersion cycles. Human mesenchymal stem cells (hMSCs) were used for evaluation of the biocompatibility of pristine as well as composite membranes for tissue engineering applications. The presence of apatite layers on the surface of beta-chitin membranes increased the cell attachment and spreading suggesting that beta-chitin-HAp composite membranes can be used for tissue engineering applications.

Synthesis of phosphorylated chitosan by novel method and its characterization

Chitosan a natural based polymer is non-toxic, biocompatible and biodegradable. Chemical modification of chitosan to generate new bifunctional materials and finally would bring new properties depending on the nature of the group introduced. In our present study, we prepared phosphorylated chitosan (P-chitosan) by using H(3)PO(4)/P(2)O(5)/Et(3)PO(4)/hexanol method. From our present method, we got high yield and high degree of substitution (DS). The prepared P-chitosan (DS-1.18) was characterized by FT IR, (13)C NMR, (31)P NMR, elemental, XRD, TGA, DTA and SEM studies. After the phosphorylation, the solubility of the polymer was improved. The P-chitosan showed less thermal stability and crystallinity than the chitosan. It was due to the phosphorylation.

Synthesis, characterization and bioactivity studies of novel β-chitin scaffolds for tissue-engineering applications

Chitin is a biopolymer and it is non-toxic, biodegradable and biocompatible. Chitin has many potential industrial applications because of its abundance, biodegradability, non-toxicity, chemical inertness. beta-Chitin scaffolds were prepared by using saturated calcium chloride alcoholic solution (CaCl(2).6H(2)O/EtOH) and then followed by dialysis with lyophilization. The prepared beta-chitin scaffolds were characterized by FT-IR, scanning electron microscopy (SEM) and thermogravimetric (TGA). The preliminary bioactivity studies of beta-chitin scaffolds were studied by using simulated body fluid (SBF) solution for 7, 14 and 21 days. We also immersed the beta-chitin scaffolds in saturated aqueous CaCl(2) and Na(2)HPO(4) solution over 12h. After 7, 14 and 21 days, the scaffolds were characterized by SEM and FT-IR studies. The SEM studies showed that there is a calcium phosphate layer in the surface as well as in the cross-section of beta-chitin scaffolds. It seems that the beta-chitin scaffolds are useful in the tissue-engineering field.

Preparation, characterization, bioactive and cell attachment studies of α-chitin/gelatin composite membranes

The chitin/gelatin composite membranes were prepared by mixing of chitin hydrogel with gelatin. The prepared composite membranes were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD), mechanical, swelling, enzymatic degradation and thermal studies. The XRD pattern of the chitin/gelatin composite membranes showed almost the same pattern as alpha-chitin. The bioactivity studies of these chitin/gelatin membranes were carried out with the simulated body fluid solution (SBF) for 7, 14 and 21 days followed by the characterization with the scanning electron microscopy (SEM) and Energy Dispersive Spectrum (EDS) studies. The SEM and EDS studies confirmed the formation of calcium phosphate layer on the surface of chitin/gelatin membranes. Biocompatibility of the chitin/gelatin membrane was assessed using human MG-63 osteoblast-like cells. After 48 h of incubation, it was found that the cells had attached and completely covered the membrane surface. Thus, the prepared chitin/gelatin membranes are bioactive and are suitable for cell adhesion suggesting that these membranes can be used for tissue-engineering applications.

Influence of amide content on the crystal structure of chitin

Abstract The crystalline structure of chitin and chitosan was investigated by X-ray diffraction (XRD). Chitosan filaments were prepared using a saturated solution of calcium chloride dihydrate in methanol (CaCl2-MeOH) as a mild solvent and the study focused on the interaction between the degree of acetylation (DA) and crystallinity. There were pronounced differences in the XRD patterns for specimens with DA values between 44.2% and 52.2%. We suggest that the crystalline structure changes from an anhydrous-type chitosan to an α-chitin type without any additives. CaCl2-MeOH is a good solvent for chitin and a poor solvent for chitosan and we found that it can regulate the distribution of N-acetyl glucosamine and glucosamine between amorphous and crystalline regions.