The basic research on functionalization of the natural polysaccharide and application to biomedical materials.

天然多糖功能化及其在生物医用材料中的应用基础研究。

• 批准号: 14350495
• 负责人: ABE Koji
• 金额: $ 9.41万
• 依托单位: Shinshu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14350495/
• 关键词: Regenerative Medicine; Artificial Organ; Biofiber; Extracellular Matrix; Cartilage Cell; Polysaccharide; Polyelectrolyte Complex; Osteoblast; 人工臓器; 胚性幹細胞; ストレス応答; 細胞内シグナル伝達

In this investigation, we have examined the control of cell specific functions (proliferation and differentiation) of the hard tissue cells (bone and chondrocyte) by polyelectrolyte complex (PEC), which are composed of polysaccharides.Osteoblasts cultured on PEC containing sulfated polysaccharide (SPEC) were proliferated similar to tissue culture dish (TCD) and exhibited spreading form. On the other hand, the proliferation of cell cultured on PEG containing carboxymethylated polysaccharide (OPEC) coated dishes was slower than those on TCD and exhibited aggregate form. Expression of ALPase activity and deposition of calcium, which were initial differentiation markers of calcification, were observed mainly at the part of cell aggregate. Further, expression of mRNA of osteoblastic differentiation markers (osteocalcin and osteopontin) were observed on OPEC earlier than on SPEC and TCD. N-cadherin, cell-cell adhesion protein, expression was strong and localized at the cell-cell interfaces in the cell aggregates.Chondrocyte (ATDC5) cultured on SPEC grew well in the similar manner to TCD with spreading form. On the other hand, the growth of ATDC5 on OPEC was remarkably suppressed and exhibited cell aggregate form. The sizes of these aggregates were made bigger with time. These phenomena are well coincident with the culture of osteoblasts on these PECs. Secretion of chondrocyte matrix and expression of mRNA of typeII collagen were promoted on the OPEC.These results suggested that the cell functions, proliferation and differentiation, are mainly controlled by the chemical structures of anionic polysaccharides ; the existence of carboxylic acid residues induces the differentiation and sulfonic acid residues promotes the proliferation of cells.In conclusion, polysaccharides-type PEC are excellent biomaterials for hard tissue cell culture.

在这项研究中，我们研究了由多糖组成的多电解质复合物（PEC）对硬组织细胞（骨和软骨细胞）的细胞特异性功能（增殖和分化）的控制。在含硫酸酸化多糖（SPEC）的PEC上培养成骨细胞，其增殖与组织培养皿（TCD）相似，呈扩散形态。另一方面，在含有羧甲基化多糖（OPEC）的PEG包被培养皿上培养的细胞增殖速度比在TCD培养皿上培养的细胞慢，并呈现聚集形态。ALPase活性的表达和钙的沉积是钙化的初始分化标志，主要在细胞聚集部分观察到。此外，成骨细胞分化标志物（骨钙素和骨桥蛋白）mRNA在OPEC的表达早于SPEC和TCD。N-cadherin是一种细胞-细胞粘附蛋白，在细胞聚集体中表达强烈，并定位于细胞-细胞界面。在SPEC上培养的软骨细胞（ATDC5）生长良好，与TCD相似，呈扩散形式。另一方面，ATDC5在OPEC上的生长受到明显抑制，呈现细胞聚集形态。这些聚集体的大小随着时间的推移而变大。这些现象与这些pec上的成骨细胞培养很好地吻合。软骨细胞基质的分泌和ii型胶原mRNA的表达均得到促进。这些结果表明，阴离子多糖的化学结构主要控制着细胞的功能、增殖和分化；羧酸残基的存在诱导细胞分化，磺酸残基的存在促进细胞增殖。综上所述，多糖型PEC是培养硬组织细胞的优良生物材料。

项目成果

Nakamura, A.: "A rapid extraction procedure of human hair proteins and identification of phosphorylated species"Biol. Pharm. Bull.. 25(5). 569-572 (2002)

Nakamura, A.：“人发蛋白的快速提取程序和磷酸化物质的鉴定”Biol。

ヒト由来の爪タンパク質の抽出とフィルム

人源指甲蛋白的提取和成膜

• 发表时间: 2005
• 期刊: 高分子論文集 62
• 作者: 藤井 敏弘

ファイバー工学 原子から感性まで紡ぐ21世紀の繊維

纤维工程：21 世纪纺织品从原子到情感

• 发表时间: 2005
• 作者: 阿部 康次(白井汪芳;山浦和男[編])
• 通讯作者: 山浦和男[編])

羊毛タンパク質の抽出とコーティング繊維の新規作製法

提取羊毛蛋白及生产涂层纤维的新方法

• 发表时间: 2003
• 期刊: 高分子論文集 60
• 作者: Y.Tsujii;藤井 敏弘
• 通讯作者: 藤井 敏弘

The response of normal human osteoblasts to anionic polysaccharide polyelectrolyte complexes

• DOI: 10.1016/j.biomaterials.2005.01.041
• 发表时间: 2005-09-01
• 期刊: BIOMATERIALS
• 影响因子: 14
• 作者: Nagahata, M;Nakaoka, R;Tsuchiya, T
• 通讯作者: Tsuchiya, T