Function of Lipid-caoted cytokines

脂质诱导的细胞因子的功能

• 批准号: 11671968
• 负责人: FUKUSHIMA Tadao
• 金额: $ 2.11万
• 依托单位: Fukuoka Dental College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671968/
• 关键词: Artificial lipid; Cytokine; Bone formation; Protein; DNA; Carrier; 生体材料; b-FGF; BMP

Didecyl-N-glucono-L-glutamate as a neutral lipid was prepared in order to coat some cytokines such as b-FGF and BMP.The lipid was allowed to react with b-FGF or BMP in phosphate buffer solution. UV analysis confirmed that obtained solid contained approximately 3% b-FGF and 2% BMP, respectively. The lipid-coated b-FGF and BMP were soluble in chloroform and isopropyl ether. Circular dichroism analysis confirmed that the conformation of lipid-coated b-FGF in organic solution was stable. The lipid-coated b-FGF and BMP were implanted in rat tibia. Photomicrograph revealed that the lipid-coated b-FGF and BMP accelerated new bone formation. L-alanine n-decyl ester (C10-L-Ala)/p-toluenesulfonic acid salt or L-alanine n-lauryl ester (C12-L-Ala) /p-toluenesulfonic acid salt were synthesized as artificial amphiphilic lipids. The DNA/C10-L-Ala and DNA/C12-L-Ala complexes were prepared from the simple mixing of DNA and synthesized amphiphilic lipids to used as new carrier for the lipid-coated cytokines. These complexes were insoluble in water, but soluble in organic solvents such as ethanol and chloroform, and were confirmed to form a double helical B-form structure even in chloroform (CHCl_3)/ethanol (EtOH) solution. The self-standing, water-insoluble DNA-lipid films were prepared by casting from CHCl_3/EtOH solution. The DNA-lipid films were almost dissolved after 3 days of subcutaneous implantation in the backs of rats. There were no inflammatory reactions or an obstacle to new tissue formation. The DNA-lipid films may be suitable materials in the dental and medical fields.

为了包裹成纤维细胞生长因子、骨形态发生蛋白等细胞因子，制备了中性脂质N-葡萄糖酸-L-谷氨酸，并在磷酸盐缓冲溶液中与成纤维细胞生长因子或骨形态发生蛋白反应。UV分析证实，所得固体中分别含有约3%的b-成纤维细胞生长因子和2%的骨形态发生蛋白。脂质包被的b-FGF和BMP可溶于氯仿和异丙醚。圆二色谱分析证实脂膜b-成纤维细胞生长因子在有机溶液中构象稳定。将b-成纤维细胞生长因子和骨形态发生蛋白脂质包埋于大鼠胫骨内。光镜观察显示，脂质包裹的b-成纤维细胞生长因子和骨形成蛋白促进了新骨的形成。合成了L-丙氨酸正十一酯(C10-L-ALA)/对甲苯磺酸盐或L-丙氨酸正十二酯(C12-L-ALA)/对甲苯磺酸盐作为人工两亲性脂类。将合成的两亲性脂类与脱氧核糖核酸进行简单混合，制备了脱氧核糖核酸/C10-L-丙氨酸和脱氧核糖核酸/C12-L-丙氨酸复合体，作为脂膜细胞因子的新载体。这些配合物不溶于水，但可溶于乙醇、氯仿等有机溶剂，在氯仿(CHCl_3)/乙醇(EtOH)溶液中仍能形成双螺旋B型结构。用CHCl_3/EtOH溶液浇铸法制备了自立、不溶于水的DNA-脂膜。大鼠背部皮下埋植3天后，DNA脂膜基本溶解。没有炎症反应，也没有新组织形成的障碍。DNA-脂膜有望成为牙科和医学领域的理想材料。