Study of cytokine-carrier material by using polymer-film made from artificial lipid coated polysaccharide

人工脂质包覆多糖聚合物膜细胞因子载体材料的研究

• 批准号: 12671917
• 负责人: INOUE Yusuke
• 金额: $ 2.18万
• 依托单位: Fukuoka College of Health Sciences
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671917/
• 关键词: cytokine; BMP; lipid; polysaccharide; polymer film; サイトカン

Sixteen kinds of artificial amphiphilic lipids were prepared from the reaction between n-alkyl alcohols (n-hexyl alcohol, n-octyl alcohol, n-decyl alcohol, n-dodecyl alcohol, n-tetradecyl alcohol, n-hexadecyl alcohol, n-octadecyl alcohol), amino acids (glycine, L-alanine, L-glutamic acid) and p-toluenesulfonic acid. Alginic acid or Heparin in 100 ml of distilled water or 50 wt% ethanolwater solution was sonicated for 5 min. Afterward, one of the prepared artificial amphiphilic lipids in 100 ml of distilled water or 50 wt% ethanol-water solution was added to 100 ml of polysaccharide suspension. The reaction mixture was stirred at 20℃ for three hours, polysaccharide-lipid complexes were collected by centrifugation, and were washed with distilled water or 50 wt% ethanol-water solutions. Finally, the white polysaccharide-lipid complex was freeze-dried. Polysaccharide-lipid complex was dissolved in chloroform /ethanol solution. The solution was evenly dropped on Teflon plate. After solvent was completely vaporated, polysaccharide-lipid complex film was obtained by removing from Teflon plate.Bone morphogenetic protein (BMP) was extracted from bovine demineralized bone matrix with 4 M guanidine hydrochloride.After concentration, an extract solution was dialyzed for several times. Finally aqueous BMP was obtained by gel-filtration. The lipid coated BMP was synthesized from the reaction between aqueous BMP solution and the artificial lipid (Didocyl-N-GluconoL-Glutamate). The polysaccharide-lipid complex films were almost dissolved 3 days after subcutaneous implantation in the backs of rats and showed no inflammatory reactions or inhibition of new tissue formation. Lipid coated BMP or the mixture of the polysaccharide-lipid complex film implanted in the femora of rat showed new bone formation, and no inflammatory reactions or inhibition of new tissue formation.

Sixteen kinds of artificial amphiphilic lipids were prepared from the reaction between n-alkyl alcohols (n-hexyl alcohol, n-octyl alcohol, n-decyl alcohol, n-dodecyl alcohol, n-tetradecyl alcohol, n-octadecyl alcohol), amino acids (glycine, L-alanine, L-glutamic acid) and p-toluenesulfonic acid.将100 mL蒸馏水或50 wt％乙醇溶液中的藻酸或肝素超声检查5分钟。之后，将一种制备的人工两亲性脂质中的一种在100 mL蒸馏水或50 wt％乙醇 - 水溶液中添加到100 mL多糖浆悬浮液中。将反应混合物在20℃加深三个小时，通过离心收集多糖脂质复合物，并用蒸馏水或50 wt％乙醇 - 水溶液洗涤。最后，将白多糖脂质复合物冷冻干燥。将多糖脂质复合物溶于氯仿 /乙醇溶液中。将溶液均匀地放在特氟龙板上。溶液完全蒸发后，通过从Teflon板上去除多糖浆脂质复合膜获得。骨形态发生蛋白（BMP）从牛缝制骨基质中提取，并用4 m鸟苷盐酸盐盐水基质。氢化盐浓度，将提取溶液透析几次。最后，BMP水是通过凝胶过滤获得的。从水溶液溶液和人造脂质（狄昔酮-N-葡萄糖醇 - 谷氨酸）之间的反应中合成了脂质涂层的BMP。皮下植入后3天溶解多糖脂质脂质复合物在大鼠的背部，没有显示出炎症反应或抑制新组织形成。植入大鼠股骨的脂质涂层BMP或多糖脂脂复合膜的混合物显示出新的骨形成，没有炎症反应或抑制新的组织形成。