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.

以正己醇、正辛醇、正癸醇、正十二醇、正十四醇、正十六醇、正十八醇为原料，甘氨酸、L-丙氨酸、L-谷氨酸为氨基酸，对甲苯磺酸为原料，合成了16种人工两亲性脂质。将海藻酸或肝素在100 ml蒸馏水或50 wt%乙醇水溶液中超声处理5 min。然后，将在100 ml蒸馏水或50 wt%乙醇-水溶液中制备的人工两亲性脂质之一加入到100 ml多糖悬浮液中。将反应混合物在20 ° C下搅拌3小时，通过离心收集多糖-脂质复合物，并用蒸馏水或50重量%乙醇-水溶液洗涤。最后，将白色多糖-脂质复合物冷冻干燥。将多糖-脂质复合物溶解在氯仿/乙醇溶液中。将溶液均匀滴在Teflon板上。用4 M盐酸胍从牛脱钙骨基质中提取骨形态发生蛋白（BMP），提取液经浓缩、多次透析，得到骨形态发生蛋白（BMP）。最后通过凝胶过滤获得水性BMP。将BMP水溶液与人工脂质（Didocyl-N-GluconoL-Glutamate）反应，合成脂质包被的BMP。在大鼠背部皮下植入后3天，多糖-脂质复合物膜几乎溶解，并且没有显示出炎症反应或抑制新组织形成。脂质包被的BMP或多糖-脂质复合物膜的混合物植入大鼠股骨中显示新骨形成，并且没有炎症反应或抑制新组织形成。