摘要
目的制备一种新型-βNGF缓释系统,通过Sandwich ELISA法建立-βNGF标准释放曲线及无细胞存在下体外释放曲线,检测缓释系统能否延缓释放-βNGF。方法利用纤维蛋白作为释放系统基质,加入肝素、肝素结合肽、-βNGF及含Ca^2＋的凝血酶制成-βNGF缓释系统,加入24孔组织培养板内,37℃,95%相对湿度,5%CO2培养箱中培育5～10 min,然后每天加500μL/孔TBS液轻轻冲洗,小心吸取冲洗液收集于EP管内,共6d,最后加100μL/孔含纤溶酶的TBS液将基质完全溶解,收集基质溶解液于EP管中,用Sandwich ELISA法测定每天冲洗液含有的-βNGF的量和最后基质溶解液中剩余-βNGF的量,建立-βNGF浓度-时间释放曲线。实验分为2组,实验组：fibrin＋heparin＋peptide＋100 ng/mL-βNGF;对照组：fibrin＋100 ng/mL-βNGF。结果第1天,对照组有98.23%的NGF释放出来,而NGF缓释系统组只释放21.37%,明显少于对照组（P〈0.01）;对照组最后的基质溶解液中无NGF剩余,而实验组仍有48.92%的NGF剩余（P〈0.01）。结论在体外无细胞存在情况下,实验组能明显延缓NGF的释放,延缓时间在6 d以上。
Objective To establish a new β-NGF delivery system for controlled release and the release profile of β-NGF using Sandwich ELISA. Methods The fibrin was selected as matrix and the NGF controlled release system concluding heparin-binding peptides and heparin and β-NGF. The experiment was divided into two groups： Experiment group： fibrin ＋ heparin ＋ peptide ＋ 100 ng/mL β-NGF;Control group：fibrin＋100 ng/mL β-NGF Fibrin matrices were washed every day with 500 μL TBS per wash. The washes from the matrices were collected and stored in -20℃. After 6 days,the matrices were degraded with plasmin. The amount of β-NGF presenting in the washes and remaining in the matrices was quantified using Sandwich ELISA. The release profile of β-NGF was established. Results Without the heparin-binding peptides and heparin,almost all of the β-NGF（98.23%）was released from the matrix within the first day. However,only 21.37% of the β-NGF initially presented in experiment group was released within the first day and 48.92% of the β-NGF still remained in the matrix after 6 days. Conclusion These results demonstrated that the release of β-NGF from β-NGF controlled system is prolonged obviously in vitro.
出处
《实用临床医学（江西）》
CAS
2008年第2期3-6,共4页
Practical Clinical Medicine
关键词
神经生长因子
缓释
纤维蛋白
肝素结合肽
nerve growth factor
controlled release
fibrin
heparin-binding peptide
