釉基质蛋白控释微球的研制及其生物学性能的初步研究

华西口腔医学杂志

釉基质蛋白控释微球的研制及其生物学性能的初步研究

陈发明1,吴织芬1,金　岩2,吴　红3,杜　岩1,王国芳1,聂　鑫2

1.第四军医大学口腔医院　牙周黏膜病科; 2.第四军医大学口腔医学院　组织工程实验中心; 3.第四军医大学基础部　化学系,陕西　西安710032

收稿日期:2005-12-25 修回日期:2005-12-25 出版日期:2005-12-20 发布日期:2005-12-20
通讯作者: 吴织芬,Tel:029-84776322
作者简介:陈发明(1972-),男,安徽人,讲师,博士,现在山东青岛解放军第401医院口腔科工作
基金资助:
“十五”国家科技攻关计划资助项目(2004BA720A26)

Preparation of Enamel Matrix Proteins Controlled Release Microspheres and Their Biological Effects on the Proliferation and Differentiation of Human Periodontal Ligament Cellsin vitro

CHENFa-ming1,WUZhi-fen1,JINYan2,WUHong3,DUYan1,WANG Guo-fang1,NIEXin2

1.Dept.ofPeriodontology and Oral Medicine,College ofStomatology,The Fourth MilitaryMedical University,Xi′an710032,China; 2.Center Lab for Tissue Engineering,College ofStomatology,The Fourth MilitaryMedical University,Xi′an710032,China; 3.Dept.ofChemistry,Faculty ofPreclinical Medicine,The Fourth Military Medical University,Xi′an710032,China

Received:2005-12-25 Revised:2005-12-25 Online:2005-12-20 Published:2005-12-20

摘要/Abstract

摘要： 目的　探讨载釉基质蛋白(EMPs)的右旋糖酐基凝胶微球(dex-MPs)的制备工艺及其理化与生物学性能。方法　采用乳化化学交联技术制备载EMPs的dex-MPs (EMPs-dex-MPs),正交设计法优化制备工艺;观察EMPs-dex- MPs形态和粒径,测定其包封率与载药量;测定微球的溶胀、降解性能,动态透析法检测其体外释药特征及其影响因素;通过EMPs-dex-MPs对体外培养的人牙周膜细胞(PDLCs)增殖和分化的影响,评价其生物学性能。结果　EMPs- dex-MPs形态规则,粒径25μm左右,分布较为均匀;EMPs载药量(32·8±1·2)%,包封率(78·9±1·0)%。体外释药实验表明EMPs-dex-MPs中80%的EMPs在前20 d释放,在葡聚糖酶存在下40 d左右可以完全降解。与单纯EMPs 相比,EMPs-dex-MPs能持续促进PDLCs的增殖和分化达12 d左右。结论　dex-MPs作为活性生长因子载体,具有确定的缓释作用,并可以通过制备工艺的改变控制其释药行为;EMPs-dex-MPs作为EMPs的一种新的给药方式,比传统给药方式可以更加有效地促进牙周组织相关细胞的增殖与分化。

关键词: 釉基质蛋白类, 右旋糖酐, 微球体, 控制释放

Abstract: Objective　To prepare enamel matrix proteins (EMPs) loaded dextran-based hydrogel microspheres (EMPs-dex- MPs), and to evaluate their EMPs controlled release property and their biological effects on the proliferation and differentiation of human periodontal ligament cells (PDLCs)in vitro.Methods　Using dimethylbenzene as the oil phase, EMPs-dex-MPs were achieved by emulsion-chemical crosslinking technique. The process of the recombination preparation was optimized by orthogonal factorization method. The configuration and size of EMPs-dex-MPs were determined by scanning electron microscope. The EMPs loading content and encapsulation rate of EMPs-dex-MPs, and their biodegradation characteristic were studied by routine analysis methods. Dynamic dialysis method was used to determine the release characteristic of EMPs-dex-MPsin vitroand its influencing factors. The proliferation of cultured PDLCswas measured byMTTmethod and the differentiation of PDLCswas measured by their alkaline phosphatase (ALP) activities.Results　The results showed that EMPs-dex-MPswere homogenous and stable with the av- erage diameter 25μm, and the EMPs loading content was (32·8±1·2)%, the encapsulation rate was (78·9±1·0)%. Under 9% physiological saline solution contained a very thimbleful quantity of dextranase EMPs-dex-MPs could be biodegraded complete- ly during about 40 days. Theinvitroexperiments showed that about 80% of EMPs could be released out in 20 days. Using EMPs- dex-MPs could enhance the proliferation responses and ALP activities of PDLCs more than 12 days.Conclusion　As a new sus- tained release system of growth factors,the dex-MPs is stable, workable and biodegradable. EMPs-dex-MPs, whose drug release can be controlled by preparation technique, may be more effective in promoting periodontal tissue regeneration.

Key words: enamel matrix proteins, dextran, microsphere, controlled release

陈发明;吴织芬;金岩;吴红;杜岩;王国芳;聂鑫. 釉基质蛋白控释微球的研制及其生物学性能的初步研究[J]. 华西口腔医学杂志.

CHENFa-ming1,WUZhi-fen1,JINYan2,WUHong3,DUYan1,WANG Guo-fang1,NIEXin2. Preparation of Enamel Matrix Proteins Controlled Release Microspheres and Their Biological Effects on the Proliferation and Differentiation of Human Periodontal Ligament Cellsin vitro[J]. West China Journal of Stomatology.