中国组织工程研究

中国组织工程研究 ›› 2014, Vol. 18 ›› Issue (52): 8405-8408.doi: 10.3969/j.issn.2095-4344.2014.52.008

• 纳米生物材料 nanobiomaterials • 上一篇    下一篇

聚羟基丁酸-羟基辛酸共聚酯载生长因子的缓释纳米微球

王晓东1，张永红2   

  1. 1山西医科大学骨外科，山西省太原市 030001；2山西医科大学第二临床医学院，山西省太原市 030001
  • 修回日期:2014-09-29 出版日期:2014-12-17 发布日期:2014-12-17
  • 通讯作者: 张永红，博士，主任医师，山西医科大学第二临床医学院，山西省太原市 030001
  • 作者简介:王晓东，男，1983年生，山西省忻州市人，汉族，2011年山西医科大学毕业，硕士，医师，主要从事组织工程方面的研究。

Preparation and performance of recombinant human bone morphogenetic protein-2-poly(hydroxybutyrate-co-hydroxyoctanoate) nanospheres

Wang Xiao-dong1, Zhang Yong-hong2   

  1. 1Department of Orthopedics, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China; 2Second Hospital of Shanxi Medical University, Taiyuan 030001, Shanxi Province, China
  • Revised:2014-09-29 Online:2014-12-17 Published:2014-12-17
  • Contact: Zhang Yong-hong, M.D., Chief physician, Second Hospital of Shanxi Medical University, Taiyuan 030001, Shanxi Province, China
  • About author:Wang Xiao-dong, Master, Physician, Department of Orthopedics, Shanxi Medical University, Taiyuan 030001, Shanxi Province, China

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摘要：

背景：骨形态发生蛋白2可增加软骨细胞和祖细胞基质的产生，可增强组织金属蛋白酶抑制因子1、sox9基因、Ⅱ型胶原以及聚集蛋白聚糖的表达，具有诱导间充质细胞迁徙、增殖、分化，最终促使软骨、骨形成的作用。

目的：制备重组人骨形态发生蛋白2聚羟基丁酸-羟基辛酸共聚酯纳米微球系统纳米微球缓释系统，观察微球生长因子形态和粒径分布、载药量、包封率、体外缓释时间及生物活性。
方法：采用复乳-干燥法制备重组人骨形成蛋白2聚羟基丁酸-羟基辛酸共聚酯纳米微球系统，体外分离培养猪软骨细胞。实验分为3组：第1组培养液不添加药物做为对照；第2组培养液中添加含20 μg/L重组人骨形态发生蛋白2；第3组培养液中添加20 μg/L重组人骨形态发生蛋白2聚羟基丁酸-羟基辛酸共聚酯纳米微球系统纳米微球；其中在第2组和第3组又将重组人骨形态发生蛋白2和重组人骨形态发生蛋白2聚羟基丁酸-羟基辛酸共聚酯纳米微球系统纳米微球分别设为55，100 μg/L两种的有效浓度，采用MTT法检测微球对软骨细胞增殖的影响，模拟体内条件观察纳米微球的体外缓释性及生物活性。

结果与结论：该纳米微球表面光滑圆整，球体大小均匀，粒径为231-415 nm，扫描电镜平均粒径323 nm。微球的包封率和载药量分别为(79.63±0.16)%和(1.92±0.16)%。根据15 d内对重组人骨形态发生蛋白2聚羟基丁酸-羟基辛酸共聚酯纳米微球的体外释药的观察，保持持续释放重组人骨形态发生蛋白2，且释放的浓度呈增长水平。该微球缓释系统有生物活性，能显著促进猪软骨细胞的增殖，其效应高于单纯重组人骨形态发生蛋白2的效应。提示重组人骨形态发生蛋白2聚羟基丁酸-羟基辛酸共聚酯纳米微球缓释系统包封率、载药量、体外释药性以及生物活性符合纳米微球的一般规律，能够满足相应的软骨缺损修复要求。


中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程


全文链接：

关键词: 生物材料, 缓释材料, 聚羟基丁酸-羟基辛酸共聚酯, 生物材料与纳米技术, 重组人骨形态发生蛋白2, 骨形态发生蛋白2, 纳米微球, 包封率, 载药量, 生物活性, 组织工程

Abstract:

BACKGROUND:Bone morphogenetic protein-2 can increase the production of chondrocytes and progenitor cell matrix, enhance tissue inhibitor of metalloproteinase-1, sox9, type II collagen and aggrecan expressions, with the induction of mesenchymal cell migration, proliferation, and differentiation, leading to cartilage and bone formation.

OBJECTIVE: Using the biodegradable copolymer of poly(hydroxybutyrate-co-hydroxyoctanoate)-based materials to fabricate recombinant human bone morphogenetic-2-poly(hydroxybutyrate-co-hydroxyoctanoate) sustained release nanospheres and to investigate its morphology, particle distribution, drug loading, encapsulation efficiency, in vitro release time, and bioactivity.
METHODS: The recombinant human bone morphogenetic-2-poly(hydroxybutyrate-co-hydroxyoctanoate) sustained release nanospheres were prepared by multiple emulsion volatilizing method. Porcine chondrocytes were isolated and cultured in vitro. There were three groups: group 1, no drugs were added into the medium serving as control group; group 2, 20 μg/L recombinant human bone morphogenetic-2 was added; group 3, 20 μg/L recombinant human bone morphogenetic-2-poly(hydroxybutyrate-co-hydroxyoctanoate) sustained release nanospheres were added. The effective concentration was 55 μg/L for recombinant human bone morphogenetic-2 in group 2, and 100 μg/L for recombinant human bone morphogenetic-2-poly(hydroxybutyrate-co-hydroxyoctanoate) sustained release nanospheres in group 3. Proliferative capacity of chondrocytes was detected using 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide assay. In vitro release profile and biological activity were observed under simulated in vivo conditions.
RESULTS AND CONCLUSION: The surface of nanospheres was smooth and rounded, and the nanosheres had uniform size and particle size of 231-415 nm. Under scanning electron microscope, the average particle size was

323 nm. Microsphere encapsulation efficiency and drug loading were (79.63±0.16)% and (1.92±0.16)%, respectively. Within 15 days, in vitro release of recombinant human bone morphogenetic protein-2 from recombinant human bone morphogenetic-2-poly(hydroxybutyrate-co-hydroxyoctanoate) sustained release nanospheres lasted and increased. The nanospheres had biological activity and could promote chondrocyte proliferation. Biological activity of recombinant human bone morphogenetic-2 was lower than that of the recombinant human bone morphogenetic-2-poly (hydroxybutyrate-co-hydroxyoctanoate) sustained release nanospheres. It prompts that the encapsulation efficiency, drug loading, in vitro release and biological activity of the recombinant human bone morphogenetic-2- poly(hydroxybutyrate-co-hydroxyoctanoate) sustained release nanospheres are basically consistent with the general requirements of cartilage tissue engineering.


中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程


全文链接：

Key words: biocompatible materials, bone morphogenetic proteins, nanotechnology, nanospheres

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