中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (25): 3750-3756.doi: 10.3969/j.issn.2095-4344.2016.25.016

• 药物控释材料 drug delivery materials • 上一篇    下一篇

3D打印多孔β-磷酸三钙负载聚乳酸-羟基乙酸共聚物抗结核药物缓释微球复合材料:构建及细胞毒性评价

孟 磊1,2,甄 平2,梁晓燕2   

  1. 1甘肃中医药大学,甘肃省兰州市  7300002解放军兰州军区兰州总医院全军骨科中心,甘肃省兰州市  730050
  • 收稿日期:2016-03-22 出版日期:2016-06-17 发布日期:2016-06-17
  • 通讯作者: 甄平,博士,主任医师,解放军兰州军区兰州总医院全军骨科中心,甘肃省兰州市 730050
  • 作者简介:孟磊,男,1983年生,安徽省蚌埠市人,汉族,甘肃中医药大学在读硕士,主要从事骨与关节损伤研究。
  • 基金资助:

     国家自然科学基金(81371983)

Three-dimensionally printed porous beta-tricalcium phosphate scaffold loading poly(lactic-co-glycolic acid)/anti-tuberculosis drug control-release microspheres: a cytotoxic evaluation

Meng Lei1, 2, Zhen Ping2, Liang Xiao-yan2   

  1. 1Gansu University of Chinese Medicine, Lanzhou 730000, Gansu Province, China; 2Military Orthopedics Center, Lanzhou General Hospital of Lanzhou Military Region of Chinese PLA, Lanzhou 730050, Gansu Province, China
  • Received:2016-03-22 Online:2016-06-17 Published:2016-06-17
  • Contact: Zhen Ping, M.D., Chief physician, Military Orthopedics Center, Lanzhou General Hospital of Lanzhou Military Region of Chinese PLA, Lanzhou 730050, Gansu Province, China
  • About author:Meng Lei, Studying for master’s degree, Gansu University of Chinese Medicine, Lanzhou 730000, Gansu Province, China; Military Orthopedics Center, Lanzhou General Hospital of Lanzhou Military Region of Chinese PLA, Lanzhou 730050, Gansu Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81371983

摘要:

 文章快速阅读:

 

文题释义:
3D打印骨组织工程材料:
在计算机CAD文件数据所创建的控制喷头轨迹的指令下,打印喷头将含有黏结剂和材料粉末的墨水通过层层堆积,最终完成骨组织工程材料的打印。在此过程中,粉末堆积密度和流动性、黏合剂的量、浆料中固体成分的亲水性、打印过程打印喷头移动的速度等多种因素共同影响着打印的分辨率和准确性。
CCK-8法检测材料的细胞毒性:CCK-8检测法是一种新型、简便、客观的细胞毒性检测方法,该试剂中含有WST28,它在电子载体12 methoxy PMS的作用下,被细胞线粒体中的乳酸脱氢酶还原为具有高度水溶性的黄色甲臢染料,所生成的甲臢物的数量与活细胞的数量呈正比,通过实时酶联免疫检测仪所测得的吸光度(A)值简便而准确地进行细胞增殖和毒性分析。与传统的MTT比色法相比,它具有操作更加安全简便、结果灵敏度更高和重现性更好的优点。

 

摘要
背景:
3D打印多孔β-磷酸三钙负载聚乳酸-羟基乙酸共聚物抗结核药物缓释微球复合材料,对细胞尤其是成骨细胞生长和增殖的影响及影响程度尚缺乏可靠的生物医学证据。
目的:构建3D打印多孔β-磷酸三钙负载聚乳酸-羟基乙酸共聚物抗结核药物缓释微球材料,检测其细胞毒性。
方法:利用3D打印技术制备出20个孔径400 μm的多孔β-磷酸三钙材料,随机抽取其中10个负载聚乳酸-羟基乙酸共聚物抗结核药物缓释微球,作为载药支架,其余10个作为非载药支架。将以上两种支架浸提液与成骨细胞共同培养72 h,通过倒置相差显微镜观察细胞形态,采用CCK-8法测得成骨细胞A值,计算细胞相对增殖率,评价支架材料的细胞毒性。
结果与结论:载药支架大小适度,结构规则,孔隙均匀。培养72 h内,载药支架浸提液中的成骨细胞呈长条形或长梭形,可见少量细胞核固缩,表现出轻微中毒表现,细胞毒性分级为1级,无明显细胞毒性。结果表明3D打印多孔β-磷酸三钙负载聚乳酸-羟基乙酸共聚物抗结核药物缓释微球无明显细胞毒性。

 

 ORCID: 0000-0003-3122-4042(甄平)

关键词: 生物材料, 材料相容性, 3D打印, β-磷酸三钙负载抗结核药物缓释微球复合材料, 生物相容性, CCK-8法, 成骨细胞, 相对增殖率, 细胞毒性分级, 国家自然科学基金

Abstract:

BACKGROUND: So far there is a lack of reliable biomedical evidence about the effects of three-dimensionally (3D) printed porous β-tricalcium phosphate (β-TCP) scaffold loading poly(lactic-co-glycolic acid) (PLGA)/anti-tuberculosis drug control-release microspheres on the growth and proliferation of cells, especially osteoblasts.

 
OBJECTIVE: To construct porous β-TCP scaffold loading PLGA/anti-tuberculosis drug control-release microspheres by 3D printing technology and to detect its cytotoxicity.
METHODS: Twenty porous β-TCP scaffolds whose aperture was 400 μm were prepared by 3D printing technology. Ten of these scaffolds were randomly selected for loading PLGA/anti-tuberculosis drug control-release microspheres, and the others were without any drugs. Then the extracts from two groups were cultured with osteoblasts for 72 hours. Afterwards, cell morphology was observed by inverted phase contrast microscope, and the absorbance value was detected using cell counting kit-8 assay. Besides, the relative growth rate of osteoblasts was calculated to evaluate the cytotoxicity of the scaffold.

RESULTS AND CONCLUSION: The drug-loaded scaffold exhibited with moderate size, regular structure and uniform pores. Within 72 hours of culture in the extracts from the drug-loaded scaffold, elongated or fusiform osteoblasts appeared, with less karyopycnosis. Moreover, the drug-loaded scaffold showed slight cytotoxicity, which was classified as grade 1. In conclusion, the 3D-pinted porous β-TCP scaffold loading PLGA/anti-tuberculosis drug control-release microspheres exhibits no obvious cytotoxicity.

 

 

 

Key words: Calcium Phosphates, Antitubercular Agents, Tissue Engineering

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