中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (6): 930-935.doi: 10.3969/j.issn.2095-4344.1546

• 材料生物相容性 material biocompatibility • 上一篇    下一篇

新型三元复合材料纳米羟基磷灰石/聚酰胺66/氧化锆的制备及体外生物相容性

李毓灵1，蒋 科1，陈 路1，蔚 芃1，陈 骞1，谯 波2，蒋电明3   

  1. 1川北医学院附属医院骨科，四川省南充市 637000；2重庆医科大学附属第一医院，重庆市 400016；3重庆医科大学附属第三医院骨与创伤中心，重庆市 401120
  • 收稿日期:2018-09-13 出版日期:2019-02-28 发布日期:2019-02-28
  • 通讯作者: 蒋电明，教授，重庆医科大学附属第三医院骨与创伤中心，重庆市 401120
  • 作者简介:李毓灵，男，1987年生，四川省南充市人，汉族，博士，主要从事生物工程材料研究。
  • 基金资助:

    国家自然科学基金青年基金(NSFC81501876)，项目负责人：谯波；四川省科技厅应用基础项目(2018JY0250)，项目负责人：李毓灵

Preparation and in vitro biocompatibility of a novel ternary biomaterial, yttria-stabilized zirconia reinforced nano-hydroxyapatite/polyamide 66

Li Yuling1, Jiang Ke1, Chen Lu1, Yu Peng1, Chen Qian1, Qiao Bo2, Jiang Dianming3   

  1. 1Department of Orthopedics, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, Sichuan Province, China; 2the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China; 3Bone and Trauma Centre, the Third Affiliated Hospital of Chongqing Medical University, Chongqing 401120, China
  • Received:2018-09-13 Online:2019-02-28 Published:2019-02-28
  • Contact: Jiang Dianming, Professor, Bone and Trauma Centre, the Third Affiliated Hospital of Chongqing Medical University, Chongqing 401120, China
  • About author:Li Yuling, MD, Department of Orthopedics, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, Sichuan Province, China
  • Supported by:

    the National Natural Science Foundation of China for the Youth, No. NSFC81501876 (to QB); the Applied Basic Project of Science & Technology Department of Sichuan Province, No. 2018JY0250 (to LYL)

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摘要：

文章快速阅读：

 

文题释义：
马氏体相变增韧：复合材料的增韧是通过氧化锆马氏体相变来实现的。氧化锆存在三种晶体构型，分别为单斜晶相、四方相及立方相，这3种构型可在不同温度下互相转换。氧化锆与纳米羟基磷灰石的复合是通过高温烧结来实现的，高温可使氧化锆由单斜晶相向四方相转变，而在降温冷却过程中又会发生四方相向单斜晶相的转变，在这个降温转变过程中，氧化锆颗粒产生体积膨胀，同时对纳米羟基磷灰石基体产生压应力，阻止裂纹扩展，即产生应力诱导的相变增韧。
生物相容性：是指机体对生物材料产生反应的一种性能，一般指宿主与材料之间的相容性。生物材料是否能应用于未来，生物相容性是评价的首要指标，生物相容性也决定了材料的性质及用途。研究中根据ISO10993-12:2007标准选用了细胞毒性实验(CCK-8)，细胞与生物材料的结合情况观察(免疫荧光技术)来评价复合材料的生物相容性，以评价验证其未来进一步研究前景。
 
 
背景：纳米羟基磷灰石/聚酰胺66复合材料具有高仿生特性，可通过与宿主骨直接结合来发挥生物活性作用，但其缺乏足够的力学强度。
目的：制备新型三元复合材料纳米羟基磷灰石/聚酰胺66/氧化锆(nano-hydroxyapatite/polyamide 66/ yttria-stabilized tetragonal zirconia，nHA/PA66/YTZ)，验证其力学特性及体外生物相容性。
方法：采用两步法制备三元复合材料nHA/PA66/YTZ，其中纳米羟基磷灰石与氧化锆的质量比分别100∶0、90∶10、80∶20、60∶40。扫描电镜观察复合材料表征，力学测试仪测试其抗弯强度、抗张强度、抗压强度、弹性模量、断裂伸长率等力学参数，评价其力学性能，筛选最佳质量比复合材料，用于以下实验。分别采用细胞培养液(空白对照组)、纳米羟基磷灰石/聚酰胺66材料浸提液(对照组)、nHA/PA66/YTZ材料浸提液(实验组)培养小鼠成骨细胞MC3T3-E1，CCK-8法检测细胞增殖；将纳米羟基磷灰石/聚酰胺66材料(对照组)、nHA/PA66/YTZ材料(实验组)分别与小鼠成骨细胞MC3T3-E1共培养，24 h后采用激光共聚焦显微镜观察MC3T3-E1细胞在复合材料表面的黏附、增殖情况。

结果与结论：①扫描电镜显示，氧化锆晶粒填充了原本纳米羟基磷灰石晶粒之间的空隙，纳米羟基磷灰石/氧化锆均匀分散在聚酰胺66基体中；②生物力学测试显示，纳米羟基磷灰石与氧化锆质量比为60∶40 nHA/PA66/YTZ材料的抗压强度、抗弯强度、抗张强度、断裂伸长率及弹性模量最高，力学性能最优，选择其进行细胞相容性实验；③CCK-8检测显示，随着时间的延长，3组细胞数量逐渐增加，3组间细胞增殖比较无差异；④激光共聚焦显微镜显示，实验组复合材料上的细胞呈现融合、团聚及分层现象，细胞内肌动蛋白丝更多；对照组复合材料上的细胞呈现单层及分散现象，细胞数量与细胞内的肌动蛋白丝较实验组少；⑤结果表明，三元复合材料nHA/PA66/YTZ在体外实验中表现出良好的力学性能、生物安全性及生物相容性。

ORCID: 0000-0003-4659-1013(李毓灵)

 

关键词: 口腔材料, 纳米羟基磷灰石, 氧化钇稳定型氧化锆, 聚酰胺66, 三元复合材料, 肌动蛋白丝

Abstract:

BACKGROUND: Nano-hydroxyapatite/polyamide 66 (nHA/PA66) composite materials possess high bionic properties and exert biological activity by directly combining with host bone, but it lacks sufficient mechanical strength.

OBJECTIVE: To prepare a novel ternary biomaterial composed of nHA/PA66/yttria-stabilized tetragonal zirconia (YTZ), and to investigate its mechanical properties and biocompatibility.
METHODS: The biomaterial was prepared by two-step approach, and the mass ratio of nano-hydroxyapatite to yttria-stabilized tetragonal zirconia was 100:0, 90:10, 80:20, and 60:40, respectively. The characterization of nHA/PA66/YTZ was observed by scanning electron microscope. The mechanical parameters of nHA/PA66/YTZ including bending strength, tensile strength, compressive strength, elastic modulus and breaking elongation were tested to evaluate its mechanical properties. The mechanical properties were evaluated by the mechanical tester to select composite materials with the optimum mass ratio applied to the following experiments. The MC3T3-E1 cells were cultured by the cell-culture medium (blank control group), nHA/PA66 material extract (control group), nHA/PA66/YTZ material extract (experimental group). The cell proliferation was detected by cell counting kit-8 assay. The nHA/PA66 material (control group) and nHA/PA66/YTZ material (experimental group) were respectively co-cultured with MC3T3-E1 cells. The adhesion and proliferation of MC3T3-E1 cells on the surface of composite materials were observed by the laser scanning confocal microscope after 24 hours.
RESULTS AND CONCLUSION: The scanning electron microscope showed that YTZ grains filled the gaps between the original nano-hydroxyapatite grains and the nHA/YTZ was evenly dispersed in the matrix of polyamide 66. The biomechanical test revealed that the compressive strength, bending strength, tensile strength, elongation at break and elastic modulus were the highest at 60:40 of the mass ratio of nHA and YTZ, while the mechanical properties were optimal, which could be chosen for the cell compatibility experiments. Cell counting kit-8 assay showed that there was no significant difference in the cell proliferation among groups as the time expended and the number of cells in each group increased. The laser scanning confocal microscope displayed that the cells showed fusion, agglomeration and stratification and more actin filaments in themselves on nHA/PA66/YTZ composite material. The cells had monolayer and dispersion on nHA/PA66 composite material. The number of cells and actin filaments in cells were less than those in the group of nHA/PA66/YTZ. These results showed that the ternary composite materials of nHA/PA66/YTZ exhibit good mechanical properties, biological safety, and biocompatibility in vitro experiment.  

Key words: Hydroxyapatites, Nylons, Materials Testing, Tissue Engineering

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