纳米级淡水珍珠粉的制备及其生物相容性初步研究

发布时间：2018-07-20 16:46

【摘要】：目的：通过对球磨制备的纳米级淡水珍珠粉进行动物体内外实验,初步了解其组织相容性和生物安全性,为其在骨修复材料方面的应用研究提供参考实验数据。 方法：采用机械球磨法制备纳米级淡水珍珠粉(立式双轴纳米陶瓷砂磨分散机,微米级淡水珍珠粉与无水乙醇以重量体积比约为1：15的比例),利用激光粒度分析仪(Laser particle size analyzer, LPSA)、扫描电镜(scanning electron microscope, SEM)、 X射线衍射仪(X-ray diffraction,XRD)、透射电镜(transmission electron microscopy, TEM)、凯氏定氮法以及参照GB/T5009.124-2003(食品中氨基酸的测定)等方法对纳米级淡水珍珠粉与微米级淡水珍珠粉进行比较和分析；参照国际公认的(GB/T)16886/ISO10993(医疗器械生物学评价)系列标准,选择细胞毒性试验、溶血试验、皮内刺激试验、急性全身毒性试验等对纳米级淡水珍珠粉进行初步的生物相容性和生物安全性实验研究。 结果：(1)纳米陶瓷砂磨分散机是一种集研磨分散一体的高效球磨机,可以将微米级珍珠粉研磨至纳米级。(2)扫描电镜、X射线衍射法、透射电子显微镜、激光粒度分析仪都可以检测纳米材料粒径,由于它们的检测原理不同以及纳米颗粒间极易发生团聚,因此,在对纳米珍珠粉粒径检测时需要结合不同检测方法进行综合分析。(3)以无水乙醇为液体介质,经湿式球磨法制备的纳米珍珠粉中蛋白质和氨基酸含量没有发生明显变化。(4)细胞毒性试验：纳米珍珠粉-DMEM (dulbecco modified eagle medium)浸提液细胞毒性为Ⅰ级,符合ISO细胞毒性标准。(5)溶血试验：纳米珍珠粉盐水浸提液溶血率1.01%,小于5%,符合ISO溶血标准。(6)皮内刺激试验：纳米珍珠粉盐水浸提液皮内刺激指数为0,皮内无刺激性。(7)急性全身毒性试验：在实验观察期内,实验组和对照组SD大鼠无中毒症状、无不良反应、无死亡,体重增长未见异常；1w后腹腔解剖肉眼观无异常,肝、肾、脾脏病理组织检查无异常。 结论：以无水乙醇为介质,应用湿式球磨法可以将微米级珍珠粉研磨成纳米级,其中蛋白质和氨基酸含量没有发生明显变化；机械球磨法制备的纳米珍珠粉具有良好的组织相容性及生物安全性。图44
[Abstract]:Objective: to investigate the histocompatibility and biological safety of nano-sized freshwater pearl powder prepared by ball milling in vivo and in vitro, and to provide reference data for its application in bone repair materials. Methods: Nano-grade fresh water pearl powder was prepared by mechanical ball milling. The ratio of micron grade fresh water pearl powder to anhydrous ethanol is about 1:15 by weight and volume), laser particle size analyzer (particle size analyzer,), scanning electron microscope (scanning electron microscope,), X-ray diffraction (XRD), transmission electron microscope (transmission electron microscopy, TEM), Kjeldahl nitrogen determination (Kjeldahl) are used. The comparison and analysis of nanometer and micron grade fresh water pearl powder were carried out by the method of GB / T 5009.124-2003 (determination of amino acid in food). Referring to the internationally recognized (GB / T) 16886% ISO 10993 (Biological Evaluation of Medical Devices) series, we select cytotoxicity tests, hemolysis tests, intradermal irritation tests, The biocompatibility and biosafety of nano-sized fresh water pearl powder were studied in acute systemic toxicity test. Results: (1) Nano-ceramic sanding and dispersing machine is a kind of high efficiency ball mill, which can grind micron pearl powder to nanometer scale. (2) scanning electron microscope (SEM) and transmission electron microscope (TEM), X-ray diffraction (SEM), transmission electron microscope (TEM), scanning electron microscope (SEM), transmission electron microscope (TEM), scanning electron microscopy (SEM), Laser particle size analyzer can be used to detect the particle size of nanomaterials. Because of their different detection principles and the easy agglomeration between nanoparticles, the particle size of nanomaterials is easy to be agglomerated. It is necessary to analyze the particle size of nanometer pearl powder in combination with different methods. (3) taking anhydrous ethanol as liquid medium, The content of protein and amino acid in the nanometer pearl powder prepared by wet ball milling method did not change obviously. (4) Cytotoxicity test: the cytotoxicity of the extract of DMEM (dulbecco modified eagle medium) was grade 鈪，

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