【摘要】：目的：通過對球磨制備的納米級淡水珍珠粉進行動物體內外實驗,初步了解其組織相容性和生物安全性,為其在骨修復材料方面的應用研究提供參考實驗數(shù)據(jù)。 方法：采用機械球磨法制備納米級淡水珍珠粉(立式雙軸納米陶瓷砂磨分散機,微米級淡水珍珠粉與無水乙醇以重量體積比約為1：15的比例),利用激光粒度分析儀(Laser particle size analyzer, LPSA)、掃描電鏡(scanning electron microscope, SEM)、 X射線衍射儀(X-ray diffraction,XRD)、透射電鏡(transmission electron microscopy, TEM)、凱氏定氮法以及參照GB/T5009.124-2003(食品中氨基酸的測定)等方法對納米級淡水珍珠粉與微米級淡水珍珠粉進行比較和分析；參照國際公認的(GB/T)16886/ISO10993(醫(yī)療器械生物學評價)系列標準,選擇細胞毒性試驗、溶血試驗、皮內刺激試驗、急性全身毒性試驗等對納米級淡水珍珠粉進行初步的生物相容性和生物安全性實驗研究。 結果：(1)納米陶瓷砂磨分散機是一種集研磨分散一體的高效球磨機,可以將微米級珍珠粉研磨至納米級。(2)掃描電鏡、X射線衍射法、透射電子顯微鏡、激光粒度分析儀都可以檢測納米材料粒徑,由于它們的檢測原理不同以及納米顆粒間極易發(fā)生團聚,因此,在對納米珍珠粉粒徑檢測時需要結合不同檢測方法進行綜合分析。(3)以無水乙醇為液體介質,經濕式球磨法制備的納米珍珠粉中蛋白質和氨基酸含量沒有發(fā)生明顯變化。(4)細胞毒性試驗：納米珍珠粉-DMEM (dulbecco modified eagle medium)浸提液細胞毒性為Ⅰ級,符合ISO細胞毒性標準。(5)溶血試驗：納米珍珠粉鹽水浸提液溶血率1.01%,小于5%,符合ISO溶血標準。(6)皮內刺激試驗：納米珍珠粉鹽水浸提液皮內刺激指數(shù)為0,皮內無刺激性。(7)急性全身毒性試驗：在實驗觀察期內,實驗組和對照組SD大鼠無中毒癥狀、無不良反應、無死亡,體重增長未見異常；1w后腹腔解剖肉眼觀無異常,肝、腎、脾臟病理組織檢查無異常。 結論：以無水乙醇為介質,應用濕式球磨法可以將微米級珍珠粉研磨成納米級,其中蛋白質和氨基酸含量沒有發(fā)生明顯變化；機械球磨法制備的納米珍珠粉具有良好的組織相容性及生物安全性。圖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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