中国组织工程研究

中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (6): 896-901.doi: 10.3969/j.issn.2095-4344.0065

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

混酸回流时间对大内径多壁碳纳米管纯化及生物相容性的影响

孟  艾1，王  剑2，隋  磊1
  

  1. 1天津医科大学口腔医院，天津市  300070；2四川大学华西口腔医院，四川省成都市  610041
  • 收稿日期:2017-09-01 出版日期:2018-02-28 发布日期:2018-02-28
  • 通讯作者: 隋磊，主任医师，天津医科大学口腔医院修复科，天津市 300070
  • 作者简介:孟艾，女，1989年生，天津市人，汉族，2015年天津医科大学毕业，硕士，医师，主要从事材料方面研究。
  • 基金资助:
    天津市自然科学基金重点项目(16JCZDJC32800)

Effect of mixed acid reflux time on purification and biocompatibility of large-inner-diameter multi-walled carbon nanotubes

Meng Ai1, Wang Jian2, Sui Lei1 
  

  1. 1Hospital of Stomatology, Tianjin Medical University, Tianjin 300070, China; 2West China Hospital of Stomatology, Sichuan University, Chengdu 610041, Sichuan province, China
  • Received:2017-09-01 Online:2018-02-28 Published:2018-02-28
  • Contact: Sui Lei, Chief physician, Hospital of Stomatology, Tianjin Medical University, Tianjin 300070, China
  • About author:Meng Ai, Master, Physician, Hospital of Stomatology, Tianjin Medical University, Tianjin 300070, China
  • Supported by:
    Tianjin Natural Science Foundation, No. 16JCZDJC32800

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摘要：

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文题释义：
混酸回流：即按体积比3∶1将浓硫酸与浓硝酸混合于容器内，置于油浴恒温加热，并连接冷凝回流装置及尾气吸收装置，以防止气体产物中的二氧化氮等对空气的污染。
大内径多壁碳纳米管：较常规碳纳米管而言，大内径多壁碳纳米管管内容纳空间大，可负载和转运更多生物分子；而且，大内径多壁碳纳米管外壁表面积较大，能够接枝多种生物分子，充分满足改性要求。但是，商品化大内径多壁碳纳米管因受生产方法限制，在其中存在大量无定形碳及金属催化剂等杂质颗粒，这些杂质粒子对细胞易产生毒性作用，导致碳管不能直接作为药物载体等应用于生物医学领域；同时，原始大内径多壁碳纳米管管壁表面具有高度疏水性，碳管间存在强大的范德华力，易缠绕团聚，导致分散性及生物相容性差，也严重限制了其在生物医学中的应用。
 
背景：商品化大内径多壁碳纳米管存在大量杂质颗粒，易对细胞产生毒性作用，且由于其管壁表面具有高度疏水性，碳管易缠绕团聚，导致分散性及生物相容性差。因此，采用适当的方法对其进行纯化改性，是开发其生物医学应用潜能所要解决的首要问题。
目的：探索不同混酸回流时间对大内径多壁碳纳米管纯化及生物相容性的影响。
方法：首先通过高温煅烧、盐酸酸洗预处理大内径多壁碳纳米管，并采用不同混酸回流时间(1，2，4 h)对大内径多壁碳纳米管进行纯化处理，通过表面形貌和分散性两个指标筛选出纯化效果较好的混酸回流时间，优化制备工艺并进行表征。采用不同质量浓度(5，10，20，40，80 mg/L)的原始大内径多壁碳纳米管悬液与不同混酸回流时间处理的不同质量浓度的(5，10，20，40，80 mg/L)纯化大内径多壁碳纳米管悬液分别培养小鼠成纤维细胞L929及人舌癌细胞CAL-27，72 h后，CCK-8法检测细胞增殖。
结果与结论：①随着混酸回流时间的延长，大内径多壁碳纳米管长度减小，分散性提高，但回流2，4 h时碳管管壁破坏明显，尤其回流4 h时碳管破坏严重，管径出现大小不均现象；混酸回流1 h后，碳管基本结构形态未发生改变，管壁表面未见明显杂质颗粒，管间具有较好的分散性；②在培养小鼠成纤维细胞L929时，在相同质量浓度下，原始大内径多壁碳纳米管悬液组的细胞存活率低于纯化大内径多壁碳纳米管悬液组；在10-80 mg/L范围内，混酸回流处理1 h的纯化大内径多壁碳纳米管悬液组细胞存活率在90%以上，高于其余各组(P < 0.05)；③在培养人舌癌细胞CAL-27时，在相同质量浓度下，原始大内径多壁碳纳米管悬液组的细胞存活率低于纯化大内径多壁碳纳米管悬液组；在20-80 mg/L范围内，混酸回流处理1 h的纯化大内径多壁碳纳米管悬液组细胞存活率在90%以上，高于其余各组(P < 0.05)；④结果表明，混酸回流处理1 h可有效纯化大内径多壁碳纳米管，降低其细胞毒性。

关键词: 碳纳米管, 大内径多壁碳纳米管, 混酸回流, 纯化, 杂质颗粒, 分散性, 生物相容性, 生物材料, 纳米材料

Abstract:

BACKGROUND: A mass of debris particles that can lead to cytotoxicity exist in commercial large-inner-diameter multi-walled carbon nanotubes (LID-MWCNTs). Because of the high hydrophobicity on the surface of the tube wall, the carbon tubes can be twisted and agglomerated, resulting in the low dispersion and poor biocompatibility. Therefore, to explore the effective methods of purifying and modifying LID-MWCNTs is the primary problem to develop its potency in the biomedical application.
OBJECTIVE: To explore the effects of different mixed acid reflux time on LID-MWCNTs purification and biocompatibility.
METHODS: After pretreatment with high temperature calcinations and hydrochloric acid pickling, LID-MWCNTs were purified under different time of mixed acid reflux time (1, 2, 4 hours). The mixed acid reflux time for best purification was chosen based on surface morphology and dispersibility, so as to optimize preparation technology and observe the characterization. The L929 cells and CAL-27 cells were treated with different concentrations of raw LID-MWCNTs (5, 10, 20, 40, 80 mg/L) and purified LID-MWCNTs by mixed acid reflux (5, 10, 20, 40, 80 mg/L). After 72 hours, cell counting kit-8 assay was employed to test the proliferation of L929 cells and CAL-27 cells.
RESULTS AND CONCLUSION: (1) With the time of mixed acid reflux, the length of LID-MWCNTs was decreased, and the dispersion was improved. However, the external surface of the tubes after mixed acid reflux 2 and 4 hours were destroyed obviously. Especially after mixed acid reflux 4 hours, the tubes were destroyed seriously and the diameter of tubes was not uniform. However, after mixed acid reflux 1 hour, the fundamental structure and morphology of the tubes were not changed, the debris particles were undetected on the tube wall surface, and the tubes had the good dispersion. (2) Under the same concentration, the survival rate of L929 cells in the raw LID-MWCNTs group was lower than that in the purified LID-MWCNTs group. At the concentration of 10-80 mg/L, the survival rate of L929 cells in the group of mixed acids reflux 1 hour was up to 90%, higher than that in the other groups (P < 0.05). (3) Under the same concentration, the survival rate of CAL-27 cells in the raw LID-MWCNTs group was lower than that in the purified LID-MWCNTs. At the concentration of 20-80 mg/L, the survival rate of CAL-27 cells in the group of mixed acids reflux 1 hour was up to 90%, higher than that in the other groups (P < 0.05). These results revealed that the raw LID-MWCNTs were purified effectively after mixed acid reflux 1 hour, and the cytotoxicity was decreased.

Key words: Biocompatible Materials, Nanotubes, Carbon, Tissue Engineering

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