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聚醚胺ED900-单宁酸构建表面微结构化复合涂层及其细胞相容性的研究

韩健美 夏雨祥 程昉 何炜

韩健美, 夏雨祥, 程昉, 等. 聚醚胺ED900-单宁酸构建表面微结构化复合涂层及其细胞相容性的研究[J]. 复合材料学报, 2022, 40(0): 1-11
引用本文: 韩健美, 夏雨祥, 程昉, 等. 聚醚胺ED900-单宁酸构建表面微结构化复合涂层及其细胞相容性的研究[J]. 复合材料学报, 2022, 40(0): 1-11
Jianmei HAN, Yuxiang XIA, Fang CHENG, Wei HE. Construction of microstructured composite coatings based on polyetheramine ED900-tannic acid and cytocompatibility study[J]. Acta Materiae Compositae Sinica.
Citation: Jianmei HAN, Yuxiang XIA, Fang CHENG, Wei HE. Construction of microstructured composite coatings based on polyetheramine ED900-tannic acid and cytocompatibility study[J]. Acta Materiae Compositae Sinica.

聚醚胺ED900-单宁酸构建表面微结构化复合涂层及其细胞相容性的研究

基金项目: 国家自然科学基金(31971255)
详细信息
    通讯作者:

    何炜,博士,教授,博士生导师,研究方向为生物医用材料  E-mail: wlhe@dlut.edu.cn

  • 中图分类号: TB332

Construction of microstructured composite coatings based on polyetheramine ED900-tannic acid and cytocompatibility study

  • 摘要: 为改善医用材料的生物相容性并赋予材料表面一定的生物学功能,本研究以单宁酸TA和聚醚胺ED900为组装单元,通过层层自组装的方法制备了复合涂层。采用纳米粒度仪、ζ电位分析仪、紫外分光光度计、红外光谱、QCM-D、扫描电子显微镜等仪器对ED900-TA复合溶液行为及复合涂层的理化性质进行表征。通过细胞实验考察了涂层对细胞行为的影响。利用DPPH法和FRAP法评价了涂层的抗氧化性。最后,分别通过琼脂糖插入实验和细胞培养液浸泡研究涂层的稳定性。结果表明,ED900-TA涂层具有良好的细胞相容性和抗氧化性,表面的微结构呈现调控亲/憎细胞的能力。此外,涂层在模拟植入的过程未出现脱落。并在细胞培养条件下,涂层形貌在21天的评价周期内无显著变化。该复合涂层为生物材料表面多功能化提供了新思路。

     

  • 1  TA (a)和ED900 (b) (x+z ~6; y ~12.5) 化学结构式

    1.  Chemical structures of TA (a) and ED900 (b) (x+z ~6; y ~12.5)

    图  1  ED900-TA水溶液复合;(a, b) 2 mg/mL ED900 与2 mg/mL TA等体积混合后pH 值对混合物粒径(a),ζ电位及浊度(b)影响;(c, d) 2 mg/mL ED900(pH 7)与2 mg/mL TA(pH 5)等体积混合时间的影响;(e, f) ED900和TA等体积混合后TA溶液浓度的影响

    Figure  1.  Complexation of ED900-TA in solution; (a, b) Effects of pH on particle size (a) and ζ potential, and turbidity of mixtures (b) of equal volume of 2 mg/mL ED900 with 2 mg/mL TA; (c, d) Effects of time on the mixtures from equal volume combination of 2 mg/mL ED900 (pH 7) and 2 mg/mL TA (pH 5); (e, f) Effects of TA concentration on the mixtures from of equal volume combination of ED900 and TA

    图  2  (a) Urea或SDS影响下ED900-TA 混合物的宏观状态及浊度随时间的变化;(b)各样品的紫外光谱

    Figure  2.  (a) Macroscopic state and turbidity changes of ED900-TA mixture as a function of time under the influence of urea or SDS; (b) UV-vis spectra of various samples

    图  3  (a) 石英基底表面组装ED900-TA八个循环过程的紫外光谱;(b) (ED900-TA)30,TA 和 ED900的红外光谱

    Figure  3.  (a) UV-vis study of assembly process for eight cycles of ED900-TA on quartz substrate; (b) FTIR spectra of (ED900-TA)30, TA and ED900

    图  4  QCM-D研究ED900-TA组装过程中频率及耗散随时间的变化,箭头表示冲洗步骤 (插图:净频率随循环周期的变化)

    Figure  4.  QCM-D study frequency and dissipation changes as a function of time during assembly of ED900-TA, arrows indicate rinsing steps (inset: net change in frequency as a function of assembly cycles)

    ΔF—Frequency change; ΔD—Dissipation change

    图  5  (ED900-TA)n (n = 4, 8, 16, 20)涂层的形貌表征:(a) 水合条件下光学显微镜图像及粒径分析;(b) 干燥条件下SEM图像及粒径分析

    Figure  5.  Surface morphology of (ED900-TA)n (n = 4, 8, 16, 20) coatings: (a) Optical microscope images and size analysis of hydrated samples; (b) SEM images and size analysis of dry samples

    图  6  (ED900-TA)n (n = 4, 8, 16, 20)样品的水接触角(a)与水滴照片及表面TA浓度量化(b)

    Figure  6.  (a) Water contact angles and water droplets photos of (ED900-TA)n (n = 4, 8, 16, 20) samples; (b) Quantification of surface concentrations of TA.

    图  7  硅片及ED900-TA涂层上培养24h的L929细胞的活/死染色结果(活细胞:绿色;受损细胞:红色)

    Figure  7.  Live/dead staining of L929 cells on silicon and ED900-TA coatings after 24 h culture (viable cells: green; damaged cells: red)

    图  8  L929细胞在ED900-TA涂层的响应:(a) 24 h时细胞SEM和荧光显微镜图;(b, c) 各个样品表面细胞在不同培养时间点的荧光显微镜图像和细胞数量化统计图 (平均值 ± std,n = 4,* p < 0.05,** p < 0.01)

    Figure  8.  L929 cell response of ED900-TA coating: (a) SEM and fluorescent images of cells cultured for 24 h; (b, c) Fluorescent images and cell number quantification at various time points (average ± std, n = 4, * p < 0.05, ** p < 0.01)

    图  9  PC12细胞在ED900-TA涂层的响应:(a) 24 h时的细胞SEM和荧光显微镜图; (b, c) 各个样品表面细胞在不同培养时间点的荧光显微镜图像和细胞数量化统计图(平均值 ± std,n = 4,** p < 0.01,*** p < 0.001)

    Figure  9.  PC12 cell response of ED900-TA coating: (a) SEM and fluorescent images of cells cultured for 24 h; (b, c) Fluorescent images and cell number quantification at various time points (average ± std, n = 4, ** p < 0.01, *** p < 0.001)

    图  10  (a) DPPH溶液分别与硅片和ED900-TA涂层作用后的紫外光谱。(b) ED900-TA涂层总抗氧化性

    Figure  10.  (a) UV-vis spectra of the DPPH solution after incubation with silicon substrates with or without various repeats of ED900-TA assembly. (b) The total antioxidant activity of silicon substrates with various modifications

    图  11  (ED900-TA)n (n = 4, 8, 16, 20)涂层插入0.2%琼脂糖凝胶的稳定性:(a) 涂层插入琼脂糖凝胶的照片;(b) 涂层插入部分的光学显微镜图片

    Figure  11.  Stability of the (ED900-TA)n (n = 4, 8, 16, 20) coating inserted into 0.2% agarose gel: (a) Photographs and (b) optical microscope images of inserted surfaces

    图  12  (ED900-TA)20在37℃细胞完全培养基中浸泡不同时间的稳定性:(a)光学显微镜图像;(b) 表面TA浓度量化

    Figure  12.  Stability of (ED900-TA)20 immersed in complete culture medium at 37°C for different times: (a) Optical microscope images; (b) Quantification of the surface concentrations of TA

  • [1] 赵鸣岐, 黄威嫔, 胡米, 等. 生物医用材料表面高分子基涂层的功能化构筑[J]. 材料导报, 2019, 33(1):27-39. doi: 10.11896/cldb.201901003

    ZHAO M Q, HUANG W B, H M, et al. Functional-polymer-based coatings for biomedical materials’ surface[J]. Materials reports,2019,33(1):27-39(in Chinese). doi: 10.11896/cldb.201901003
    [2] 郭志君, 邹琴, 王立军, 等. 钛表面聚氨酯涂层的生物矿化及其细胞生物学响应[J]. 复合材料学报, 2014, 31(6):1612-1617.

    GUO Z J, ZOU Q, WANG L J, et al. Biomineralization of Polyurethane-coated Titanium Surface and cell behavior[J]. Acta Materiae Compositae Sinica,2014,31(6):1612-1617(in Chinese).
    [3] WANG H P, GONG X C, MIAO Y L, et al. Preparation and characterization of multilayer films composed of chitosan, sodium alginate and carboxymethyl chitosan-ZnO nanoparticles[J]. Food Chemistry,2019,283(15):397-403.
    [4] 朱继翔, 李树祎, 阳范文, 等. 负载淫羊藿苷的丝蛋白/β-磷酸三钙复合骨修复材料制备及性能[J]. 复合材料学报, 2017, 34(11):2580-2585.

    ZHU J X, LI S Y, YANG F W, et al. Preparation and properties of icariin loaded silk fibroin/β-tricalcium phosphate bone repair composite[J]. Acta Materiae Compositae Sinica,2017,34(11):2580-2585(in Chinese).
    [5] CHUNG K T, WONG T Y, WEI C I, et al. Tannins and Human Health: A Review[J]. Critical Review in Food Science and Nutrition,1998,38(6):421-464. doi: 10.1080/10408699891274273
    [6] WANG J, TIAN L L, CHEN N, et al. The cellular response of nerve cells on poly-L-lysine coated PLGA-MWCNTs aligned nanofibers under electrical stimulation[J]. Materials Science & Engineering C,2018,91(1):715-726.
    [7] VANCHA A R, GOVINDARAJU S, PARSA K V L, et al. Use of polyethyleneimine polymer in cell culture as attachment factor and lipofection enhancer[J]. BMC Biotechnology,2004,4(1):23-34. doi: 10.1186/1472-6750-4-23
    [8] SU J M, SUN Y, LI Z S, et al. Effect of tannic acid on lysozyme activity through intermolecular noncovalent binding[J]. Journal of Agriculture and Food Research,2019,1(10):4-11.
    [9] SAGLE L B, ZHANG Y J, LITOSH V A, et al. Investigating the hydrogen-bonding model of urea denaturation[J]. Journal of the American Chemical Society,2009,131(26):9304-9310. doi: 10.1021/ja9016057
    [10] DUARTE A, JONG E, KOEHORST R, et al. Conformational studies of peptides representing a segment of TM7 from H+-VO-ATPase in SDS micelles[J]. Biophysics of Structure and Mechanism,2010,39(4):639-6460.
    [11] EREL I, SUKHISHVILI S A. Hydrogen-Bonded Multilayers of a Neutral Polymer and a Polyphenol[J]. Macromolecules,2008,41(11):3962-3970. doi: 10.1021/ma800186q
    [12] FEI L, KOZLOVSKAYA V, ZAVGORODNYA O, et al. Encapsulation of anticancer drug by hydrogen-bonded multilayers of tannic acid[J]. Soft Matter,2014,10(46):9237-9247. doi: 10.1039/C4SM01813C
    [13] HOOK F, RODAHL M, KASEMO B, et al. Structural changes in hemoglobin during adsorption to solid surfaces: Effects of pH, ionic strength, and ligand binding[J]. Proceedings of the National Academy of Sciences. U. S. A.,1998,95(21):12271-12276. doi: 10.1073/pnas.95.21.12271
    [14] PENG L H, CHENG F, ZHENG Y N, et al. Multilayer Assembly of Tannic Acid and an Amphiphilic Copolymer Poloxamer 188 on Planar Substrates toward Multifunctional Surfaces with Discrete Microdome-Shaped Features[J]. Langmuir,2018,34(36):1982-1990.
    [15] HAN J M, XIA Y X, CHENG F, et al. Mechanistic Understanding of the Discrete Morphology Formed by Multi-cycle Assembly of Tannic Acid with Poloxamer 188 on Silicon Using QMC-D[J]. Colloids and Surfaces A:Physicochemical and Engineering Aspects,2021,628(5):127302-127310.
    [16] LAMPIN M, WAROCQUIER-CLEROUT R, LEGRIS C, et al. Correlation between substratum roughness and wettability, cell adhesion, and cell migration[J]. Journal of Biomedical Materials Research,1997,36(1):99-108. doi: 10.1002/(SICI)1097-4636(199707)36:1<99::AID-JBM12>3.0.CO;2-E
    [17] BACAKOVA L, FILOVA E, PARIZEK M, et al. Modulation of cell adhesion, proliferation and differentiation on materials designed for body implants[J]. Biotechnology Advances,2011,29(6):739-767. doi: 10.1016/j.biotechadv.2011.06.004
    [18] ZHAO L Z, MEI S L, CHU P K, et al. The influence of hierarchical hybrid micro/nano-textured titanium surface with titania nanotubes on osteoblast functions[J]. Biomaterials,2010,31(19):5072-5082. doi: 10.1016/j.biomaterials.2010.03.014
    [19] NGUYEN A T, SATHE S R, YIM E K F. From nano to micro: topographical scale and its impact on cell adhesion, morphology and contact guidance[J]. Journal of Physics Condensed Matter An Institute of Physics Journal,2016,28(18):183001-183016. doi: 10.1088/0953-8984/28/18/183001
    [20] LI J G, ZHANG K, YANG P, et al. Human vascular endothelial cell morphology and functional cytokine secretion influenced by different size of HA micro-pattern on titanium substrate[J]. Colloids and Surfaces B:Biointerfaces,2013,110(1):199-207.
    [21] DALBY M J, RIEHLE M O, JOHNSTONE H, et al. Investigating the limits of filopodial sensing: a brief report using SEM to image the interaction between 10 nm high nano-topography and fibroblast filopodia[J]. Cell Biololgy International,2004,28(3):229-236. doi: 10.1016/j.cellbi.2003.12.004
    [22] KANG K, CHOI S E, JANG H S, et al. In vitro developmental acceleration of hippocampal neurons on nanostructures of self-assembled silica beads in filopodium-size ranges[J]. Angewandte Chemie,2012,124(12):2839-2839. doi: 10.1002/ange.201108840
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  • 收稿日期:  2021-12-01
  • 录用日期:  2022-01-15
  • 修回日期:  2022-01-09
  • 网络出版日期:  2022-02-21

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