不同形貌无定形二氧化硅的可控合成及其生物矿化意义
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摘要
生物矿化是一个非常活跃的研究领域,正在引起人们越来越多的关注。以二氧化硅为基质的硅藻细胞壁,英文名为frustules,是生物体产生的纳米艺术品中最优秀的范例。这些华丽的外壳通常展现出高度分化的纳米级和微米级图案,这些图案是由孔有规则排列产生的。它们具有独特的形态和层次结构,迷人的机械性能和在许多领域中潜在的应用价值,并因此成为许多领域的研究热点。要想了解生物矿化,就需要对其内在的细胞和分子生物学过程进行研究。一方面,人们已经从硅藻细胞壁提取物中成功地分离并鉴定出多种有机和生物分子,如蛋白质、多糖、有机胺和脂质。另一方面,通过对硅藻细胞壁的系统研究,发现硅藻细胞壁的硅质结构是由零维纳米颗粒通过一维的绳状结构组装成具有孔排列的二维微米级图案。与此同时,还观察到一些有趣的结构细节。例如,在硅藻细胞壁发育早期,硅藻产生了拉长硅质结构,而且这些结构上具有局部膨胀的部位。另一个例子是在硅藻的网眼状孔隙形成以后,出现的网眼状孔隙壁的不对称发育。然而,很少有人知道它们的生长机制。本论文中,在仿生矿化条件下,采用几种不同的沉淀二氧化硅的方法,制备具有特定形貌的硅质矿物。然后,采用多种表征方法对所得产物进行分析,其中包括粉末X-射线衍射(XRD)、场发射扫描电子显微镜(FESEM)、(高分辨)透射型电子显微镜(TEM和HRTEM),傅里叶变换红外光谱(FTIR),热重差热分析(TG-DTA),固态29Si核磁共振,动态光散射(DLS)以及氮气吸附脱附等温线等。基于上述检测结果,可以全面研究样品的组成、结构特征以及表面形貌。与此同时,我们也进行了各种单变量实验,以了解所添加的有机质对二氧化硅沉淀的影响。结果,我们针对不同的硅质结构(包括局部膨胀的拉长结构,碟状不对称结构,二维硅质筛板以及具有穿透孔的二氧化硅带)提出了相应的生长机制。因此,我们的研究结果可以为进一步了解生物硅化机制提供有用信息,并且展示出体外合成类似于生物成因二氧化硅形貌的硅质结构的可能性。下面对论文的主要内容进行简要描述。
     1.采用正硅酸四乙酯[TEOS, Si(OCH2CH3)4]为硅源,引入磷脂(PL)和十二胺(DA)为有机添加剂,研究这些添加剂对矿化过程中产生的二氧化硅形貌的影响。FESEM和TEM分析表明,在DA浓度不变的情况下,PL的加入会导致硅质拉长结构的出现。而PL浓度的进一步增大会导致拉长结构进一步生长,并出现局部膨胀现象,这些都展示出硅藻壳面发育早期的一些特征。另外,我们用无机氨替代有机胺,进行了一系列PL浓度不同的平行实验。但是并没有得到任何拉长结构,这表明拉长硅质结构的产生是在PL和DA分子协同作用情况下得到的。由于有机胺(例如,长链聚胺)和膜脂(例如,硅质膜鞘)在硅藻和海绵体内生物硅化过程的特殊重要性,我们的结果可能指示,膜脂参与了生物体内有机聚集体的形成,并因此而影响到胺诱导的二氧化硅沉积。
     2.以正硅酸四乙酯[TEOS, Si(OCH2CH3)4]为硅源,通过控制磷脂(PL)和十二胺(DA)等有机组元的相分离过程,影响二氧化硅的沉淀过程。结果显示,有机组元的相分离过程会导致不对称非球形二氧化硅结构的形成。而且,可以通过调节PL和DA的浓度来控制不对称结构的横纵比。在时间序列实验的基础上,我们尝试着提出了不对称二氧化硅形貌形成的机制。因此,相分离过程不仅与生物二氧化硅的多级多孔图案的产生有关,而且可能有利于硅质结构在特定方向上的分异生长。由于有机胺(例如,长链聚胺)、磷脂(例如,硅质膜鞘)以及相分离过程都与硅藻的生物硅化密切相关,当前结果可能拓展了对生物硅化的机制的了解。
     3.大孔多孔二氧化硅筛板(silica sieve plate, SSP)是硅藻壳面结构的重要特征。正电性的多肽natSil-lA和负电性的natSil-2形成的有机聚集体是形成硅藻硅质细胞壁的模板,而且富含羟基和羧基的natSil-2被认为是大孔多孔硅质结构形成过程中的调节剂。为了进一步了解这种生物二氧化硅筛板结构矿化机制,特别的选择阴离子型表面活性剂十二烷基硫酸钠(SDBS)和阳离子表面活性剂十六烷基三甲基溴化铵(CTAB)分别模拟正电性的多肽natSil-1A和负电性的多肽natSil-2,控制形成复合模板;以同时带有羧基和羟基的L-酒石酸模拟natSil-2结构中的羧基和羟基,调节硅化产物多空结构的形成。结果在阴、阳离子型表面活性剂复合模板和L-酒石酸调节剂存在下,一种类似生物硅藻壳面的二维二氧化硅筛板结构被成功地合成。同时,一系列不同浓度添加剂L-酒石酸存在下的实验结构揭示羟基和羧基共存的酒石酸分子对于SSPs的形成至关重要。因此,实验结果有助于加深对生物硅藻壳面大孔多孔二氧化硅筛板成因机制的了解。
     4.受生物硅化作用的启发,在L-(+)-酒石酸、十二烷基苯磺酸钠(SDBS)和NaSCN存在的情况下,我们得到了一种带有穿透孔的二氧化硅纳米带(silica ribbons with through-holes, SRTHs)。FESEM和TEM照片表明,这种SRTH结构是由小的球形颗粒组成的,穿透孔结构的出现是由于这些纳米颗粒的组装受限导致的。基于一系列的表征结果,我们全面讨论了SRTHs的形成机制。在合成过程中,预先制得的针状酒石酸钠晶体是带状二氧化硅沉积产生的硬模板。在NaSCN的协助下,阴离子表面活性剂SDBS在酒石酸盐晶体表面吸附,这对于穿透孔结构的形成至关重要。另外,我们通过进行时间序列实验,揭示了二氧化硅从颗粒到SRTH结构的生长过程。
Biomineralization is an active research field of increasing interest. Frustule, the silica-based cell wall of diatoms, is one of the most outstanding nanostructured artworks created by living organisms. These ornate shells usually exhibit highly differentiated nano-and micropatterns with regular arrays of holes, which has attracted much attention because of their unique morphologies and hierarchical structures, fascinating mechanical properties and potential applications in many fields. Understanding biomineralization requires elucidation of the underlying cellular and molecular biological processes. On the one hand, a variety of organic and biological molecules, such as proteins, polysaccharides, organic amines and lipids, have been successfully separated and identified from cell-wall extracts of diatoms. One the other hand, the formation process of cell wall in diatoms has been studied systematically. The hierarchical assembly from zero-dimensional nanoparticles through one-dimensional rope-like structures to the two-dimensional micropatterns with regular arrays of holes can been well discerned. Moreover, some interesting structure details were observed. For example, elongated siliceous structures and their localized enlargement occurred at the early stage of frustule development. Another example is the asymmetrical development of areolae walls after the formation of areolae structure in diatoms. However, little is known about their growth mechanism. In this dissertation, various mineralization methods of silica deposition under biomimetic conditions were utilized to prepare siliceous minerals with different shapes, asymmetrical particles with different aspect ratios and assembled silica architectures with several levels of hierarchy. The obtained products were analyzed with several characterization methods, including the powder X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM),(high-resolution) transmission electron microscope (TEM and HRTEM), Fourier transform infrared spectra (FTIR), thermogravimetric and differential thermal analysis (TG-DTA), solid-state29Si NMR, dynamic light scattering (DLS) and nitrogen physisorption. Based on the above determinations, the sample compositions, textural characteristics and surface morphologies can be comprehensively investigated. Meanwhile, various univariate experiments were also carried out to understand the influence of the added organics on silica formation. As a result, different growth mechanisms were proposed to illustrate the formation of the corresponding siliceous structures, such as the elongated siliceous structures with localized enlargement, the disk-like asymmetrical particles, the two-dimensional silica sieve plates and the silica ribbons with through-holes. Therefore, our results may provide available information for further understanding the mechanism of biosilicification. Some new possibilities have also been exhibited on the fabrication of silica structures whose morphologies are similar to the biosilica formed in organism. The details of dissertation are described below.
     1. Using tetraethyl orthosilicate [TEOS, Si(OCH2CH3)4] as a silica source, phospholipid (PL) and dodecylamine (DA) were introduced as model organic additives to investigate their influence on the formation and morphology of silica in the mineralization process. The FESEM and TEM analyses demonstrate that increasing PL concentrations at constant DA content leads to the formation of siliceous elongated structures. Localized enlargement can also be observed during further growth of elongated structures, displaying some features of the earliest recognizable stage of valve development in diatoms. In addition, in the presence or absence of PL, a series of control experiments using,ammonia instead of DA show that no elongated structures are obtained, suggesting that the formation of elongated silica structures results from the cooperative interactions between PL and DA molecules. Because both organic amines (e.g., long-chain polyamines, LCPA) and phospholipid membranes (e.g., silicalemma) are of special importance for biosilicification in diatoms and sponges, our results imply that phospholipids are involved in the formation of organic aggregates, and thus influence the amines-mediated silica deposition.
     2. Using tetraethyl orthosilicate [TEOS, Si(OCH2CH3)4] as silica precursor, phospholipid (PL) and dodecylamine (DA) were introduced to initiate phase separation of organic components and influence silica precipitation. Morphology, structure and composition of the mineralized products were studied. The results demonstrate that the phase separation process of the organic components leads to the formation of asymmetrically non-spherical silica structures, and the aspect ratios of the asymmetrical structures can be well controlled by varying the concentration of PL and DA. On the basis of the time-dependent experiments, a tentative mechanism is also proposed to illustrate the asymmetrical morphogenesis. Therefore, our results imply that in addition to explaining the hierarchical porous nanopatterning of biosilica, the phase separation process may be also responsible for the growth differentiation of siliceous structures in specific directions. Because organic amine (e.g., long-chair polyamines), phospholipids (e.g., silicalemma) and the phase separation process are associated with the biosilicification of diatoms, our results may provide a new insight into the mechanism of biosilicification.
     3. Macroporous silica sieve plate (SSP) is an important characteristic of the valve structures in diatoms. The organic aggregates formed by opposite-charged peptides natSil-lA and natSil-2can act as a structure template during siliceous cell wall formation in diatoms, and the peptide natSil-2modified by hydroxyl-rich carbohydrate and anionic groups is considered to be a regulator of macroporous siliceous architectures. Inspired by these biological design principles, we provide a simple pathway for the fabrication of two-dimensional silica sieve plates by use of catanionic surfactants as templates and L-tartrate with hydroxyl and carboxyl groups as a regulator. The present synthesis method has the advantage of being a single-step self-assembly process. The formation mechanism of the hierarchical SSPs was also discussed in details, and the hydroxyl and carboxyl groups incorporated in one tartrate molecule are crucial to the formation of the SSPs. In this regard, our results may provide a novel pathway towards a deeper insight into biosilicification, and such well-designed conception can potentially be applied to the synthesis of other oxide sieve plate structures.
     4. We present a facile method for the preparation of silica ribbons with through-holes (SRTHs) in the presence of L-(+)-tartaric acid, SDBS and NaSCN. The FESEM and TEM analyses demonstrate that the SRTH structures with rough surfaces are constituted by small spherical particles, and the appearance of through-holes is due to the limited assembly of these nano-particles. The formation mechanism of SRTHs is systematically discussed based on the characterization results. In the synthetic process, the preformed needle-like crystals of sodium tartrate can act as the hard template of ribbon-like silica deposition. With the help of NaSCN, the adsorption of anionic surfactant (SDBS) on the surfaces of tartrate needles is considered to be important for the formation of through-hole structures. And the time-dependent experiments exhibit the formation process from silica particles to SRTH structures.
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