Gerasimenko Alexander Yu, Ten Galina N, Ryabkin Dmitry I, Shcherbakova Natalia E, Morozova Elena A, Ichkitidze Levan P
National Research University of Electronic Technology MIET, Shokin Square, Zelenograd, Moscow, 124498, Russia; I.M. Sechenov First Moscow State Medical University, bld. 2-4, Bolshaya Pirogovskaya street, Moscow, 119991, Russia.
Saratov State University, 83 Astrakhanskaya street, Saratov, 410012, Russia.
Spectrochim Acta A Mol Biomol Spectrosc. 2020 Feb 15;227:117682. doi: 10.1016/j.saa.2019.117682. Epub 2019 Oct 23.
The results of the study of composites based on bovine serum albumin (BSA) and single-walled carbon nanotubes (SWCNT) are presented. Nanocomposites were created by evaporation of the water-albumin dispersion with nanotubes using diode laser with temperature control. Two types of nanotubes were used. SWCNT I were synthesized using the electric arc method, SWCNT II were synthesized using the gas phase method. SWCNT I had a diameter and length less than SWCNT II. The mechanism of interaction between BSA and SWCNT in solid nanocomposites is considered. An experimental and theoretical studies of the interaction between aspartic (Asp) and glutamic (Glu) amino acids located on the outer surface of BSA and nanotubes using of vibrational spectroscopy (Fourier-transform infrared (FTIR) and Raman spectroscopy) was carried out. The possibility of nanotubes functionalization by oxygen atoms of negative amino acid residues Asp and Glu, which are on the outer surface of BSA, is shown by molecular modeling. The formation of covalent bonds between BSA and SWCNT in nanocomposites with different concentrations of nanotubes (0.01, 0.1 and 1 g/l) was confirmed by vibrational spectra. The covalent interaction between BSA with SWCNT under the laser irradiation leads to the conformational changes in the secondary and tertiary structures of albumin. This is confirmed by a significant decrease in the intensity of the absorption bands in the high-frequency region. The calculation of the vibrational spectra of the three Glycine:Glycine, Glutamic acid:Threonine and Aspartic acid:Lysine complexes, which take into account hydrogen, ion-dipole and ion-ion bonds, showed that a disturbance in the intermolecular interaction between amino acid residues led to significant decrease in the intensity of absorption bands in the region of stretching vibrations bonds OH and NH. From the Raman spectra, it was found that a significant number of defects in SWCNT is caused by the covalent attachment of oxygen atoms to the graphene surface of nanotubes. An increase in the diameter of nanotubes (4 nm) has practically no effect on the absorption spectrum of nanocomposite, while measuring the concentration of SWCNT affects the FTIR spectra. This confirmed the hydrophobic interaction between BSA and SWCNT. Thus, it was shown that BSA solid nanocomposites with CNTs can interact either with the help of hydrophobic forces or with the formation of covalent bonds, which depends on the diameter of the used nanotubes. The viability of connective fibroblast tissue cells on nanocomposites with both types of SWCNT was demonstrated. It was found that nanocomposites based on SWCNT I provide slightly better compatibility of their structure with fibroblasts. It allows to achieve better cell adhesion to the nanocomposite surface. These criteria make extensive use of scaffold nanocomposites in biomedicine, depending on the requirements for their quality and application.
本文展示了基于牛血清白蛋白(BSA)和单壁碳纳米管(SWCNT)的复合材料的研究结果。通过使用带温度控制的二极管激光器蒸发含纳米管的水 - 白蛋白分散液来制备纳米复合材料。使用了两种类型的纳米管。SWCNT I采用电弧法合成,SWCNT II采用气相法合成。SWCNT I的直径和长度小于SWCNT II。研究了固体纳米复合材料中BSA与SWCNT之间的相互作用机制。利用振动光谱(傅里叶变换红外(FTIR)和拉曼光谱)对位于BSA外表面的天冬氨酸(Asp)和谷氨酸(Glu)氨基酸与纳米管之间的相互作用进行了实验和理论研究。分子建模表明,位于BSA外表面的带负电荷氨基酸残基Asp和Glu的氧原子可使纳米管功能化。通过振动光谱证实了在不同纳米管浓度(0.01、0.1和1 g/l)的纳米复合材料中BSA与SWCNT之间形成了共价键。激光照射下BSA与SWCNT之间的共价相互作用导致白蛋白二级和三级结构发生构象变化。高频区域吸收带强度的显著降低证实了这一点。对三种甘氨酸:甘氨酸、谷氨酸:苏氨酸和天冬氨酸:赖氨酸复合物的振动光谱进行计算,其中考虑了氢键、离子 - 偶极键和离子 - 离子键,结果表明氨基酸残基之间分子间相互作用的干扰导致OH和NH伸缩振动键区域吸收带强度显著降低。从拉曼光谱发现,纳米管中大量的缺陷是由氧原子共价附着在纳米管的石墨烯表面引起的。纳米管直径增加到4 nm对纳米复合材料的吸收光谱几乎没有影响,而测量SWCNT的浓度会影响FTIR光谱。这证实了BSA与SWCNT之间的疏水相互作用。因此,研究表明,含CNT的BSA固体纳米复合材料可以借助疏水力相互作用,也可以通过形成共价键相互作用,这取决于所用纳米管的直径。证明了两种类型SWCNT的纳米复合材料对结缔组织成纤维细胞的活力。发现基于SWCNT I的纳米复合材料在其结构与成纤维细胞的兼容性方面略好。这使得细胞能够更好地粘附在纳米复合材料表面。根据对其质量和应用的要求,这些标准使得支架纳米复合材料在生物医学中得到广泛应用。