Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education, School of Biological Science and Medical Engineering, Beihang University, Beijing 100083, China.
J Mater Chem B. 2019 Aug 7;7(31):4821-4832. doi: 10.1039/c9tb00894b.
Preparation of stable and effective artificial oxygen carriers (AOCs) is a promising strategy to temporarily replace transfused blood and solve tissue hypoxia. Developing hemoglobin (Hb) loaded particles is one of the main ways to prepare suitable AOCs. Particles with a hierarchical micro/nanostructure can be loaded with plenty of proteins and have attracted great attention. Therefore, multiwall carbon nanotubes (MWCNTs) were chosen to fabricate AOCs. To improve the Hb-loading capacity of MWCNTs, functionalized MWCNTs, including carboxyl-functionalized MWCNTs (MWCNT-COOH), amino-functionalized MWCNTs (MWCNT-NH2), and heparin-conjugated MWCNTs (MWCNT-Hep), were prepared. Then, in this study, Hb was coupled to the functionalized MWCNTs to fabricate the AOCs. The functionalized MWCNTs and the AOCs were characterized by FTIR, SEM, TEM, and zeta potential analysis. The oxygen/Hb-loading capacity of the AOCs was also measured. The adverse effects of the AOCs on human umbilical vein endothelial cells (HUVECs) and human red blood cells (RBCs) were evaluated. The results showed that (1) the functional groups were grafted on the surface of the MWCNTs, and Hb was bound to the functionalized MWCNTs, thus the AOCs were successfully prepared; (2) MWCNT-Hep-Hb had the most stable dispersibility (i.e., the most negative zeta potential) in 0.9 wt% NaCl solution (MWCNT-Hep-Hb < MWCNT-COOH-Hb < MWCNT-Hb < MWCNT-NH2-Hb < 0); (3) MWCNT-Hep had the best Hb-loading capability, which was three times that of purified MWCNTs; (4) with concentrations increased up to 400 μg mL-1, MWCNT-Hep-Hb still had the highest cell viability (97.63% > 80%, ISO 10993-5:2009) and excellent blood biocompatibility. Therefore, MWCNT-Hep-Hb might be a satisfactory candidate as a blood substitute.
制备稳定有效的人工氧载体（AOCs）是一种有前途的策略，可以暂时替代输注的血液并解决组织缺氧问题。负载血红蛋白（Hb）的颗粒的开发是制备合适的 AOCs 的主要方法之一。具有分级微/纳结构的颗粒可以负载大量蛋白质，因此受到了极大的关注。因此，选择多壁碳纳米管（MWCNTs）来制备 AOCs。为了提高 MWCNTs 的 Hb 负载能力，制备了功能化的 MWCNTs，包括羧基化 MWCNTs（MWCNT-COOH）、氨基化 MWCNTs（MWCNT-NH2）和肝素接枝 MWCNTs（MWCNT-Hep）。然后，在这项研究中，将 Hb 偶联到功能化的 MWCNTs 上以制备 AOCs。通过傅里叶变换红外光谱（FTIR）、扫描电子显微镜（SEM）、透射电子显微镜（TEM）和 Zeta 电位分析对功能化的 MWCNTs 和 AOCs 进行了表征。还测量了 AOCs 的氧/Hb 负载能力。评估了 AOCs 对人脐静脉内皮细胞（HUVECs）和人红细胞（RBCs）的不良影响。结果表明：（1）功能基团接枝在 MWCNTs 表面，Hb 结合在功能化的 MWCNTs 上，从而成功制备了 AOCs；（2）在 0.9wt%NaCl 溶液中，MWCNT-Hep-Hb 具有最稳定的分散性（即最负的 Zeta 电位）（MWCNT-Hep-Hb<MWCNT-COOH-Hb<MWCNT-Hb<MWCNT-NH2-Hb<0）；（3）MWCNT-Hep 具有最佳的 Hb 负载能力，是纯化 MWCNTs 的三倍；（4）浓度高达 400μg mL-1 时，MWCNT-Hep-Hb 仍具有最高的细胞活力（97.63%>80%，ISO 10993-5:2009）和出色的血液生物相容性。因此，MWCNT-Hep-Hb 可能是一种令人满意的血液替代品候选物。
