Khajavinia Amir, Varshosaz Jaleh, Dehkordi Abbas Jafarian
Faculty of Pharmacy and Novel Drug Delivery Systems Research Centre, Department of Pharmaceutics Isfahan University of Medical Sciences, Isfahan, Iran.
Nanotechnology. 2012 Oct 12;23(40):405101. doi: 10.1088/0957-4484/23/40/405101. Epub 2012 Sep 14.
The aim of the present study was to evaluate the diverse properties of transferrin (Tf)-conjugated nanostructured lipid carriers (NLCs) prepared using three different fatty amines, including stearylamine (SA), dodecylamine (DA) and spermine (SP), and two different methods for Tf coupling. Etoposide-loaded NLCs were prepared by an emulsion-solvent evaporation method followed by probe sonication. Chemical coupling of NLCs with Tf was mediated by an amide linkage between the surface-exposed amino group of the fatty amine and the carboxyl group of the protein. The physical coating was performed in a Ringer-Hepes buffer medium. NLCs were characterized by their particle size, zeta potential, polydispersity index, drug entrapment percentage, drug release profiles and Tf-coupling efficiency. The cytotoxicity of NLCs on K562 acute myelogenous leukaemia cells was studied by MTT assay, and their cellular uptake was studied by a flow cytometry method. SA-containing NLCs showed the lowest particle size, the highest zeta potential and the largest coupling efficiency values. The drug entrapment percentage and the zeta potential decreased after Tf coupling, but the average particle size increased. SP-containing formulations released their drug contents comparatively slower than SA- or DA-containing NLCs. Unconjugated NLCs released moderately more drug than Tf-NLCs. Flow cytometry studies revealed enhanced cellular uptake of Tf-NLCs compared to unconjugated ones. Blocking Tf receptors resulted in a significantly higher cell survival rate for Tf-NLCs. The highest cytotoxic activity was observed in the chemically coupled SA-containing nanoparticles, with an IC(50) value of 15-fold lower than free etoposide.
本研究的目的是评估使用三种不同脂肪胺（硬脂胺（SA）、十二烷基胺（DA）和精胺（SP））制备的转铁蛋白（Tf）偶联纳米结构脂质载体（NLCs）的多种特性，以及两种不同的Tf偶联方法。通过乳化溶剂蒸发法然后探针超声处理制备载有依托泊苷的NLCs。NLCs与Tf的化学偶联是通过脂肪胺表面暴露的氨基与蛋白质的羧基之间的酰胺键介导的。物理包被在林格 - 赫佩斯缓冲介质中进行。NLCs通过其粒径、zeta电位、多分散指数、药物包封率、药物释放曲线和Tf偶联效率进行表征。通过MTT法研究NLCs对K562急性髓性白血病细胞的细胞毒性，并通过流式细胞术方法研究其细胞摄取情况。含SA的NLCs显示出最低的粒径、最高的zeta电位和最大的偶联效率值。Tf偶联后药物包封率和zeta电位降低，但平均粒径增加。含SP的制剂比含SA或DA的NLCs释放药物内容物相对较慢。未偶联的NLCs比Tf - NLCs释放的药物略多。流式细胞术研究表明，与未偶联的NLCs相比，Tf - NLCs的细胞摄取增强。阻断Tf受体导致Tf - NLCs的细胞存活率显著更高。在化学偶联的含SA纳米颗粒中观察到最高的细胞毒性活性，其IC(50)值比游离依托泊苷低15倍。
