Delavari Behdad, Bigdeli Bahareh, Mamashli Fatemeh, Gholami Mahdi, Bazri Behrouz, Khoobi Mehdi, Ghasemi Atiyeh, Baharifar Hadi, Dehghani Sadegh, Gholibegloo Elham, Amani Amir, Riahi-Alam Nader, Ahmadian Shahin, Goliaei Bahram, Asli Naisana S, Rezayan Ali Hossein, Saboury Ali Akbar, Varamini Pegah
Division of Nanobiotechnoloy, Department of Life Science Engineering, Faculty of New Sciences & Technologies, University of Tehran, Tehran, 1417466191, Iran.
Institute of Biochemistry and Biophysics, University of Tehran, Mailbox 13145-1384, Tehran, Iran.
ACS Biomater Sci Eng. 2019 Oct 14;5(10):5189-5208. doi: 10.1021/acsbiomaterials.9b01236. Epub 2019 Sep 23.
A nanotheranostic system was developed using α-lactalbumin along with FeO nanoparticles as an magnetic resonance imaging (MRI) contrast agent for medical imaging and doxorubicin as the therapeutic agent. α-lactalbumin was precipitated and cross-linked using poly(ethylene glycol) and glutaraldehyde. Besides, polyethylenimine was applied to increase the number of amine groups during cross-linking between α-lactalbumin and FeO nanoparticles. Interestingly, 90% of the initial protein used for the coaggregation process was incorporated in the prepared 130 nm nanocomposites, which facilitated the 85% doxorubicin loading. Formation of pH-sensitive imine bonds between glutaraldehyde and amine groups on α-lactalbumin and polyethylenimine resulted in higher release of doxorubicin at acidic pHs and consequently development of a pH-sensitive nanocarrier. The designed nanocomposite was less immunogenic owing to stimulating the production of less amounts of C3a, C5a, platelet factor 4, glycoprotein IIb/IIIa, platelet-derived β-thromboglobulin, interleukin-6, and interleukin-1β compared to the free doxorubicin. Furthermore, 1000 μg/mL nanocomposite led to 0.2% hemolytic activity, much less than the 5% standard limit. The void nanocarrier induced no significant level of cytotoxicity in breast cancer and normal cells following 96 h incubation. The doxorubicin-loaded nanocomposite presented higher cytotoxicity, apoptosis induction, and doxorubicin uptake in cancer cells than free doxorubicin. Conversely, lower cytotoxicity, apoptosis induction, and doxorubicin uptake were observed in normal cells treated with the doxorubicin-loaded nanocarrier compared to free doxorubicin. In line with the results of in vitro experiments, in vivo studies on tumor-bearing mice showed more suppression of tumor growth by the doxorubicin-loaded nanocomposite compared to the free drug. Moreover, the pharmacokinetic study revealed slow release of doxorubicin from the nanocomposite. Besides, in vitro and in vivo MRI studies presented a higher / ratio and comparable contrast to the commercially available DOTAREM, respectively. Our findings suggest that this new nanocomposite is a promising nanotheranostic system with promising potential for cancer therapy and diagnosis.
开发了一种纳米诊疗系统，该系统使用α-乳白蛋白以及FeO纳米颗粒作为用于医学成像的磁共振成像（MRI）造影剂，并使用阿霉素作为治疗剂。α-乳白蛋白通过聚乙二醇和戊二醛沉淀并交联。此外，在α-乳白蛋白与FeO纳米颗粒交联过程中，使用聚乙烯亚胺来增加胺基数量。有趣的是，用于共聚集过程的初始蛋白质中有90%被掺入所制备的130 nm纳米复合材料中，这有利于85%的阿霉素负载。戊二醛与α-乳白蛋白和聚乙烯亚胺上的胺基之间形成pH敏感的亚胺键，导致阿霉素在酸性pH值下有更高的释放量，从而开发出一种pH敏感的纳米载体。与游离阿霉素相比，所设计的纳米复合材料由于刺激产生的C3a、C5a、血小板因子4、糖蛋白IIb/IIIa、血小板衍生的β-血小板球蛋白、白细胞介素-6和白细胞介素-1β量较少，因此免疫原性较低。此外，1000 μg/mL的纳米复合材料导致0.2%的溶血活性，远低于5%的标准限值。在孵育96小时后，空白纳米载体在乳腺癌细胞和正常细胞中未诱导出显著水平的细胞毒性。负载阿霉素的纳米复合材料在癌细胞中表现出比游离阿霉素更高的细胞毒性、凋亡诱导和阿霉素摄取。相反，与游离阿霉素相比，用负载阿霉素的纳米载体处理的正常细胞中观察到较低的细胞毒性、凋亡诱导和阿霉素摄取。与体外实验结果一致，对荷瘤小鼠的体内研究表明，与游离药物相比，负载阿霉素的纳米复合材料对肿瘤生长的抑制作用更强。此外，药代动力学研究表明阿霉素从纳米复合材料中缓慢释放。此外，体外和体内MRI研究分别显示出更高的/比值和与市售的多他灵相当的对比度。我们的研究结果表明，这种新型纳米复合材料是一种有前途的纳米诊疗系统，在癌症治疗和诊断方面具有广阔的潜力。
