Mutlu Esra Cansever, Kaya Özge, Wood Matthew, Mager Imre, Topkara Kübra Çelik, Çamsarı Çağrı, Birinci Yildirim Arzu, Çetinkaya Ayhan, Acarel Diğdem, Odabaşı Bağcı Jale
Department of Biomedical Engineering, Faculty of Engineering and Architecture, Beykent University, Sarıyer, 34398 Istanbul, Turkey.
Scientific Industrial and Technological Application and Research Center, BETUM, Bolu Abant Izzet Baysal University, 14030 Bolu, Turkey.
Materials (Basel). 2020 Jul 27;13(15):3344. doi: 10.3390/ma13153344.
Immature dendritic cells (IDc), 'dexosomes', are promising natural nanomaterials for cancer diagnose and therapy. Dexosomes were isolated purely from small-scale-up production by using t25-cell-culture flasks. Total RNA was measured as 1.43 ± 0.33 ng/10 cell. Despite the fact that they possessed a surface that is highly abundant in protein, this did not become a significant effect on the DOX loading amount. Ultrasonication was used for doxorubicin (DOX) loading into the IDc dexosomes. In accordance with the literature, three candidate DOX formulations were designed as IC50 values; dExoIII, 1.8 µg/mL, dExoII, 1.2 µg/mL, and dExoI, 0.6 µg/mL, respectively. Formulations were evaluated by MTT test against highly metastatic A549 (CCL-185; ATTC) cell line. Confocal images of unloaded (naïve) were obtained by CellMask membrane staining before DOX loading. Although, dexosome membranes were highly durable subsequent to ultrasonication, it was observed that dexosomes could not be stable above 70 °C during the SEM-image analyses. dExoIII displayed sustained release profile. It was found that dynamic light scattering (DLS) and nanoparticle tracking analysis (NTA) results were in good agreement with each other. Zeta potentials of loaded dexosomes have approximately between -15 to -20 mV; and, their sizes are 150 nm even after ultrasonication. IDcJAWSII dexosomes can be able to be utilized as the "BioNanoMaterial" after DOX loading via ultrasonication technique.
未成熟树突状细胞（IDc）来源的“外泌体”是用于癌症诊断和治疗的很有前景的天然纳米材料。通过使用T25细胞培养瓶从小规模放大生产中纯分离出外泌体。测得总RNA为1.43±0.33 ng/10个细胞。尽管它们具有蛋白质高度丰富的表面，但这对阿霉素（DOX）的负载量没有显著影响。使用超声处理将阿霉素（DOX）负载到IDc外泌体中。根据文献，设计了三种候选DOX制剂作为IC50值；分别为dExoIII，1.8 µg/mL；dExoII，1.2 µg/mL；dExoI，0.6 µg/mL。通过MTT试验评估这些制剂对高转移性A549（CCL-185；ATTC）细胞系的作用。在DOX负载之前，通过CellMask膜染色获得未负载（天然）外泌体的共聚焦图像。尽管超声处理后外泌体膜具有高度耐久性，但在扫描电子显微镜图像分析中观察到外泌体在70℃以上不稳定。dExoIII显示出缓释曲线。发现动态光散射（DLS）和纳米颗粒跟踪分析（NTA）结果彼此吻合良好。负载外泌体的zeta电位约在-15至-20 mV之间；并且，即使经过超声处理，它们的尺寸仍为150 nm。通过超声技术负载DOX后，IDcJAWSII外泌体能够用作“生物纳米材料”。
