In vivo Non-invasive Imaging of Radio-Labeled Exosome-Mimetics Derived From Red Blood Cells in Mice.
In vivo Non-invasive Imaging of Radio-Labeled Exosome-Mimetics Derived From Red Blood Cells in Mice.
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DOI：
10.3389/fphar.2018.00817
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发表时间：
2018
影响因子：
5.6
通讯作者：
Ahn BC
中科院分区：
文献类型：
--
作者：
Gangadaran P;Hong CM;Oh JM;Rajendran RL;Kalimuthu S;Son SH;Gopal A;Zhu L;Baek SH;Jeong SY;Lee SW;Lee J;Ahn BC
外泌体是天然的纳米大小的膜囊泡，由于其作为药物输送载体的潜力而引起了人们的兴趣。尽管外泌体是有效的药物载体，但其产生和体内生物分布尚未完全阐明。我们分析了红细胞 (RBC) 外泌体模拟物 (EM) 的产生以及用于体内成像的 RBC-EM 的放射性标记。通过一步挤出法大规模生产来自红细胞的工程化 EM，并通过密度梯度离心进一步纯化。 RBC-EM 用 technetium-99m (99mTc) 标记。对于非侵入性成像，将 99mTc（游离）或 99mTc-RBC-EM 注射到小鼠体内，并通过伽马相机成像分析它们的生物分布。处死动物，收集器官用于进一步的生物分布分析。 RBC-EM 具有与 RBC 外泌体相似的特征，但就颗粒数量而言，产量高出 130 倍。 99mTc-RBC-EMs 的放射化学纯度在 2 小时前几乎为 100%，在 3 小时时降至 97%。放射性标记不影响 RBC-EM 的大小和形态。与游离 99mTc 相比，小鼠体内 99mTc-RBC-EM 的体内成像显示肝脏和脾脏的摄取量较高，而甲状腺中没有摄取。离体成像证实了体内发现。此外，荧光成像证实了核成像结果。免疫荧光成像显示，肝脏对 RBC-EM 的摄取显着由库普弗细胞（常驻肝巨噬细胞）介导。我们的结果证明了一种新型 RBC-EM 的简单而大规模的生产方法，该方法可以用 99mTc 有效标记，并通过核成像进行体内监测。 RBC-EM 可用作体内药物递送载体。
Exosomes are natural nano-sized membrane vesicles that have garnered recent interest owing to their potential as drug delivery vehicles. Though exosomes are effective drug carriers, their production and in vivo biodistribution are still not completely elucidated. We analyzed the production of exosome mimetics (EMs) from red blood cells (RBCs) and the radio-labeling of the RBC-EMs for in vivo imaging. Engineered EMs from RBCs were produced in large-scale by a one-step extrusion method, and further purified by density-gradient centrifugation. RBC-EMs were labeled with technetium-99m (99mTc). For non-invasive imaging, 99mTc (free) or 99mTc-RBC-EMs were injected in mice, and their biodistribution was analyzed by gamma camera imaging. Animals were sacrificed, and organs were collected for further biodistribution analysis. RBC-EMs have similar characteristics as the RBC exosomes but have a 130-fold higher production yield in terms of particle numbers. Radiochemical purity of 99mTc-RBC-EMs was almost 100% till 2 h reduced to 97% at 3 h. Radio-labeling did not affect the size and morphology of RBC-EMs. In contrast to free 99mTc, in vivo imaging of 99mTc-RBC-EMs in mice showed higher uptake in the liver and spleen, and no uptake in the thyroid. Ex vivo imaging confirmed the in vivo findings. Furthermore, fluorescent imaging confirmed the nuclear imaging findings. Immunofluorescent imaging revealed that the hepatic uptake of RBC-EMs was significantly mediated by kupffer cells (resident hepatic macrophages). Our results demonstrate a simple yet large-scale production method for a novel type of RBC-EMs, which can be effectively labeled with 99mTc, and feasibly monitored in vivo by nuclear imaging. The RBC-EMs may be used as in vivo drug delivery vehicles.
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影响因子：
3.8
作者：
Alam H;Bhate AV;Gangadaran P;Sawant SS;Salot S;Sehgal L;Dange PP;Chaukar DA;D'cruz AK;Kannanl S;Gude R;Kane S;Dalal SN;Vaidya MM
通讯作者：
Vaidya MM
间充质干细胞衍生的外泌体作为肝病的新治疗策略。
影响因子：
12.8
作者：
Lou G;Chen Z;Zheng M;Liu Y
通讯作者：
Liu Y
影响因子：
16
作者：
Imai T;Takahashi Y;Nishikawa M;Kato K;Morishita M;Yamashita T;Matsumoto A;Charoenviriyakul C;Takakura Y
通讯作者：
Takakura Y
影响因子：
8
作者：
Banizs AB;Huang T;Dryden K;Berr SS;Stone JR;Nakamoto RK;Shi W;He J
通讯作者：
He J
影响因子：
10.8
作者：
Gangadaran, Prakash;Rajendran, Ramya Lakshmi;Ahn, Byeong-Cheol
通讯作者：
Ahn, Byeong-Cheol