Intracellular delivery of liposome-encapsulated Finland trityl radicals for EPR oximetry.
Intracellular delivery of liposome-encapsulated Finland trityl radicals for EPR oximetry.
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DOI：
10.1039/d0an00108b
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发表时间：
2020-06
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影响因子：
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通讯作者：
Xing Wang;Chao Peng;K. He;Kaiyun Ji;Xiaoli Tan;Guifang Han;Yanxia Liu;Yangping Liu;Yuguang Song
中科院分区：
文献类型：
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作者：
Xing Wang;Chao Peng;K. He;Kaiyun Ji;Xiaoli Tan;Guifang Han;Yanxia Liu;Yangping Liu;Yuguang Song
四硫代三芳基甲基（TAM，三苯甲基）自由基在电子顺磁共振（EPR）血氧测定中有着广泛的应用。然而，TAM自由基的生物医学应用仅限于细胞外区域，这是由于其带负电荷的性质。TAM自由基的细胞内递送仍然是一个挑战。在目前的工作中，我们报告了一种基于脂质体的方法来封装水溶性芬兰三苯甲基自由基CT-03的细胞内交付。采用脂质薄膜水化法，以DSPC/胆固醇/DOTAP为原料制备了CT-03脂质体，其平均粒径为167.5 ± 2.4 nm，zeta电位为27.8 ± 0.8 mV。EPR结果表明，CT-03包封在脂质体中，并仍表现出良好的氧（O2）敏感性。此外，使用负载CT-03的脂质体成功地将CT-03递送到HepG 2细胞和HUVECs中。重要的是，脂质体包封的自由基CT-03和其他TAM自由基CT 02-H的组合使得能够同时测量HepG 2细胞中的细胞内和细胞外O2浓度和O2消耗速率。我们目前的研究提供了一种新的途径，细胞内传递TAM自由基，并可能显着扩大其生物医学应用。
Tetrathiatriarylmethyl (TAM, trityl) radicals have found wide applications in electron paramagnetic resonance (EPR) oximetry. However, the biomedical applications of TAM radicals were exclusively limited to an extracellular region owing to their negatively charged nature. The intracellular delivery of TAM radicals still remains a challenge. In the present work, we report a liposome-based method to encapsulate the water-soluble Finland trityl radical CT-03 for its intracellular delivery. Using the thin lipid film hydration method, CT-03-loaded liposomes were prepared from DSPC/cholesterol/DOTAP with a mean size of 167.5 ± 2.4 nm and a zeta potential of 27.8 ± 0.8 mV. EPR results showed that CT-03 was entrapped into the liposomes and still exhibited good oxygen (O2) sensitivity. Moreover, CT-03 was successfully delivered into HepG2 cells and HUVECs using the CT-03-loaded liposomes. Importantly, the combination of the liposome-encapsulated radical CT-03 and the other TAM radical CT02-H enabled simultaneous measurements of the intracellular and extracellular O2 concentrations and O2 consumption rates in HepG2 cells. Our present study provides a new approach for intracellular delivery of TAM radicals and could significantly expand their biomedical applications.