Osteoblast-Derived Extracellular Vesicles Are Biological Tools for the Delivery of Active Molecules to Bone
Osteoblast-Derived Extracellular Vesicles Are Biological Tools for the Delivery of Active Molecules to Bone
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DOI：
10.1002/jbmr.3332
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发表时间：
2018-03-01
影响因子：
6.2
通讯作者：
Teti, Anna
中科院分区：
文献类型：
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作者：
Cappariello, Alfredo;Loftus, Alexander;Teti, Anna
细胞外囊泡是近年来备受关注的组织稳态调节因子，也是细胞间通讯的一种手段。报道已经调查了EV及其货物在细胞调节中的作用，并试图微调其生物技术用途，但迄今为止对其在骨生物学中的功能知之甚少。为了研究EV介导的骨细胞间通讯的相关性，我们从原代小鼠成骨细胞中分离EV，并通过透射电子显微镜（TEM）和荧光激活细胞分选（FACS）评估膜的完整性、大小和结构。EV主动将负载的荧光染料穿梭到成骨细胞、单核细胞和内皮细胞。此外，成骨细胞EV含有与供体细胞共享的mRNA。已知成骨细胞通过前骨细胞因子、核因子-B配体的受体活化剂（Rankl）调节破骨细胞生成、破骨细胞存活和破骨细胞功能。成骨细胞EV在Rankl中富集，其在PTH处理后增加。这些EV具有生物活性，支持破骨细胞存活。从rankl（-/-）成骨细胞分离的EV失去了这种促破骨细胞功能，表明其Rankl依赖性。它们离体整合到小鼠颅骨中，并且在体内EV全身给药后，EV穿梭荧光染料迅速被骨吸收。Rankl（-/-）小鼠缺乏破骨细胞谱系，并且其特异性标志物抗酒石酸酸性磷酸酶（TRAcP）为阴性。用野生型成骨细胞EV处理rankl（-/-）小鼠以EV密度依赖性方式诱导TRAcP阳性细胞的出现。最后，成骨细胞EV内化并穿梭抗破骨细胞药物（唑来膦酸盐和达沙替尼），在体外和体内抑制破骨细胞活性。我们的结论是，成骨细胞EV参与骨细胞之间的细胞间通讯，有助于Rankl pro-osteocytes效应，并穿梭抗破骨细胞药物，代表了一种潜在的靶向治疗手段。(c)2017作者骨与矿物质研究杂志由Wiley Periodicals Inc.出版。
Extracellular vesicles (EVs) are newly appreciated regulators of tissue homeostasis and a means of intercellular communication. Reports have investigated the role of EVs and their cargoes in cellular regulation and have tried to fine-tune their biotechnological use, but to date very little is known on their function in bone biology. To investigate the relevance of EV-mediated communication between bone cells, we isolated EVs from primary mouse osteoblasts and assessed membrane integrity, size, and structure by transmission electron microscopy (TEM) and fluorescence-activated cell sorting (FACS). EVs actively shuttled loaded fluorochromes to osteoblasts, monocytes, and endothelial cells. Moreover, osteoblast EVs contained mRNAs shared with donor cells. Osteoblasts are known to regulate osteoclastogenesis, osteoclast survival, and osteoclast function by the pro-osteoclastic cytokine, receptor activator of nuclear factor -B ligand (Rankl). Osteoblast EVs were enriched in Rankl, which increased after PTH treatment. These EVs were biologically active, supporting osteoclast survival. EVs isolated from rankl(-/-) osteoblasts lost this pro-osteoclastic function, indicating its Rankl-dependence. They integrated ex vivo into murine calvariae, and EV-shuttled fluorochromes were quickly taken up by the bone upon in vivo EV systemic administration. Rankl(-/-) mice lack the osteoclast lineage and are negative for its specific marker tartrate-resistant acid phosphatase (TRAcP). Treatment of rankl(-/-) mice with wild-type osteoblast EVs induced the appearance of TRAcP-positive cells in an EV density-dependent manner. Finally, osteoblast EVs internalized and shuttled anti-osteoclast drugs (zoledronate and dasatinib), inhibiting osteoclast activity in vitro and in vivo. We conclude that osteoblast EVs are involved in intercellular communication between bone cells, contribute to the Rankl pro-osteoclastic effect, and shuttle anti-osteoclast drugs, representing a potential means of targeted therapeutic delivery. (c) 2017 The Authors. Journal of Bone and Mineral Research Published by Wiley Periodicals Inc.