A novel collagen-nanohydroxyapatite microRNA-activated scaffold for tissue engineering applications capable of efficient delivery of both miR-mimics and antagomiRs to human mesenchymal stem cells
A novel collagen-nanohydroxyapatite microRNA-activated scaffold for tissue engineering applications capable of efficient delivery of both miR-mimics and antagomiRs to human mesenchymal stem cells
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DOI：
10.1016/j.jconrel.2014.12.034
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发表时间：
2015-02-28
影响因子：
10.8
通讯作者：
O'Brien, Fergal J.
中科院分区：
文献类型：
--
作者：
Castano, Irene Mencia;Curtin, Caroline M.;O'Brien, Fergal J.
通过使用microRNAs （miRNAs）来操纵基因表达为组织工程领域提供了巨大的潜力。然而，缺乏足够的位点特异性和生物活性传递系统严重阻碍了基于mirna的治疗的临床转化。在这项研究中，我们为miRNA模拟物和拮抗剂开发了一种新的非病毒生物活性递送平台，以实现广泛的治疗应用。通过将纳米羟基磷灰石（nHA）颗粒与报告miRNAs （nanomiRs）和胶原-纳米羟基磷灰石支架结合，本研究引入了迄今为止第一个非病毒、非脂质平台，能够将成熟的miRNA分子有效地传递到人间充质干细胞（hMSCs）中，hMSCs是一种特别难以有效转染的细胞类型，并且具有最小的治疗相关细胞毒性。首先，mirna以单层形式被成功递送到hMSCs中，纳米mir -mimics和纳米拮抗剂的内化效率分别为17.4%和39.6%，纳米mir -mimics和纳米拮抗剂在单层中7天内的持续干扰活性均大于90%。当应用于3D支架时，在7天的时间内，纳米mir模拟物的RNA干扰率为20%，纳米拮抗剂的RNA干扰率为88.4%，没有细胞毒性问题。总之，内部合成的非病毒nHA颗粒在单层和支架上都有效地传递了报告mirna，这表明了这种创新的mirna激活支架系统在组织工程应用中的巨大潜力。(C) 2014 Elsevier B.V.版权所有
Manipulation of gene expression through the use of microRNAs (miRNAs) offers tremendous potential for the field of tissue engineering. However, the lack of sufficient site-specific and bioactive delivery systems has severely hampered the clinical translation of miRNA-based therapies. In this study, we developed a novel non-viral bioactive delivery platform for miRNA mimics and antagomiRs to allow for a vast range of therapeutic applications. By combining nanohydroxyapatite (nHA) particles with reporter miRNAs (nanomiRs) and collagen-nanohydroxyapatite scaffolds, this work introduces the first non-viral, non-lipid platform to date, capable of efficient delivery of mature miRNA molecules to human mesenchymal stem cells (hMSCs), a particularly difficult cell type to transfect effectively, with minimal treatment-associated cytotoxicity. Firstly, miRNAs were successfully delivered to hMSCs in monolayer, with internalisation efficiencies of 17.4 and 39.6% for nanomiR-mimics and nanoantagomiRs respectively, and both nanomiR-mimics and nanoantagomiRs yielded sustained interfering activity of greater than 90% in monolayer over 7 days. When applied to 3D scaffolds, significant RNA interference of 20% for nanomiR-mimics and 88.4% for nanoantagomiRs was achieved with no cytotoxicity issues over a 7 day period. In summary, in-house synthesised non-viral nHA particles efficiently delivered reporter miRNAs both in monolayer and on scaffolds demonstrating the immense potential of this innovative miRNA-activated scaffold system for tissue engineering applications. (C) 2014 Elsevier B.V. All rights reserved.