A hyaluronic acid-based hydrogel enabling CD44-mediated chondrocyte binding and gapmer oligonucleotide release for modulation of gene expression in osteoarthritis
A hyaluronic acid-based hydrogel enabling CD44-mediated chondrocyte binding and gapmer oligonucleotide release for modulation of gene expression in osteoarthritis
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DOI：
10.1016/j.jconrel.2017.03.004
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发表时间：
2017-05-10
影响因子：
10.8
通讯作者：
Howard, Kenneth A.
中科院分区：
文献类型：
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作者：
Cai, Yunpeng;Lopez-Ruiz, Elena;Howard, Kenneth A.
透明质酸（HA）是一种有吸引力的生物材料，用于骨关节炎（OA）治疗，由于固有的功能和兼容性特性，作为一种内源性膝关节组件。在这项工作中，我们描述了一种基于HA的水凝胶，其具有增加的CD44依赖性软骨细胞结合和用于无辅助细胞进入和随后的基因沉默活性的gapmer反义寡核苷酸的受控释放的双重功能。采用Schiff碱介导的凝胶法制备了一组不同的水凝胶，其中明胶修饰的HA（900 kDa）与壳聚糖的比例为3：7、5：5和7：3，用于鉴定显示OA患者来源的表达CD44的软骨细胞的最佳结合的设计。发现细胞结合和CD44表达之间存在相关性，HA/壳聚糖比例为7：3时表现出最大结合，比CD44阴性MCF-7细胞对照细胞高181%。通过共聚焦和流式细胞术分析，证实了与裸siRNA相比，具有侧翼锁核酸（LNA）gapmer设计的荧光13-mer DNA寡核苷酸向OA软骨细胞的无转染剂摄取。发现7：3水凝胶在5天内持续和完全释放，相比之下，5：5和3：7水凝胶在相同时间内分别释放60%和43%的负载缺口聚物。设计具有最大软骨细胞基因沉默的考克斯-2特异性缺口聚物（与错配缺口聚物序列相比，在500 nM下类似于70%的沉默效率）导致在接种有OA软骨细胞的水凝胶中14天内有效的考克斯-2沉默，在第3天和第10天之间表现出显著差异。这项工作介绍了一种新的HA为基础的CD44介导的细胞结合和gapmer控制释放平台，以调节细胞基因表达。(C)2017爱思唯尔B.V.保留所有权利。
Hyaluronic acid (HA) is an attractive biomaterial for osteoarthritis (OA) treatment due to inherent functional and compatibility properties as an endogenous knee joint component. In thiswork, we describe a HA-based hydrogel with the dual functionality of increased CD44-dependent chondrocyte binding and controlled release of gapmer antisense oligonucleotides for unassisted cellular entry and subsequent gene silencing activity. A Schiff base-mediated gelationmethodwas used to produce a panel of hydrogels varying in the aldehyde-modifiedHA(900 kDa) to chitosan ratios (3: 7, 5: 5 and 7: 3) for identifying designs displaying optimal engagement of OA patient-derived CD44-expressing chondrocytes. Correlation was found between cell binding and CD44 expression, with maximal binding exhibited at a HA/chitosan ratio of 7: 3, that was 181% higher than CD44-negative MCF-7 cell control cells. Transfection agent-free uptake into OA chondrocytes of fluorescent 13-mer DNA oligonucleotides with a flanked locked nucleic acid (LNA) gapmer design, in contrast to naked siRNA, was demonstrated by confocal and flow cytometric analysis. A sustained and complete release over 5 days was found with the 7: 3 hydrogel, in contrast, the 5: 5 and 3: 7 hydrogel released 60% and 43% of loaded gapmers, respectively over the same period. A COX-2-specific gapmer designed with maximal chondrocyte gene silencing (similar to 70% silencing efficiency at 500 nM compared with a mismatch gapmer sequence) resulted in effective COX-2 silencing over 14 days in hydrogels seeded with OA chondrocytes, with significant difference exhibited between day 3 and 10. This work introduces a novel HA-based CD44-mediated cellular binding and gapmer controlled release platform to modulate cellular gene expression. (C) 2017 Elsevier B.V. All rights reserved.