A hyaluronic acid-based hydrogel enabling CD44-mediated chondrocyte binding and gapmer oligonucleotide release for modulation of gene expression in osteoarthritis

JOURNAL OF CONTROLLED RELEASE

卷:253

Article

Cai, Yunpeng¹  Lopez-Ruiz, Elena^1,2  Wengel, Jesper³  Creemers, Laura B.⁴  Howard, Kenneth A.¹ 

[1] Aarhus Univ, Interdisciplinary Nanosci Ctr iNANO, Dept Mol Biol & Genet, DK-8000 Aarhus C, Denmark

[2] Univ Jaen, Dept Hlth Sci, E-23071 Jaen, Spain

[3] Univ Southern Denmark, Nucle Acid Ctr, Dept Phys Chem & Pharm, DK-5230 Odense M, Denmark

[4] Univ Med Ctr Utrecht, Dept Orthopaed, NL-3584 CX Utrecht, Netherlands

关键词: Hyaluronic acid;    Hydrogel;    Osteoarthritis;    CD44 receptor;    Gapmer;    Antisense;   

DOI  :  10.1016/j.jconrel.2017.03.004

来源: Elsevier

【 摘 要 】

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.

【 授权许可】

Free