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

作者：Cai Yunpeng; Lopez Ruiz Elena; Wengel Jesper; Creemers Laura B; Howard Kenneth A

来源：Journal of Controlled Release, 2017, 253: 153-159.

DOI：10.1016/j.jconrel.2017.03.004

• Hyaluronic acid
• Hydrogel
• Osteoarthritis
• CD44 receptor
• Gapmer
• Antisense

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.

• 出版日期2017-5-10