Quantitative Biosciences and Bioengineering, Colorado School of Mines, Golden, Colorado, USA.
Chemical and Biological Engineering, Colorado School of Mines, Golden, Colorado, USA.
J Biomed Mater Res A. 2024 Dec;112(12):2346-2357. doi: 10.1002/jbm.a.37784. Epub 2024 Aug 15.
The growth plate is a cartilage structure at the end of long bones which mediates growth in children. When fractured, the formation of bony repair tissue known as a "bony bar" can occur and cause limb deformities. There are currently no effective clinical solutions for the prevention of the bony bar formation or regeneration of healthy growth plate cartilage after a fracture. This study employs previously developed alginate/chitosan polyelectrolyte complex (PEC) hydrogels as a sustained release vehicle for the delivery of short-interfering RNA (siRNA). Specifically, the siRNA targets the p38-MAPK pathway in mesenchymal stem cells (MSCs) to prevent their osteogenic differentiation. In vitro experimental findings show sustained release of siRNA from the hydrogels for 6 months. Flow cytometry and confocal imaging indicate that the hydrogels release siRNA to effectively knockdown GFP expression over a sustained period. MAPK-14 targeting siRNA was used to knockdown the expression of MAPK-14 and correspondingly decrease the expression of other osteogenic genes in MSCs in vitro over the span of 21 days. These hydrogels were used in a rat model of growth plate injury to determine whether siMAPK-14 released from the gels could inhibit bony bar formation. No significant reduction of bony bar formation was seen in vivo at the one concentration of siRNA examined. This PEC hydrogel represents a significant advancement for siRNA sustained delivery, and presents an interesting potential therapeutic delivery system for growth plate injuries and other regenerative medicine applications.
生长板是长骨末端的软骨结构,介导儿童的生长。骨折时,可能会形成称为“骨条”的骨修复组织,导致肢体畸形。目前,尚无有效的临床解决方案来预防骨条形成或骨折后健康生长板软骨的再生。本研究采用先前开发的藻酸盐/壳聚糖聚电解质复合物(PEC)水凝胶作为一种持续释放载体,用于输送小干扰 RNA(siRNA)。具体来说,该 siRNA 靶向骨髓间充质干细胞(MSCs)中的 p38-MAPK 通路,以防止其成骨分化。体外实验结果表明,水凝胶中的 siRNA 可在 6 个月内持续释放。流式细胞术和共聚焦成像表明,水凝胶在一段时间内有效释放 siRNA 以持续敲低 GFP 表达。MAPK-14 靶向 siRNA 用于敲低 MAPK-14 的表达,并相应减少 MSC 中其他成骨基因在 21 天内的表达。这些水凝胶在生长板损伤的大鼠模型中被用于确定凝胶中释放的 siMAPK-14 是否可以抑制骨条形成。在体内,在所检查的 siRNA 浓度下,未见骨条形成有明显减少。该 PEC 水凝胶代表了 siRNA 持续递送的重大进展,并为生长板损伤和其他再生医学应用提供了一种有趣的潜在治疗性递药系统。
J Biomed Mater Res B Appl Biomater. 2020-8