Division of Cardiology, The Children's Hospital of Philadelphia, Abramson Research Center, and Department of Pediatrics, Perelman School of Medicine of the University of Pennsylvania, Philadelphia, PA 19104, USA.
Biomaterials. 2013 Sep;34(28):6938-48. doi: 10.1016/j.biomaterials.2013.05.047. Epub 2013 Jun 15.
The use of arterial stents and other medical implants as a delivery platform for surface immobilized gene vectors allows for safe and efficient localized expression of therapeutic transgenes. In this study we investigate the use of hydrolyzable cross-linkers with distinct kinetics of hydrolysis for delivery of gene vectors from polyallylamine bisphosphonate-modified metal surfaces. Three cross-linkers with the estimated t1/2 of ester bonds hydrolysis of 5, 12 and 50 days demonstrated a cumulative 20%, 39% and 45% vector release, respectively, after 30 days exposure to physiological buffer at 37 °C. Transgene expression in endothelial and smooth muscles cells transduced with substrate immobilized adenovirus resulted in significantly different expression profiles for each individual cross-linker. Furthermore, immobilization of adenoviral vectors effectively extended their transduction effectiveness beyond the initial phase of release. Transgene expression driven by adenovirus-tethered stents in rat carotid arteries demonstrated that a faster rate of cross-linker hydrolysis resulted in higher expression levels at day 1, which declined by day 8 after stent implantation, while inversely, slower hydrolysis was associated with increased arterial expression at day 8 in comparison with day 1. In conclusion, adjustable release of transduction-competent adenoviral vectors from metallic surfaces can be achieved, both in vitro and in vivo, through surface immobilization of adenoviral vectors using hydrolyzable cross-linkers with structure-specific release kinetics.
作为表面固定基因载体的递送平台,动脉支架和其他医疗植入物允许安全有效地局部表达治疗性转基因。在这项研究中,我们研究了使用具有不同水解动力学的可水解交联剂从聚烯丙基双膦酸盐修饰的金属表面递送电 载体。三种交联剂的酯键水解半衰期估计分别为 5、12 和 50 天,在 37°C 生理缓冲液中暴露 30 天后,分别释放了 20%、39%和 45%的载体。用基质固定的腺病毒转导的内皮细胞和平滑肌细胞中的转基因表达导致每个交联剂的表达谱显著不同。此外,腺病毒载体的固定有效地延长了其转导效果,超过了释放的初始阶段。腺病毒结合支架在大鼠颈动脉中的转基因表达表明,交联剂水解速度越快,支架植入后第 1 天的表达水平越高,但在第 8 天下降,而相反,较慢的水解与第 8 天的动脉表达水平高于第 1 天有关。总之,通过使用具有结构特异性释放动力学的可水解交联剂将腺病毒载体表面固定,可以在体外和体内实现具有转导能力的腺病毒载体的可调节释放。