Division of Trauma, Burns and Critical Care, Department of Surgery, 200 W. Arbor Dr., University of California-San Diego, San Diego, CA 92103-8236, USA.
Biomaterials. 2009 Dec;30(35):6788-93. doi: 10.1016/j.biomaterials.2009.07.069. Epub 2009 Sep 24.
Because there are few reports using gene delivery in clinically-approved synthetic matrices, we examined the feasibility of using a noninvasive imaging system to study the kinetics of luciferase gene expression when delivered in an adenoviral vector. Using a mouse model of full thickness injury, we quantified the kinetics of gene expression, determined the optimal dose of particle delivery, and established the temporal importance of drug delivery in obtaining optimal gene expression. Specifically, we found that the ideal time to deliver adenovirus to a graft is during the early phase of graft wound closure (days 0-3 post-operatively) for a peak of gene expression to occur 7 days after delivery. Under these conditions, there is a saturating dose of 6 x 10(8) adenoviral particles per graft. In light of these findings, we examined whether the efficacy of delivery could be increased by modulating the composition of the grafts. When a collagen gene-activated matrix (GAM) containing basic fibroblast growth factor (FGF2) was compared to matrix alone, a significant increase in gene expression is observed when identical amounts of vector are delivered (p<0.05). Taken together, these results show how a noninvasive and quantitative assessment of gene expression can be used to optimize gene delivery and that the composition of matrices can dramatically influence gene expression in the wound bed.
由于使用基因传递在临床批准的合成基质中的报道较少，我们研究了使用非侵入性成像系统研究在腺病毒载体中传递时荧光素酶基因表达的动力学的可行性。使用全层损伤的小鼠模型，我们量化了基因表达的动力学，确定了颗粒传递的最佳剂量，并确定了在获得最佳基因表达时药物传递的时间重要性。具体来说，我们发现将腺病毒递送至移植物的理想时间是在移植物伤口闭合的早期阶段（术后 0-3 天），以便在递送后 7 天发生峰值基因表达。在这些条件下，每个移植物的腺病毒颗粒的饱和剂量为 6x10(8)。鉴于这些发现，我们研究了通过调节移植物的组成是否可以提高传递的功效。当含有碱性成纤维细胞生长因子 (FGF2)的胶原基因激活基质 (GAM)与基质单独比较时，当传递相同量的载体时，观察到基因表达的显著增加（p<0.05）。总之，这些结果表明如何使用非侵入性和定量评估基因表达来优化基因传递，并且基质的组成可以极大地影响伤口床中的基因表达。
