Green Jordan J, Shi Julie, Chiu Eugene, Leshchiner Elizaveta S, Langer Robert, Anderson Daniel G
Division of Biological Engineering, Massachusetts Institute of Technology, Cambridge, 02139, USA.
Bioconjug Chem. 2006 Sep-Oct;17(5):1162-9. doi: 10.1021/bc0600968.
Endothelial cells are an important cell type to both cardiovascular disease and cancer, as they play critical roles in vascular function and angiogenesis. However, effective and safe gene delivery to primary endothelial cells in the presence of serum proteins is known to be particularly challenging. A library of biodegradable poly(beta-amino esters) was synthesized for use as potential vectors. Promising vectors were optimized for high efficacy and low cytotoxicity to human umbilical vein endothelial cells (HUVECs) in serum. Vector parameters including polymer type, polymer weight, and DNA loading were varied, and biophysical properties including particle size, zeta potential, and particle stability over time were studied. While many of the poly(beta-amino ester) vectors have similar biophysical properties in the presence of buffer, their biophysical properties changed differentially in the presence of serum proteins, and the properties of these serum-interacting particles correlated to transfection efficacy. Leading poly(beta-amino ester) vectors were found to transfect HUVECs in the presence of serum significantly higher (47 +/- 9% positive, n = 10) than the best commercially available transfection reagents including jetPEI (p < 0.001) and Lipofectamine 2000 (p < 0.01). These results demonstrate the potential of a new class of biomaterials, poly(beta-amino esters), for effective human endothelial cell gene therapy.
内皮细胞对心血管疾病和癌症而言都是一种重要的细胞类型，因为它们在血管功能和血管生成中发挥着关键作用。然而，在存在血清蛋白的情况下，向原代内皮细胞进行有效且安全的基因递送尤其具有挑战性。合成了一个可生物降解的聚（β-氨基酯）文库用作潜在载体。对有前景的载体进行了优化，以使其在血清中对人脐静脉内皮细胞（HUVECs）具有高效能和低细胞毒性。改变了包括聚合物类型、聚合物重量和DNA负载在内的载体参数，并研究了包括粒径、zeta电位和颗粒随时间的稳定性在内的生物物理性质。虽然许多聚（β-氨基酯）载体在缓冲液存在下具有相似的生物物理性质，但它们在血清蛋白存在下的生物物理性质发生了不同的变化，并且这些与血清相互作用的颗粒的性质与转染效率相关。发现领先的聚（β-氨基酯）载体在血清存在下转染HUVECs的效率显著高于包括jetPEI（p < 0.001）和Lipofectamine 2000（p < 0.01）在内的最佳市售转染试剂（阳性率为47 +/- 9%，n = 10）。这些结果证明了一类新型生物材料聚（β-氨基酯）在有效的人类内皮细胞基因治疗中的潜力。
