Holladay Carolyn, Keeney Michael, Greiser Udo, Murphy Mary, O'Brien Timothy, Pandit Abhay
Network of Excellence in Functional Biomaterials, National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland.
J Control Release. 2009 Jun 19;136(3):220-5. doi: 10.1016/j.jconrel.2009.02.006. Epub 2009 Feb 20.
Non-viral gene delivery suffers from a number of limitations including short transgene expression times and low transfection efficiency. Collagen scaffolds have previously been investigated as in vitro DNA reservoirs, which allow sustained release of genetic information. Efficient viral gene-transfer from these scaffolds has previously been demonstrated. However, due to concerns about the safety of viral gene therapy, the use of non-viral vectors may be preferable. In this study a DNA-dendrimer complex embedded in a cross-linked collagen scaffold was investigated as a reservoir for non-viral delivery. Elution from the scaffolds and transfection of seeded rat mesenchymal stem cells were used to evaluate the scaffold's ability to act as a reservoir for the complexes. Elution from the scaffolds was minimal after 2 days with a total of 25% of the complexes released after 7 days. Extended transgene expression after DNA-dendrimer complex delivery from the scaffolds in comparison to direct delivery to cells was observed. The elongated transfection period and relatively high levels of reporter gene expression are significant advantages over other non-viral gene therapy techniques. This platform has the potential to be an effective method of scaffold-mediated gene delivery suitable for in vitro and in vivo applications.
非病毒基因递送存在许多局限性，包括转基因表达时间短和转染效率低。胶原蛋白支架先前已被研究作为体外DNA储存库，可实现遗传信息的持续释放。先前已证明从这些支架进行高效的病毒基因转移。然而，由于对病毒基因治疗安全性的担忧，使用非病毒载体可能更可取。在本研究中，研究了嵌入交联胶原蛋白支架中的DNA-树枝状聚合物复合物作为非病毒递送的储存库。通过从支架上洗脱以及对接种的大鼠间充质干细胞进行转染，来评估支架作为复合物储存库的能力。2天后支架的洗脱量极少，7天后总共释放了25%的复合物。与直接递送至细胞相比，观察到从支架递送DNA-树枝状聚合物复合物后转基因表达延长。延长的转染期和相对较高水平的报告基因表达是相对于其他非病毒基因治疗技术的显著优势。该平台有可能成为一种适用于体外和体内应用的支架介导基因递送的有效方法。
