Department of Chemical and Biological Engineering, Northwestern University, Evanston, Illinois 60208-3120, USA.
Mol Ther. 2010 Apr;18(4):700-6. doi: 10.1038/mt.2009.300. Epub 2010 Jan 5.
Hydrogels can provide a controllable cell microenvironment for numerous applications in regenerative medicine, and delivery of gene therapy vectors can be employed to enhance their bioactivity. We investigated the delivery of lentiviral vectors from hydrogels, and employed the immobilization of lentivirus to hydroxylapatite (HA) nanoparticles as a means to retain and stabilize vectors within hydrogels, and thereby increase delivery efficiency. Entrapment of the vector alone within the hydrogel maintained the activity of the virus more effectively compared to the absence of a hydrogel, and release was slowed with an increasing solid content of the hydrogel. Association of the lentivirus with HA increased the activity of the complexes, with HA increasing the virus activity for 72 hours. Cells seeded onto lentivirus-HA-loaded hydrogels had a decreased number of infected cells outside of the hydrogel relative to the absence of HA. In vivo studies with collagen hydrogels loaded with lentivirus and HA-lentivirus demonstrated sustained and localized transgene expression for at least 4 weeks, with increased expression using the lentivirus-HA complex. This strategy of nanoparticle immobilization to stabilize and retain vectors is broadly applicable to hydrogels, and may provide a versatile tool to combine gene therapy and biomaterials for applications in regenerative medicine.
水凝胶可为再生医学中的众多应用提供可控的细胞微环境，并且可以通过基因治疗载体的传递来提高其生物活性。我们研究了从水凝胶中传递慢病毒载体，并采用将慢病毒固定在羟基磷灰石（HA）纳米颗粒上来保留和稳定水凝胶内的载体，从而提高传递效率。与没有水凝胶相比，将载体单独包埋在水凝胶中更有效地保持了病毒的活性，并且随着水凝胶固体含量的增加，释放速度减慢。慢病毒与 HA 的结合增加了复合物的活性，HA 使病毒的活性增加了 72 小时。与不存在 HA 相比，接种在负载有慢病毒-HA 的水凝胶上的细胞在水凝胶外的感染细胞数量减少。用负载有慢病毒和 HA-慢病毒的胶原水凝胶进行的体内研究表明，至少在 4 周内持续且局部表达转基因，使用慢病毒-HA 复合物可增加表达。这种固定纳米颗粒以稳定和保留载体的策略广泛适用于水凝胶，并且可能为将基因治疗和生物材料结合应用于再生医学提供一种通用工具。
