Lentivirus Immobilization to Nanoparticles for Enhanced and Localized Delivery From Hydrogels

Abstract

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.

Scaffold-mediated lentiviral transduction for functional tissue engineering of cartilage

“…Our findings extend previous work showing biomaterialmediated gene delivery using viral vectors (20,(40)(41)(42)(43)(44)(45)(46)(47)(48)(49). The majority of this work demonstrated the ability to specify the spatial location of delivered vectors, rather than the induction of tissuespecific differentiation.…”

Dynamic transcription factor activity and networks during ErbB2 breast oncogenesis and targeted therapy

“…Lentivirus for each TFr was produced by co-transfecting HEK-293T cells with one of the transfer vector constructs and three packaging plasmids (pMDL-GagPol, pRSV-Rev, and pIVS-VSV-G) 48 using techniques described previously 49 . Number of physical particles (PP) for each virus batch was determined using an HIV-1 p24 Antigen ELISA kit (ZeptoMetrix, Buffalo, NY).…”

Harnessing the Potential of Biomaterials for Brain Repair after Stroke

“…Biomaterials can be used to address these concerns by sequestering viral particles, increasing local concentrations of the vector and extending the length of virus activity to increase transfection efficiency of infiltrating cells. Retaining the viral particles within the gel limits transfection to cells at the boundary and to infiltrating cells, controlling reprogramming to cells solely within the glial scar-bounded infarct and reducing diffuse non-specific exposure (Shin and Shea, 2010;Seidlits et al, 2013). Recent developments have found that cocktails of small molecules can achieve the same reprogramming as viral vectors, without xenobiotic concerns, in astroglial cells (Zhang et al, 2015) and fibroblasts (Hu et al, 2015;Li et al, 2015).…”