Lentivirus delivery of IL-10 to promote and sustain macrophage polarization towards an anti-inflammatory phenotype.

Gene delivery from biomaterials can create an environment that promotes and guides tissue formation. However, the immune response induced upon biomaterial implantation can be detrimental to tissue regeneration. Macrophages play a central role in mediating early phases of this response, and functional "polarization" of macrophages towards M1 (inflammatory) or M2 (anti-inflammatory) phenotypes may bias the local immune state at the implant site. Since gene delivery from biomaterial scaffolds can confer transgene expression in macrophages in vivo, we investigated whether transduction of macrophages with an IL-10 encoding lentivirus can (1) induce macrophage polarization toward an M2 phenotype even in an pro-inflammatory environment, and (2) prevent a shift in polarization from M2 to M1 following exposure to pro-inflammatory stimuli. IL-10 lentivirus delivery to pre-polarized M1 macrophages reduced TNF-α production 1.5-fold when compared to cells treated with either a control virus or a bolus delivery of recombinant IL-10 protein. IL-10 lentivirus delivery to naïve macrophages reduced the amount of TNF-α produced following an inflammatory challenge by 2.5-fold compared to cells treated with both the control virus and recombinant IL-10. At a mechanistic level, IL-10 lentivirus delivery mediated sustained reduction in NF-κB activation and, accordingly, reduced transcription of TNF-α. In sum, lentiviral delivery of IL-10 to macrophages represents a promising strategy for directing and sustaining macrophage polarization towards an M2 phenotype in order to promote local immune responses that facilitate tissue engineering.

A PLG/HAp composite scaffold for lentivirus delivery.

Gene delivery from tissue engineering scaffolds provides the opportunity to control the microenvironment by inducing expression of regenerative factors. Hydroxyapatite (HAp) nanoparticles can bind lentivirus, and we investigated the incorporation of HAp into poly(lactide-co-glycolide) (PLG) scaffolds in order to retain lentivirus added to the scaffold. PLG/HAp scaffolds loaded with lentivirus enhanced transgene expression over 10-fold in vitro relative to scaffolds without HAp. Following in vivo implantation, PLG/HAp scaffolds promoted transgene expression for more than 100 days, with the level and duration enhanced relative to control scaffolds with lentivirus/HAp complexes added to PLG scaffolds. The extent of HAp incorporated into the scaffold influenced transgene expression, in part through its impact on porous architecture. Expression in vivo was localized to PLG/HAp scaffolds, with macrophages the primary cell type transduced at day 3, yet transduction of neutrophils and dendritic cells was also observed. At day 21 in PLG/HAp scaffolds, non-immune cells were transduced to a greater extent than immune cells, a trend that was opposite results from PLG scaffolds. Thus, in addition to retaining the virus, PLG/HAp influenced cell infiltration and preferentially transduced non-immune cells.