Controllable release of interleukin-4 in double-layer sol–gel coatings on TiO 2 nanotubes for modulating macrophage polarization

Abstract Classically activated M1 macrophages and alternatively activated M2 macrophages play key roles in regulating immune responses. M1 macrophages initiate angiogenesis in the early stages of wound healing or after implantation. However, their prolonged activation can lead to chronic inflammation. We speculated that biomedical implants with specific properties can induce a shift from M1 to M2 macrophages at a specific time point to promote tissue repair and wound healing. To investigate this possibility, drug-loaded double-layer sol–gel coatings were fabricated on TiO 2 nanotubes (TNTs), which were used to modulate the switch from the M1 to the M2 phenotype by controlled release of interleukin (IL)-4. The lower sol–gel layer with IL-4 consisted of a carboxymethyl chitosan (CMCS) hydrogel, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, and N-hydroxysuccinimide as a crosslinker (IL4/TNT). The upper layer fabricated on the IL4/TNT sample was another type of CMCS hydrogel that used genipin (GP) as a crosslinker (GP/IL4/TNT). We found that IL-4 was released from GP/IL4/TNTs in a controlled manner, with the greatest release occurring after 72 h. GP/IL4/TNT stimulated the polarization of macrophages from the M1 to M2 phenotype after the macrophage polarization from the M0 to M1 phenotype. This provides a template for the fabrication of biomaterials that can direct macrophage polarization and stimulate tissue regeneration following the initial inflammatory response to implants.