Macrophage Polarization Profiling on Native and Regenerated Silk Biomaterials

We investigated the plasticity and polarization of THP-1 cells on native and regenerated silk-based biomaterials to address the basic paradigm of immune response. Here, we report redox kinetics, adhesion morphology, and nitric oxide release patterns to identify specific subtypes of macrophages at different time points. Water-annealed silk film and native fibrous braids from Bombyx mori silkworms showed higher anti-inflammatory cytokine profiles or M2 subtypes (as evidenced by the enhanced expression of interleukin-10, interleukin-13, and interleukin-4). Ethanol-treated Bombyx mori silk films and Antheraea mylitta braids exhibited higher levels of pro-inflammatory cytokines or the M1 subtype (as evidenced by enhanced expression of interleukin-1, interleukin-6, interleukin-8, interferon-γ, TNF-α, and GM-CSF) in contact with healthy THP monocytes for 14 days; such a long study is unprecedented. Cytokine microarray analysis revealed the transition (M0–M1, M1–M2), plasticity, and stable phenotype of THP-1 cells in a variable stage in contact with different physicochemical properties of silk-based biomaterials. The detailed immunogenicity in the context of the physicochemical properties of native and regenerative silk-based biomaterials will enable us to accurately predict the possibility of a pro-/anti-inflammatory response. It will helps to predict the in vivo reprogramming and avoid fibrosis formation to enhance their clinical translational potential.