Redefining Post-detoxification with Novel Surface Coatings: Proteomic Insights into the Early Host Response

Conventional peri-implantitis treatments often rely on detoxification protocols to reduce surface-associated biofilms. However, biofilm removal alone may be insufficient, as successful outcomes ultimately depend on subsequent cellular and molecular responses that support tissue healing. Cellular interactions after detoxification are influenced by proteins at the implant-tissue interface, and strategies to engineer a proregenerative protein profile following detoxification remain unexplored. This study investigated the potential of antimicrobial and lubricative dicatonic imidazolium-based ionic liquid (IL) coating to modulate protein adsorption following clinically relevant detoxification protocols. The interfacial proteome on titanium (Ti) surfaces was evaluated at 24 h using a subcutaneous rat model after exposure to a mixed-species oral biofilm, detoxification, and subsequent IL application. Experimental groups included implants detoxified with citric acid (CA 40%), chlorhexidine (CHX 1%), or saline (SA 0.9%), as well as combination groups with IL (CAIL, CHXIL, and SAIL). Liquid chromatography-tandem mass spectrometry analysis recovered 318,659 peptides representing 5084 proteins. Detoxification monotherapies (CA, CHX, and SA) were associated with distinct interfacial proteomic profiles, including patterns with pro-inflammatory signaling. To directly evaluate the effect of IL after each detoxification treatment, we compared CAIL vs CA, CHXIL vs CHX, and SAIL vs SA. The strongest direct protein-level IL-associated effect was observed following CA treatment, whereas no proteins met the pre-specified significance criteria of |log2FC| > 0.58 and false-discovery rate < 0.05 for CHXIL vs CHX or SAIL vs SA. Pathway analysis showed a treatment-dependent pattern consistent with NRF2-mediated cytoprotective signaling, Th2-associated immune modulation, tissue repair, cell adhesion, and inflammatory responses; we interpreted these pathway findings as predicted molecular associations rather than direct evidence of pathway activation or suppression. Altogether, these results suggest that post-detoxification IL application may further modify the early interfacial proteomic response in a treatment-dependent manner, supporting further investigation of IL coating as a strategy to improve biomaterial-host interactions following implant-surface detoxification.