Localized heat generation by the thermo-plasmonic effect of metal nanoparticles has great potential in biomedical engineering research. Precise patterning of the nanoparticles using inkjet printing can enable the application of the thermo-plasmonic effect in a well-controlled way (shape and intensity). However, a universally applicable inkjet printing process that allows good control in patterning and assembly of nanoparticles with good biocompatibility is missing. Here we developed inkjet-printing-based biofunctional thermo-plasmonic interfaces that can modulate biological activities. We found that inkjet printing of plasmonic nanoparticles on a polyelectrolyte layer-by-layer substrate coating enables high-quality, biocompatible thermo-plasmonic interfaces across various substrates (rigid/flexible, hydrophobic/hydrophilic) by induced contact line pinning and electrostatically assisted nanoparticle assembly. We experimentally confirmed that the generated heat from the inkjet-printed thermo-plasmonic patterns can be applied in micrometer resolution over a large area. Lastly, we demonstrated that the patterned thermo-plasmonic effect from the inkjet-printed gold nanorods can selectively modulate neuronal network activities. This inkjet printing process therefore can be a universal method for biofunctional thermo-plasmonic interfaces in various bioengineering applications.