The increased interest in printed electronics necessitates the development of suitable sustainable substrates for them. In this study, the suitability of acetylated cellulose nanofiber (ACNF) films as substrates for printed electronics were examined through (I) the ink-substrates interaction, (II) print quality, and (III) electrical and (IV) mechanical properties of the printed pattern on the ACNF substrates. The results have been compared with cellulose nanofiber (CNF) and commercial reference material (CRM) substrates. The wetting of the silver nanoparticle (AgNP) ink on the ACNF substrate was found out to be excellent. The thickness of the printed pattern increased and the hole area fraction decreased with each consecutive layer of ink. The comparative investigations demonstrated that the electrical properties of the printed patterns were almost as good for 4 layers of ink on the ACNF substrate (1.6 Ω resistance) as the ones on the CRM substrate (1.2 Ω resistance). Additionally, the printed pattern on the ACNF substrate endured the adhesion and bending tests significantly better compared to the CRM substrate. Therefore, this study demonstrates that ACNF substrates could be a suitable candidate for printed electronics, which are more sustainable yet with similar functionalities in comparison with the conventional fossil based substrates.