Bio-based adhesives have gained considerable attention in the last years as more sustainable and healthier alternatives to the formaldehyde-based adhesives used today in wood-based panel manufacturing. In this study, dialdehyde starch (DAS) with various aldehyde contents was prepared by using sodium metaperiodate as an oxidizing agent. Characterizations were performed by employing Fourier-transform infrared spectroscopy, nuclear magnetic resonance, and thermal stability analysis. Different adhesive compositions were used for making medium-density fiberboard (MDF) panels. They were based on DAS (12 wt% based on fiber), emulsifiable diphenylmethane diisocyanate (eMDI, 2-4 wt% based on DAS), and microfibrillated cellulose (MFC, 0.5-1.0 wt% based on DAS). Fibers and the adhesive components were mixed with a combination of dry mixing and wet spraying. The physical and mechanical properties of MDF panels bonded with different DAS-based adhesives were compared with those of melamine urea-formaldehyde (MUF) adhesive and sole eMDI. The results showed that the MDF panels made with DAS-MFC-eMDI of 99.52% bio-based content showed comparable properties to standard panels with a commercial MUF adhesive. It was implied that DAS in the presence of small amount of eMDI can create strong bonds with wood fibers, while an additional positive effect on bonding was due to the contact surface enlargement of MFC.