Increasing total solids in anaerobic digestion can reduce the methane yield by highly complex bio-physical–chemical mechanisms. Therefore, understanding those mechanisms and their main drivers becomes crucial to optimize this waste treatment biotechnology. In this study, seven batch experiments were conducted to investigate the effects of increasing the initial total solids in high-solids anaerobic digestion of the organic fraction of municipal solid waste. With inoculum-to-substrate ratio = 1.5 g VS/g VS and maximum total solids ≤ 19.6%, mono-digestion of the organic fraction of municipal solid waste showed a methane yield = 174–236 NmL CH4/g VS. With inoculum-to-substrate ratio ≤ 1.0 g VS/g VS and maximum total solids ≥ 24.0%, mono-digestion experiments acidified. Co-digestion of the organic fraction of municipal solid waste and beech sawdust permitted to reduce the inoculum-to-substrate ratio to 0.16 g VS/g VS while increasing total solids up to 30.2%, though achieving a lower methane yield (117–156 NmL CH4/g VS). At each inoculum-to-substrate ratio, higher total solids corresponded to higher ammonia and volatile fatty acid accumulation. Thus, a 40% lower methane yield for mono-digestion was observed at a NH3 concentration ≥ 2.3 g N–NH3/kg reactor content and total solids = 15.0%. Meanwhile, co-digestion lowered the nitrogen content, being the risk of acidification exacerbated only at total solids ≥ 20.0%. Therefore, the biodegradability of the substrate, as well as the operational total solids and inoculum-to-substrate ratio, are closely interrelated parameters determining the success of methanogenesis, but also the risk of ammonia inhibition in high-solids anaerobic digestion.