Several studies have shown the benefits of using timber in the construction sector in terms of climate change, which has received the support of decision-makers. However, few studies focus on other environmental indicators. The environmental impacts (i.e., such as human health, ecosystem quality, and resources) of material substitutions, consequence of the increasing use of wood, are still lacking in the literature. Studies that compare different structural materials focus either on a single building or on one structural system that would be representative on a larger scale. However, the environmental benefits of some projects are not easily extrapolated to the scale of a given region with a variety of architectures. Higher-order effects, beyond the building scale, can indeed imply changes in the resource availability according to the unconstrained suppliers and resources. The objective of this paper is to assess the life cycle environmental consequences of wood substitutions at a regional scale for the non-residential construction sector. The increasing use of wood structures for non-residential buildings in the province of Quebec (Canada) is used as a case study. The method includes the development of material substitution factors by comparing several structures. The substitution factors show that, on average, wood can replace, simultaneously, 0.59 and 4.54 times the weight of steel and concrete, respectively. However, steel substitution has more advantages than concrete substitution. Among the tested parameters, the variability of the material substitution factors implies more uncertainties in the results. The originality of our study lies in the limitations that may change the conclusions drawn from the substitution at a large scale. With three substitution scenarios for four impact categories, the results showed an advantage of using wood in seven of the twelve combinations. The ecosystem quality indicator is the most unfavorable and shows opposite trends to climate change, human health, and resource indicators. Considering average material substitution factors combined with the maximum potential use of wood in non-residential (NR) structures in Quebec in 2050, wood can contribute to avoid 2.6 Mt of carbon dioxide equivalent (CO2 eq.). This amount is equivalent to 3.5% of Quebec’s CO2 eq. emission reduction target by 2050 compared to 1990.