Mechanical pretreatment is an effective process for chemical or biochemical conversion of woody biomass. The deconstruction features of the wood cell wall play an important role in its chemical or biochemical processing. In this work, we evaluated the wood cell wall fracture in the early stage of mechanical pretreatment process conducted with various initial moisture contents. Electronic microscopy (i.e., SEM and TEM) and confocal laser scanning microscopy (CLSM) were used to visualize the cellular structure changes due to cell wall fractures. Results reveal that the enzymatic digestibility of micronized wood produced from different initial moisture contents was improved by 2-6 folder than that of the raw material. The types of cell wall fractures after mechanical pretreatment were distinguished by the initial moisture contents of wood. In wood samples with lower moisture content, interwall fracture occurred predominantly at the middle lamella region, while intrawall fracture occurred primarily at inner cell wall layers, with sever breakage in wood fibers for high moisture content samples.Differences in the distribution of surface chemical composition also resulted from different cell wall fractures. Lignin preferentially covered the fracture surface of low-moisture content samples, while carbohydrates were more predominate in high-moisture content samples. These morphological and structural alternation contributed to improving enzymatic digestibility of micronized wood. Findings from this study demonstrate how mechanical pretreatment modifies the fracture features of wood cell wall for further chemical/biochemical reactions.