A facile and environmentally friendly method was introduced to incorporate montmorillonite (MMT) as an inorganic phase into quaternized hemicelluloses (QH) for forming hemicellulose-based films. Two fillers, polyvinyl alcohol (PVA) and chitin nanowhiskers (NCH), were added into the hemicelluloses/MMT hybrid matrices to prepare hybrid films, respectively. The hybrid films were nanocomposites with nacre-like structure and multifunctional characteristics including higher strength and good oxygen barrier properties via the electrostatic and hydrogen bonding interactions. The addition of PVA and NCH could induce changes in surface topography, and effectively enhance mechanical strength, thermal stability, transparency, and oxygen barrier properties. The tensile strengths of the composite films FPVA(0.3), FPVA(0.5), and FNCH(0.8) were 53.7, 46.3, and 50.1 MPa, respectively, which were 171%, 134%, and 153% larger than the FQH-MMT film (19.8 MPa). The tensile strength, and oxygen transmission rate of QH-MMT-PVA film were better than those of quaternized hemicelluloses/MMT films. Thus, the proper filler is very important for the strength of the hybrid film. These results provide insights into the understanding of the structural relationships of hemicellulose-based composite films in coating and packaging application for the future.