This paper introduces anisotropic nickel yttrium oxide (NYO) film formed by the brush coating technique. X-ray photoelectron spectroscopy confirmed well-formed NYO film after the curing process, and the morphology of the surface was investigated using atomic force microscopy. The shear stress driven from brush hair movements caused the nano/micro-grooved anisotropic surface structure of NYO. This anisotropic surface induced uniform liquid crystal (LC) alignment on the surface, which was confirmed by pre-tilt angle analysis and polarized optical microscopy. The contact angle measurements revealed an increase in hydrophilicity at higher temperature curing, which contributed to homogenous LC alignment. The NYO film achieved good optical transmittance and thermal stability as an LC alignment layer. In addition, the film demonstrated good electro-optical properties, stable switching, and significantly enhanced operating voltage performance in a twisted-nematic LC system. Therefore, we expect that this brush coating method can be applied to various inorganic materials to achieve an advanced LC alignment layer.