This work aims to enhance the mechanical properties, oil resistance, and thermal properties of acrylonitrile butadiene rubber (NBR) by using the Nile tilapia fish scales as a filler and using bis(triethoxysilylpropyl)tetrasulfide (TESPT) as a coupling agent (CA). The prepared fish scale particles (FSp) are B-type hydroxyapatite and the particle shape is rod-like. The filled NBR with FSp at 10 phr increased tensile strength up to 180% (4.56 ± 0.48 MPa), reduced oil absorption up to 155%, and increased the decomposition temperature up to 4 °C, relative to the unfilled NBR. The addition of CA into filled NBR with FSp at 10 phr increased tensile strength up to 123% (5.62 ± 0.42 MPa) and percentage of elongation at break up to 122% relative to the filled NBR with FSp at 10 phr. This work demonstrated that the prepared FSp from the Nile tilapia fish scales can be used as a reinforcement filler to enhance the NBR properties for use in many high-performance applications.