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Background: the clinical application of orthopedic or dental implants improves the quality of the lives of patients. However, the long-term use of implants may lead to implant loosening and related complications. The purpose of this study is to deposit titanium (Ti)-niobium (Nb)-zirconium (Zr)-tantalum (Ta) alloys on the surface of Ti-6Al-4V to increase structural strength and biocompatibility for the possible future application of implants. Materials and methods: Ti, Nb, Zr, and Ta served as the materials for the surface modification of the titanium alloy. TiNbZr and TiNbZrTa coatings were produced using cathodic arc evaporation, and a small amount of nitrogen was added to produce TiNbZrTa(N) film. Annealing and oxidation were then conducted to produce TiNbZrTa-O and TiNbZrTa(N)-O coatings. In this study, biological tests and finite element analyses of those five alloy films, as well as uncoated Ti-6Al-4V, were performed. Human osteosarcoma cells (MG-63) and mouse fibroblast cells (L-929) were used to analyze cytotoxicity, cell viability, and cell morphology, and the bone differentiation of MG-63 was evaluated in an alkaline phosphatase experiment. Furthermore, for measuring the gene expression level of L-929, reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) was conducted. The three-dimensional (3D) computational models of the coated and uncoated sample films were constructed using images of transmission electron microscopy and computer-aided design software and, then, the stress distributions of all models were evaluated by finite element analysis. Result: the cytotoxicity test revealed that the surface treatment had no significant cytotoxic effects on MG-63 and L-929 cells. According to the results of the cell viability of L-929, more cell activity was observed in the surface-treated experimental group than in the control group; for MG-63, the cell viability of the coated samples was similar to that of the uncoated samples. In the cell morphology analysis, both MG-63 and L-929 exhibited attached filopodia and lamellipodia, verifying that the cells were well attached. The alkaline phosphatase experiment demonstrated that the surface treatment did not affect the characteristics of early osteogenic differentiation, whereas RT-qPCR analysis showed that surface treatment can promote better performance of L-929 cells in collagen, type I, α1, and fibronectin 1. Finally, the results of the finite element analysis revealed that the coated TiNb interlayer can effectively reduce the stress concentration inside the layered coatings. Conclusions: TiNbZrTa series films deposited using cathodic arc evaporation had excellent biocompatibility with titanium alloys, particularly in regard to soft tissue cells, which exhibited an active performance. The finite element analysis verified that the TiNb interlayer can reduce the stress concentration inside TiNbZrTa series films, increasing their suitability for application in biomedical implants in the future. [ABSTRACT FROM AUTHOR]