Objective: Polyetheretherketone (PEEK) is widely used as an orthopedic implant material owing to its good biocompatibility and mechanical strength; however, PEEK implants are biologically inert, resulting in suboptimal cellular responses after implantation. The aim of this study was to enhance the biological activity of PEEK through sulfonation treatment. Methods: In this study, distal phalangeal implants of PEEK were customized by fused deposition modeling (FDM) printing technology and soaked in concentrated sulfuric acid at different times to obtain sulfonated PEEK (SPEEK). The groups were divided into five groups according to the sulfonation time as follows: 0 min (control group), 1 min (group SPEEK1), 2 min (group SPEEK2), 4 min (group SPEEK4), and 8 min (group SPEEK8). Then the physicochemical characteristics of implants were determined by SEM, XRD, EDS, etc. The implants were co-cultured with stem cells from human exfoliated deciduous teeth (SHED), and then the cell proliferation, adhesion, alkaline phosphatase (ALP) activity, and alizarin red staining were performed to detect the biological activity, biocompatibility, and osteogenic activity of the SPEEK implants. Results: The sulfonation time range of 1 to 8 min could promote the formation of micropores on the surface of PEEK implants, while slightly affecting the composition and compression performance of the implants. Compared with the control group, the hydrophilicity of PEEK materials was not improved after sulfonation treatment. Tests for adhesion and proliferation of SHED indicated that SPEEK2 showed superior biocompatibility. Furthermore, ALP activity and semi-quantitative analysis of Alizarin red staining showed that the osteogenic activity of SPEEK2 phalanges exhibited significantly stronger osteogenic activity than the other groups. Conclusion: The method presented here provides a promising approach to improve the surface bioactivity of PEEK implants prepared by FDM, providing a shred of primary evidence to support the application of SPEEK in orthopedics.