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OBJECTIVES The objective of this study is to investigate leaching residual monomer and biological effects of four types of conventional and computer-aided design/computer-aided manufacturing (CAD/CAM) dental polymers on human gingival fibroblasts (HGFs). MATERIALS AND METHODS A total of 540 disk-shaped specimens were fabricated from four different materials (n=135 per group): compression-molding polymethylmethacrylate (PMMA) (conventional denture polymer), CAD/CAM PMMA (CAD/CAM denture polymer), bis-acrylic composite resin (conventional temporary polymer), and CAD/CAM PMMA (CAD/CAM temporary polymer). Specimens were eluted in cell culture medium for 72 h at 37°C, and the residual monomer in eluates subsequently was measured by high-performance liquid chromatography (HPLC). The biological effects of material eluates on HGFs were analyzed by CCK-8 assay, flow cytometry, real-time quantitative PCR, Western blotting, and enzyme-linked immunosorbent assay (ELISA) to identify cell death patterns and its biological mechanism. RESULTS Methyl methacrylate (MMA) was detected only in compression-molding PMMA, and by-products were detected in bis-acrylic composite resin. The cell proliferation of CAD/CAM denture polymer or CAD/CAM temporary polymer was greater than that of compression-molding PMMA or bis-acrylic composite resin at 72 h in culture. No apoptosis and necrosis were detected in CAD/CAM dental polymers. Apoptosis was detected only in bis-acrylic composite resin and further confirmed by the upregulation of Bax and cleaved Caspase-3, as well as the downregulation of Bcl-2 gene. And no significant variation in inflammatory cytokines secretion was observed in all materials. CONCLUSIONS CAD/CAM dental polymers (including temporary and denture polymers) have favorable biocompatibility due to lower residual monomer, which provides scientific evidence to the controversy of biocompatibility of conventional and CAD/CAM dental polymers. CLINICAL RELEVANCE The use of CAD/CAM dental polymers is recommended in the fabrication of temporary restorations and dentures due to their favorable biocompatibility.