Ti6Al4V printing particles have been recently used for fabricating orthopedic implants. Removing these particles completely from fabricated implants is challenging. Furthermore, recycled particles are commonly used in fabrication without additional analysis. Ti6Al4V wear particles derived from orthopedic implants are known to induce inflammatory responses and osteolysis. However, the biosafety of printing particles remains unknown. Here, we investigated the proinflammatory and osteolysis-inducing effects of commonly used original and recycled Ti6Al4V printing particles in vitro and in vivo. Our results indicated that although less serious effects were induced compared to wear particles, inflammatory responses and osteoclast-mediated bone resorption were induced by the original printing particles in a particle size-dependent manner. Recycled particles were found to more strongly stimulate bone resorption and inflammatory responses than the original particles; the in vivo effect was enhanced with an increase in particle concentration. Furthermore, the results of our in vitro experiments verified that the printing particles activate macrophages to secrete inflammatory cytokines and promote osteoclastogenesis, which is closely related to particle size and concentration. Taken together, our findings provide a valuable reference for the use of raw printing materials and examination of recycling procedures for implant fabrication.