Backgrounds: 125I brachytherapy is effective in relieving cancer pain due to osteolytic bone metastases. However, fewer studies focused on painful osteogenic bone metastases (OBMs), we conducted a retrospective study to evaluate the efficacy of 125I brachytherapy for the treatment of painful OBMs.Methods: From April 2017 to April 2019, 65 patients of OBMs were treated with CT/Cone-beam CT (CBCT)-guided 125I brachytherapy. Technical success, visual analog scale scores (VAS), daily morphine consumption (DMC), relief of pain (RoP), quality of life (QoL), local tumor control (LCR), progression-free survival (PFS), and overall survival (OS) were recorded and analyzed. The Logistic regression analysis and the Cox regression models were performed to predict RoP and the PFS, respectively. Results: Technical success rates were 100%. VAS and DMC continuously decreased significantly at 2 weeks, 6 weeks, and 10 weeks (all P<0.05). The RoP at 6 weeks was 84.62%. QoL including sleep, appetite, spiritual state, and fatigue showed improvement at 6 and 10 weeks. LCR was 100% at 10 weeks. The PFS and OS were 10.40 and 29.80 months, respectively. Two factors were significantly associated with the RoP: max diameter (MD< 3 cm vs. ≥ 3 cm, P=0.028) and serum levels of bone alkaline phosphatase (B-ALP) (< 100 U/L vs. ≥ 100 U/L, P=0.038). Three independent risk factors were identified for PFS: D90 < 110 Gy (HR 2.89, P=0.012), B-ALP ≥ 100 U/L (HR 3.39, P=0.009), and MD ≥ 3 cm (HR 5.86, P=0.005). Conclusions: 125I brachytherapy is an effective treatment in relieving painful OBMs and improving patients’ QoL.