The invention discloses a sustained-release oxygen microsphere as well as a preparation method and application thereof. The sustained-release oxygen microsphere is a microsphere using gelatin as a matrix; the interior of the microsphere contains peroxidated inorganic particles, and preferably, the peroxidated inorganic particles are selected from one or multiple of CaO2 (calcium peroxide), MgO2 (magnesium peroxide), NaCO3 (sodium carbonate), SrO2 (strontium peroxide), BaO2 (barium peroxide), K2O2 (potassium peroxide), and Na2O2 (sodium peroxide). The sustained-release oxygen microsphere has the advantages that the size is controllable, the oxygen release time is long and controllable, the sudden release rate is low, the cell toxicity is low, the immunogenicity is low, the preparation process is relatively simple, the industrialized production is easily realized, and the like; the larger application prospect is realized in clinical application of bone defect repair.