The invention discloses a preparation method of gentamicin sulphate-loading porous hydroxyapatite/PLGA (poly(DL-lactide-co-glycolide) microspheres. The preparation method comprises the following steps: adding PLGA microspheres in an acetone/alcohol solution, adding a diammonium hydrogen phosphate solution, pouring in a calcium nitrate solution, adjusting pH, ageing, washing, drying, calcining, and grinding to obtain porous hydroxyapatite; placing the porous hydroxyapatite powder in a water solution containing gentamicin sulphate/PEG (polyethylene glycol), and carrying out freeze drying to obtain gentamicin sulphate-loading porous hydroxyapatite; and uniformly mixing the PLGA solution and the gentamicin sulphate-loading porous hydroxyapatite, pouring in a polyvinyl alcohol solution, stirring, washing, and carrying out freeze drying to obtain the gentamicin sulphate-loading porous hydroxyapatite/PLGA (poly(DL-lactide-co-glycolide) microspheres. The gentamicin sulphate-loading porous hydroxyapatite/PLGA (poly(DL-lactide-co-glycolide) microspheres prepared by adopting the preparation method are high in medicine loading rate, small in burst release, good in biocompatibility, simple in preparation process, easily available in raw materials, and low in cost; and industrialization is easily realized.