This article was aimed at preparation and characterization of drug delivery carriers made from biodegradable polyhydroxyalkanoates (PHAs) for slow release of tetracycline (TC) for periodontal treatment. Four PHA variants; polyhydroxybutyrate (PHB), poly(hydroxybutyrate-co-hydroxyvalerate) with 5, 12, and 50% hydroxyvalerate were used to formulate TC-loaded PHA microspheres by double emulsion-solvent evaporation method. We also compared the effect of different molecular weight (M w ) of polyvinyl alcohol (PVA) acting as surface stabilizer on particle size, drug loading, encapsulation efficiency, and drug release profile. The TC-loaded PHA microspheres exhibited microscale and nanoscale spherical morphology under scanning electron microscopy. Among formulations, TC-loaded PHB:low M w PVA demonstrated the highest TC loading with slow release behavior. Our results showed that the release rate from PHA microspheres was influenced by both the type of PHA and M w of PVA stabilizer. Lastly, TC-loaded PHB microspheres showed efficient killing activity against periodontitis-causing bacteria, suggesting its potential application for treating periodontal disease.