The purpose of this study was to investigate the effect of thermal denaturation on albumin nanoparticles (NPs) conjugated with fatty acid for the controlled release of poorly water-soluble drug. Albumin - oleic nanoparticle (AON) was prepared as follows. Sodium oleate was covalently conjugated to the backbone of human serum albumin (HSA) based on 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) reaction to form self-assembled fattigation-platform nanoparticles (NPs) using a simple nanoprecipitation method. Optimization of the stability of AONs was carried out using different concentrations of glutaraldehyde from which 1% concentration was selected. Cilostazol (CLT) was chosen as a model hydrophobic drug and loaded during the process of preparation to NPs by in process method. The formed AON was then dispersed in different heating condition for denaturation of albumin. Effect of mainly two factors, i.e., heating temperature and heating time, were evaluated for drug release from CLT-loaded AONs. Circular dichroism spectropolarimeter (CD) was used to investigate the structural change of human serum albumin after denaturation. Other physicochemical properties such as particle size and shape, CLT drug loading (%), loading efficiency (%), and release profile were also characterized. For in vitro release studies, thermally modified AON were injected into a dialysis bag containing 0.2 % (w/v) sodium lauryl sulfate (10 mL) in pH 7.4 buffer at 37°C using an incubation shaker at 100 rpm. The concentration of CLT was determined by RP-HPLC at 257 nm. CD spectra exhibited change in HSA secondary structure by heating denaturation. In addition, the degree of denaturation increased as heating intensity became higher. The sustained release effect was proportional to the degree of secondary structural modification of HSA. Moreover, the solubilizing effect of amphiphilic AONs was visible in all the samples. The thermal modification of amphiphilic AON could provide a versatile tool to modulate solubility and release rate of poorly water-soluble drugs.