Presently, hydroxyapatite-based nanocomposites are widely employed in treating bone infections. Due to its excellent biocompatibility, hydroxyapatite (HAP) nanocomposites with drugs act as a bone substitute for bone-related disorders and anticancer studies. This work reports the synthesis of HAP nanoparticles through the hydrothermal method and fabricates nanocomposite with polyvinyl alcohol (PVA) and vitamin C (Vit-C). Later, it was loaded with amoxicillin, namely, HAP-PVA-Vit-C-amoxicillin. The phase purity of these nanocomposites was confirmed using XRD, whereas functional groups were confirmed using FT-IR and Raman spectroscopy. The particle’s size, surface stability, and surface charge were investigated by DLS and ZETA potential measurements. The morphological analysis was investigated using SEM and TEM analysis. The drug release analysis shows that the drug release follows the diffusion mechanism, and the drug release profiles correspond with the zero order, Hickson cross well model, and Kors Myer-peppers kinetics models. The synthesized HAP-PVA-Vit-C and HAP-PVA-Vit-C-amoxicillin nanocomposites showed quite good antimicrobial activity and biocompatibility. The results clearly showed that HAP-PVA-Vit-C nanocomposites are suitable for biomedical applications.