A scaffold that replicates the physicochemical composition of bone at the nanoscale level is a promising replacement for conventional bone grafts such as autograft, allograft, or xenograft. However, its creation is still a major challenge in bone tissue engineering. The fabrication of a fibrous PVA-HA/Sr matrix made of strontium (Sr)-substituted hydroxyapatite from the shell of Pomecea canaliculate L. (golden apple snail) is reported in this work. Since the fabrication of HAp from biogenic resources such as the shell of golden apple snail (GASs) should be conducted at very high temperature and results in high crystalline HAp, Sr substitution to Ca was applied to reduce crystallinity during HAp synthesis. The resulted HAp and HA/Sr nanoparticles were then combined with PVA to create fibrous PVA-HAp or PVA-HA/Sr matrices in 2 or 4 mol % Sr ions substitution by electrospinning. The nanofiber diameter increased gradually by the addition of HAp, HA/Sr 2 mol %, and HA/Sr 4 mol %, respectively, into PVA. The percentage of the swelling ratio increased and reached the maximum value in PVA-HA/Sr-4 mol %, as well as in its protein adsorption. Furthermore, the matrices with HAp or HA/Sr incorporation exhibited good bioactivity, increased cell viability and proliferation. Therefore, the fibrous matrices generated in this study are considered potential candidates for bone tissue engineering scaffolds. Further in vivo studies become an urgency to valorize these results into real clinical application.