Objective(s): To evaluate the potential of a novel alginate/chitosan/Zinc-strontium hydroxyapatite (AG/CS/SHA) composite scaffold for periodontal tissue regeneration by assessing its biocompatibility, bioactivity, and degradation characteristics. Materials and Methods: An AG/CS/SHA composite scaffold was fabricated using a freeze-drying technique. ray Diffraction (XRD) to analyze crystallinity, Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) to investigate microstructure and surface morphology. In vitro studies assessed biocompatibility using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, cell viability with Acridine Orange/Propidium Iodide (AO/PI) staining, alkaline phosphatase (ALP) activity, and calcium deposition to evaluate osteogenic differentiation. Results: FTIR and XRD confirmed SHA incorporation. SEM/TEM revealed a porous structure with uniform SHA distribution. The composite exhibited controlled swelling and degradation. MTT assay and AO/PI staining demonstrated good cell viability. ALP activity (20% higher) and ARS staining (35% increase) were significantly higher in Zn and Sr in HA within AG/CS (AG/CS/SHA) scaffolds compared to AG/CS (control), indicating enhanced osteogenic differentiation and mineralization. Conclusion: The results demonstrate that the AG/CS/SHA composite possesses favorable characteristics for bone tissue engineering applications, including excellent biocompatibility, suitable mechanical properties, and the ability to promote osteogenic differentiation. These findings suggest that the AG/CS/SHA scaffold holds significant promise as a promising biomaterial for periodontal tissue regeneration, providing a supportive environment for cell growth, differentiation, and new tissue formation.