Naturally derived polysaccharide biopolymer-based nanoparticles with their size and drug release potentials have appeared as promising biomaterials for osteogenic differentiation. A metallic nanoparticle (GS-AgNP) prepared from a sulfated polygalactan characterized as -> 3)-2-O-methyl-O-6-sulfonato-beta-D galactopyranosyl-(1 -> 4)-2-O-methyl-3,6-anhydro-alpha-D-galactopyranose-(1 -> isolated from the marine macroalga Gracilaria salicornia exhibited a prospective osteogenic effect. Upon treatment with the studied GS-AgNP, alkaline phosphatase activity (88.9 mU/mg) was significantly elevated in human mesenchymal osteoblast stem cells (hMSCs) compared to that in the normal control (33.7 mU/mg). A mineralization study of GS-AgNPs demonstrated an intense mineralized nodule formation on the hMSC surface. A fluorescence-activated cell sorting study of osteocalcin and bone morphogenic protein-2 (BMP-2) expression resulted in an increased population of osteocalcin (78.64%) and BMP-2-positive cells (46.10%) after treatment with GS-AgNPs (250 mu g/mL) on M2 macrophages. A time-dependent cell viability study of GS-AgNPs exhibited its non-cytotoxic nature. The studied polygalactan-built nanoparticle could be developed as a promising bioactive pharmacophore against metabolic bone disorder and the treatment for osteogenesis therapy.