Bone related problems due to numerous reasons are global concerns and increasing recently. Although bone tissues have an ability to regenerate themselves, bone become unable to completely heal without external intervention when severe defects occur. It is required to develop new biomaterials termed as a scaffold which can support and induce bone tissue regeneration. A biomaterial scaffold for bone repair possesses the requirements of non-toxicity, biocompatibility, osteoconductivity, and non-immunogenicity. Among natural osteogenic compounds, tamarind seed polysaccharide (TSP) is a desirable agent for fabrication of scaffolds for bone tissue engineering due to its suitable bio-chemical properties. In the previous study, sulfated derivative of TSP was synthesized and characterized. It is proven that this TSP sulfate (TSPS) is an osteogenic compound by inducing osteoblastic cell differentiation and bone mineralization in vitro. In the current study, osteogenic activities of TSPS were further investigated in vivo. Our data showed that TSPS supplemented orally at a dose of 62.5 mg/kg rescued bone loss in osteoporotic mice. Furthermore, histological images of the solid and spongy bone areas indicated that TSPS treated group improved the density and cell morphology of osteocytes and osteoblasts. From positive results observed in vivo, we further continued to study effects of TSPS on HA/Collagen/TSPS composite on bone formation by evaluating alkaline phosphatase (ALP) activity, and bone mineralization. It is showed that TSPS induced cell adherence and proliferation in HA/Collagen (HA/Col) composite. TSPS 100 µg/mL supplemented onto the culture medium and coated onto HA/Col composites resulted in increased of ALP activity to 258 ± 13% and 539 ± 38%, respectively; and of bone mineralization to 135±28.8% and 119±23.5%, respectively. Overall, TSPS was proved to be a good osteogenic agent for bone formation and could be a highly potential candidate in pharmaceutical industry or biomaterial scaffold for bone tissue engineering.