Bone related disorders affect 75 million individual through elaborating significant bone fracture situation throughout Europe, USA and Japan. The possible treatment methods are notably expensive. The hydrogel based bioactive composite scaffold can be a suitable approach due to its cost-effective production attributes. The primary focus of the work is the biological characterization (i.e. cell growth study after 5 days of incubation with sample through optical microscopy and bone marker analysis or alkaline phosphate expression study) of the novel inorganic calcium filled bacterial cellulose (BC) based hydrogel composite scaffolds in the context of their application in bone tissue engineering application. BC based hydrogel scaffolds were developed by applying the homogeneous suspension of BC (holding 99% water) with carboxymethyl cellulose (CMC), polyvinylpyrrolidone (PVP). β-tricalcium phosphate (β-TCP) and hydroxyapatite (HA) were incorporated to achieve the calcium phosphate filled hydrogel scaffolds (termed as “BC-PVP-β-TCP/HA” and “BC-CMC-β-TCP/HA”). These hydrogel scaffolds were in vitro bio-mineralized to incorporate CaCO3 (termed as “BC-PVP-β-TCP/HA-CaCO3” and “BC-CMC-β-TCP/HA-CaCO3”). BC-PVP and BC-CMC hydrogel scaffolds were used as control set. Human diploid fibroblast cells, Lep-3 and mouse bone explant cell line (BEC) were used in our study. The cell viability was found significant for BC-PVP-β-TCP/HA with both cell lines. The ALP expression in BEC cell culture of BC-PVP-β-TCP/HA and BC-CMC-β-TCP/HA-CaCO3 hydrogel scaffold is also found significant. These results suggest the BC-PVP-β-TCP/HA hydrogel composite scaffold has the notable potentiality for the soft bone tissue engineering application.