Cytotoxicity and Biomineralized Nodule Formation Induced by Tinospora crispa Crude Extract in Human Bone Cells

Abstract

Objective

          To determine the properties of Tinospora crispa crude extract on the cell toxicity and biomineralized nodule formation in human bone cells in Vitro.

Materials and Methods

          To test the cytotoxicity of Tinospora crispa crude extract, human bone cells were cultured in 96-well plates and divided into 3 separated groups including 1)the control in alpha-MEM, 2) the control in media plus dimethyl sulfoxide, and 3) the experimental group in media supplemented with different concentrations of Tinospora crispa. Cells were treated for 24 hours before investigating the cell viability by MTT assay.

          To examine the induction of biomineralized nodule formation of Tinospora crispa crude extract, human bone cells were cultured in 24-well plates and divided into 3 groups as follows; 1) the negative control cultured in alpha-MEM, 2) the positive control cultured in media plus beta-glycerophosphate and ascorbic acid, 3) the experimental group cultured in media plus Tinospora crispa extract. Cells were treated for 5 weeks before examining by Alizarin Red S staining to observe the orange-red positive stain of biomineralized nodules.

Results

          MTT assay showed that human bone cells cultured in Tinospora crispa crude extract with the concentration less than 0.35% (w/v) survived more than 80%. After cells were treated with the Tinospora crispa crude extract for 5 weeks cells were induced to generate biominerlized nodules. Cells in both positive control and Tinospora crispa treated groups were able to form the nodules which were positively stained for Alizarin Red S.

Conclusion

          Tinospora crispa crude extract which is less than 0.25% (w/v) is non-toxic to human bone cells. The extract at 0.025% (w/v) can enhance the human bone cells to form biomineralized nodules.