Extracellular matrix calcification in human dental pulp stem cells treated with ethanol extract of <i>Cissus quadrangularis</i> – An <i>in vitro</i> study

Objectives: To induce osteoblastic differentiation in HDP-SC using ethanolic extract of C. quadrangularis and evaluate extracellular matrix calcification using von Kossa staining. Materials and Methods: An alcoholic extract of C. quadrangularis extract was prepared using the Soxhlet apparatus. HDP-SCs were procured from HiMedia laboratories in Mumbai and subcultured. On confluence, the cells were subjected to 100, 200, 300, 400 and 500 µg/mL of ethanolic extract of C. quadrangularis . 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (MTT) assay was used to ascertain cell proliferation and cell cytotoxicity. The cells were then treated with the desired concentration of C. quadrangularis extract and incubated for 5 days. The presence of matrix calcification was determined using von Kossa staining on the 5 th , 10 th and 15 th day. Results: The MTT assay revealed that at concentrations of 100, 200 and 300 µg/mL, the percentage of viable HDPSCs were 99.1%, 98.3% and 95.7%, respectively. The comparison of mean value between all the concentrations (100, 200, 300, 400 and 500 µg/mL) was established to be significant, showing P &lt; 0.05. The calcification assay showed extracellular matrix calcification as brown to black nodules on the 10 th and 15 th day for 100 µg/dl concentration of C. quadrangularis extract, on the 5 th , 10 th and 15 th day for 200 and 300 µg/dL concentration of C. quadrangularis extract. Conclusion: The current research is an attempt to evaluate the regenerative potential of plant-based products on HDP-SC. C. quadrangularis has shown potential for osteoblastic differentiation on HDP-SC and subsequent matrix formation and calcification and, hence, can be utilised as a therapeutic agent in bone regeneration therapies.