The Effects Of Piper Sarmentosum Aqueous Extract On MC3T3-E1 Differentiation

Abstract

Piper sarmentosumis an herbaceous plant under the Piperaceae family. It is also known as ‘kaduk’ among Malaysians. The plant extracts contain various compounds such as flavonoids and benzoic acid derivatives that are believed to repair and maintain bone strength. The aim of this study was to determine the capability of P. sarmentosum aqueous extract in inducing osteoblast differentiation of MC3T3-E1 cell line.The MC3T3-E1 cells were treated with specific concentrations of P. sarmentosumaqueous extract which were 1 mg/mL, 2 mg/mL, 3 mg/mL and 4 mg/mL and were cultured for 14 days. Three analyses were used to determine the differentiation potential of MC3T3-E1 cell into osteoblast, i.e., biochemical assay, gene expression and morphological analysis. Biochemical assay was carried out by determining the specific activity of alkaline phosphatase (ALP). The results showed an increase in ALP activity in cells treated with P. sarmentosumaqueous extract with the highest activity was on day 14. Analysis of molecular biology marker expression showed the expression of Alp, Runx2 and Col 1 genes which were osteoblast genes. The von Kossa staining showed formation of calcium nodules through brownish and black colour on day 14 of culture. In conclusion, P. sarmentosumaqueous extract through ALP activity, osteoblast genes expression and calcium mineralization were shown able to induce MC3T3-E1 cell differentiation into osteoblasts.