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Morinda citrifolia Leaf Enhances In Vitro Osteogenic Differentiation and Matrix Mineralization by Human Periodontal Ligament Cells

Kanitsak Boonanantanasarn,Kajohnkiart Janebodin*,Prapan Suppakpatana,Tawepong Arayapisit,Panjit Chunhabundit,Jit-Aree Rodsutthi ,Orapin Termvidchakorn,Wanida Sripairojthikoon

Morinda citrifolia L. (noni) has been reported to have a variety of potentially therapeutic effects such as antibacterial, antiviral, antifungal, antitumor, antihelmin, analgesic, hypotensive and anti-inflammatory, as well as immune enhancing properties. In this study, we aimed to investigate a potential effect of Morinda citrifolia leaf aqueous extract to induce osteogenic differentiation and matrix mineralization of human periodontal ligament (hPDL) cells in vitro. The optimal biocompatible dose of noni leaf aqueous extract was determined by a cell proliferation assay. The hPDL cells were cultured in growth medium alone, or medium supplemented with noni leaf aqueous extract, or β-glycerophosphate and ascorbic acid. The early osteogenic differentiation marker, alkaline phosphatase activity, was assayed using a biochemical analysis. Matrix mineralization was examined after 6 weeks in vitro by calcium staining and energy dispersive x-ray analysis. The noni leaf aqueous extract group significantly increased alkaline phosphatase activity, compared to either growth medium alone or β-glycerophosphate and ascorbic acid groups. Nodules containing mineralized matrices were formed only when the hPDL cells were cultured in the presence of noni leaf aqueous extract. These nodules were positively stained with Alizarin Red and were rich in calcium and phosphorus as revealed by the energy dispersive x-ray analysis. These data indicate the potential effect of noni leaf aqeous extract to promote osteogenic differentiation as well as matrix mineralization in hPDL cells and suggest that noni leaf has therapeutic benefits in bone and periodontal tissue regeneration.