The cytotoxicity of three extracts (petroleum ether, ethyl acetate and n-butanol) from a plant used in folk medicine, Marchantia convoluta, to human non-small cell lung carcinoma (H1299) and liver carcinoma (HepG2) cell lines was tested. After 72-h incubation of lung and liver cancer cell cultures with varying concentrations of extracts (15 to 200 µg/mL), cytotoxicity was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and reported in terms of cell viability. The extracts that showed a significant cytotoxicity were subjected to gas chromatography-mass spectrometry analysis to identify the components. The ethyl acetate, but not the petroleum ether or n-butanol extract, had a significant cytotoxicity against lung and liver carcinoma cells with IC 50 values of 100 and 30 µg/mL, respectively. A high concentration of ethyl acetate extract (100 µg/mL) rapidly reduced the number of H1299 cells. At lower concentrations of ethyl acetate extract (15, 30, and 40 µg/mL), the numbers of HepG2 cells started to decrease markedly. Gas chromatography-mass spectrometry analysis of the ethyl acetate extract revealed the presence of several compounds such as phytol (23.42%), 1,2,4-tripropylbenzene (13.09%), 9-cedranone (12.75%), ledene oxide (7.22%), caryophyllene (1.82%), and caryophyllene oxide (1.15%). HPLC analysis result showed that there were no flavonoids in ethyl acetate extract, but flavonoids are abundant in n-butanol extract. Further studies are needed regarding the identification, toxicity, and mechanism of action of active compounds. Correspondence
A three-layer structure, including a columnar layer (CL), a buffer layer, and unaffected intact enamel, was successively formed from the outer to the inner part of artificial enamel lesions (AEL) by topical applications of a high-concentration acidic sodium fluoride solution (10,000 ppm, pH 5.6) in vitro. The AEL was produced in bovine enamel that was decalcified for 5 days in a lactic acid gel system. The morphological observations by using scanning electron microscopy showed that the CL was made of columnar deposits of small globules about 0.5 μm in diameter. It was observed for the first time that small globules filled the demineralized interprismatic regions in the buffer layer. The unaffected intact enamel was protected from further demineralization under the acidic condition. Structure and composition of the CL were investigated by using X-ray diffraction and X-ray photoelectron spectroscopy. In the CL the atomic ratio was Ca:P = 12.6 and Ca:F = 0.75, and the small globules were mainly a mixture of polycrystalline calcium fluoride and hydroxyapatite. For comparison, the sound enamel and the AEL attained by applications of 0 and 100 ppm acidic sodium fluoride solutions (pH 5.6) were also investigated. The formation mechanism of the three-layer structure and the related cariostatic effects are discussed.
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