4-oxo-4H-pyran-2.6-dicarboxylic acid (chelidonic acid, ChA) in the native state and in the complex with calcium [Ca(ChA)(H2O)3], named saucalchelin (CaChA), was isolated from the extract of Saussurea controversa leaves for the first time for the Asteraceae family. The structure of ChA was determined by NMR, MS and confirmed by X-ray analysis of its monomethyl ester, and CaChA was described by IR, ICP-MS, CHN analysis. The yield of ChA and CaChA was 45 mg/g and 70 mg/g of extract, respectively. The osteogenic activity of ChA, n-monobutyl ester of chelidonic acid, and CaChA has been studied in vitro in a 21-day culture of human adipose-derived multipotent mesenchymal stromal cells (hAMMSCs) in a standard nutrient medium without osteogenic supplements. CaChA significantly stimulated the growth of cell mass and differentiation of hAMMSCs into osteoblasts with subsequent mineralization of the culture and it may be a promising substance for accelerating bone tissue regeneration and engineering.
Five flavonoids were isolated from leaves of Saussurea controversa, one new flavonoid (5), three described for the first time from the genus Saussurea (1, 2, and 4), and one described previously (3). A number of flavonol glycosides can be easily isolated from the butanol fraction of the ethanol extract from S. controversa. As a result of separating these using column chromatography on silica gel and microcrystalline cellulose, five glycosides of quercetin were isolated in the proportional relationship of 2 (1): 8 (2): 10 (3): 1 (4): 4 (5). The structure of these substances was determined using the method of NMR by conducting various 1H-1H and 1H-13C correlation spectroscopy experiments and by comparing their spectroscopic data with published values, 5,3/,4/-trihydroxyflavone-7-О-α-L-rhamnopyranoside-3-О-β-D-glucopyranosidе (1), 5,7,3/,4/-tetrahydroxyflavone-3-О-β-D-glucopyranoside-(1→6)-β-D-glucopyranoside-(1→6)-О-α-L-rhamnopyranoside (2), 5,7,3/,4/-tetrahydroxyflavone-3-О-β-D-glucopyranoside-(1→6)-О-α-L-rhamnopyranoside (3), 5,3/,4/-trihydroxyflavone-7-О-α-L-rhamnopyranoside-3-О-β-D-xylopyranoside (4), and 5,3/,4/-trihydroxyflavone-7-О-β-D-glucopyranoside-3-O-α-L-rhamnopyranoside-(1→2)-О-β-D-glucopyranoside (5). It was established for the first time that flavonol glycosides from S. controversa stimulate granulopoiesis and lymphopoiesis in the bone marrow of rats, and enhance the reparative process in the affected area in experimental osteomyelitis.
A chromatographically inseparable crystalline mixture of the previously undescribed quercetin-4′-O-β-galactopyranoside and the known quercetin-3-O-β-glucopyranoside (isoquercitrin) in a 7:5 ratio was isolated from the extract of the aerial part of Filipendula ulmaria (L.) Maxim.
A model of traumatic osteomyelitis in rats is created. The disease development was evaluated by bone marrow myelograms, results of scintigraphy, and histological studies.
The osteogenic, cytotoxic, and antibacterial activities of polysaccharide (PS-SC) and flavonoid (F-SC) fractions isolated from the leaves extract of Saussurea controversa were studied in vitro. F-SC consists of the five quercetin glycosides in the ratio 2:8:10:1:4, which were isolated from the leaves extract of S. controversa and have been characterized previously. PS-SC was first isolated from the leaves extract of S. controversa and has been described. PS-SC consists in 30 compounds is characterized by a high degree of heterogeneity with a heterogeneity index of 19.74. The Mw and Mn of PS-SC were 108.6 and 5.5 kDa, respectively. Structural fragments are represented by galactose, arabinose, xylose, glucose, uronic acids, mannose, and rhamnose in a 10.1:3.3:2.2:2.1:1.7:0.9:0.5 molar ratio. F-SC as compared with PS-SC showed in vitro microbicidal (50 g/L) and better bacteriostatic (6.25 g/L versus 25 g/L of PS-SC) effects against the 24-h growth of Staphylococcus aureus strain 209 P and a 21-day absence of cytotoxicity on human adipose-derived multipotent mesenchymal stromal cells (hAMMSCs). Both fractions (PS-SC > F-SC) at doses of 10–50 mg/L stimulated differentiation of hAMMSCs into secreting osteoblasts accompanied by local mineralization of extracellular matrix. These fractions of S. controversa and especially F-SC, might be promising peroral drugs in the complex treatment of bone fractures and for prophylaxis of their infectious complications.
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