Application of ruthenium complex modified nano-selenium in promoting osteogenic differentiation of mesenchymal stem cells
A ruthenium complex and mesenchymal stem cell technology, applied in the application field of ruthenium complex modified nano-selenium in promoting osteogenic differentiation of mesenchymal stem cells, to achieve the effect of increasing drug intake, simple product system, and inhibiting adipogenic differentiation
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Embodiment 1
[0036] Embodiment 1: the preparation of the nano selenium (Cit-RuSe) of bipyridyl ruthenium complex-citric acid modification
[0037] [Ru(bpy) 2 (dhipH 3 )] (ClO 4 ) 2 Preparation of: cis-[Ru(bpy) 2 Cl 2 ]·2H 2 O (Guangzhou Dongzheng Huabo Instrument Co., Ltd.) (0.156g, 0.3mmol) and 3,4-dihydroxy-imidazo[4,5-f][1,10]-phenanthroline (dhipH 3 ) (Guangzhou Dongzheng Huabo Instrument Co., Ltd.) (0.098g, 0.3mmol) was dissolved in 10mL of ethylene glycol, and heated at 120°C for 6h under the protection of argon. The solution gradually turned bright red. After cooling to room temperature, add saturated NaClO dropwise while stirring 4 , immediately produced a large amount of red precipitate. After stirring and standing still, the solution was suction-filtered, and the crude product was washed and dried, and then separated with a neutral alumina (200 mesh) column. The eluent is a mixed solvent of V (toluene): V (acetonitrile) = 1:4, the main red band components are collected,...
Embodiment 2
[0046] Embodiment 2: The experiment that bipyridyl ruthenium complex-citric acid modified nano-selenium (Cit-RuSe) promotes osteogenic differentiation of stem cells in vitro
[0047] Isolation and culture of human umbilical cord mesenchymal stem cells (HUMSCs): According to the hospital's regulations and with the consent of the family members, the umbilical cord tissues of normal full-term healthy infants were taken from the operating table and soaked in DMEM. Take out the umbilical cord in the ultra-clean workbench, fully wash the residual blood on the umbilical cord with D-Hanks’ balance solution, remove the umbilical vein, umbilical artery and umbilical cord adventitia, peel off the Wharton glue, and cut it into pieces of about 1mm 3 organization block. Move the tissue block into collagenase II (1g / L), add a little DMEM, and digest in a constant temperature shaker at 37°C until Wharton's glue is completely digested. Aspirate the digestive solution containing cells into a s...
Embodiment 3
[0051] Embodiment 3: The experiment that bipyridyl ruthenium complex-citric acid modified nano-selenium (Cit-RuSe) inhibits stem cell adipogenic differentiation in vitro
[0052] Human umbilical cord mesenchymal stem cells (5×10 per well 5 cells) were seeded in 24-well tissue culture plates in 5% CO 2 , 37°C incubator for 3 days. Remove the basal medium and replace it with an induction solution containing adipogenic inducing components (i.e. adipogenic inducing agent AS, DMEM (L)+10% FBS+10 μmol / L dexamethasone+200 μmol / L indomethacin+0.5mmol / L3-isobutyl-1-methylxanthine (IBMX)+10mg / L insulin), and add respectively the bipyridyl ruthenium complex modified nano-selenium (final concentration is 5 μ g / mL) or NaF ( The final concentration is 1mmol / L), induced for 9 and 18 days. Adding the ruthenium complex of the present invention to modify nano-selenium in the medium is the experimental group, and NaF is the positive control group. Fat droplets within stem cell-differentiate...
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