Method for preparing nanocluster gel of hydroxyl-like phosphorite component
A hydroxyapatite-like and nano-cluster technology is applied in non-active ingredients medical preparations, inorganic non-active ingredients, pharmaceutical formulations, etc., and can solve the problems of complicated preparation steps, high cost, poor biocompatibility, etc. Achieve the effect of low cost, high yield and excellent degradable properties
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Embodiment 1
[0023] Example 1 AuATP-Ca
[0024] (1) Preparation of precursor gold clusters of AuATP gel: Weigh 200 μL of 0.1 mol / L chloroauric acid solution and 4 mL of 10 mmol / L adenosine triphosphate, mix the two well, and allow them to fully react for 15 minutes (the two are mixed The color of the final solution is light yellow to nearly colorless), the ratio of gold to adenine nucleotide series is 1:2; then add ultrapure water to the mixed solution, set the volume to 15mL, after 0.5h, add 0.2M 5mL of citric acid buffer solution, the molar ratio of citric acid to gold is 50:1; Mix 20mL of absolute ethanol with the same volume as the above reaction solution, mix and shake well; finally, centrifuge 40mL of the turbid solution in a centrifuge at 8000rpm for 15min to precipitate, take out the precipitate and dry it overnight, and weigh the sample after overnight drying, according to Proportionally add ultra-pure water and mix to form a solution of 10 μg / mL, and store it in freezer at -20°C...
Embodiment 2
[0028] Example 2 AuAMP-Ca
[0029] The basic steps are the same as in Example 1, except that adenosine monophosphate is used for the adenine nucleotide series.
Embodiment 3
[0030] Example 3 AuADP-Ca
[0031] The basic steps are the same as in Example 1, except that adenosine diphosphate is used in the adenine nucleotide series.
[0032] Cytotoxicity Assay
[0033] HepG2 cells in the logarithmic phase of rapid growth and normal liver cells L02 were selected, collected after trypsinization, added 100 μL of medium to each well of a 96-well plate, and inoculated at a density of 10,000 cells / well. The cells were placed in a constant temperature incubator with 5% carbon dioxide concentration at 37°C and cultured for 10 hours to complete attachment. Add AuAMP-Ca, AuADP-Ca and 100 μL of AuATP-Ca medium, after continuing to culture for 24 h, add 20 μL of 5 mg / mL MTT solution to each well, continue to culture for 4 h, discard the supernatant in the well, add 150 μL of dimethyl sulfoxide to each well, Shake horizontally for 15 minutes to completely dissolve the MTT product crystals in the wells. Use a microplate reader to measure the light absorption va...
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