Preparation and Application of Lipid Calcium Phosphate Nanoparticle Encapsulating Cell Growth Factor
A growth factor and nanoparticle technology, used in skin care preparations, cosmetic preparations, cosmetic preparations, etc., can solve the problems of inability to exert biological activity and low transdermal absorption rate of macromolecular FGF.
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Embodiment 1F
[0058] The preparation of embodiment 1FGF lipid calcium phosphate nanoparticles (method one)
[0059] (1) Mix 2.52 mL of polyethylene glycol octylphenyl ether (Triton X-1), 1.68 mL of n-hexanol, and 2.8 mL of cyclohexane to form 7 mL of an oil phase ternary system. Mix 100 μL of 250 mM calcium chloride aqueous solution, 100 μL of 250 mM disodium hydrogen phosphate and 25 μL of 0.1 mg / mL aFGF or bFGF to make a water phase, slowly add the water phase to the oil phase and stir well to make a water-in-oil microemulsion, After standing at room temperature for 15 minutes, calcium phosphate precipitates, and 125 μL of 15 mM sodium citrate solution is added dropwise to the microemulsion, stirred and mixed evenly, and the system returns to clarity and transparency. Pass the microemulsion through 1 g of silica gel column filled with 60-200 mesh silica gel, and elute with ethanol / water at a volume ratio of 3:1. The eluent is evaporated to dryness with a rotary evaporator and dissolved in...
Embodiment 2F
[0062] The preparation of embodiment 2FGF lipid calcium phosphate nanoparticles (method two)
[0063] (1) Take 300 μL of 500 mM calcium chloride aqueous solution, 100 μL of 2 mg / mL aFGF or bFGF solution, and 15 mL of cyclohexane / nonylphenol polyoxyethylene ether (71:29) and mix them as solution A. Mix 300 μL of 25 mM disodium hydrogen phosphate solution (pH=9.0), 15 mL of cyclohexane / nonylphenol polyoxyethylene ether (71:29) and add 200 μL of 20 mg / mL DOPA (purchased from Xi’an Ruixi Biotechnology Co., Ltd. ) and stirred evenly as solution B. Mix solution A and solution B for 20 minutes, add 30 mL of ethanol, centrifuge at 12000 g for 15 minutes, wash with ethanol 2 to 3 times, evaporate the ethanol, and dissolve with 1 mL of deionized water. The calcium phosphate nanoparticle suspension containing FGF was obtained.
[0064](2) Mix DOTAP and cholesterol (both purchased from Xi’an Ruixi Biotechnology Co., Ltd.) at a mass ratio of 4:1, dissolve in 5 mL of absolute ethanol, and...
Embodiment 3a
[0066] The preparation of embodiment 3aFGF lipid calcium phosphate nanoparticle gel
[0067] Prepare the lipid calcium phosphate nanoparticle freeze-dried powder containing aFGF according to Example 1 for subsequent use, and prepare the gel according to the following method:
[0068] The mass percent of each component is as follows (mass %):
[0069] aFGF Lipid Calcium Phosphate Nanoparticles 1%
[0070] carbomer 0.5% glycerin 8% Potassium sorbate 0.1% Triethanolamine 2% Deionized water margin
[0071] Dissolve 10 g of aFGF lipid calcium phosphate nanoparticle freeze-dried powder in distilled water for later use, add 1 g of carbomer, 8 g of glycerin, and 0.11 g of potassium sorbate, stir and dissolve, add 2 g of triethanolamine to adjust the pH value to 6-7, and add distilled water to The whole amount was stirred evenly, and 100 g of gel was made to obtain the lipid calcium phosphate nanoparticle gel containing aFGF.
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