Transfersome-exosome membrane fusion preparation with transdermal enhancement effect as well as preparation method and application thereof
A technology of enhancement and exosomes, which is applied in the field of biomedicine, can solve problems such as difficulty in ensuring the stability and effectiveness of multiple whitening ingredients, poor drug loading effects, and insufficient carrier advantages, etc., to achieve good development and application prospects, Improve the effect of poor self-stability and excellent transdermal effect
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Embodiment 1
[0044] Preparation of a transfersome-exosome membrane fusion preparation with enhanced transdermal effect:
[0045] (1) Extraction, separation and purification of exosomes from mesenchymal stem cells:
[0046] in DMEM containing 10% fetal bovine serum at 37°C in 5% CO 2 Human umbilical cord-derived mesenchymal stem cells were cultured to the 4th passage under the conditions, and the cells adhered to the wall and grew to 90% of the culture dish (about 1×10 7 1), replace with serum-free DMEM medium and continue to culture at 37°C for 24h, collect the medium, use an ultra-high-speed centrifuge at 4°C, 2000g for 10min to remove cell debris, take the supernatant through a 0.22μm filter membrane, and centrifuge at 100000g After 70 minutes, exosomes were obtained and stored at -80°C for later use;
[0047] (2) Preparation of small molecule peptide transfer body:
[0048] Weigh oligopeptide-1 (5mg), dissolve it with 5mL PBS (pH7.4), and use it as a hydration solution for later use....
Embodiment 2
[0056] Preparation of a transfersome-exosome membrane fusion preparation with enhanced transdermal effect:
[0057] (1) Extraction, separation and purification of exosomes from mesenchymal stem cells, as in Example 1;
[0058] (2) Preparation of small molecule peptide transfer body:
[0059] Weigh tripeptide-1 (4mg), dissolve it with 5mL PBS, and set aside. Weigh EPC (44 mg), cholesterol (6 mg), Tween 80 (8 mg) in a 100 mL eggplant-shaped bottle, add 5 mL of absolute ethanol to dissolve it ultrasonically, form a film under reduced pressure at 25 ° C to remove the organic solvent, dry it in vacuum overnight, add three Peptide-1 in PBS solution 5mL, rotate and hydrate at 25°C and 120rpm for 20min to obtain a transfersome suspension, use a dialysis bag (mW=30kD) to dialyze overnight to remove unencapsulated free peptide in the transfersome suspension , to obtain the transfersome solution, which was stored at 4°C for future use;
[0060] (3) Preparation of transfersome-exosome ...
Embodiment 3
[0066] Preparation of a transfersome-exosome membrane fusion preparation with enhanced transdermal effect:
[0067] (1) Extraction, separation and purification of exosomes from mesenchymal stem cells, as in Example 1;
[0068] (2) Preparation of small molecule peptide transfer body:
[0069] Weigh tripeptide-1 (5mg), dissolve it with 5mL PBS, and set aside. Weigh SPC (87.50mg), cholesterol (12.50mg), Tween 80 (8mg), sodium deoxycholate (7mg) in a 100mL eggplant-shaped bottle, add 5mL absolute ethanol to dissolve it ultrasonically, and remove it under reduced pressure at 25°C to form a film Organic solvent, vacuum-dry overnight, add 5 mL of PBS solution containing tripeptide-1, rotate and hydrate at 25°C, 120 rpm for 20 min to obtain a transfersome suspension, and use a dialysis bag (mW=30kD) to dialyze overnight to remove the transfersome mixture. The unencapsulated free peptide in the suspension was obtained as a transfersome solution, which was stored at 4°C for later use;...
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