Ultrasound triggered liposome payload release
A payload and liposome technology, applied in liposome delivery, ultrasonic introduction, inorganic non-active ingredients, etc., can solve problems such as unfavorable biomedical applications
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Embodiment 1
[0253] Example 1: Ultrasonic-triggered gelation of fibrinogen hydrogels
[0254] figure 1 Schematic diagrams of ultrasound-triggered enzymatic catalysis and hydrogels are shown. exist figure 1 In a: Ultrasound applied to calcium-loaded liposomes to release Ca 2+ ions and activate transglutaminase. The active transglutaminase is then able to catalyze the reaction between the protein substrate and dansylcadaverine. This conjugation process results in a shift in the wavelength of maximum fluorescence emission and an increase in fluorescence at 505 nm. exist figure 1 In b: A similar process is used to catalyze the cross-linking of soluble fibrinogen molecules. Intermolecular cross-linking leads to the formation of fibrinogen hydrogels.
[0255] Liposome formulation
[0256] Calcium-loaded liposomes were formulated using an established staggered fusion vesicle method (Biochim. Biophys. Acta-Biomembr. 1994, 1195, 237). Briefly, 99 mol% DPPC and 1 mol% DSPE-PEG were prepared...
Embodiment 2
[0290] Example 2: Ultrasonic-triggered gelation of fibrinogen hydrogels using liposome-microbubble conjugates
[0291] Microbubble formulation and size
[0292] Microbubbles were formulated using a method adapted from a previously reported protocol (Small 2014, 10, 3316). 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), DSPE-PEG in a molar ratio of 86:9:5 with 0.6M NaCl 2000 and DSPE-PEG 2000 Lipid membrane hydration of biotin to a final lipid concentration of 6.32 mg mL -1 . The lipid suspension was vortexed for 15 seconds and heated at 75°C for 2 minutes, then vortexed and heated again. The perfluorohexane / air mixture was pumped into the lipid suspension and the samples were sonicated using a Vibra Cell probe sonicator (20 kHz, 40% amplitude, 100% duty cycle, 3 seconds). Four centrifugation washes (100 g, 3 min) were performed to remove excess lipids. Samples were imaged in brightfield mode on an Olympus IX71 inverted microscope using a 60× oil immersion objective. ...
Embodiment 3
[0306] Example 3: Ultrasound-triggered gelation of alginate hydrogels
[0307] Calcium-loaded liposomes were also shown to induce alginate hydrogelation. Alginate is an anionic polysaccharide that can be transported by divalent cations such as Ca 2+ ) cross-linking and are widely used in cell in vitro studies and human clinical trials.
[0308] The same liposome formulation described in the discussion of ultrasound-triggered enzymatic gelation was used. Briefly, liposomes were formulated by the staggered fusion vesicle method (DPPC: DSPE PEG biotin = 99:1 molar ratio). Total encapsulated calcium was measured by o-cresolphthalein (o-CPC) assay after solubilization of liposomes with Triton X-100 and was 32.4 mM ± 0.8 mM.
[0309] temperature-dependent calcium release
[0310] Temperature-dependent release experiments were performed prior to ultrasound exposure experiments. Here, 50 μL of sample was placed in a 500 μL tube immersed in a water bath set at the desired T. The ...
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