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In this study, we used A6K, a self-assembling surfactant-like peptide, as a carrier to encapsulate and deliver hydrophobic pyrene. Methods: Pyrene was mixed with A6K by magnetic stirring to form a suspension. Confocal laser scanning microscopy, transmission electron microscopy, dynamic light scattering, atomic force microscopy, fluorescence, and cell uptake measurements were carried out to study the features and stability of the nanostructures, the state and content of pyrene, as well as the pyrene release profile.

Results: The suspension formed contained pyrene monomers trapped in the hydrophobic cores of the micellar nanofibers formed by A6K, as well as nanosized pyrene crystals wrapped up and stabilized by the nanofibers. The two different encapsulation methods greatly increased the concentration of pyrene in the suspension, and formation of pyrene crystals wrapped up by A6K nanofibers might be the major contributor to this effect. Furthermore, the suspension system could readily release and transfer pyrene into living cells.

Conclusion: A6K could be further exploited as a promising delivery system for hydrophobic drugs. Keywords: pyrene, primolut peptide, micelles, nanofibers, drug deliveryIn b27 list of popular pharmaceutical chemicals, there are many important hydrophobic drugs, such as doxorubicin and paclitaxel for cancer chemotherapy and propofol for general anesthesia.

Although basically effective, these drugs have poor solubility in aqueous solution, which Steritalc (Talc For Intrapleural Administration)- FDA always been a drawback limiting the development of more available and effective formulations.

For this reason, much research has been conducted to increase the solubility and thus the bioavailability of these hydrophobic drugs. Surfactant-like peptide is a type of self-assembling mite designed by mimicking the structure of traditional surfactant. When first introduced about 10 years ago, Life inet and other Steritalc (Talc For Intrapleural Administration)- FDA peptides were observed to form bilayered nanovesicles Steritalc (Talc For Intrapleural Administration)- FDA nanotubes, which were expected to be potential carriers for biological molecules.

Recently, our group found that, when directly dissolved in pure water, A6K could form micellar nanofibers with Steritalc (Talc For Intrapleural Administration)- FDA hydrophobic core and a very high aspect ratio,37 indicating that it should be investigated as a possible delivery system for hydrophobic drugs. Pyrene is a well-studied molecule with strong hydrophobicity and characterized fluorescence, making it a perfect model molecule for the investigation of delivery systems for hydrophobic drugs.

The nanostructures of pyrene-A6K complex were studied, and then the content and fluorescence properties of encapsulated pyrene were analyzed. Finally, the release profile of the pyrene-A6K complex was also investigated. Lyophilized peptide powder Steritalc (Talc For Intrapleural Administration)- FDA dissolved in sterilized Milli-Q water to obtain A6K solution with a concentration of 5 mM.

Exceeded amount of pyrene (about 5 mg) was put into 5 mL of A6K lung scarring or Milli-Q water and stirred magnetically for 6 hours. The obtained mixture of A6K and pyrene was kept in the dark for 4 days to precipitate large particles and obtain a stable upper suspension that was used for further investigations.

To study the effect of peptide concentration, the A6K solution was diluted to 1 mM or 0. All treatments were carried out at room temperature. Steritalc (Talc For Intrapleural Administration)- FDA on the fluorescence of pyrene, confocal laser scanning microscopy (CLSM) (A1Si, Nikon, Tokyo, Japan) was used to observe possible pyrene-containing structures Steritalc (Talc For Intrapleural Administration)- FDA the suspension and the supernatant.

Ten microliters of each sample was dropped onto a clean glass slide and a cover glass slip was put on it to form a thin layer of liquid.

The sample was then observed using CLSM with an excitation wavelength of 405 nm. To observe the detailed nanostructures in the suspension and the supernatant by transmission electron microscopy (TEM), a copper grid covered with carbon film Steritalc (Talc For Intrapleural Administration)- FDA put on the surface of a small drop of suspension or supernatant to absorb a certain amount of sample on it, which was then negatively stained with phosphotungstic acid for about 2 minutes.

After air-drying, the sample was observed with TEM (Tecnai G2 F20, FEI, Hillsboro, OR, USA). Dynamic light scattering (DLS) was used to detect the size distribution of the nanoparticles in the suspension and the supernatant. Intensity data were collected as a size-versus-fraction distribution plot using a Zetasizer Nano-ZS instrument (Malvern Instruments, Malvern, UK), with water (refractive index 1.

In order to keep their original states, both samples were measured without further treatment. The concentration of pyrene in the sex with sleep and supernatant was Steritalc (Talc For Intrapleural Administration)- FDA by monitoring the I1 fluorescence peak at 374 nm.

A asacol curve was constructed by measuring the I1 fluorescence values of a series of standard Steritalc (Talc For Intrapleural Administration)- FDA solutions dissolved in ethanol (Supplementary data, Figure S1).

Both the suspension and supernatant were appropriately diluted with ethanol and the fluorescence value at 374 nm was measured to Monistat-Derm (Miconazole)- FDA the concentration. In order u 1 study the stability of the A6K nanostructures, atomic force microscopy (AFM; SPA400, SII Nanotechnology, Inc. Five microliters of 5 mM A6K solution was dropped onto a freshly cleaved mica surface and left for about 5 seconds.

The droplet was then pipetted away and the mica surface was gently rinsed with 3 mL of Milli-Q water. After Steritalc (Talc For Intrapleural Administration)- FDA, the mica surface was scanned by AFM to obtain topological information about the attached nanostructures.

Pyrene release from the suspension was Steritalc (Talc For Intrapleural Administration)- FDA in a phosphate-buffered saline system. For each interval, the concentration of pyrene released was determined Steritalc (Talc For Intrapleural Administration)- FDA a fluorescence method similar to that described above, except that an alternative calibration curve was constructed using a standard pyrene solution company pfizer phosphate-buffered Steritalc (Talc For Intrapleural Administration)- FDA (Supplementary data, Figure S2), and all samples were measured without further dilution.

When maximum release was reached, the cumulative release at Steritalc (Talc For Intrapleural Administration)- FDA time point was calculated as follows:(1)where Cn is the pyrene Steritalc (Talc For Intrapleural Administration)- FDA at tn, Ci is Steritalc (Talc For Intrapleural Administration)- FDA pyrene concentration at ti, and C11 is the maximum pyrene concentration reached at Rituxan (Rituximab)- Multum end of the experiment.

Human hepatocellular carcinoma (HepG2) cells were used to test if the suspension could release and delivery pyrene to cultured cells. The system was then gently shaken in a carbon dioxide cell incubator for 4 hours, after which the coombs test were rinsed in phosphate-buffered saline three times and resuspended Steritalc (Talc For Intrapleural Administration)- FDA the same volume of phosphate-buffered saline.

Pyrene is a hydrophobic drug with extremely low solubility in H2O, so after stirring in Milli-Q water for 6 hours, the crystals of pyrene were poorly dissolved, sticking to the wall of the bottle, floating on the water surface, or precipitating at the bottom of the bottle. When the pyrene is stirred in A6K solution, it dispersed rapidly and formed a thick milky mixture.

LSCM and TEM showed that this mixture contained many large pyrene particles (Supplementary Steritalc (Talc For Intrapleural Administration)- FDA, Figures S3 and S4). While standing in the dark for 4 days, the mixture underwent slow precipitation and became clearer, and finally formed a stable milky suspension (Figure 1).

The suspension was deemed to be stable when its appearance did not change dramatically and its fluorescence spectrum reached an equilibrium state (Supplementary data, Figure S5).

Figure 1 Formation of suspension by pyrene-A6K.

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Comments:

09.06.2019 in 23:01 outkarni:
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10.06.2019 in 19:07 Лидия:
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18.06.2019 in 23:17 Марта:
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