Biologically active peptides, including growth factors and cytokines, participate in various biological processes in human skin. They could provide a great advantage of maintaining healthy skin. Many peptide growth factors like epidermal growth factor (EGF) and human growth hormone (hGH) have been used in cosmetic formulations. The delivery of peptide growth factors across the Stratum corneum, however, seems not sufficient because of their physical properties such as high molecular weight and hydrophilicity. So increasing the penetration of growth factors of interest into skin would be a major concern for ensuring their maximum biological efficacy. In this study, we have identified several skin penetration-enhancing peptides which facilitate delivery of growth factors, when fused at N-terminus of the target protein, into skin. For efficient and rapid screening, we constructed a skin-penetrating assay system using Franz cell and porcine skin. Next, we carried out phage display screening using M-13 bacteriophage with random 12 -amino acid library on its coat protein P3 on that system. After several selection rounds, peptide sequences facilitate the penetration of phages through the porcine skin were identified from a large population of phages. We found that phages with the most potent peptide (S3-2, NGSLNTHLAPIL) could penetrate the porcine skin eight times more than those with control peptide (12 mino acids scrambled peptide). Furthermore, growth factors conjugated with S3-2 peptide penetrate porcine skin three to five times efficiently than non-conjugated growth factors. In conclusion, our data shows that the skin penetration-enhancing peptide we have characterized could increase the delivery of growth factors and is useful for cosmeceutical application.
An, Gyu Min;Park, Su In;Kim, Min Gi;Heo, Soo Hyeon;Shin, Moon Sam
Korean Chemical Engineering Research
/
v.58
no.1
/
pp.21-28
/
2020
The stem extract from Pyrus serotina var has natural antioxidant ability, but the extraction method does not result in a soluble compound in cosmetic formulations. This study investigated the cosmetic efficacy of the Pyrus serotina var stem extract and its epidermis permeation ability when combined with polymer micelles and a cell penetrating peptide. The total concentration of polyphenol compounds was determined to be 103.1644 ± 1.38 mg/g in the ethanol extract and 78.97 ± 1.45 mg/g in the hydrothermal extract. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging effects were 55.94 ± 0.22% in the ethanol extract at 1,000 mg/L. Superoxide dismutase (SOD) activity rates were 104.05 ± 3.28% in the ethanol extract at 62.5 mg/L. The elastase inhibition rate was 67.21 ± 2.72% in the ethanol extract at 1,000 mg/L. An antimicrobial effect was observed in the Propionibacterium acnes strain. In the epidermal permeability experiment, it was confirmed that formulation of the polymer micelle containing the Pyrus serotina var stem extract and cell penetrating peptide (R6, hexa-D-arginine) showed small particle size and much better skin permeability. The cumulative amount of total Pyrus serotina var stem extract that penetrated to the skin over time increased over 24 hours in three formulations. The three formulations showed 51.61 ㎍/㎠ (Formulation 0), 75.97 ㎍/㎠ (Formulation 1) and 95.23 ㎍/㎠ (Formulation 2) skin penetration, respectively. Therefore, it was confirmed that the ethanol extracts of Pyrus serotina var stem showed good cosmetic efficacy and excellent epidermis permeation ability when combined with a polymer micelle and cell penetrating peptide. Thus, this extract has the potential to be used as a safe and natural cosmetic material in the future.
Heo, Soo Hyeon;Park, Su In;Lee, Jinseo;Kim, Miok;Shin, Moon Sam
International Journal of Advanced Culture Technology
/
v.8
no.1
/
pp.226-235
/
2020
In this paper, we present to evaluate physiological activity of Robinia pseudo-acacia leaf and its skin penetration using liposome and cell penetrating peptide. After extraction with Robinia pseudo-acacia leaf using the distilled water and supercritical, various physiological activities were examined. In antioxidants experiments, the total concentration of polyphenol compounds was determined to be 56.88 mg/g in hydrothermal extract, 45.07 mg/g in supercritical extract. The DPPH radical scavenging ability at 1,000 ㎍/mL was 33.97% in supercritical extract. The scavenging effect on SOD experiment at 500 ㎍/mL was 76.41% in supercritical extract. In the antimicrobial experiments, the hydrothermal extract had no effect, but supercritical extract represented maximum clear zone of 14.00 mm in Staphylococcus aureus strain. Liposome containing the RSE (Robinia pseudo-acacia leaf supercritical extract) reduced particle size and stabilized zeta potential. In the epidermal permeability experiment, it was confirmed that the permeation of liposome containing the RSE and cell penetrating peptides was remarkable.
Heo, Soo Hyeon;Park, Su In;An, Gyu Min;Shin, Moon Sam
The Journal of the Convergence on Culture Technology
/
v.5
no.3
/
pp.271-282
/
2019
This study was conducted to evaluate physiological activity of Robinia pseudo-acacia leaf and its skin penetration using polymer micelles and skin penetrating peptide. After extraction with Robinia pseudo-acacia using the ethanol and distilled water, various physiological activities were examined. The total concentration of polyphenol compounds was determined to be 47.42 mg/g (ethanol extract), 56.88 mg/g (hydrothermal extract) and DPPH radical scavenging ability at $1,000{\mu}g/mL$ was 44.24% in ethanol extract and it is higher than value(41.50%) in hydrothermal extract. The elastase inhibitory assay showed concentration dependence and elastase inhibition of Robinia pseudo acacia leaf ethanol extract was 54.09%, which was the highest at $500{\mu}g/mL$. In the SOD-like experiments, the concentration-dependent results were showed and the SOD-like activity of the Robinia pseudo-acacia leaf ethanol extract was higher than that of the Robinia pseudo acacia leaf hydrothermal extract at all concentrations. At a concentration of $500{\mu}g/mL$, Robinia pseudo acacia leaf ethanol extract showed the highest SOD-like activity of 76.41%. The tyrosinase inhibition at $20{\mu}g/mL$ was determined to be 56.47% (ethanol extract), 23.05% (hydrothermal extract). In the antimicrobial experiments, the hydrothermal extract had no effect, but ethanol extract represented maximum clear zone of 11.00 mm in Propionbacterium acnes strain and maximum clear zone of 10.50 mm. in Bacillus subtilis strain. To solve the problem of insolubility and to improve skin penetration, PCL-PEG polymer micelles containing Robinia pseudo-acacia leaf ethanol extracts and 1.0% cell permeable peptide, hexa-D-arginine (R6) were successfully prepared with particle size of 108.23 and 126.47 nm and excellent skin permeation effects could be showed.
Park, Su In;An, Gyu Min;Kim, Min Gi;Heo, Soo Hyeon;Shin, Moon Sam
Journal of the Korean Applied Science and Technology
/
v.36
no.3
/
pp.853-865
/
2019
In this study, the skin permeability was measured by adding skin penetrating peptides, arginine oligomers R4(tetra-D-arginine), R6(hexa-D-arginine) to little skin-permeable wrinkle improvement peptides GHK, GHK-Cu, and Pal-GHK liposomes, and the results were analyzed by the following six cases. (1) In cases where only wrinkle improvement peptides GHK, GHK-Cu, and Pal-GHK were contained liposomes; the final cumulative permeations in 24 hours were 6.05%, 7.4%, and 8.83% respectively. (2) In cases where arginine oligomers R4, R6 were added to GHK liposomes; the final cumulative permeations in 24 hours were 13.63% and 7.68%. (3) In cases where R4, R6 were added to GHK-Cu liposomes; the final cumulative permeations in 24 hours were 15.46% and 8.64%. (4) In cases where R4, R6 were added to Pal-GHK liposomes; the final cumulative permeations in 24 hours were 16.9% and 10.67%. (5) In cases where R4 were added to GHK, GHK-Cu, and Pal-GHK liposomes; the final cumulative permeations in 24 hours were 13.63%, 15.46%, and 16.9% respectively. (6) In cases where R6 were added to GHK, GHK-Cu, and Pal-GHK liposomes; the final cumulative permeations in 24 hours were 7.68%, 8.64%, and 10.67% respectively. This experiment showed that skin absorption of GHK was increased by copper ion (Cu2+) and palmitic acid and skin absorption of wrinkle improvement peptides was enhanced by cell penetrating peptides, and R4 showed higher effect than R6 in GHK, GHK-Cu and Pal-GHK. Through this process, we propose broad use and application in wrinkle improvement functional cosmetics by presenting the optimal conditions for increasing skin absorption of GHK, GHK-Cu, thus maximizing its efficacy.
Park, Su In;An, Gyu Min;Kim, Min Gi;Heo, Soo Hyeon;Shin, Moon Sam
Korean Chemical Engineering Research
/
v.58
no.1
/
pp.29-35
/
2020
In this study, the skin permeability was measured by adding cell penetrating peptides, arginine oligomers; (tetra-D-arginine (R4) and hexa-D-arginine (R6)) to little skin-permeable anti-wrinkle peptides (GHK, GHK-Cu, and Pal-GHK), and the results were analyzed by the following six cases. First, in cases where only anti-wrinkle peptides were contained, copper ions (Cu2+) and palmitic acid enhanced the transdermal permeability. Second, when arginine oligomers (R4, R6) were added to GHK, arginine oligomers (R4, R6) increased percutaneous permeability, and R4 showed better percutaneous permeability. Third, the addition of R4 and R6 to GHK-Cu resulted in increased percutaneous transmittance, followed by R6 < R4 percutaneous transmittance. Fourth, when R4 and R6 were added to Pal-GHK, the percutaneous permeability increased with results in R6 < R4 order. Fifth, when R4 was added to GHK, GHK-Cu, and Pal-GHK, the transdermal permeability increased in the order of GHK+R4 < GHK-Cu+R4 < Pal-GHK+R4. Finally, the addition of R6 to GHK, GHK-Cu and Pal-GHK also resulted in increased percutaneous transmittance in the order of GHK+R4 < GHK-Cu+R4 < Pal-GHK+R4. This study provides optimal conditions for enhancing skin absorption of anti-wrinkle peptides GHK, GHK-Cu, and Pal-GHK, and propose a wide range of applications in anti-wrinkle functional cosmetics by suggesting ways to maximize their efficacy.
An, Gyu Min;Park, Su In;Kim, Min Gi;Heo, Soo Hyeon;Shin, Moon Sam
Journal of the Korean Applied Science and Technology
/
v.36
no.3
/
pp.685-699
/
2019
Antioxidant, antibacterial, and skin penetration tests were conducted to investigate the skin absorption of Pyrus serotina var leaf extracts using polymer micelles and their applicability to cosmetic ingredients. Total polyphenol content was found to be $118.83{\pm}9.39mg/g$ in Pyrus serotina var leaf ethanol extract and $106.89{\pm}4.45mg/g$ in Pyrus serotina var leaf hydrothermal extract. The DPPH radical scavenging activity was found to be the highest radical scavenging activity of $74.39{\pm}7.48%$ of the Pyrus serotina var leaf ethanol extract at the concentration of 500 mg/L. The SOD-like activity was $91.62{\pm}0.43%$, the highest value at the concentration of 1,000 mg/L in the hydrothermal extract. After the experiment, antioxidation, wrinkle improvement and whitening activity were confirmed, and the Pyrus serotina var leaf extract was highly likely to be realized as antioxidant and antibacterial material. In the skin penetration experiment with the Pyrus serotina var leaf ethanol extract, the permeation amount of total accumulated tannic acid was found to be Formulation 2 ($55.45{\mu}g/cm^2$), Formulation 1 ($46.43{\mu}g/cm^2$), Formulation 0 ($34.36{\mu}g/cm^2$). In the liposome's skin penetration experiment containing pear leaf hydrothemal extract, the total amount of accumulated tannic acid permeation was found to be Formulation 5 ($75.01{\mu}g/cm^2$), Formulation 4 ($64.01{\mu}g/cm^2$) and Formulation 3 ($36.60{\mu}g/cm^2$). Through this study, we confirmed the possibility of antioxidant and wrinkle effects of Pyrus serotina var leaf extract. In addition, as a result of skin penetration through the production of polymer micelles and liposome containing Pyrus serotina var leaf extract, It will be more usable in cosmetic industry.
Park, Young-Mi;Kang, Myung-Joo;Moon, Ki-Young;Park, Sang-Han;Kang, Mean-Hyung;Choi, Young-Wook
Journal of Pharmaceutical Investigation
/
v.41
no.4
/
pp.205-210
/
2011
Genistein (GT), a major isoflavone found in soybeans, has a potent antioxidant effect that protects the skin from UV-induced damages and malignant melanoma. In order to enhance the cellular uptake of GT, liposome/Tat complexes were prepared by an electrostatic interaction of anionic liposome (DMPC/DCP, 9:1 in molar ratio) with Tat peptide (0.02 to 0.08 mole), one of the well-known cell penetrating peptide (CPP). As the amount of Tat increased, the size increased but the zeta potential decreased. In vitro release study with dialysis membrane elicited GT release from liposomal preparations in a controlled manner. The addition of Tat increased GT release, especially for the initial period. In the cellular uptake study by incubating B16 melanoma cells with various liposomal preparations containing GT, B16 melanoma cells demonstrated a time-dependent increase of drug accumulation. Compared to the aqueous GT suspension, intracellular uptake was substantially enhanced by anionic liposomal formulation and further increased by the complex formulation. Therefore, liposome/ Tat complex might be a good candidate for facilitating intracellular drug delivery.
Kang, Jin Sun;La, Ha Na;Bak, Sun Uk;Eom, Hyo Jung;Lee, Byung Kyu;Shin, Hee Je
Journal of the Society of Cosmetic Scientists of Korea
/
v.45
no.2
/
pp.175-184
/
2019
The epidermal growth factor (EGF) has a intrinsic function of inducing growth and proliferation of cells through interacting with cell membrane receptors in human epidermis and dermis layer. These functions of EGF are used as a main ingredient for wound healing medicines and anti-aging cosmetics. As a cosmetic ingredient, the EGF has a problem in exhibiting its natural efficacy due to the lack of the ability to penetrate through the stratum corneum, which is known as the skin barrier. In this study, a recombinant human epidermal growth factor ($MTD_{151}-EGF$) fused with the macromolecule transduction domain $(MTD)_{151}$ with the skin penetration ability was developed to improve the skin penetration efficiency of the EGF. Expression of $MTD_{151}-EGF$ was performed in E. coli transformed with a vector encoding the $MTD_{151}-EGF$ gene and then purified. The purified $MTD_{151}-EGF$ was evaluated using cell proliferation assay, cytotoxicity test and skin penetration test by franz diffusion cell assay and artificial skin. Cell proliferation activity of $MTD_{151}-EGF$ purified to high purity of 99% or above was equivalent to the EGF or better, and cytotoxicity was not observed. In addition, the $MTD_{151}-EGF$ showed an excellent penetration efficiency compared to the EGF in the skin penetration test with EGF and $MTD_{151}-EGF$ labeled by FITC in an artificial skin penetration model. Based on the quantitative analysis of the penetrating substance using franz diffusion cell assay, the amount of penetration was about 16 times more than that of EGF. These results can be regarded as an effective alternative to improve the existing physical transdermal penetration method related to the use of various active ingredients for cosmetics.
An, Gyu Min;Park, Su In;Kim, Min Gi;Heo, Soo Hyeon;Shin, Moon Sam
Journal of the Society of Cosmetic Scientists of Korea
/
v.45
no.3
/
pp.265-275
/
2019
This study was conducted to investigate physiological activity and its skin permeability of Petroselinum crispum extract using polymer micelles and cell penetrating peptide. In the antioxidant test, the total concentrations of polyphenol compounds were determined to be $121.68{\pm}2.49mg/g$ (for ethanol extract and), $72.42{\pm}1.52mg/g$ (for hydrothermal extract.). The DPPH radical scavenging ability was $90.48{\pm}0.46%$ (for ethanol extract) and $83.92{\pm}0.13%$ (for hydrothermal extract) at 2000 mg/L. ABTS radical scavenging ability was $91.08{\pm}0.14%$ for ethanol extract ethanol extract, which is higher than that of hydrothermal extract at 800 mg/L ($69.63{\pm}0.55%$). In the SOD experiments, the P. crispum ethanol extract showed higher SOD activity than that of the P. crispum hydrothermal extract at all concentrations.. At a concentration of 16,000 mg/L, P. crispum ethanol extract showed the highest SOD activity of $128.45{\pm}0.70%$. The elastase inhibitory assay also showed concentration dependence and elastase inhibition of P. crispum ethanol extract was $99.99{\pm}1.54%$, which was the highest at 2,000 mg/L. To solve the problem of insolubility and to improve skin permeability of the extract, PCL-PEG polymer micelle containing P. crispum ethanol extracts and 1% cell permeable peptide, hexa-D-arginine (R6) were successfully prepared with a particle size of 40.10 nm. In the results of 24 hours of skin permeation experiment, total accumulated beta-carotene amounts showed $37.99{\mu}g/cm^2$ in Petroselinum crispum extracts and $68.38{\mu}g/cm^2$ (1.8 times) in P. crispum extract of the particles.
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