• Tissue Engineering and Regenerative Medicine
• /
• v.15 no.6
• /
• pp.711-719
• /
• 2018

BACKGROUND: Collagen organization within tissues has a critical role in wound regeneration. Collagen fibril diameter, arrangements and maturity between connective tissue growth factor (CTGF) small interfering RNA (siRNA) and mismatch scrambled siRNA-treated wound were compared to evaluate the efficacy of CTGF siRNA as a future implement for scar preventive medicine. METHODS: Nanocomplexes of CTGF small interfering RNA (CTGF siRNA) with cell penetrating peptides (KALA and $MPG^{{\Delta}NLS}$) were formulated and their effects on CTGF downregulation, collagen fibril diameter and arrangement were investigated. Various ratios of CTGF siRNA and peptide complexes were prepared and down-regulation were evaluated by immunoblot analysis. Control and CTGF siRNA modified cells-populated collagen lattices were prepared and rates of contraction measured. Collagen organization in rabbit ear 8 mm biopsy punch wound at 1 day to 8 wks post injury time were investigated by transmission electron microscopy and histology was investigated with Olympus System and TS-Auto software. CONCLUSION: CTGF expression was down-regulated to 40% of control by CTGF siRNA/KALA (1:24) complexes (p<0.01) and collagen lattice contraction was inhibited. However, down-regulated of CTGF by CTGF $siRNA/MPG^{{\Delta}NLS}$ complexes was not statistically significant. CTGF KALA-treated wound appeared with well formed-basket weave pattern of collagen fibrils with mean diameter of $128{\pm}22nm$ (n = 821). Mismatch siRNA/KALA-treated wound showed a high frequency of parallel small diameter fibrils (mean $90{\pm}20nm$, n = 563). CONCLUSION: Controlling over-expression of CTGF by peptide-mediated siRNA delivery could improve the collagen orientation and tissue remodeling in full thickness rabbit ear wound.

• Journal of Veterinary Science
• /
• v.25 no.1
• /
• pp.12.1-12.10
• /
• 2024

Background: Staphylococcus aureus and S. pseudintermedius are the major etiological agents of staphylococcal infections in humans, livestock, and companion animals. The misuse of antimicrobial drugs has led to the emergence of antimicrobial-resistant Staphylococcus spp., including methicillin-resistant S. aureus (MRSA) and methicillin-resistant S. pseudintermedius (MRSP). One novel therapeutic approach against MRSA and MRSP is a peptide nucleic acid (PNA) that can bind to the target nucleotide strands and block expression. Previously, two PNAs conjugated with cell-penetrating peptides (P-PNAs), antisense PNA (ASP)-cmk and ASP-deoD, targeting two essential genes in S. aureus, were constructed, and their antibacterial activities were analyzed. Objectives: This study analyzed the combined antibacterial effects of P-PNAs on S. aureus and S. pseudintermedius clinical isolates. Methods: S. aureus ATCC 29740 cells were treated simultaneously with serially diluted ASP-cmk and ASP-deoD, and the minimal inhibitory concentrations (MICs) were measured. The combined P-PNA mixture was then treated with S. aureus and S. pseudintermedius veterinary isolates at the determined MIC, and the antibacterial effect was examined. Results: The combined treatment of two P-PNAs showed higher antibacterial activity than the individual treatments. The MICs of two individual P-PNAs were 20 and 25 µM, whereas that of the combined treatment was 10 µM. The application of a combined treatment to clinical Staphylococcus spp. revealed S. aureus isolates to be resistant to P-PNAs and S. pseudintermedius isolates to be susceptible. Conclusions: These observations highlight the complexity of designing ASPs with high efficacy for potential applications in treating staphylococcal infections in humans and animals.

• 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.

• 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.

• BMB Reports
• /
• v.51 no.5
• /
• pp.207-208
• /
• 2018

Chitinase-Like Proteins (CLPs) are an evolutionarily conserved protein which lose their enzymatic activity for degrading chitin macromolecules. Chitinase-3-like-1 (Chi3l1) is a type of CLP that is highly expressed in epithelial cells, macrophages, etc., and is known to have correlations with type 2 inflammation and cancer. Although the increased level of Chi3l1 in the blood was reported in various disease patients, the function of Chi3l1 in adaptive immunity has been totally unknown. Recently, we found that Chi3l1 is expressed in T cells and has a negative regulatory role in T-cell activation and proliferation. A genetic ablation study of Chi3l1 in T cells showed hyperresponsiveness to TcR stimulation, which increased proliferation and Th1 differentiation. A significant increase of $IFN{\gamma}$ signaling in Chi3l1-deficient T cells synergistically increased Th1 and CTL functions against melanoma cells in vitro and in vivo. In addition, targeted knockdown by Chi3l1 siRNA complexed with the cell-penetrating peptide dNP2, which showed decreased pulmonary melanoma metastasis with increased infiltration of Th1 and CTL in the lung. This study first suggests that Chi3l1 is a novel regulator of Th1/CTL responses and could be a target for treating cancer to increase tumor immunity.

• Molecules and Cells
• /
• v.27 no.2
• /
• pp.159-166
• /
• 2009

Myocardial ischemia-reperfusion injury is a medical problem occurring as damage to the myocardium following blood flow restoration after a critical period of coronary occlusion. Oxygen free radicals (OFR) are implicated in reperfusion injury after myocardial ischemia. The antioxidant enzyme, Cu, Zn-superoxide dismutase (Cu, Zn-SOD, also called SOD1) is one of the major means by which cells counteract the deleterious effects of OFR after ischemia. Recently, we reported that a PEP-1-SOD1 fusion protein was efficiently delivered into cultured cells and isolated rat hearts with ischemia-reperfusion injury. In the present study, we investigated the protective effects of the PEP-1-SOD1 fusion protein after ischemic insult. Immunofluorescecnce analysis revealed that the expressed and purified PEP-1-SOD1 fusion protein injected into rat tail veins was efficiently transduced into the myocardium with its native protein structure intact. When injected into Sprague-Dawley rat tail veins, the PEP-1-SOD1 fusion protein significantly attenuated myocardial ischemia-reperfusion damage; characterized by improving cardiac function of the left ventricle, decreasing infarct size, reducing the level of malondialdehyde (MDA), decreasing the release of creatine kinase (CK) and lactate dehydrogenase (LDH), and relieving cardiomyocyte apoptosis. These results suggest that the biologically active intact forms of PEP-1-SOD1 fusion protein will provide an efficient strategy for therapeutic delivery in various diseases related to SOD1 or to OFR.

• 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.

• 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.

• 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.

• 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.