• Biomolecules & Therapeutics
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• v.25 no.3
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• pp.296-307
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• 2017

In spite of frequent usage of primary human foreskin keratinocytes (HFKs) in the study of skin biology, senescence-induced block-age of in vitro proliferation has been a big hurdle for their effective utilization. In order to overcome this passage limitation, we first isolated ten HFK lines from circumcision patients and successfully immortalized four of them via a retroviral transduction of high-risk human papillomavirus (HPV) E6 and E7 oncogenes. We confirmed expression of a keratinocyte marker protein, keratin 14 and two viral oncoproteins in these immortalized HFKs. We also observed their robust responsiveness to various exogenous stimuli, which was evidenced by increased mRNA expression of epithelial differentiation markers and pro-inflammatory genes in response to three reactive chemicals. In addition, their applicability to cytotoxicity assessment turned out to be comparable to that of HaCaT cells. Finally, we confirmed their differentiation capacity by construction of well-stratified three dimensional skin cultures. These newly established immortalized HFKs will be valuable tools not only for generation of in vitro skin disease models but also for prediction of potential toxicities of various cosmetic chemicals.

• Journal of Microbiology and Biotechnology
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• v.28 no.8
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• pp.1391-1400
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• 2018

Angelica tenuissima root has historically been used as a traditional medicine in Korea. Previous studies have identified the anti-melanogenic effects of the extract of A. tenuissima root fermented by Aspergillus oryzae (FAT). This study investigated the protective effects of FAT against ultraviolet light B exposure (UVB; $30mJ/cm^2$) in HaCaT (human keratinocyte) or Hs68 (human foreskin fibroblast) skin cells. FAT treatment was able to stimulate wound healing rate at the basal condition. FAT also favored the maintenance and/or improvement of extracellular matrix impairment caused by UVB irradiation through: 1) upregulation of procollagen Type-1 synthesis and secretion; 2) suppression of MMP-1 and elastase expression. FAT was able to play a role in the attenuation of inflammatory responses caused by UVB irradiation via upregulation of photo-protective hemeoxygease-1 and suppression of proinflammatory cyclooxygenase-2 expression. After further verification of the anti-photoaging potential of FAT, it could be utilized as an effective ingredient in anti-aging and anti-wrinkle cosmetics.

• The Korean Journal of Physiology and Pharmacology
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• v.6 no.5
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• pp.275-279
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• 2002

Nerve growth factor (NGF) is an important autocrine growth factor and also a survival factor for keratinocytes. NGF may act in the hyperproliferative condition, psoriasis. Clinically, the combination of psoralen and UVA (PUVA) has been used in the treatment of a wide variety of cutaneous disorders, such as psoriasis and vitiligo. However, the precise therapeutic mechanism of PUVA on the dermatologic diseases remains unclear. The purpose of this study was to examine whether the expression of NGF in cultured keratinocytes is influenced by PUVA. Thus, normal human keratinocytes were isolated from neonatal foreskin, and the third to fifth-passaged cells were used in this study. The cells were exposed to various doses of UVA (30, 60, 120 $mJ/cm^2)$ after adding 8-methoxypsoralen (8-MOP) to examine the expression of NGF mRNA. The RNA and protein of the cells were extracted at various time points (1, 8, 24 hours) after UVA irradiation to examine the expression of NGF mRNA and production of NGF protein. In keratinocytes, there were no differences in the expression of NGF mRNA between the different doses of UVA irradiation, however, the expression of NGF mRNA in UVA and PUVA groups tended to increase as the time increased. The expression of NGF mRNA was the highest in PUVA group, followed by UVA group and the lowest in 8-MOP group. The expressions of NGF protein at 1 and 8 hours after UVA irradiation were lower in the PUVA group than in the other groups. This study showed that the expression level of NGF protein in keratinocytes was relatively lower in the PUVA groups than in the other groups, suggesting that the therapeutic mechanism of PUVA in psoriasis is related to the decrease of NGF protein.

• Applied Biological Chemistry
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• v.46 no.1
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• pp.60-65
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• 2003

MMP-1 inhibitory compounds were isolated from 120 Korean traditional edible plants. UP- 1 activity significantly increased linearly with increasing UVB dose in normal human foreskin fibroblast HS68 cell, showing maximum activity at approximately 35 $mJ/cm^2$, whereas in HaCaT cell, normal human keratinocyte, no increase was observed. Maximum secretion of MMP-1 after UVB treatment occurred around 36-48 k after treatment. MMP-1 inhibitory compound isolated from cold-water fraction of Cataegus pinnatifida Bunge showed the mort potent activity. The MMP-1 inhibitory compound was deduced as a peptide based on the fact that pronase digestion decreased the activity whereas periodate oxidation did not. The most potent UP- 1-inhibitory protein, CP-2Va-2, showing an activity of 88.5% against MMP-1, was isolated through sequential column chromatography on DEAE-Toyopearl 650C, Butyl-Toyopearl 650M, and Bio-Gel P-30. Molecular weight of CP-2Va-2 determined through high performance liquid chromatography and SDS PACE was 19 and 20 kDa. respectively, signifying a monomeric structure.