본 연구에서는 미세먼지에서 유도된 염증 발현 효과를 확인하고 이에 대한 측정방법을 정립하였으며, 지모추출물, 베타글루칸, 폴리감마글루탐산의 3종 혼합물에 대해 안티폴루션 효능에 대해 세포 독성 (LDH assay), 세포 생존율 (MTT assay), 자유라디칼 소거능 (DPPH assay) 및 활성산소 억제 효과 (DCF-DA assay)를 측정하였고 미세먼지로 유도된 자극에서 3종 혼합물의 안티폴루션 화장품 소재로의 활용 가능성을 피부 수분량, 피부 각질량, 가려움증 개선 임상시험을 통해 확인하였다.
지모추출물, 베타글루칸, 폴리감마글루탐산의 3종 혼합물은PM1648a로 농도별로 자극을 유발시켰을 때 자유라디칼 소거능을 유의 있게 증가시켰고, 모든 혼합물 농도에서 6.5% 이하의 세포 독성을 확인하였고, 91.25% 이상의 세포 생존율을 확인하였으며, HaCaT cell에서 미세먼지로 자극한 후 혼합물 농도별로 처리하여 유의 있는 활성산소 생성 억제 효과를 확인하였다.
1.0% 지모추출물, 0.4% 베타글루칸, 0.5% 폴리감마글루탐산 혼합물을 함유한 로션 제형 안티폴루션 화장품으로 4주 동안 임상시험을 실시한 결과, 피부 수분 함유량 증가, 피부 각질 감소, 가려움증 감소가 통계적으로 유의한 결과를 나타내었다.
미세먼지 자극 유도에 의한 1.0% 지모추출물, 0.4% 베타글루칸, 0.5% 폴리감마글루탐산 혼합물에 대한 세포실험과 임상시험의 안티폴루션 결과를 종합해 볼 때, 본 혼합추출물은 세포 독성 (LDH)과 세포 생존율 (MTT)에서 효과적인 효능을 지니며, 미세먼지 자극에 의한 항산화력 (DPPH), 세포 내 활성산소 (ROS) 억제 효과에서도 유의적인 효과를 지니고 있으며, 피부건조증이 있는 시험대상자의 임상시험에서도 피부 수분량, 피부 각질, 피부 가려움증 개선에 유의적인 효과를 나타내어 안티폴루션화장품으로 활용가능성을 확인할 수 있었다.
This study characterized Prostaglandin E2 (PGE2) production from Raw264.7 cells stimulated with particulate matter (PM1648a) and tested anti-pollution capacity of mixtures of Anemarrhena asphodeloides root extract, β-glucan and poly- γ-glutamic acid (the complex). In order to assess cytotoxicity, we treated the Raw264.7 cells with the complex using the lactase dehydrogenase (LDH) assay method. To measure cell viability, we treated HaCaT cells with the complex using the MTT assay method. To test scavenging activity, the DPPH assay method was used. To test the anti-oxidant effect of the complex, we treated HaCaT cells with the pro-inflammatory mediator particulate matter (PM1648a) and with PM1648a and the complex, then measured the concentration of reactive oxygen species (ROS) within the cells using the dichlorofluorescin diacetate (DCF-DA) assay method. The complex had cytotoxicity below 6.5% and cell viability above 91.3% at a concentration of 0.05%. Also, the complex showed scavenging activity above 50% at a concentration of 1.0%. In addition, ROS production at the 0.05% complex concentration was reduced to 61.0%. In a clinical experiment, the subjects who had skin urtication used a cosmetic product containing mixtures of 1.0% Anemarrhena asphodeloides root extract, 0.4% β-glucan and 0.5% poly-γ-glutamic acid for 4 weeks. The experimental group showed a statistically significant improvement in skin humidity, desquamation index and skin urtication. Based on the results with in vitro test and clinical experiment, mixtures of Anemarrhena asphodeloides root extract, β-glucan and poly-γ-glutamic acid could be used as cosmetic anti-pollution materials.
This study characterized Prostaglandin E2 (PGE2) production from Raw264.7 cells stimulated with particulate matter (PM1648a) and tested anti-pollution capacity of mixtures of Anemarrhena asphodeloides root extract, β-glucan and poly- γ-glutamic acid (the complex). In order to assess cytotoxicity, we treated the Raw264.7 cells with the complex using the lactase dehydrogenase (LDH) assay method. To measure cell viability, we treated HaCaT cells with the complex using the MTT assay method. To test scavenging activity, the DPPH assay method was used. To test the anti-oxidant effect of the complex, we treated HaCaT cells with the pro-inflammatory mediator particulate matter (PM1648a) and with PM1648a and the complex, then measured the concentration of reactive oxygen species (ROS) within the cells using the dichlorofluorescin diacetate (DCF-DA) assay method. The complex had cytotoxicity below 6.5% and cell viability above 91.3% at a concentration of 0.05%. Also, the complex showed scavenging activity above 50% at a concentration of 1.0%. In addition, ROS production at the 0.05% complex concentration was reduced to 61.0%. In a clinical experiment, the subjects who had skin urtication used a cosmetic product containing mixtures of 1.0% Anemarrhena asphodeloides root extract, 0.4% β-glucan and 0.5% poly-γ-glutamic acid for 4 weeks. The experimental group showed a statistically significant improvement in skin humidity, desquamation index and skin urtication. Based on the results with in vitro test and clinical experiment, mixtures of Anemarrhena asphodeloides root extract, β-glucan and poly-γ-glutamic acid could be used as cosmetic anti-pollution materials.
