• 한국응용과학기술학회지
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• 제37권4호
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• pp.820-829
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• 2020

본 연구에서는 기능성 화장품의 소재로서의 금전초(Lysimachia christinae Hance)의 항산화와 항주름 효과를 조사하였다. 최근 천연물의 주름 개선 개발의 연구가 지속적인 관심을 받고 있어 본 연구를 통해 활성산소종(reactive oxygen species, ROS) 생성과 pro-collagen 합성 및 MMPs의 연관성에 대해 알아보았다. 금전초는 70% 에탄올(LcHE)과 열수(LcHW)로 각각 추출하여 실험을 진행하였다. HaCaT cells에서 LcHE가 LcHW보다 ROS 저해효능이 더 우수하고 세포독성 결과 250 ㎍/mL 농도까지 독성을 보이지 않아 LcHE를 선택하여 주름 개선 소재연구를 진행하였다. pro-collagen 합성실험을 통하여 UVB에 의해 감소된 type-1 pro-collagen의 합성 활성을 유의미하게 확인하였다. Western blot 실험을 통하여 피부세포에서 UVB에 의해 유도된 MMPs 중 MMP-1 -3 -9의 증가를 억제함을 확인하였으며, Real time PCR을 통하여 상위단계인 mRNA levels에서도 MMP-1, MMP-2, MMP-3, MMP-9의 mRNA levels가 농도 의존적으로 유의미한 감소를 보여 추출물의 효능을 확인하였다. 위의 실험결과에 따라 UVB에 의한 주름생성과 피부 광노화를 효과적으로 예방할 수 있는 화장품의 천연소재로서의 이용이 기대된다.

• 한국식품영양과학회지
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• 제21권3호
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• pp.241-246
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• 1992

L-Ascorbic acid (AsA) 생합성이 불가능한 guinea pig을 실험 동물로 하여 collagne 함량이 높은 조직인 폐 및 피부중의 AsA함량과 동일조직중 collagne 함량이 높은 조직인 폐 및 피부중의 AsA 함량과 동일조직중 collagen의 proline잔기의 수산화율을 조사하여 collagne 생합성에 대한 조직중 AsA의 영향에 대해서 알아보았다. Guinea pig(체중 약 250g)를 AsA 무투여군(A), 투여군(B), 300mg/day 투여군(C)으로 나눠 14일간 사육한 후, 마취하에서 개복하여 복부 대동맥으로부터 채혈함과 동시에 간장과 폐를 적출하였으며, 등부위의 피부를 채취하여 분석용 시료로 하였다. 이들 시료로부터 혈청중 alkaline phosphatase (ALP)활성과 각조직중의 AsA 함량, proline 함량 및 그 수산화율, (1-$^{14}$ C) proline 의 incorporation 양을 측정하였다. 그 결과, AsA 토여군인 B, C군의 경우 순조로운 체중증가와 함께 혈청중 ALP활성도 정상값을 나타냈으며 현저한 ALP활성 저하가 관찰되었다. 한편, AsA 함량이 높을수록 (1-14C) proline 의 incorporation 양이 많고 collagne 중의 hydroxyproline 함량도 증가하는 것으로 나타나, 조직중의 collagen합성량과 AsA함량과의 사이에는 높은 상관관계가 존재함이 확인되었다.

• Archives of Plastic Surgery
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• 제36권6호
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• pp.679-684
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• 2009

Purpose: Human lipoaspirate cells are relatively easy to obtain in large quantities without cell culture. The aim of this in vitro pilot study was to determine the effect of cell therapy using uncultured lipoaspirate cells on cell proliferation and collagen synthesis of diabetic fibroblasts, which are the major contributing factors in wound healing. Methods: In order to get diabetic fibroblasts, dermis tissues were obtained from foot skin of diabetic patients who underwent debridements or toe amputations(n = 4). In order to isolate lipoaspirate cells, the same diabetic patients' abdominal adipose tissues were obtained by liposuction. The diabetic fibroblasts were co - cultured with or without autogenous lipoaspirate cells using porous culture plate insert. Initial numbers of the lipoaspirate cells and diabetic fibroblasts seeded were 15,000 cells/well, respectively. For cell proliferation assay, two treatment groups were included. In group I, diabetic fibroblasts were cultured with the insert having no cells, which serves as a control. In group II, the lipoaspirate cells were added in the culture plate insert. For collagen synthesis assay, one additional group(group III), in which diabetic fibroblasts were not seeded in the well and only lipoaspirate cells inside the insert were incubated without diabetic fibroblasts, was included for a reference. Results: One hundred to one hundred sixty thousand lipoaspirate cells were isolated per ml of aspirated adipose tissue. After 3 - day incubation, the mean cell numbers in group I and II were 17,294/well and 22,163/well. The mean collagen level in group I, II, and III were 29, 41, and 2 ng/ml, respectively. These results imply that both cell proliferation and collagen synthesis in the lipoaspirate cell treatment group were 28 and 44 percents higher than in the control group, respectively(p < 0.05). Conclusion: Uncultured lipoaspirate cell autografts may stimulate the wound healing activity of diabetic fibroblasts.

• Journal of Periodontal and Implant Science
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• 제27권2호
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• pp.291-303
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• 1997

Periodontal ligament cells may have a role in the regulation of hard and soft periodontal tissues, but their specific function has not yet to be determined. To evaluate further their role in periodontal regeneration, they were examined for osteoblast-like behavior. Periodontal ligament cells and gingival fibroblasts were primarily cultured from extracted premolar with non-periodontal diseases. Cells were cultured with DMEM at $37^{\circ}C$, 5% $CO_2$, 100% humidity incubator, and as a measure of cell characterization, it was examined that the morphology, alkaline phosphatase activity, collagen synthesis, and immunocytochemistry for osteonectin, osteocalcin, and collagen type I. Healthy periodontal ligament cells has more osteoblastic-like cell property in alkaline phosphatase activity. and collagen synthesis than gingival fibroblast. Immunocytochemistry localization explained that calcitonin were expressed in periodontal ligament cells only, and osteonectin and type I collagen were produced in both cells simultaneously. This results indicate that the growth characteristics of periodontal ligament cells and gingival fibroblasts exhibit some differences in proliferative rates and biochemical synthesis. The differences may help to calrify the role such cells play in the regenearation of periodontal tissues.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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• pp.659-659
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• 2003

In this study, we have investigated the potent anti-aging effect of DA-3711, a cosmetic ingredient derived from artificial skin culture. The artificial skin was originally developed as a skin replacement for the treatment of chronic skin wounds. To produce DA-3711, neonatal human fibroblasts were seeded into biocompatible collagen/chitosan/glycosaminoglycan (GAG) scaffolds and cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with fetal bovine serum and nonessential amino acids. Analysis of the culture broth (DA-3711) showed that growth factors such as VEGF, TGF-$\beta$, KGF were present at significantly higher levels that in the culture broth of fibroblasts cultured in monolayer. The biological activity of DA-3711 was assessed by measuring in vitro cell proliferation and collagen synthesis of normal human fibroblasts. Fibroblasts treated with 10％ DA-3711 showed a 2-fold higher proliferation and 2 to 4-fold higher collagen synthesis than untreated cells. DA-3711 also exhibited anti-oxidative effects, since cells under peroxide-induced oxidative stress showed a 30％ higher viability in DA-3711-containing medium than in medium without DA-3711 addition. The results suggest that DA-3711 may have anti-aging effects by stimulating skin regeneration and protecting against oxidative stress.

• Archives of Plastic Surgery
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• 제32권4호
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• pp.529-532
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• 2005

Expanding cells ex-vivo is very important in tissue-engineering. Culture medium is usually supplemented with fetal bovine serum(FBS) in most of the experiments. However, cells grown in bovine serum media may posses the possibilities of disseminating bovine diseases and/or stimulating the patient's immune reactions. To overcome these problems, autologous or homologous serum should be used instead of the FBS. The purpose of this study is to compare cell proliferation and collagen synthesis depending on the kind of sera mixed on media and to provide a guideline on applying established experimental data to clinical cases. Human dermal fibroblasts were obtained from four patients. Five thousand cells per well in 96-well plates were incubated DMEM/F-12 Nutrient with varying serum mixture; 10% autologous serum, 10% homologous serum, and 10% FBS. Five days after incubation fibroblast proliferation and collagen production were determined by MTT assay and CICP enzyme immunoassay. The mean cell number were; $3.95{\times}10^4/well$, $2.97{\times}10^4/well$ and $2.30{\times}10^4/well$, respectively. The average amounts of collagen synthesized were; 238.13 ng/ml, 204.88 ng/ml, and 163.88 ng/ml in each. These results show that the use of human serum mixture may contribute to, not only preventing disseminated infection of bovine diseases. but also increase cell proliferation and collagen synthesis without simulating the patient's immune reactions.

• Journal of Applied Biological Chemistry
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• 제58권2호
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• pp.183-187
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• 2015

시호 추출물로부터 자외선에 의한 주름개선 효과를 확인하여 화장품 소재로서의 가능성을 검증하였다. 주름 활성 검증을 위하여 전자공여능, elastase, pro-collagen 생합성, Matrix metalloprotease-1 (MMP-1)의 활성을 측정하였다. 시호 추출물의 세포 독성을 측정하기 위하여 MTT assay를 하여 세포 독성이 100%에 가까운 농도인 5, 10, $50{\mu}g/mL$의 농도에서 pro-collagen 생합성과 MMP-1의 활성 검증하였다. 시호 추출물의 전자 공여능과 elastase 활성을 측정한 결과 $1,000{\mu}g/mL$의 농도에서 각각 80, 52%의 저해 활성을 가졌다. 또한 pro-collagen 생합성 역시 UVB를 조사한 후 시호 추출물을 농도 별로 처리 한결과 농도의존적으로 증가하는 것을 확인하였다. 주름생성에 관련되어져 있는 MMP-1의 발현을 알아본 결과 시호 추출물에 의해 total protein 양이 또한 감소되는 것을 확인 할 수 있었다. 따라서 시호는 주름활성을 개선시킬 수 있는 기능성 소재로 활용 될 수 있을것으로 사료된다.

• Molecules and Cells
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• 제26권6호
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• pp.625-630
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• 2008

This study was undertaken with the aim of developing an easy and quick means of analyzing the effect of various compounds on the synthesis and secretion of human type I collagen at the protein level. A modification of the ELISA method was used on HFF-1 cells. For the proof of concept, we used thirteen compounds most of which are known to be antioxidants. Each compound was tested at concentrations of 0, 10 and $100{\mu}m$ on HFF-1 cells for 24 h. Thirteen sets of experiments for each compound were performed in ANOVA with three replicates. Duncan multiple range test (DMRT) was used to compare the mean values obtained from the treatment groups. From the results it was concluded that Vitamin C, undecylenic acid, conjugated linoleic acid, glycolic acid, and citric acid at $100{\mu}m$ concentration could be used for anti-wrinkling or protection from premature aging, which requires enhancement of collagen synthesis. Lactic acid, EGCG, resveratrol, and retinol that can inhibit collagen synthesis effectively in a dose-dependent manner may be used for anti-fibrosis treatment purposes.

• 한방안이비인후피부과학회지
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• 제21권1호
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• pp.1-15
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• 2008

Objective : Skin aging is commonly observed in patients with diabetes mellitus, which can be accessed by the amount of skin collagen and matrix metalloproteinase-1 (MMP-1). In the present study, anti-skin-aging effects of Root Cortex of Paeonia Suffruticosa Andrews (PSA), which has been widely used to treat diabetes mellitus, are investigated. Methods : Streptozotocin (STZ) was intraperitoneally injected to rats to induce diabetes. Body weights, feed intake, organ weights, blood glucose, and other biochemical index are determined in both normal and diabetic rats. In order to study the effect of PSA on skin aging, the amount of skin collagen was measured in diabetic rats after PSA treatments. Also, MMP-1 synthesis in UVB-irradiated human skin fibroblasts was investigated. Results : 1. When PSA was administered to STZ-induced diabetic rats, feed intake was significantly increased and blood glucose and total cholesterol were decreased in a dose-dependent manner. However, there are no differences in individual organ weights, GOT, and GPT. 2. A decrease of skin collagen in diabetic rats was significantly suppressed when PSA was treated. 3. PSA also inhibited MMP-1 synthesis in UVB-irradiated normal human skin fibroblasts, similar to retinoid, a well-known effective anti-skin-aging substance. Conclusion: PSA suppressed a collagen decrease in diabetic rats and inhibited MMP-1 synthesis in UVB-irradiated human skin fibroblasts. Therefore, the treatment of PSA is very effective to slow down the skin aging process.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2003년도 추계학술대회
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• pp.120-120
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• 2003

Synthesis and distribution of extracellular matrix (ECM) components are dynamically altered in response to the pathophysiological processes including infection, inflammation and apoptosis. In particular, the levels of hyaluronan (HA) change with concomitant increases in the levels of collagen (e.g. type I collagen) and fibronectin in chronic liver diseases.(omitted)