• Restorative Dentistry and Endodontics
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• 제21권2호
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• pp.546-558
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• 1996

The purpose of this vitro study was to evaluate the activity of human pulpal cells to adhesive glycoprotein-coated and non-coated culture dishes. Well known adhesive glycoproteins were used, such as type I collagen, type IV collagen, fibronectin, laminin, and vitronectin. Each adhesive glycoproteins applied onto the culture dishes. In this study, the protein coated and non-coated dishes were classified as each groups. Human pulpal cells cultured onto each groups. After 24 hours, 48 hours, 72 hours incubation time, radioactivity with scintillation counter for evaluation of the activity of human pulpal cells. The results as follows : 1. After 24 hours incubation time, activity of human pulpal cells were best in laminin-coated group among groups. Then fibronectin, type I collagen group were better, and all proteins were better than control. 2. After 48 hours incubation time, activity of human pulpal cells were best in fibronectin coated group. 3. After 72 hours incubation time, activity of human pulpal cells were not significantly different in all of adhesive glycoproteins. 4. After 24 hours incubation time, activity of human pulpal cells were best in fibronectin and laminin coated group. Activity of human pulpal cells in type I collagen coated group were better after 24 hours incubation time then 48 hours incubation time.

• Nutrition Research and Practice
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• 제1권4호
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• pp.279-284
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• 2007

Solar ultraviolet (UV) irradiation leads to distinct changes in the skin connective tissues by degradation of collagen, which is a major structural component in the extracellular matrix. UV irradiation induces the production of matrix metalloproteinases (MMP) capable of attacking native fibrillar collagen and responsible for inhibiting the construction of collagenous extracellular matrix. In this study, we attempted to investigate the protective actions of Rubus coreanus ethanol extract (RCE) on the MMP production and the consequent procollagen/collagen degradation in UV-B-irradiated human dermal fibroblasts. The analytical data showed that Rubus coreanus ethanol extract was mostly comprised of cyanidin 3-rutinoside. Pre-treatment of fibroblasts with this extract inhibited UV-B-induced production of MMP-1, MMP-8 and MMP-13 in dose-dependent manners. In addition, Western blot analysis and immunocytochemical staining assay revealed that RCE markedly augmented the cellular levels of procollagen/collagen declined in UV-B-exposed dermal fibroblasts. These results demonstrate that RCE blocks UV-B-induced increase of the collagen degradation by inhibiting MMP production. Thus, RCE may act as an agent inhibiting excessive dermal collagen degradation leading to the skin photoaging.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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• pp.89-89
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• 2003

본 연구에서는 1.7kb 및 3.1kb bovine $\beta$-casein promoter의 유전자 발현 조절능력을 알아보기 위해 1 7kb 및 3.1kb bovine $\beta$-casein promoter에 human Type II Collagen 유전자를 연결해서 DNA microinjection으로 형질전환생쥐를 생산하였다. 총 8마리의 founder생쥐(1.7kb collagen : 5마리, 3.1kb collagen 3마리)를 생산하였고 이 founder생쥐와 wild type 생쥐를 mating시켜서 $F_1 및 F_2$ 새끼를 얻었다. $F_1 및 F_2$새끼들에서 human Type II collagen 유전자의 transmission rate는 약 50%로 Mendel의 법칙에 따라 분리되어 안정적으로 유전자가 염색체에 정착되어 있음을 확인하였다. 이들 $F_1 및 F_2$새끼 중 암컷들을 임신시켜 분만 후 5-10 일경에 유선조직을 포함하여 여러 조직으로부터 RNA를 추출하여 Northern blotting 및 RT-PCR 방법을 이용하여 Type II collagen mRNA의 발현을 분석하였다. 유선에서의 발현은 1 7 kb 및 3.1 kb line별로 각각 1 line씩 발현되지 않았고, 그 외 line에서는 모두 발현되는 것으로 확인되었다. 유선에서의 Type II collagen mRNA 발형양은 1.7 kb 및 3.1 kb bovine $\beta$-casein promoter사이에서는 큰 차이를 나타내지 않았으나 1.7 kb promoter 형질전환생쥐의 경우 유선 이외 조직에서도 발현되는 양상을 나타내었고, 3.1kb promoter line에서는 유선특이적으로 발현시키는 양상을 나타내었다. 그러므로 bovine $\beta$-casein promoter의 1.7 kb와 3.1 kb 사이에 유선특이적 발현을 유도하는 조절부위가 있을 것으로 추정된다.

• Tuberculosis and Respiratory Diseases
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• 제71권3호
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• pp.172-179
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• 2011

Background: Statins can regulate the production of pro-inflammatory cytokines and inhibit MMP production or activation in a variety of types of cells. This study evaluated whether statins would inhibit MMP release from human lung fibroblasts, which play a major role in remodeling processes. Methods: This study, using an in-vitro model (three-dimensional collagen gel contraction system), evaluated the effect of cytokines (tumor necrosis factor-${\alpha}$, TNF-a and interleukin-$1{\beta}$, IL-1b) on the MMP release and MMP activation from human lung fibroblasts. Collagen degradation induced by cytokines and neutrophil elastase (NE) was evaluated by quantifying hydroxyproline. Results: In three-dimensional collagen gel cultures (3D cultures) where cytokines (TNF-a and IL-1b) can induce the production of MMPs by fibroblasts, it was found that simvastatin inhibited MMP release. In 3D cultures, cytokines together with NE induced collagen degradation and can lead to activation of the MMP, which was inhibited by simvastatin. Conclusion: Simvastatin may play a role in regulating human lung fibroblast functions in repair and remodeling processes by inhibiting MMP release and the conversion from the latent to the active form of MMP.

• 대한화장품학회지
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• 제47권2호
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• pp.147-153
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• 2021

콜라겐 가수 분해물(collagen hydrolysate, CH)은 피부 진피 섬유아세포를 자극하여 콜라겐과 엘라스틴 같은 세포 외 기질 합성을 촉진함으로써 피부 노화 방지에 도움을 주는 것으로 알려져 있다. 최근 피부 노화를 일으키는 다양한 요인 중 최종당화산물(advanced glycation end products, AGEs)이 주목을 받고 있음에도 불구하고, 아직까지 CH가 AGE축적에 미치는 영향은 연구된 바 없다. CH는 피부 구조단백질의 생성을 촉진하여 AGE 축적에 영향을 줄 수 있으므로, 이를 확인하기 위해 트리펩타이드25%, Gly-Pro-Hyp 4%를 함유한 CH인 저분자콜라겐펩타이드(low molecule weight collagen peptide, LMCP)를 이용하여 임상시험을 수행하였다. 피부 조직 내 AGE 축적량을 평가하기 위해 AGE reader를 사용하여 피부자가형광(skin autofluorescence, SAF) 값을 측정하였다. 0.5%와 1.0% LMCP 용액을 8 주 동안 피험자의 전박에 도포한 결과, 시험부위의 SAF값이 대조부위에 비해 유의하게 감소하였다. 추가적으로, LMCP에 의한 피부섬유아세포의 콜라겐 합성 촉진을 평가하기 위해 CCD-986sk를 이용하여 in vitro test를 수행하였다. 그 결과, 800 ㎍/mL의 LMCP는 CCD-986sk의 human pro-collagen Iα1(COL1A1) 합성을 증가시켰다. 본 연구를 통해 LMCP도포가 콜라겐 합성을 촉진하여 항당화 효과에 도움을 줄 수 있다는 것을 확인하였으며, 이는 LMCP가 노화 방지 화장품 원료로써 잠재력이 있음을 시사한다.

• Preventive Nutrition and Food Science
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• 제16권4호
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• pp.291-298
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• 2011

Yam extracts (Dioscorea batatas) have been reported to possess a variety of functions. However, studies on its osteogenic properties are limited. In this study, we investigated the effect of ethanol and water extracts on osteoblast proliferation and bone matrix protein synthesis, type I collagen and alkaline phosphatase (ALP), using osteoblastic MC3T3-E1 cell model. MC3T3-E1 cells were cultured with yam ethanol and water extracts (0~30 mg/L) within 39 days of osteoblast differentiation period. Cell proliferation was measured by MTT assay. Bone matrix proteins were assessed by the accumulation of type I collagen and ALP activity by staining the cell layers for matrix staining. Also, the secreted (media) matrix protein concentration (type I collagen) and enzyme activity (ALP) were measured colorimetrically. Yam ethanol and water extracts stimulated cell proliferation within the range of 15~30 mg/L at 15 day treatment. The accumulation of type I collagen in the extracellular matrix, as well as secreted collagen in the media, increased with increasing doses of yam ethanol (3~15 mg/L) and water (3~30 mg/L) extracts. ALP activity was not affected by yam ethanol extracts. Our results demonstrated that yam extracts stimulated osteoblast proliferation and enhanced the accumulation of the collagenous bone matrix protein type I collagen in the extracellular matrix. These results suggest that yam extracts may be a potential activator for bone formation by increasing osteoblast proliferation and increasing bone matrix protein type I collagen. Before confirming the osteogenic action of yam, further studies for clarifying how and whereby yam extracts can stimulate this ostegenesis action are required.

• 대한의용생체공학회:의공학회지
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• 제15권3호
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• pp.341-346
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• 1994

To develop an artificial bone substitute that is gradually degraded and replaced by the regenerated natural bone, the authors designed a composite that is consisted of calcium phosphate and collagen. To use as the structural matrix of the composite, collagen was purified from human umbilical cord. The obtained collagen was treated by pepsin to remove telopeptides, and finally, the immune-free atel- ocollagen was produced. The cross linked atelocollagen was highly resistant to the collagenase induced collagenolysis. The cross linked collagen demonstrated an improved tensile strength.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book II
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• pp.239-240
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• 2003

Type I (collagen) and procollagen are reduced in aged human skin. This reduction could result from increased degration by metalloproteinases and from reduced procollagen synthesis and skin collagenase is required for initiation of the degration of type I collagen. In the present study, we study on assay the collagen and collagenase in natural plants using the fibroblast human skin cell. We select the 15 kind of plants used to herbal and 4 kind of fraction(by methylene chloride, ethyl acetate, n-butanol, water). Among these extract, the ethyl acetate fraction from benincasa hispida/prunus persica, trichosanthes kiriowii, trogopterus xanthipes and methylene chloride fractions from benincasa hispida/prunus persica, torilis japonica and n-butanol fraction from cnidium officinale, chrysanthemum sibiricum were selected for further experiments as they exhibited distinctive amount of collagen compared to other natural extracts. These extracts were again subjected to collagenase assay test. Benincasa hispida/prunus persica extract was shown to have exellent collagen synthesis activity from result of the collagen assay test and the other extract was shown to have over 130% of collagen synthesis activity. But, in the study of collagenase assay test just only trogopterus xanthipes extract was shown to have collagenase inhibition.

• 한국간담췌외과학회지
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• 제27권4호
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• pp.342-349
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• 2023

Backgrounds/Aims: Liver organoids have emerged as a powerful tool for studying liver biology and disease and for developing new therapies and regenerative medicine approaches. For organoid culture, Matrigel, a type of extracellular matrix, is the most commonly used material. However, Matrigel cannot be used for clinical applications due to the presence of unknown proteins that can cause immune rejection, batch-to-batch variability, and angiogenesis. Methods: To obtain human primary hepatocytes (hPHs), we performed 2 steps collagenase liver perfusion protocol. We treated three small molecules cocktails (A83-01, CHIR99021, and HGF) for reprogramming the hPHs into human chemically derived hepatic progenitors (hCdHs) and used hCdHs to generate liver organoids. Results: In this study, we report the generation of liver organoids in a collagen scaffold using hCdHs. In comparison with adult liver (or primary hepatocyte)-derived organoids with collagen scaffold (hALO_C), hCdH-derived organoids in a collagen scaffold (hCdHO_C) showed a 10-fold increase in organoid generation efficiency with higher expression of liver- or liver progenitor-specific markers. Moreover, we demonstrated that hCdHO_C could differentiate into hepatic organoids (hCdHO_C_DM), indicating the potential of these organoids as a platform for drug screening. Conclusions: Overall, our study highlights the potential of hCdHO_C as a tool for liver research and presents a new approach for generating liver organoids using hCdHs with a collagen scaffold.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book II
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• pp.519-523
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• 2003

Alpha hydroxy acid (AHA) includes a group of organic acids found in natural foods such as sugarcane (glycolic acid), milk (lactic acid), apples (malic acid) and oranges (citric acid). Earlier studies demonstrated the effect of AHAs on the skin by diminishing the adhesiveness of the corneal layer and increasing the viable epidermal thickness. Recent data suggest that AHAs have some effects on the dermal component of skin and even affect the aging process of the skin. A previous study revealed increased collagen production by treatment with glycolic acid among AHAs in vitro. However, the mechanism of the regulation of collagen production by glycolic acid was unclear. In present study, we tried to demonstrate the effect of glycolic acid on human dermal fibroblasts and to unveil the mechanism of regulation of collagen production by glycolic acid in human dermal fibroblasts: proliferation of fibroblasts and collagen synthesis and degradation by collagenases in fibroblasts. Our results suggested that glycolic acid had no effect on proliferation and cytotoxicity of adult human dermal fibroblasts. However, glycolic acid not only induced the increase of the collagen synthesis in human dermal fibroblasts at lower concentration than 0.1 % but also inhibited MMP-2 activity of human dermal fibroblast in the range between 0.01 and 0.4% and MMP-9 activity of human dermal fibroblast in the range between 0.06 and 0.09%. In summary, our results suggest that glycolic acid may increase wrinkle reduction partially by both increase in collagen synthesis and decrease in collagen degradation.