• 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.

• 한국표면공학회:학술대회논문집
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• 한국표면공학회 2013년도 춘계학술대회 논문집
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• pp.38-38
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• 2013

The mechanical cues that adherent cells derive from the extracellular matrix (ECM) can effect dramatic changes in cell migration, proliferation, and differentiation. Using a thin film of Type I collagen fibrils comprised of 100 nm to 200 nm collagen fibrils overlaying a bed of smaller fibrils, changes in cellular response to systematically controlled changes in mechanical properties of collagen was investigated. Further, an experimental and modeling approaches to calculate the elastic modulus of individual collagen fibrils, and thereby the effective stiffness of the entire collagen thin film matrix, from atomic force microscopy force spectroscopy data was performed. These results demonstrate an approach to analysis of fundamental properties of thin, heterogeneous, organic films, and add further insights into the mechanical properties of collagen fibrils that are of relevance to cell response to the ECM.

• 한국산학기술학회논문지
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• 제6권2호
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• pp.134-143
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• 2005

포유동물에서 중요한 구조단백질인 콜라겐은 다양한 조직내 단백질 조성과 사슬간 복잡한 가교결합의 존재로 인해 그 구조가 복잡하고 다양하여, 직접적으로 분석할 적당한 방법이 없었다. 본 연구에서는 간단한 전처리만으로 다수의 생체분자 시료에 대해 정확한 분자량 측정이 가능한 매트릭스 보조 레이저 탈착/이온화 비행시간 질량분석법(MALDI-TOF MS)을 이용하여 콜라겐과 조각화된 콜라겐을 분석하고 이의 아미노산 서열을 사중극자-비행시간 직렬 질량분석법(Q-TOF MS/MS)으로 확인하여, 시료 중의 콜라겐의 종류에 대한 정보를 확인하여, MALDI-TOF 질량분석을 이용한 콜라겐 분석에서 콜라겐의 종류를 쉽게 예측할 수 있는 방법을 제시하고자 하였다. 쥐의 꼬리에서 분리한 콜라겐을 SDS-PAGE로 분리한 결과 10개의 band를 얻을 수 있었는데, 같은 시료를 MALDI-TOF MS로 확인하여 각 band의 정확한 분자량을 결정할 수 있었다. SDS-PACE상의 10개의 분리된 band에 대해 각각 tryptic digestion후 MALDI-TOF 질량분석을 수행한 결과 4개의 band에서 type I 콜라겐의 $\alpha_1$-chain내의 fragment인 Gly1056-Arg1073을 확인할 수 있었고, 5개의 band에서 type I 콜라겐의 $\alpha_2$-chain내의 fragment인 Gly985-Arg1002을 확인하였다. 잔재한 콜라겐의 가교결합으로 인해 예상되는 fragment중 둘만이 확인되었지만, 확인된 fragment를 통해 적어도 7개의 band에서는 type I콜라겐이 존재함을 확인할 수 있었다. 확인된 두 콜라겐 flagment의 아미노산 서열을 Q-TOF MS/MS로 분석한 결과 MALDI-TOF MS에서의 예측과 일치함을 확인할 수 있었으며, 이를 통해 확인된 두 fragment에 의한 peak을 지문으로 하여 MALDI-TOF MS측정시에 시료내의 type I 콜라겐의 존재를 쉽게 확인할 수 있음을 볼 수 있었다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.429-432
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• 2000

생체 적합성, 생분해성, 항균성 등의 특징을 갖는 키토산 지지체는 type I -p collagen과 bFGF 또는 fibronectin을 함께 코팅함으로써 세포적합성을 향상시켜 섬유아세포의 증식과 ECM의 분비를 증가시킬 수 있으며, 인공피부를 위한 적합한 지지체로 사용될 수 있다고 사료된다.

• Archives of Plastic Surgery
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• 제32권3호
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• pp.335-342
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• 2005

This study was undertaken to investigate possibility of the allogenic type I collagen inducing osteoinduction or osteoconduction at critical sized bone defect in the rabbit. Twenty Newzealand white rabbit, weighted from 2.8 kg to 3.5 kg, were used in this study. The skull was exposed and two bony defects were created with diameter of 10 mm. Group I(n=10), the bony defects was grafted from the other side bone. Group II(n=10), the bony defects was grafted by the allogenic type I collagen with bone morphogenic protein(BMP). Group III(n=10), the bony defects was grafted by the allogenic type I collagen only. Group IV(n=10), the bony defects was lefted with no grafts. The grafted bones and allogenic type I collagen were investigated with radiologic densitometry, histologic analysis and immunohistochemistry after 12 weeks. No major difference was observed in the gross finding between Group I, II, III, but dura mater was exposed in bony defect,the Group IV. The radiologic study demonstrated more bony opacity in the Group I, but the other groups did not demonstrate a significant difference. In the histologic study, grafted bone edge was completely consolidated with original bone in group I and new bone ingrew into the grafted allogenic type I collagen(group II, III),but there is no bone regeneration from the original bony edge in the group IV. The percent of the new bone formation by cross-sectional area was considered statistically significant at a p value of less than 0.05(p<0.05). In the immunohistochemistry study about BMP antibodies, the group IV demonstrated osteogenic activity in front of advancing original bone edge, in which the osteoblast stained strongly for BMP antibodies, but other group does not demonstrated any osteoblastic expression. There was no immunologic rejection. In conclusion, this results do not demonstrate that the allogenic type I collagen is useful for bone substitute, but the characters of the collagen, such as pliability, easy-handling, sponge-like structure, are useful in interpositional bone graft substitutes. The further evaluation of long term results about the resorption, immunologic tissue reaction, response of applied tissue growth factor to the allogenic collagen is needed.

• 통합자연과학논문집
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• 제9권3호
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• pp.166-170
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• 2016

Collagen plays a vital role in the maintenance of structure and function of a human body. It has been widely applied in various fields including biomedical, cosmeceutical, food, pharmaceutical and tissue engineering. In the present study, the docking behaviour of type I collagen with 15 different ligands namely hydroxymethylfurfural, methylglyoxal, methylsyringate, O-methoxyacetophenone, 3-phenyllactic acid, 4-hydroxybenzoic acid, kojic acid, lumichrome, galangin, artoindonesianin F, caffeic acid, 4-coumaric acid, origanol A, thymoquinone and quercetin was evaluated along with their putative binding sites using Discovery Studio Version 3.1. Docking studies and binding free energy calculations revealed that origanol A has maximum interaction energy (-40.48 kcal/mol) and quercetin with the least interaction energy (-15.44 kcal/mol) as compared to the other investigated ligands. Three ligands which are galangin, methylsyringate and origanol A were shown to interact with Asp21 amino acid residue of chain B (type I collagen). Therefore, it is strongly suggested that the outcomes from the present study might provide new insight in understanding these 15 ligands as potential type I collagen modulators for the prevention of collagen associate disorders.

• Archives of Plastic Surgery
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• 제34권4호
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• pp.413-419
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• 2007

Purpose: In this study, porous type I collagen scaffolds were cross-linked using dehydrothermal(DHT) treatment and/or 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide(EDC), in the presence and absence of chondroitin-6-sulfate(CS) and cultured autologous chondrocytes(Chondro) for cartilage regeneration. Methods: Cartilage defects were created in the proximal part of the ear of New Zealand rabbits. Four prepared types of scaffolds(n=4) were inserted. The groups included Chondro-Collagen-DHT(Group 1), Chondro- Collagen-DHT-EDC(Group 2), Chondro-CS-Collagen- DHT(Group 3), and Chondro-CS-Collagen-DHT-EDC (Group 4). Histomorphometric analysis and cartilage-specific gene expression of the reconstructed tissues were evaluated 4, 8, and 12 weeks after implantation. Results: EDC cross-linked groups 2 and 4 regenerated more cartilage than other groups. However, calcification was observed in the 4th week after implantation. CS did not increase chondrogenesis in all groups. Cartilage-specific type II collagen mRNA expression increased in the course of time in all groups.Conclusion: EDC cross-linking methods maintain the scaffold and promote extracellular matrix production of chondrocytes.

• Journal of Ginseng Research
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• 제36권1호
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• pp.61-67
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• 2012

In the present study, effects of sun ginseng (SG) on the collagen synthesis and the proliferation of dermal fibroblast were investigated. Collagen synthesis was measured by assaying procollagen type I C-peptide production. In addition, the level of matrix metalloproteinase (MMP)-1 was assessed by western blot analysis. SG suppressed the MMP-1 protein level in a dose-dependent manner. In contrast, SG dose-dependently increased tissue inhibitors of MMP (TIMP)-1 production in fibroblasts. SG increased type I collagen production directly and/or indirectly by reducing MMP-1 and stimulating TIMP-1 production in human dermal fibroblasts. SG dose-dependently induced fibroblast proliferation and this, in turn, can trigger more collagen production. These results suggest that SG may be a potential pharmacological agent with anti-aging properties in cultured human skin fibroblast.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제34권1호
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• pp.36-48
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• 2008

To repair bone defects in the oral and maxillofacial field, bone grafts including autografts, allografts, and artificial bone are used in clinical dentistry despite several disadvantages. The purpose of this study was to evaluate new bone formation and healing in rat calvarial bone defects using hydroxyapatite (HA, $Ca_{10}[PO_4]_6[OH]_2,\;Bongros^{(R)}$, Bio@ Co., KOREA) and tricalcium phosphate (${\beta}-TCP,\;Ca_3[PO_4]_2$, Sigma-Aldrich Co., USA) mixed at various ratios. Additionally, this study evaluated the effects of type I collagen (Rat tail, BD Biosciences Co., Sweden) as a basement membrane organic matrix. A total of twenty, 8-week-old, male Sprague-Dawley rats, weighing 250-300g, were divided equally into a control group (n=2) and nine experimental groups (n=2, each). Bilateral, standardized transosseous circular calvarial defects, 5.0 mm in diameter, were created. In each experimental group, the defect was filled with HA and TCP at a ratio of 100:0, 80:20, 70:30, 60:40, 50:50, 40:60, 30:70, 20:80, and 0:100 with or without type I collagen. Rats were sacrificed 4 and 8 weeks post-operation for radiographic (standardized plain film, Kodak Co., USA), histomorphologic (H&E [Hematoxylin and Eosin], MT [Masson Trichrome]), immunohistochemical staining (for BMP-2, -4, VEGF, and vWF), and elementary analysis (Atomic absorption spectrophotometer, Perkin Elmer AAnalyst $100^{(R)}$). As the HA proportion increased, denser radiopacity was seen in most groups at 4 and 8 weeks. In general radiopacity in type I collagen groups was greater than the non-collagen groups, especially in the 100% HA group at 8 weeks. No new bone formation was seen in calvarial defects in any group at 4 weeks. Bridging bone formation from the defect margin was marked at 8 weeks in most type I collagen groups. Although immunohistochemical findings with BMP-2, -4, and VEGF were not significantly different, marked vWF immunoreactivity was present. vWF staining was especially strong in endothelial cells in newly formed bone margins in the 100:0, 80:20, and 70:30 ratio type I collagen groups at 8 weeks. The calcium compositions from the elementary analysis were not statistically significant. Many types of artificial bone have been used as bone graft materials, but most of them can only be applied as an inorganic material. This study confirmed improved bony regeneration by adding organic type I collagen to inorganic HA and TCP mixtures. Therefore, these new artificial bone graft materials, which are under strict storage and distribution systems, will be suggested to be available to clinical dentistry demands.

• Journal of Periodontal and Implant Science
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• 제30권2호
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• pp.389-405
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• 2000

The purpose of this study is to evaluate the effect of IGF-I for DNA synthetic activity and the mRNA expression of bone matrix protein, type I collagen and osteopontin in prolifetation and differentiation of MC3T3-E1 cells. To evaluate DNA synthetic activity, cells were seeded at $2{\times}10^4cells/ml$ in 24 well plates and to evaluate mRNA of type I collagen and osteopontin cells were seeded at $5{\times}10^5cells/ml$ in 100mm culture dishes. These cells were cultured in alpha-minimum essential medium(${\alpha}-MEM$) containing 10% fetal bovine serum at $37^{\circ}C$, 5% $CO_2$ incubator. For DNA synthetic activity test 1, 10, 100ng/ml IGF-I were added to the cells which had been cultured for 3 days before 24 hours. For type I collagen mRNA expression 1, 10ng/ml IGF-I were added to the cells which had been cultured for 5, 10 days and for osteopontin mRNA expression 0.1, 1, 10ng/ml IGF-I were added to the cells which had been cultured for 5, 15, 20 days. Cell proliferaton was measured by the incorporation of [$^3H$]-thymidine into DNA and expression for type I collagen and osteopontin were measured by northern blot analysis. The results were as follows : DNA synthetic activity were generally higher in experimental group than control group. Expressions of type I collagen mRNA were higher at 5 day group and much lower at 10 day group in the control groups. In the experimental groups, mRNA expressions were slightly increased when 1 ng/ml IGF-I were added to 5 day group and decreased in all experimental 10 day groups. Expressions of osteopontin mRNA were higher at 20 day groups and lower at 15 day groups than the control groups. In the experimental groups, mRNA expressions were incereased when 0.1, 1 ng/ml IGF-I were added to 5 day group and in all the 15 day groups, but decreased when 0.1, 1, 10 ng/ml IGF-I were added to 20 day groups. IGF-I stimulated DNA synthetic activity of MC3T3-E1 cells during proliferation stage significantly, did not greatly changed effects on type I collagen mRNA expression and stimulated osteopontin mRNA expression at 15 day especially. In conclusion, we suggests that IGF-I have a tendency of stimulation effect of DNA synthetic activity but do not stimulate type I collagen mRNA in proliferation stage of MC3T3-E1 cell cultures, and stimulate osteopontin mRNA in differentiation stage of MC3T3-E1 cell cultures.