Keloid 세포의 Collagen 합성에 대한 Phorbol ester, Okadaic acid 및 Steroid의 작용

김원태,김인산,박낭운,손건영,조준승 慶北大學校 醫科大學 1995 慶北醫大誌 Vol.36 No.1

목적 : 이 연구는 Keloid 병변부위와 그 인접부위 그리고 정상부위의 세포에서의 collagen 합성에 대한 phorbol myristate acetate (PMA), okadaic acid (OKA) 및 hydrocortisone (HC)의 영향을 조사·비교 한 것이다. 재료 및 방법 : Keloid 병변부위와 그 인접부위 및 정상피부조직의 작은 절편을 배양하여 섬유아세포가 자라나와 밀생상태로 된후 trypsin 처리로써 섬유아세포만을 분리·계대 배양하였으며, 이 세포배양액에 PMA(100ng/㎖), OKA(10ng/㎖) 또는 HC(1.5μM)틀 부여, 24시간 배양해서 Peterkofsky 등의 방법에 따라 총단백질 및 collagen 합성능을 측정하여 대조군의 그것과 비교하였다. 결과 : Keloid 병변부위 및 그 인접부위의 세포는 정상부위의 세포보다 collagen 합성능이 높았다. 그 중에도 인접부위에서 가장 높았다. 그리고 PMA, OKA 및 HC는 다같이 비슷하게 또 부위에 관계없이 총단백질에 대한 collagen 합성비율을 억제하였다. 이들 약제중 PMA는 주로 noncollagen protein(NCP)을 많이 증가시켰고, OKA는 collagen 합성을 감소시켰다. 결론 : Keloid 병변부위와 그 인접부위의 세포에서 collagen 합성능이 높았다. 그리고 인접부위에선 더욱 PMA, OKA 및 HC에 의해 keloid 병변부위와 정상부위와는 달리 섬유아세포에서의 collagen 합성이 억제되었다. Keloid is a disease characterized by excessive deposition of various connective tissue components, such as collagen. In this study, three types of cells originate from keloid, keloid-adjacent and normal site of keloid patients were used to evaluate collagen synthesis by phorbol myristate acetate (PMA), okadaic acid (OKA) and hydrocortisone (HC). Average collagen synthetic rate by fibroblasts derived form keloid and keloid-adjacent cells was higher than that of normal skin fibroblasts. Keloid-adjacent cell has the most active collagen synthetic rate. PMA, OKA and HC inhibit % collagen synthesis in all three types of cells. In terms of noncollagen protein and collagen synthesis, PMA mainly increased noncollagen protein synthesis on the contrary OKA usually inhibit collagen synthesis in three types of cells. PMA, OKA and HC also inhibit % collagen synthesis in keloid and normal cells derived from the same patient similarity. Also there was no difference % collagen synthesis between keloid cells and keloid-adjacent cells derived from the same patient by three above mensioned reagents. Taken together, collagen synthetic rate was higher in keloid cells, when compared to the normal fibroblasts. In addition, there was no difference between keloid and normal cells in inhibitory effect of PMA, OKA and HC on collagen synthesis.

배양액 내의 교원성 기질이 피부 섬유모세포의 DNA 및 단백질 합성에 미치는 영향

이종원,곽승일,심형곤,이종건,변준희 大韓成形外科學會 1997 Archives of Plastic Surgery Vol.24 No.2

Collagen is the major component of scar tissue. Considerable progress of fibroblast growth kinetics and of collagen synthesis has been achieved in the past decade. We have been interested in fibroblasts activities as they are expressed by cells cultured in collagen substrate. This study is to examine the Effects of collagen substrate and peptide growth factors in culture medium on DNA and protein synthesis of human dermal fibroblast. Collagen, interleukin-1(IL-1) and transforming growth factor-β(TGF-β) were added to fibroblast culture media according to the designed experiment model and DNA and protein synthesis were measured by [³H]-thymidine, [³H]-leucine, and [³H]-proline incorporation method the morphological features of fibroblasts were observed by light microscape. The results were as follows; 1) There were significant deceases of DNA and protein synthesis of cultured fibroblasts in the presence of collagen substrate compared with those in control groups ( p<0.01 ) 2) DNA and protein synthesis were decreased as dose dependent manner of collagen density in culture media. 3) Morphological features of fibroblasts became less stellate and flat, more spindle-like in the presence of collagen. 4) In responsiveness to IL-1, Collagen non-treated groups responded to IL-1 but collagen treated groups were unresponsive to IL-1 (P<0.05). 5) Cells in collagen non-treated groups responded to TGF-β as dose-related manner ( P<0.01 ). Collagen treated groups responded to TGF-β but did not show TGF-β dose-dependent relationship. In conclusion, Collagen substrate in the culture medium could lower the DNA and protein synthesis of fibroblasts. Cells in Collagen substrate were unresponsive or less responsive to peptide growth factors than those in non-collagen substrate.

콜라겐과 난백 및 식초의 복합식이가 혈액 성분 및 손톱의 아미노산에 미치는 영향

신은정 ( Eun Jung Shin ),강상모 ( Sang Mo Kang ) 한국미용학회 2012 한국미용학회지 Vol.18 No.4

This study was performed to examined the variation of contents of collagen, albumin, ferritin in blood and the change of amino acid content of nails of female adults, aged 40 to 60 who presumed to be hypochlorhydria and to be easily broken in their finger nails because aging began. This study was conducted for 12 weeks dietary experiment to 11 people in each group by categorizing into Complex Diet group of Collagen+Egg White+Vinegar, Single Diet group of Collagen+Egg White+Vinegar. The results showed that Complex Diet group of Collagen+Egg White+Vinegar(C+EW+V), increased 5.7 times more increased variation of collagen level in blood of Collagen(C) group, 1.2 times of Collagen+Vinegar(C+V) group, 8.1 times that of Egg White(EW) group, 3.0 times that of Collagen+ Egg White(C+EW) group, and 1.3 times that of Egg White+Vinegar(EW+V) group. As for a albumin level and ferritin levels in blood, Complex Diet group of Collagen+Egg White+Vinegar(C+EW+V) was the highest. The results showed that Complex Diet group of Collagen+Egg White+Vinegar (C+EW+V) increased 3.3 times increased variation of nail amino acid of Collagen(C) group, 1.7 times that of Collagen+Vinegar group(C+V), 3.8 times that of Egg White(EW) group, 2.7 times that of Collagen+Egg White group(C+EW), and 2.1 times that of Egg White+Vinegar group(EW+V), which found that increased amount of amino acid in Complex Diet group of Collagen+Egg White+Vinegar(C+EW+V) was the most.

방사선 조사가 백서 피부 Fibroblast의 Collagen 합성에 미치는 영향

이영호,유재덕,나동균 大韓成形外科學會 1992 Archives of Plastic Surgery Vol.19 No.4

Keloids are predominantly fibrous sKin tumors that take the form, variable pruritic or tender growth near the site of an injury. Histopathologically, they are rich in various connective tissue components such as collagen. The exact mechanism of keloid formation is at pressure, local injection of syntheic steroids and radiation are now clinically used in the therapy of these lesions. Although radiation is widely used in keloid treatment at present, its mechanism is unknown at the molecular level. In the present study, cultured fibroblasts of rat skin were treated with various radiation doses and the cell grouth and the amounts of collagen and collagen mRNA were measured. Fibroblasts were isolated and cultured from 2-day-old rat skin, and the subcultured cells were irradiated with 300rads at 24hour intervals(one time with 300 rads, 2 times for a total of 600 rads, and 3 times for a total of 900 rads)from 24 hours after the subculture. The thymidine incoration in the culture flasks treated with 300, 600, and 900 rads was 54.6, 29.0, and 23.9% of the control at 2 hours, and 59.8, 40.1, and 34.6% at 8 hours respectively, indicating that radiation severely ingibits the cell growth. The effects of radiation on the amount of collagen were examined by measuring the hydroxyproline, which is a specific amino acid in collagen. The amounts of hydroxyproline in the 300 and 600 rad radiation groups were decreased to 44.2 and 33.3% of that of the control group, and that of the 900 rad radiation group could not be detected. To discriminate whether-these decreases in collagen content by radiation were due only to inhibition of cell growth or whether the radiation had a selective inhibitory effect on collagen content by radiation were due only to inhibition of cell growth or whether the radiation had a selective inhibitory effect on collagen synthesis, we counted the radioactivities of the peptide released by collagenase with the quantitation of DNA. The newly synthesized total protein if the 300, 600 and 900 rad radiation groups was decreased to 66.8, 48.2 and 40.3% of that the control group, respectively, but the total protein per ㎍ DNA was increased to 137.2, 132.3 and 109.7% respectively. The amounts of newly synthesized collagen were also decreased to 66.2, 50.2 and 40.1% of that of the control, but the amount of collagen per ㎍ DNA was increased to 136.1, 138.5 and 40.1% of that of the control, but the amount of collagen per ㎍ DNA was increased to 136.1, 138.5 and 109.3%, respectively, which was a similar pattern to the observed changes of total protein. the radiation did not inhibit the transcription of collagen α1(I)mRAN, according to the result of RAN dot hybridization. These results indicate that radiation has no selective inhibitory effect on collagen synthesis and the decrease of collagen amounts by radiation is due only to its inhibition of cell growth.

Fibrillar assembly and stability of collagen coating on titanium for improved osteoblast responses

Kim, Hae-Won,Li, Long-Hao,Lee, Eun-Jung,Lee, Su-Hee,Kim, Hyoun-Ee Wiley Subscription Services, Inc., A Wiley Company 2005 Journal of biomedical materials research. Part A Vol.a75 No.3

<P>Collagen, as a major constituent of human connective tissues, has been regarded as one of the most important biomaterials. As a coating moiety on Ti hard-tissue implants, the collagen has recently attracted a great deal of attention. This article reports the effects of fibrillar assembly and crosslinking of collagen on its chemical stability and the subsequent osteoblastic responses. The fibrillar self-assembly of collagen was carried out by incubating acid-dissolved collagen in an ionic-buffered medium at 37°C. The degree of assembly was varied with the incubation time and monitored by the turbidity change. The differently assembled collagen was coated on the Ti and crosslinked with a carbodiimide derivative. The partially assembled collagen contained fibrils with varying diameters as well as nonfibrillar aggregates. On the other hand, the fully assembled collagen showed the complete formation of fibrils with uniform diameters of ∼100–200 nm with periodic stain patterns within the fibrils, which are typical of native collagen fibers. Through this fibrillar assembly, the collagen coating had significantly improved chemical stability in both the saline and collagenase media. The subsequent crosslinking step also improved the stability of the collagen coating, particularly in the unassembled collagen. The fibrillar assembly and the crosslinking of collagen significantly influenced the osteoblastic cell responses. Without the assembly, the collagen layer on Ti adversely affected the cell attachment and proliferation. However, those cellular responses were improved significantly when the collagen was assembled to fibrils and the assembly degree was increased. After crosslinking the collagen coating, these cellular responses were significantly enhanced in the case of the unassembled collagen but were not altered much in the assembled collagen. Based on these observations, it is suggested that the fibrillar assembly and the crosslinking of collagen require careful considerations in the collagen administration as a coating moiety. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res, 2005</P>

Collagen Scaff olds in Cartilage Tissue Engineering and Relevant Approaches for Future Development

Vincent Irawan,Tzu-Cheng Sung,Akon Higuchi,Toshiyuki Ikoma 한국조직공학과 재생의학회 2018 조직공학과 재생의학 Vol.15 No.6

BACKGROUND: Cartilage tissue engineering (CTE) aims to obtain a structure mimicking native cartilage tissue through the combination of relevant cells, three-dimensional scaffolds, and extraneous signals. Implantation of ‘matured’ constructs is thus expected to provide solution for treating large injury of articular cartilage. Type I collagen is widely used as scaffolds for CTE products undergoing clinical trial, owing to its ubiquitous biocompatibility and vast clinical approval. However, the long-term performance of pure type I collagen scaffolds would suffer from its limited chondrogenic capacity and inferior mechanical properties. This paper aims to provide insights necessary for advancing type I collagen scaffolds in the CTE applications. METHODS: Initially, the interactions of type I/II collagen with CTE-relevant cells [i.e., articular chondrocytes (ACs) and mesenchymal stem cells (MSCs)] are discussed. Next, the physical features and chemical composition of the scaffolds crucial to support chondrogenic activities of AC and MSC are highlighted. Attempts to optimize the collagen scaffolds by blending with natural/synthetic polymers are described. Hybrid strategy in which collagen and structural polymers are combined in non-blending manner is detailed. RESULTS: Type I collagen is sufficient to support cellular activities of ACs and MSCs; however it shows limited chondrogenic performance than type II collagen. Nonetheless, type I collagen is the clinically feasible option since type II collagen shows arthritogenic potency. Physical features of scaffolds such as internal structure, pore size, stiffness, etc. are shown to be crucial in influencing the differentiation fate and secreting extracellular matrixes from ACs and MSCs. Collagen can be blended with native or synthetic polymer to improve the mechanical and bioactivities of final composites. However, the versatility of blending strategy is limited due to denaturation of type I collagen at harsh processing condition. Hybrid strategy is successful in maximizing bioactivity of collagen scaffolds and mechanical robustness of structural polymer. CONCLUSION: Considering the previous improvements of physical and compositional properties of collagen scaffolds and recent manufacturing developments of structural polymer, it is concluded that hybrid strategy is a promising approach to advance further collagen-based scaffolds in CTE.

Protein Phosphatase의 억제와 Protein Kinase C의 활성이 Collagen 합성 및 유전자 발현에 미치는 영향

권오종,김인산,박낭운,조준승 慶北大學校 醫科大學 1995 慶北醫大誌 Vol.36 No.2

목적 : 이 연구는 섬유아세포에서 collagen유전자 발현을 조절하는 기전을 밝히는 노력의 일환으로 세포내 단백질의 인산화에 중요한 역할을 하는 protein phosphatase의 길항제이며 non-phorbol ester tumor promoter인 okadaic acid(OKA)와 protein kinase C의 활성제인 phorbol myristate acetate(PMA)가 정상섬유아세포를 비롯한 수종의 서로 다른 특성의 세포에서 collagen합성 및 유전자 발현에 어떤 영향을 미치는가를 조사하였다. 재료 및 방법 : 각종 섬유아세포 즉 사람 폐섬유아세포인 HEL 299 세포, 골아세포인 MC3T3-El세포, 생쥐 섬유아세포인 NIH3T3 세포, 사람피부섬유아세포인 HSF30세포, keloid 섬유아세포를 비롯하여 마우스 정상 섬유아세포인 Balb/c 3T3 세포를 virus로 형질전환시킨 K-balt세포, SV-T2 세포 및 M-MSV-BALB 세포 등을 적당한 조건하에 배양하여 일정한 상태로 만들고 배양액에 protein Phosphatase의 억제제인 OKA와 protein kinase C의 활성제인 PMA를 일정한 조건으로 첨가하고서 ^3H-proline을 넣어서 총 단백질과 collagen에 편입되는 방사능의 정도를 측정하여 이들의 collagen 합성에 미치는 영향을 산출하였다. 또 이들의 영향이 유전자 수준에서 작용하는지를 조사하기 위하여 Chomczgynski와 Sacchi법에 의한 총 RNA의 수출, Thu등과 Feinberg 및 Vogelstein법에 따라 northern blot hybridization을 실시하고, Labarca 및 Paigen의 방법을 이용하여 DNA양을 측정하였다. 결과 : OKA와 PMA는 세포마다 정도의 차이는 있지만 collagen합성을 농도에 비례하여 억제되었으며, 그리고 OKA의 효과는 PMA에 의해 증대되었다. OKA의 collagen합성억제 효과는 반응 12시간 뒤에 나타났으며 PMA의 효과는 반응 3시간째에 나타났다. 이것은 OKA와 PMA의 collagen합성에 대한 작용기전의 다름을 시사한다. 또한 OKA와 PMA은 정상 섬유아세포의 α_1,(Ⅰ)procollagen mRNA치를 현저히 감소시켰으며 이것은 OKA와 PMA의 collagen합성 억제작용이 유전자 발현의 억제에 의한 것임을 뜻한다. Collagen 합성이 정상 보다 현저히 증가된 keloid세포에서도 OKA와 PMA는 모두 상대적 collagen합성을 감소시켰다. 바이러스로 형질전환시킨 세포에서는 OKA와 PMA에 대한 반응이 이질적으로 나타나 형질 전환과정에서 생긴 세포내의 변화가 OMA와 PMA의 collagen합성조절 기전에 영향을 주었음을 시사한다. 결론 : 이상의 결과를 종합하면 OMA와 PMA는 둘다 결합조직세포의 α_1,(Ⅰ)procollagen 유전자발현을 억제함으로써 collagen합성을 감소시켰으나 그 작용양상은 달라서 서로 다른 기전으로 작용한다 할 수 있다. 또한 형질전환세포에서 OKA와 PMA의 collagen합성에 대한 작용이 이질적으로 나타나 이들의 collagen합성에 대한 작용이 세포의 형질전환고정에서 역시 서로 다른 영향을 받는다고 할 수 있다. In this study, effects of two well known chemicals to affect intracellular signal transduction pathways, okadaic acid (OKA), a protein phosphatase inhibitor, and phorbol myristate acetate (PMA), a protein kinase C activator, on collagen synthesis and mRNA levels in various cell lines were evaulated. Both of OKA and PMA inhibited collagen synthesis in a dose-dependent manner in several fibro blasts and osteoblast. In human fibroblasts, a suboptimal dose of PMA potentiated the inhibitory effect of OKA on collagen synthesis. Time couse study, however, showed that OKA inhibited collagen synthesis at 12 h and PMA at 3 h after treatment indicating that OKA and PMA may affect colla gen sythesis by different pathways. Northern blot anaysis showed that both of OKA and PMA decreased α_1[Ⅰ]procollagen mRNA levels in human fibroblasts and in this effect, they did not require a new protein synthesis. OKA and PMA also decreased collagen sythesis in several keloid fibroblasts of which collagen synthetic acitivities were elevated. In virally transformed fibroblasts, however, OKA and PMA showed quite different effects on collagen synthesis suggesting that intracellualr changes in the processes of viral transformation might effect the regulatory mechanisms of collagen synthesis and thus affect collagen synthesis in responses to OKA and PMA. Taken together, both of OKA and PMA inhibited collagen synthesis in several different cell lines but their actions were different mechanisms.

Skeletal myotube formation enhanced through fibrillated collagen nanofibers coated on a 3D-printed polycaprolactone surface

Chae, SooJung,Lee, JiUn,Kim, GeunHyung Elsevier 2019 Colloids and Surfaces B Vol.181 No.-

<P><B>Abstract</B></P> <P>This work focused on considering the cellular responses of the growth and differentiation of myoblasts, C2C12, on fibrillated collagen-coated poly(ε-caprolactone) (PCL) surfaces. Through a fibrillation processing window using NaCl and collagen weight fractions, collagen fibril coating density can be controlled. Three different collagen-fibril densities coated on PCL strut were used to investigate the effects of the collagen fibril on the myoblast activities. After physical and cellular analyses of the scaffolds, such as surface morphology, fibronectin absorption, wettability, and mechanical properties, the rate of cell growth and the proficiency of the myoblasts to develop skeletal myotubes were evaluated. Based on the results, although the coated collagen nanofibers were randomly distributed, the fibrillated collagen layer with the appropriate density on the PCL surface promoted a greater myotube formation than that of the control, which had no fibrillated collagen. In particular, relatively higher densities of collagen fibril showed significantly greater myotube formation than those of the control (not-fibrillated collagen-coated on the PCL surface) and lower density of collagen fibril.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A fibrillated collagen-coated polycaprolactone surface was prepared. </LI> <LI> The proper fibrillation condition of the coated collagen was selected through a processing window. </LI> <LI> Faster myotube formation on fibrous collagen surface was achieved. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

쥐의 신장에서 부분 요관폐색에 의한 Collagen의 변화

이경준,이충범,윤재영,조대행,황태곤 大韓泌尿器科學會 1999 Korean Journal of Urology Vol.40 No.4

어피 콜라겐을 이용한 전기방사 용액 및 나노섬유 제조방법

오상연,정구,김의화 한국섬유공학회 2020 한국섬유공학회지 Vol.57 No.3

Collagen is a natural, biocompatible, and biodegradable material with low immunogenicity, which is useful for tissue and organ formation and in multiple cellular functions. Moreover, Type I collagen is known to be a suitable material for medical applications. In this study, acid-soluble Type I collagen isolated from fish skin was used to make nanofibers through electrospinning using a high concentration of collagen solutions prepared by dissolving the collagen to 18 wt% in 20 wt% acetic acid at 20 °C. The collagen and collagen nanofiber were Type I collagen subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. The nonaqueous collagen nanofiber was heat-treated, which yielded a chemical crosslinking agent, such as polyacrylic acid, besides collagen.