• 한방재활의학과학회지
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• 제26권3호
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• pp.17-30
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• 2016

Objectives The purpose of this study was to find out the effects of ChondroT on arthralgia of the Collagenase-induced osteoarthritis in rats. Methods Osteoarthritis was induced into rat by injecting Collagenase in its knee joint. Rats are divided into a total of 8 groups (n=6). Normal group was not induced for osteoarthritis whereas control groups were induced for osteoarthritis by Collagenase. Positive-A (Indomethacin) was injected with Collagenase and after 8 days, 2 mg/kg of Indomethacin was medicated. Positive-B (JOINS TAB) was injected with Collagenase and after 8 days, 20 mg/kg of JOINS TAB was medicated. Experimental groups (Chondro T) at three dose levels (50, 100 and 200 mg/kg) were injected with Collagenase and after 8days they were medicated with 10 ml/kg. Indomethacin, JOINS TAB and ChondroT were medicated each substances once a day for 10 days. Thereafter, the changes in plantar withdrawal response of osteoarthritis rats by dynamic plantar aesthesiometer were observed and then RT-PCR analysis was done to investigate the expression of related proteins. Results 1. ChondroT significantly decreased withdrawal response of mechanical allodynia compared with control group in all of the experimental groups (ChondroT-A, ChondroT-B, ChondroT-C). 2. ChondroT significantly reduced Bax/Bcl-2 ratio in all of the experimental groups (ChondroT-A, ChondroT-B, ChondroT-C). 3. ChondroT significantly reduced the expression of INF-${\gamma}$ compared with control group in group ChondroT-B, ChondroT-C. Conclusions This results suggest that ChondroT may be meaningful for suppressing the pain of osteoarthritis. Further study is needed to conduct a rigorous clinical research.

• The Korean Journal of Physiology and Pharmacology
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• 제6권5호
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• pp.269-274
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• 2002

A large number of factors such as osteotropic hormones, cytokines, or growth factors are related to the bone remodeling which is characterized by the coupling of osteoclast-mediated bone resorption and osteoblast-mediated bone formation. Recent investigations have indicated that cytokines such as $interleukin-1{\beta}\;(IL-1{\beta})$ and tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ play a potential role in the bone resorption associated with a variety of pathological conditions such as inflammatory osteolytic disease. Collagen is the most abundant protein of the extracellular matrix of bone, and the participation of collagenase in bone resorption has been widely investigated. In this study, effects of $IL-1{\beta}$ and $TNF-{\alpha}$ on the release of collagenase from osteoblastic cells were measured. The gelatinase activity was also measured by gel substrate analysis (zymography) after electrophoresis of conditioned media of osteoblastic cell culture. $IL-1{\beta}$ increased the collagenase activity in ROS17/2.8 and HOS cell culture. $TNF-{\alpha}$ also increased the collagenase activity of osteoblastic cells. When two kinds of cytokines were treated simultaneously in the culture of osteoblastic cells, synergistic increase of collagenase activity was seen in ROS17/2.8 cells. $IL-1{\beta}$ and $TNF-{\alpha}$ significantly increased the collagenase activity after 6 hour treatment in the osteoblastic cell culture, and there was no additional increase according to the culture period. Osteoblastic cells released the gelatinase and molecular weight of this enzyme was measured about 70 KDa as assessed by zymogram. $IL-1{\beta}$ and $TNF-{\alpha}$ showed increase of the gelatinase activity produced by ROS17/2.8 and HOS cells. Taken together, this study suggested that $IL-1{\beta}$ and $TNF-{\alpha}$ can modulate bone metabolism, at least in part, by increased release of collagenase and gelatinase from osteoblasts.

• 생명과학회지
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• 제16권2호
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• pp.245-252
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• 2006

피부 트러블을 가진 남,여 40명의 피부에서 분리한 collagenase를 생산하는 균주를 분리, 동정한 결과 Staphylococcus aureus로 판명되었으며 이를 S. aureus JJ-11이라 명명하였다. S. aurells JJ-11 균주의 collagenase의 최적 생산 조건은 1.5%(w/v) gelatin, 1%(w/v) yeast extract, 0.4% (w/v) $K_2HPO_4$, 0.005%(w/v) $NiSO_4{\cdot}6H_2O$를 함유한 배지 (pH 7.0)에서 $37^{\circ}C$, 200 rpm으로 18시간 동안 배양하는 것이다. 분리 균주가 생산하는 collagenase를 정제하기 위해서 amberlite IRA-900과 sephacryl S-300 HR columns를 이용하였고, 6.66-folds로 정제되었다. S. allreus JJ-11 균주가 생산하는 collagenase를 정제한 결과 분자량은 약 62 kDa이었으며, pH 7.0과 $37^{\circ}C$에서 각각 최대의 활성을 가졌고, pH와 온도에 대한 안정성은 pH 4.0-8.0, $40^{\circ}C$까지 100%의 활성이 있었다. 금속이온에 대해서는 $Fe^{2+},\;Co^{2+},\;Ba^{2+}$ 존재 하에서는 5 mM 농도에서도 활성을 유지하였다. $Sr^{2+},\;Hg^{2+}$에서는 30% 이상이 저해를 받는 것으로 확인되었다. 또한 EDTA와 O-phenanthroline에 의해 65% 이상이 저해되는 일 반적인 collagenase의 특정인 metalloproteinase의 특정을 보였으며, 그리고, 여러 가지 기절에 대해 효소활성을 비교한 결과, insoluble collagen (type I)에 대해 효소 활성이 가장 높았다.

• 대한소아치과학회지
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• 제34권2호
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• pp.285-291
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• 2007

상아질-레진 접착강도에 대한 collagenase와 esterase의 영향을 살펴보기 위해, 소구치의 교합면 상아질에 Single Bond 2와 Clearfil SE Bond로 접착을 시행하고 미세 시편을 제작하여 PBS collagenase 용액, esterase 용액에 4주간 보관한 후 미세인장결합강도를 측정, 비교하여 다음과 같은 결론을 얻었다. 1. 모든 보관 용액에서 Single Bond 2의 미세인장결합강도는 Clearfil SE Bond보다 유의하게 낮았다(p<0.05). 2. Single Bond 2의 미세인장결합강도는 collagenase군이 PBS군, esterase군보다 낮았다(p>0.05). 3. Clearfil SE Bond의 미세인장결합강도는 esterase군이 PBS군에 비해 낮았으나(p>0.05), collagenase군보다는 높았다(p>0.05). Collagenase군은 PBS군에 비해 유의하게 낮은 미세인장결합강도를 보였다(p<0.05).

• Archives of Pharmacal Research
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• 제28권10호
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• pp.1152-1155
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• 2005

We examined the inhibitory activities of various flavonoids, including the flavanones, flavones/isoflavones and flavonols, on collagenase from Clostridium histolyticum to establish their therapeutic potential against skin inflammation and photoaging. In general, the flavonols were stronger inhibitors than the flavones/isoflavones, and this indicated the importance of the C-3 hydroxyl substitution. Quercetin was the most active flavonoid among those tested, and it showed an $IC_{50}$ of 286 ${\mu}M$. These novel results suggest that certain flavonoids, particularly the flavonols, may prevent collagen breakdown by inhibiting collagenase in inflamed skin as well as photoaged skin.

• Advances in environmental research
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• 제11권1호
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• pp.1-16
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• 2022

Amongst 27 isolates from deteriorated leather samples, Arthrobacter creatinolyticus KP015744 zzx28 was found to be an efficient collagenase producer. Collagenase production of 13.33 µmoles/min was shown at an optimum temperature at 37℃ after 72h and at pH 7.5 by using 2 ml/dL inoculum in 10 mg/ml collagen peptide type I as a substrate. In presence of Hg²⁺, EDTA and 𝛽-mercaptoethanol the collagenase production by the isolate was strongly inhibited however Fe²⁺, Ca²⁺and DMSO enhanced production of the enzyme. Specific activity was found to be 19.46×10³ U/mg and molecular weight 66 kD by SDS PAGE. Isolate also has potential to hydrolyze keratin which is another important protein found in leather. Experimental results propose that collagenase can be effectively used as a tool for collagen and keratin rich solid waste treatment.

• 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.

• 대한치과교정학회지
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• 제28권1호
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• pp.165-174
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• 1998

교정력이 치아에 가해지면 치주인대의 재생과 치조골의 개조가 일어난다. 치주인대 섬유아세포는 collagenase와 TIMP-1을 분비하여 치주조직의 교원질의 분해와 합성을 담당한다. 본 연구에서는 치주인대 섬유아세포예 기계적 자극과 interleukin-$1{\beta}$를 가해 collagenase와 TIMP-1의 발현을 RT-PCR과 면역조직화학 염색을 사용하여 알아보았다. 4명의 10대 남자 교정환자에게서 아무런 병소가 없는 제1소구치를 발치후 치주인대 섬유아세포를 배양하여 4-6세대의 세포를 사용하였다. 대조군, $Petriperm dish^{\circledR}$ 바닥의 표면적을 $5\%$ 증가시킨 기계적 자극을 가한 군, interleukin-$1{\beta}$를 1.0 ng/ml를 가한 군과 기계적 자극과 interleukin-$1{\beta}$를 같이 가한 군으로 나누어 4명의 환자에서 얻은 세포군을 각 군별로 2, 4, 8시간 후 RT-PCR을 시행하여 그 산물을 반정량하여 대조군에 대한 각 실험군의 상대적인 증감을 나타내었고, 24시간후 면역조직화학 염색을 시행하여 다음과 같은 결과를 얻었다. 1. 광학 현미경으로 세포의 형태를 관찰한 결과 대조군에서는 전형적인 별모양과 길쭉한 모양을 함께 보였으나 기계적 자극과 interleukin-$1{\beta}$를 각각 혹은 동시에 준 군들에서는 별모양의 세포가 사라지고 모양이 더욱 길어졌다. 2. collagenase는 대조군에 비해 기계적 자극과 interleukin-$1{\beta}$를 각각 혹은 동시에 준 군들에서 증가하였고, 실험 8시간 후에서는 interleukin-$1{\beta}$를 준 군, 기계적 자극과 interleukin-$1{\beta}$를 동시에 준 군에서 뚜렷한 증가를 보였다. 3. TIMP-1은 세포 자극 2, 4시간 후에는 대조군에 비해 기계적 자극과 interleukin-$1{\beta}$를 각각 혹은 동시에 준 군들에서 감소하였지만, 실험 8시간 후에서는 증가를 보였다. 4. 면역조직화학 염색을 통해 collagenase와 TIMP-1이 대조군에 비해 기계적 자극과 interleukin-$1{\beta}$를 각각 혹은 동시에 준 군들에서 더욱 강한 염색상을 나타내었다. 본 실험의 결과 섬유아세포는 외부 자극이 가해지면 collagenase와 TIMP-1의 발현 조절을 통해 치주인대 재생과 치조골의 개조에 영향을 미쳐 항상성을 유지하려고 함을 알 수 있었다.

• 대한치과교정학회지
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• 제22권2호
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• pp.297-308
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• 1992

The effect of orthodontic force on the collagenase and phosphatase activities of the adjacent alveolar bone was evaluated. Maxillary canines of male cats were treated orthodontically with closed coil spring so as to exert about 80g force. Sixteen cats were equally divided into one control group and seven experimental groups (12 hrs, 24 hrs, 36 hrs, 2 days, 3 days, 5 days and 7 days after orthodontic treatment). After sacrificing all animals on experimental intervals, collagenase, acid phosphatase (ACP) and alkaline phosphatase (ALP) activities were determined in the pressure and tension sides of alveolar bones. ACP activities increased in both the pressure and tension sides, but significantly increased in the pressure side continuously. ALP activities increased in the tension side at early stage (1-2 days after treatment), but changed small amount in the pressure side. Collagenase activities increased in the pressure side, especially at late stage (5-7 days after treatment). These results suggest that (1) orthodontic fore force increases the ACP, ALP and collagenase activities generally and (2) activities of ACP and collagenase increase in the pressure side, but that of ALP in the tension side and (3) activities of ACP and ALP increase at early stage, but that of collagenase at late stage after orthodontic treatment. Therefore it is shown that there are time differences in the formation and destruction of organic and inorganic components in the bone metabolism of alveolus with application of the orthodontic forces.

• 대한화장품학회지
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• 제44권3호
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• pp.335-341
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• 2018

본 연구는 버드나무 속 식물의 화장품 소재로써의 활용 가능성을 평가하기 위해 수행하였다. 버드나무 속 식물 중 갯버들, 능수버들 및 키버들 70% 에탄올 추출물의 DPPH radical 소거능 분석을 통한 항산화 활성을 평가한 결과 모든 추출물이 대조군에 비해 DPPH radical 소거능이 유의적으로 증가하였다. 또한 3종의 버드나무 속 식물 추출물 처리는 RAW 264.7 세포의 nitric oxide (NO) 생성을 유의적으로 억제하여 항염 활성이 있는 것으로 확인되었으며, 모두 콜라게나제 저해 활성이 있는 것으로 확인되었다. 그 중 갯버들 추출물이 가장 높은 콜라게나제 저해활성을 나타내어 갯버들 추출물의 용매 분획물에 대한 추가적인 콜라게나제 저해활성을 분석한 결과, 물 및 부탄올 분획물이 가장 높은 콜라게나제 저해활성이 있는 것으로 확인되었다. 결과적으로 버드나무 속 식물 중 갯버들은 높은 콜라게나제 저해활성이 있는 것으로 확인되어 향후 효과적인 기능성 화장품 소재로 활용 가능 할 것으로 사료된다.