• Title/Summary/Keyword: Collagenase

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Analgesic Effects of ChondroT in Collagenase-induced Osteoarthritis Rat Model (Collagenase로 유발된 동물모델에서 ChondroT의 진통효과에 관한 연구)

  • Won, Jeong-Yoon;Jeong, Ji-Won;Na, Chang-Su;Kim, Seon-Jong
    • Journal of Korean Medicine Rehabilitation
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    • v.26 no.3
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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.

Effects of Interleukin-1${\beta}$ and Tumor Necrosis $Factor-{\alpha}$ on the Release of Collagenase and Gelatinase from Osteoblasts

  • Eun, Jong-Gab;Baek, Dong-Heon;Kim, Se-Won
    • The Korean Journal of Physiology and Pharmacology
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    • v.6 no.5
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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.

Purification and Characterization of Collagenase Produced by Staphylococcus aureus JJ-11 Isolated from the Human Skin (피부에서 분리한 Staphylococcus aureus JJ-11이 생산하는 collagenase의 정제 및 특성)

  • Lee Jin-Kyoung;Kim Hae-Nam;Kang Ho-Young;Jun Hong-Ki
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.245-252
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    • 2006
  • A bacterial strain, identified as Staphylococcus aureus JJ-11, producing collagenase was isolated out of 40 persons having skin troubles. S. aureus JJ-11 produced collagenase optimally in the media containing 1.5%(w/v) gelatin, 1%(w/v) yeast extract, 0.4%(w/v) $K_2HPO_4$, 0.005%(w/v) $NiSO_4{\cdot}6H_2O$ at $37^{\circ}C$ for 18 hrs. The collagenase produced by Staphylococcus aureus JJ-11 was purified at 6.66-folds purity through application of chromatography with Amberlite IRA-900 and Sephacryl S-300 HR columns. The molecular weight of the partially purified enzyme was estimated to be 62 kDa by SDS-PAGE. The protein exhibited optimum enzymatic activity at pH 7.0, and showed a stable activity at pH 4-8. The optimum temperature for collagenase was at $37^{\circ}C$, and activity was maintained upto $40^{\circ}C$. The enzyme activity was slightly elevated in the presence of divalents such as, $Fe^{2+},\;Co^{2+}\;and\;Ba^{2+}$ However, the activity was inhibited in the presence of $Sr^{2+}\;or\;Hg^{2+}$. The inhibition of activity by O-phenanthroline and EDTA suggested that the enzyme may contain metal which is required for activity. The enzyme showed the highest activity when insoluble collagen (type I) was, used as a substrate.

THE EFFECTS OF COLLAGENASE AND ESTERASE ON THE MICROTENSILE BOND STRENGTH IN DENTIN BONDING (상아질 접착에서 collagenase와 esterase가 미세인장결합강도에 미치는 영향)

  • Jung, Young-Jung;Hyun, Hong-Keun;Kim, Young-Jae;Kim, Jung-Wook;Lee, Sang-Hoon;Kim, Chong-Chul;Hahn, Se-Hyun;Jang, Ki-Taeg
    • Journal of the korean academy of Pediatric Dentistry
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    • v.34 no.2
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    • pp.285-291
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    • 2007
  • The purpose of this study was to evaluate the effect of collagenase and esterase on the microtensile bond $strength({\mu}TBS)$ in dentin bonding. After resin composites were bonded to occlusal dentin. ${\mu}TBS$ specimens were formed and stored in PBS, collagenase, or esterase solution After 4-week storage, ${\mu}TBS$ was determined and, the results were as follows : 1. ${\mu}TBS$ values of Single Bond 2 were lower than those of Clearfil SE Bond for all storage medium (p<0.05). 2. In Single Bond 2 group, collagenase solution lowered bond strength more than PBS and esterase solution (p>0.05). 3. In Clearfil SE Bond group, esterase solution lowered bond strength more than PBS(p>0.05). Collagenase solution lowered bond strength more than esterase solution(p>0.05) and PBS(p<0.05).

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Inhibition of Collagenase by Naturally-Occurring Flavonoids

  • Sin, Bo-Young;Kim, Hyun-Pyo
    • Archives of Pharmacal Research
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    • v.28 no.10
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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.

Metallo-collagenase production by Arthrobacter creatinolyticus KP015744

  • Savita A. Kate;Madhuri Sahasrabudhe;Archana Pethe
    • Advances in environmental research
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    • v.11 no.1
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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.

The study on collagen sysnthesis and collagenase inhibition assay in natural plants

  • W. J. Yang;S. J. Yang;Kim, W. H.;T. B. Kang;Park, S. S.
    • Proceedings of the SCSK Conference
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    • 2003.09b
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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.

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Effects of mechanical stress and interleukin-$1{\beta}$ on collagenase and TIMP-1 expression in human periodontal ligament fibroblasts (기계적 자극과 interleukin-$1{\beta}$가 치주인대 섬유아세포의 collagenase와 TIMP-1의 발현에 미치는 영향)

  • Kim, Myung-Lip;Bae, Chang
    • The korean journal of orthodontics
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    • v.28 no.1 s.66
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    • pp.165-174
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    • 1998
  • The turnover of collagen is controlled by the balance between collagen synthesis and degradation. The production of collagenase (matrix metalloproteinase-1) and its inhibitor, tissue inhibitor of matrix metallopmteinase-1 (TIMP-1) are one of the substances which regulate this balance. The periodontal ligament fibroblast plays an important role in collagen metabolism during orthodontic treatment and is believed to be an origin of the osteoblast in the alveolar bone. The collagenase secreted by the periodontal ligament fibroblast and the osteoblast initiates the bone resorption by removing the osteoid layer in the alveloar bone. The interleukin-$1{\beta}$ is secreted by the macrophage during orthodontic treatment. The present study was undertaken to assess the effect of mechanical stress and interleukin-$1{\beta}$ on the expression of collagenase and TIMP-1 in the periodontal ligament fibroblasts using reverse transcription polymerase chain reaction and immunohistochemical staining. The periodontal ligament fibroblasts were stitched by placing the $Petriperm dish^{\circledR}$ dish on the top of spheroidal convex watch glass ($5\%$ surface increase) and tented with interleukin-$1{\beta}$ (1.0 ng/ml), or treated with both of them. Treatment with mechanical stress and/or interleukin-$1{\beta}$ resulted in increased collagenase mRNA expression. The mechanical stress treated group (1.61, 1.62, 1.37 fold increase), the interleukin-$1{\beta}$, tented group (1.68, 1.60, 3.78 fold increase), the mechanical stress and interleukin-$1{\beta}$ treated group (1.89, 1.72, 5.48 fold increase) induced increases in collagenase mRNA compared with the control group after 2, 4, 8 hours respectively. But TIMP-1 mRNA expressions at experimental groups were decreased after 2, 4 hours and increased after 8 hours. The mechanical stress treated group (0.16, 0.49 fold decrease and 3.77 fold increase), the interleukin-$1{\beta}$ treated group (0.15,0.44 fold decrease and 4.46 fold increase), the mechanical stress and interleukin-$1{\beta}$ tented group (0.15, 0.69 fold decrease and 4.81 fold increase) induced changes in TIMP-1 mRNA compared with the control group after 2, 4, 8 hours, respectively. Immunohistochemical stain showed that increased collagenase and TIMP-1 staining of the mechanical stress tented group, the interleukin-$1{\beta}$ treated group, and the mechanical stress and interleukin-$1{\beta}$ treated group compared with that of the control group after 8 hours. These findings suggest that mechanical stress and interleukin-$1{\beta}$ regulate expression of collagenase and TIMP-1.

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EFFECT OF ORTHODONTIC FORCE ON THE ENZYME ACTIVITIES IN ALVEOLAR BONE (교정력이 치조골의 효소활성에 미치는 영향)

  • Kim, Hyung-Soo;Nahm, Dong-Seok
    • The korean journal of orthodontics
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    • v.22 no.2 s.37
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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.

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Studies on Antioxidant, Anti-Inflammation, and Collagenase Inhibitory Effects of Extracts from Plants of The Salix genus (버드나무 속 식물 추출물의 항산화, 항염 및 콜라게나제 저해 활성 연구)

  • Jeong, Yong Un;Park, Young Jin
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.3
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    • pp.335-341
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    • 2018
  • This study was carried out to evaluate the possibility of willow plants (the genus Salix) as a cosmetic material. DPPH radical scavenging abilities of 70% ethanol extracts of S. gracilistyla, S. pseudolasiogyne, and S. koriyanagi were significantly increased compared to control. In addition, the treatment of three species of willow plant extracts significantly inhibited the production of nitric oxide (NO) in RAW 264.7 cells, indicating that they had anti-inflammatory activity, and all of them had collagenase inhibitory activity. Among them, the extracts of S. gracilistyla extracts exhibited the highest collagenase inhibitory activity. As a result of analyzing the collagenase inhibitory activity against the solvent fraction of S. gracilistyla extracts, water and butanol fractions showed the highest collagenase inhibitory activity. These results suggested that S. gracilistyla among the willow plants had high collagenase inhibitory activity, and thus it can be utilized for cosmetics as an effective functional cosmetic material in the future.