• Journal of Life Science
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• v.13 no.3
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• pp.308-313
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• 2003

The collagenase gene from Vibrio parahaemolyticus 04 was subcloned into an expression vector pET-29b. The recombinant collagenase was expressed in Escherichia coli BL2l(DE3) and partially purified by Hi-Trap affinity and Sephacryl S-100 size exclusion chromatographies. The recombinant enzyme was purified by 43.7-fold and the yield was 73%. SDS-PAGE revealed that the molecular weight of the enzyme was approximately 35 kDa. Substrate specificity study of the enzyme displayed that the enzyme showed the highest activity with the type I collagen and the synthetic peptide, Z-GPGGPA, indicating that the enzyme was indeed a collagenase. The enzyme showed broad pH optimum around pH 6-12 and was stable between pH 5.5 and 11.5. The optimum temperature for the type I collagen degradation was $35^{\circ}C$. The thermostability measurement of the enzyme indicated that the enzyme was stable up to $55^{\circ}C$, but the activity was diminished quickly above $60^{\circ}C.$

• Maxillofacial Plastic and Reconstructive Surgery
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• v.32 no.6
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• pp.497-503
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• 2010

Purpose: The purpose of this study is to induce artificial arthritis on rabbit TMJ by injecting collagenase. Materials and Methods: An experimental animal model of arthritis induced by surgical method or intraarticular injection of chemical agent like LDH, papain, ketorolac. Surgical method is complex and needs a long time in inducing arthritis. Intra-articular injection of chemical agent like LDH, papain, ketorolac is simple. But chemical agent like LDH, papain, ketololac needs multiple injections to induce arthritis and mechanism inducing arthritis was known. Collagenase destroys helical domain of type II collagen in extracellular matrix produced by chondrocyte and then induces arthritis. We injected collagenase (0.5, 1.0, 2.0 mg) into the temporomandibular joint of rabbit. In the control group saline was intra-articularly injected. The condylar cartilage, disk and synovia were histologically examined at 1, 2, 4, 6 weeks after the initiation of collagenase injections. Results: Four weeks after injection of 2.0 mg collagenase, we could see histologic change like arthritis. In other groups, we couldn't see arthritis-like change. Conclusion: In our study, we produce arthritis on temporomandibular joint of rabbit by using injection of collagenase in temporomandibular joint of rabbit. And this experimental osteoarthritis is a useful animal model.

• Journal of Yeungnam Medical Science
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• v.4 no.1
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• pp.17-23
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• 1987

New born rat heart cells were dissociated using trypsin and/or collegenase to elucidate the dissociation efficiency of these two enzymes. And the effect of dimethyl sulfoxide during and immediately after cell dissociation was also investigated to clarify the so-called protective activity of dimethyl sulfoxide on cell performance. The results can be summarized as follows. 1. Cold trypsin 18 hours pretreatment followed by warm collagenase treatment resulted best cell viability and cell yield. 2. Single, warm trypsin treatment gave the poorest result. 3. Dimethyl sulfoxide did not seem to play any protective role during or immediately after rat heart cell dissociation. It had very damaging effect on rat heart cells.

• Herbal Formula Science
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• v.17 no.1
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• pp.153-162
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• 2009

Panax ginseng C.A. Meyer, Korean herb medicine, has been widely used in China and Japan for fatigue and enhancement of resistance to many diseases. In this study, we investigated the synergistic effect of Korean red ginseng and Fagopyrum esculentum extracts (RGFE) on dermal bioactive properties. RGFE treatment significantly increased electron donating ability, nitrite scavenging activity and collagenase inhibitory activity compared to red ginseng-treated group. Superoxide dismutase (SOD) activity in RGFE-treated group was similar to that of red ginseng. However, tyrosinase activity as indicator of melanin synthesis was not affected by RGFE or red ginseng. The results indicate that RGFE has anti-oxidative property and inhibitory effect of collagenase, and it may serve as a effective ingredient for beauty food.

• The Korea Journal of Herbology
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• v.30 no.6
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• pp.1-6
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• 2015

Objectives : Draconis Resina (DR), the resin of Daemonorops draco Bl., is used to circulate the blood and to stop bleeding. It also has been used to generate flesh including ulceration. The present study investigated the effects of DR extract on collagen metabolism in human fibroblasts and tyrosinase activity in mushroom tyrosinase.Methods : The effect of DR extract on type I procollagen production (collagen type I synthesis) and collagenase (matrix metalloproteinase-1, henceforth referred as MMP-1) activity in human normal fibroblasts cell line. Hs68 cells after ultraviolet B (UVB, 312 nm) irradiation was measured using the enzyme - linked immunosorbent assay (ELISA). The tyrosinase activity was also measured to find out the whitening effects in mushroom tyrosinase by ELISA method.Results : There was no cytotoxicity at DR extract at concentrations of 10 μg/ml, 30 μg/ml, and 100 μg/ml. DR extract significantly inhibited the increase of collagenase activity, whereas it did not show on the reduction of type I procollagen in UVB damaged Hs68 cells. DR extract did not reduce the L - DOPA oxidation. However, it significantly reduced the tyrosinase activity by DR extract at concentraions of 0.1 mg/ml, 1 mg/ml and 10 mg/ml.Conclusions : In conclusion, DR showed the anti-wrinkle and whitening effects via the inhibition of collagenase production and the tyrosinase activity. These results suggest that DR may have potential as an anti-aging ingredient in cosmetic herb markets.

• Proceedings of the SCSK Conference
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• 2003.09a
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• pp.804-819
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• 2003

It has been known that adenine is a very important material in living cells. Because, adenine is a member of nucleotide base, so it takes part in DNA, RNA and ATP synthesis. There are many reports that adenine participated in ingredients, especially DNA, RNA, NADH and ATP, affect on the cell. As well adenosine, conjugated adenine to glycoside, was known to anti-wrinkle compound. But there is no report whether adenine shows a good effect on the skin, especially anti-wrinkle. So, in this study, we tested whether adenine affects cell proliferation, collagen synthesis, collagenase synthesis inhibition in human dermal fibroblasts. In addition, we performed clinical study with adenine cream. Cell proliferation effect was tested by MTT assay. Collagen and collagenase synthesis were measured by Immunoassay with ELISA kit. Clinical study was performed by IECK according to KFDA Functional Cosmetic method. The results of cell proliferation show that 10$^{-6}$ ~10$^{-8}$ ％ of adenine increases cell proliferation about 50 ％ compare with non-treated control. At 10$^{-7}$ ~10$^{-10}$ ％, adenine increases type I collagen synthesis about 50％, decreases type I collagenase about 22％ compare with non-treated control. The results of clinical study show that 0.05％ adenine treated group reduces wrinkle significantly compare with placebo treated group. Therefore adenine may be a new anti-wrinkle candidate, through increases cell proliferation and collagen synthesis dramatically. And it decreases collagenase synthesis. So adenine could be used as a new anti-wrinkle agent.

• The Korean Journal of Nuclear Medicine
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• v.30 no.3
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• pp.332-337
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• 1996

Purpose : It has been known that the activity of extracellular matrix degradative enzymes such as collagenase correlate well with the metastatic potential of various tumor cells in experimental study. This study was aimed at comparing the activities of type IV collagenase with bone scan findings in patients with breast cancer. Materials and Methods : We retrospectively correlated bone scan findings with the results of immunohistochemical staining for 92kDa, 72kDa type IV collagenase in 28, and 30 patients with metastatic breast cancer, respectively, as well as 23, and 27 patients with primary breast cancer, respectively. The immunohistochemical staining was performed with tissue specimens obtained from primary or metastatic breast tumor lesions. The amounts of the enzyme were graded from 0 to 4 and scored by multiplication with the percentage of tumor cells. The confidence of bone scan interpretation for metastasis was also scored from 1 to 5 with increasing probability. Results : There was a significant difference in enzyme scores between patients with and without metastasis. In patients with primary breast cancer group, the frequency of patients with enzyme score of less than 170 were 96%(26/27) and 100%(26/26) with 92kDa and 72kDa collagenase, respectively. In contrast, in patients with metastatic breast cancer group, the frequency of patients with enzyme score of more than 200 were 93%(28/30) and 87%(26/30) with 92kDa and 72kDa collagenase, respectively. All patients with each enzyme score of less than 170 show no active bony metastasis, however, there were variable bone scan findings in patients with each enzyme score of more than 200. Conclusion : Bone scan is useful to confirm, localize or follow up of bony metastasis in patients with each enzyme scores of more than 200. Acitve metastatic lesions were hardly seen on the bone scintigraphy in patients with collagenase scores of less than 170.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.3
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• pp.528-532
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• 2011

This study was designed to investigate the collagen metabolism and tyrosinase activity of Meretrix extracts (ME). The effect of ME on type I procollagen production and collagenase activity in human normal fibroblasts HS68 after UVB (312 nm) irradiation was measured by ELISA method. The tyrosinase activity after treatment of ME was measured as well. Type I procollagen production was recovered by ME in UVB damaged HS68 cells. The increased collagenase activity after UVB damage was significantly recovered by ME. The tyrosinase activity and L-DOPA oxidation were significantly reduced as well. However, the effects on tyrosinase activity and L-DOPA oxidation were not powerful enough to be used as whitening agents. ME showed the anti-wrinkle effects and some whitening effects in vitro. These results suggest that ME may be a useful drug as an anti-wrinkle treatments.

• Journal of Korean Medicine Rehabilitation
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• v.15 no.1
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• pp.89-98
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• 2005

Objectives : This study was to investigate the effects of Clematidis Radix on Proteoglycan(PG) degradation by measuring of body weight, Glycosaminoglycan(GAG), Interleukin-$1{\beta}$($IL-1{\beta}$) in synovial fluid, and PG content of articular cartilage of femur in collagenase-induced arthritis in rats. Methods : Arthritis was induced by injection of collagenase(0.1 ml) into knee joint of rats. Arthritic rats were divided into control(n=8) and treated(n=8) group. Control group was taken normal saline for 15 days and treated group was taken extracts of Clematidis Radix for same duration. Normal group(n=8) was injected with normal saline and was taken normal saline for 15 days. Body weight was measured at 0, 10, 15 days after injection. GAG, $IL-1{\beta}$ in synovial fluid were measured with ELISA kit at 15 days after injection. PG content of articular cartilage of femur, represented by safranine O staining, was measured at 15 days after injection. Histopathological study on the articular cartilage of knee joint was investigated at 15 days after injection. Results : Body weight, PG of treated group, taken Clematidis Radix, were significantly increased, and GAG was significantly decreased compared with control group. But $IL-1{\beta}$ was not significantly decreased. Conclusions : On the basis of these results, we concluded that Clematidis Radix has inhibiting effects on the proteoglycan degradation in collagenase-induced rat osteoarthritis model.

• Journal of Haehwa Medicine
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• v.21 no.2
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• pp.63-72
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• 2013

To develop a new anti-skin aging cosmetics or functional foods by using antioxidative activity and collagenase inhibitor, a potent collagenase or elastase inhibitor was screened from various extracts of medicinal plants and its optimal extraction condition was investigated. And antioxidative activity, antimicrobial activity and inhibitory of effect against collagenase activity were investigated. In the these results, we selected the Sanguisorba (Sanguisorba officinalis L.) that presents a potential biological activities. Sanguisorba which is very rich in triterpenoid saponin and tannins was recently reported its anti-oxidant activities and phytoestogenic activities in vivo test and many clinical studies. The experiments were carried out in vitro to determine anti-oxidant activities of Sanguisorba extracts on DPPH radical scavenging activity assay, Superoxidase scavenging activity assay, Elastase and collagenase activity assay. It show that the Sanguisorba extracts have the most significant anti-oxidant on free radical scavenging activity assay, and also inhibited significantly activities of elastase, collagenase. Further, Sanguisorba extracts are activated Type I collagen protein expression in CCD-986sk cells. These result suggest that the Sanguisorba extracts on DPPH radical scavenging activity assay, Superoxidase scavenging activity assay, elastase and collagenase activity assay, Type I collagen protein expression in CCD-986sk cells effected could be developed cosmetic ingredients for anti-aging.