• 생명과학회지
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• 제21권11호
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• pp.1501-1510
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• 2011

최근에 해양자원에 있는 동물, 해조류 곰팡이 세균에서 신규 잠재적인 후보약효제가 조사되어 왔다. 본 연구에서는 치료제를 탐색하기 위하여 암전이, 관절염, 만성염증 및 주름형성에 주요한 역할을 하는 기질금속단백질분해효소(s) (MMPs)를 목적효소로 이용하였다. 5종의 녹조류, 18종의 홍조류, 4종의 갈조류를 포함한 다양한 해조류가 사람피부섬유아세포 및 섬유아육종세포로부터 유래된 기질금속단백질효소에 미치는 영향을 gelatin zymography를 이용하여 조사하였다. 사람피부섬유아세포에서는 홍조류중에서 Laurencia okamurae, Polysiphonia japonica, Grateloupia lanceolate 및 Sinkoraena lancifolia에서 MMP-2 억제효과가 관찰되었다. 반면에 녹조류의 Enteromorpha compressa와 Enteromorpha linza, 갈조류의 Peltaronia bighamiae and Sargassum thunbergii에서는 MMP-2 활성화가 관찰되었다. 사람섬유아육종세포에서는 MMP-9 활성화가 갈조류인 Sargassum thunbergii, 홍조류의 Polysiphonia japonica, 녹조류의 Enteromorpha compressa와 Enteromorpha linza의 존재 하에서는 감소되었다. 본 연구에서 흥미로운 발견은 녹조류의 E. compressa와 E. linza 및 갈조류의 S. thunbergii는 정상세포에서는 MMP-2에 대하여 활성화 효과를 나타내었으나, 암세포에서는 MMP-9응 억제하는 효과를 나타낸 것이다. 이러한 결과는 녹조류의 E. compressa와 E. linza 및 갈조류의 S. thunbergii는 항암 효능을 발휘할 수 있는 성분을 함유하고 있다는 것을 암시하고 있다.

• Toxicological Research
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• 제22권4호
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• pp.349-356
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• 2006

Diallyl disulfide (DADS), an oil-soluble organosulfur compound in garlic has been reported to suppress tumor growth and to induce apoptosis in cancer. In the present study, we investigated the effects of DADS on pulmonary metastasis of B16-F10 murine melanoma cells. DADS (i.p. 40 mg/kg) significantly (p<0.05) reduced the number of pulmonary metastatic nodules (48%) in experimental pulmonary metastasis assay. We also found that DADS inhibited adhesion, invasion and migration of B16-F10 melanoma cells in a dose-dependent manner. To study the antimetastatic potential of DADS, we performed the effects of DADS on matrix metalloproteinase activity. DADS significantly inhibited the expression of matrix metalloproteinase-2 activity in B16-F10 cells by gelatin zymography. These results suggest that DADS prevent metastasis in part through suppression of migration of B16-F10 melanoma cells by Inhibiting matrix metalloproteirase-2 responsible for degradation of extracellar matrix.

• 한국발생생물학회지:발생과생식
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• 제15권2호
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• pp.99-111
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• 2011

본 실험은 transforming growth factor-${\beta}1$(TGF-${\beta}1$)이 첨가된 chondrogenic induction medium(CIM)을 이용하여 인간지방조직에서 유래된 중간엽 줄기세포(human adipose tissue-derived mesenchymal stem cells, ATMSCs)의 연골형성능과 gelatin-chondroitin-glucosamine scaffold(GCG-scaffold)에 접종시킨 ATMSCs의 연골형성능을 알아보고자 수행하였다. ATMSCs와 생쥐 chondrocyte를 기본배양액과 TGF-${\beta}1$이 첨가되지 않은 CIM1 및 TGF-${\beta}1$이 첨가된 CIM2에서 배양하여 연골형성능을 비교하였다. ATMSCs의 연골형성은 gelatin scaffold(G-scaffold)와 GCG-scaffold를 사용하여 glycosaminoglycan(GAG) 합성과 조직화학적 염색으로 연골기질형성 여부를 확인하였다. 펠렛 배양에서 ATMSCs와 chondrocyte의 GAG합성은 대조군에서 14일의 배양기간 동안 약간 증가하였으나, CIM1과 CIM2배양군은 배양 14일에 대조군에 비해크게 증가하였으며, CIM2 배양군에서 가장 높았다. 그러나 연골기질은 배양 14일에 CIM2 배양군에서만 Safranin O와trichrome에 의해 염색되었다. Well plate에서 배양된 ATMSCs의 증식은 모든 배양군에서 배양 10일까지 계속적인 세포증식이 일어났으며, 대조군에 비해 CIM 배양군이 높았다. ATMSCs 접종 후 플라스크나 scaffold의 세포부착률은 배양시간이 길어짐에 따라 모든 배양군에서 증가하였고, 플라스크에 비해 scaffold에서 더욱 높게 나타났다. Scaffold에 ATMSCs 접종후 GAG 합성은 28일의 배양기간 동안 대조군에서는 별 변화가 없었으나, CIM1, CIM2 배양군은 대조군보다 GAG합성이 증가되었다. CIM2 배양군에서 GAG 합성이 매우 높게 나타났다. 또한 G-scaffold보다 GCG-scaffold에서 GAG 합성이 약간 높게 나타났다. 조직화학적으로 관찰한 ATMSCs의 연골기질형성도 CIM2 배양군의 GCG-scaffold에서 가장높게 나타났다. 본 실험의 결과들을 종합해 볼 때, 펠렛 배양에서 ATMSCs는 생쥐 chondrocyte보다 낮은 연골형성능을 보였고, CIM2 배양군에서만 연골기질이 형성된 것으로 보아 TGF-${\beta}1$이 연골분화에 중요한 요소로 작용한 것이라 사료된다. G-scaffold는 효과적인 연골분화 환경을 제공해 줌으로써 ATMSCs의 세포부착률과 GAG 합성을 증가시키는 것으로보여지며, G-scaffold보다 GCG-scaffold에서 많은 연골기질이 형성된 것은 GCG-scaffold에 첨가된 chondroitin과 glucosamine이연골기질 형성을 촉진시키는 물질로 작용한 것으로 사료된다.

• 동의생리병리학회지
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• 제22권6호
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• pp.1470-1474
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• 2008

This study was performed to examine the anti-metastatic effect of ethanol extract of Samguikoeui-Tang (SGKE), a formula consisting of four oriental herbs, in highly-metastatic HT1080 human fibrosarcoma cells. SGKE significantly inhibited the adhesion of HT1080 cells to matrigel at nontoxic concentrations in a dose-dependent manner, while it did not exert cytotoxicity against HT1080 cells up to the concentration of 100 ${\mu}g$/ml. Also, SGKE depressed the activity of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) by gelatin zymography. However, SGKE did not affect the mRNA expression of MMP-2 and TIMP-2, an inhibitor of MMP-2, by RT-PCR analysis. In addition, the effect of SGKE on HT1080 cell invasion was determined using Boyden chamber assay. SGKE suppressed the invasion of HT1080 cells in a dose-dependent manner. Taken together, these results suggest that SGKE has an anti-metastatic effect via inhibition of MMP-2 and -9 activities.

• 한국독성학회:학술대회논문집
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• 한국독성학회 1997년도 국제 심포지움 및 춘계 학술대회
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• pp.57-60
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• 1997

Aloe의 추출 시료들 중 혈관생성을 촉진하는 물질이 G1M1D1M1 분획에 있음을 chorioallantoic-membrane(CAM) assay, in vitro angiogenesis assay, u-PA, PAI, u-PAR, 및 matrix metalloprotease(MMP)의 유전자 발현조사, gelatin zymogram assay, modified CAM assay를 하여 확인 하였다. 그런 다음 G1M1D1M1 분획을 다시 분리하여 단일 성분인 NY932와 3종의 분획들을(U>3,000, 3,000>U>1,000, U<1,000) 얻었다. 이들 각각에 대한 혈관 생성 촉진 작용을 CAM assay 방법으로 조사한 결과 순수단일 성분인NY932가 혈관 생성 촉진 활성을 가장 높게 나타났다. 즉 NY932의 농도를 100ng, 2, 10, 35, 60 $\mu$g으로 투여하였을 때, 각각 2/18($11\%$), 4/10($49\%$), 16/22($73\%$), 15/17($88\%$), 9/9($100\%$)의 혈관생성 촉진 효과를 보였다. 따라서 순수 단일 성분인 NY932의 혈관 생성 촉진 작용기전을 규명하기 위하여 혈관 생성에 관련된 효소들[matrix metalloprotease(MMP)s, urokinase-plasminogen activator(uPA) 등]과 그 저해제들(TIMPs, PAI)의 유전자 발현을 조사하였으며, wounding migration assay등을 수행하였다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제38권6호
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• pp.321-325
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• 2012

Collagen is widely used for regenerative therapy and pharmaceutical applications as one of the most useful scaffolds. Collagen is the most abundant protein in vertebrates and the natural substrate of various types of animal cells. Bone and dentin are mineralized tissues and almost similar in chemical components. They consist of collagen (18%), non-collagenous proteins (2%), hydroxyapatite (70%) and body fluid (10%) in weight volume. Pepsin-digested, type I collagen (atelocollagen) and heat-denatured collagen (gelatin) are basic collagenous materials for medical use. Demineralized dentin matrix (DDM) and demineralized bone matrix (DBM) belong to acid-insoluble group, and vital tooth-derived DDM is a unique dentin material including cementum and growth factors. In this review, collagen-based materials will be introduced and discussed for bone regenerative surgery.

• 한국막학회:학술대회논문집
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• 한국막학회 1995년도 춘계 총회 및 학술발표회
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• pp.50-51
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• 1995

The swelling behavior of hyddrogels has been interested in many applications of drug carriers. These gels show reversible swelling changes in response to pH, electric currcnt, and temperature. Among others, the temperature-responsive behavior of poly(N-isopropylacrylanxide) (p(NIPAAm)) was studied, because a lower critical solution temperature(LCST) is in the vicinity of 32$\circ$C, and remarkable temperature-response can be obtained. We propose a novel composite membrane, which is appropriate for transporting drug ingredients above the transition temperature. Our object was to design a high permeation system above the shrinking temperature of p(NIPAAm). The membrane was composed of a matrix polymer and thermosensitive p(NIPAAm) hydrogel. The flux pattern of 4-acctamidophen through membrane in response of temperature was opposite to that of p(NIPAAm) membrane.

• 동의생리병리학회지
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• 제20권6호
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• pp.1629-1635
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• 2006

The migration of vascular smooth muscle cells (VSMC) and the production of matrix metallopreteinases-9 (MMP-9) may play a key role in the development of atherosclerosis. In this study, we have more extensively investigated the inhibitory effect of UR on MMP-9 activity and TNF-${\alpha}$ induced human aortic smooth muscle cells (HASMC) migration. The result from gelatin zymography showed that UR inhibited MMP-9 activity in a dose-dependent manner (IC50 = 55 g/ml). In addition, UR strongly inhibited the migration of HASMC induced by TNF-treatment (IC50 = 125 g/ml), although it has very low cytotoxic effect on HASMC (IC50 > 500 g/ml). These results suggest that UR is a potential anti-atherosclerotic agent through inhibition of MMP-9 activity and VSMC migration.

• Asian Pacific Journal of Cancer Prevention
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• 제15권3호
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• pp.1105-1110
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• 2014

Objective: Metastases and invasion are the main reasons for oncotherapy failure. Momordica cochinchinensis (Mu Bie Zi in Chinese) had been used for a variety of purposes, and shown anti-cancer action. In this article, we focused on effects on regulation of breast cancer cell ZR-75-30 metastases and invasion by extracts of Momordica cochinchinensis seeds (ESMCs). Methods: Effect of ESMCs on ZR-75-30 human breast cancer cells proliferation were evaluated by MTT assay and on invasion and migration by wound-healing and matrigel invasion chamber assays. Expression and protease activity of two matrix metalloproteinases (MMPs), MMP-2 and MMP-9, were analyzed by Western blotting and gelatin zymography, respectively. Results: ESMC revealed strong growth inhibitory effects on ZR-75-30 cells, and effectively inhibited ZR-75-30 cell invasion in a dose-dependent manner. Western blot and gelatin zymography analysis showed that ESMC significantly inhibited the expression and secretion of MMP-2 and MMP-9 in ZR-75-30 cells. Conclusions: ESMC has the potential to suppress the migration and invasion of ZR-75-30 cancer cells, and it might prove to of interest in the development of novel inhibitors for breast cancer.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제20권4호
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• pp.341-354
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• 1998

GMI (Fibrel${(R)}$) is one of the dermal filling substances which have been successfully used for the treatment of depressed cutaneous scar and wrinkles. It's major components are; Gelatin powder, which provides a framework for the clot to form and remains stable under the scar, and ${\varepsilon}$-aminocaproic acid, which inhibits the production of fibrinolysin, and Plasma, which provides the necessary ingredients for collagen synthesis. GMI has advantages of low immunogenicity and increased longevity. It has been known to induce fibroblast activity and promote new collagen synthesis. We used 34 Sprague-Dawley rats which were bred under the same condition and duration. 18 of experimental animals were undergone cardiac puncture, and their blood were collected, centrifugated, and stored in freezer. Out of 16 animals, control group were injected with 2ml plasma into the subcutaneous tissue of Lt. scapular, while experimental group were implanted of 2 ml GMI into the Rt. same area. Experimental animals were sacrificed at the 3rd day, 5th day, 1st week and 2nd week respectively after implantation of GMI. To observe the histopathologic change of GMI and surrounding tissue reaction of GMI, we had examined with H&E staining, immunohistochemical staining with vimentin, ${\alpha}$-SMA, S-100 under LM and SEM. The obtained results were as follows ; 1. In LM study, the inflammatory cell infiltrations and granulation tissue formation were observed, and muscle tissues were well attached with adipose tissues in the control group. In the experimental group, inflammatory cell infiltrations had been observed by the 2nd week and irregular adipiose tissues and well differentiated mesenchymal tissues were examined. 2. In immunohistochemical study, the experimental group of ${\alpha}$-SMA study, there were a prominent positive response on endothelial development of granulation tissues and mesenchymal tissues compare with the control group. In vimentin study, positive response on mescenchymal fibroblast continued to 2nd week, but negative in the control group. In S-100 study, both groups were positively responded on irregular adipose tissues. 3. In SEM study, collagen fibers were embedded by the plasma by the 5th day in the control group, and in the 3rd day experiment GMI were resorved but communited with collagen fiber till the 1st week. Collagen fibers were infilt-rated into GMI at the 2nd week and the infilltrated GMI were conglomerated with the mature adipose cells and the collagen fibers. From the above results, GMI implantation in the subcutaneous tissue of Sprague-Dawley rat, the mild infiltration of inflammatory cells were showed till 2nd week and the granulation tissues were observed. GMI were nearly resorbed till 2nd week, but well attached with adipose tissue and collagen fibers. The endothelium and fibroblasts were actively proliferated. Adipose tissues and mesenchymal tissue cells were observed. As already expressed, GMI showed resorptive change in course of time without any early immune reaction, and seemed to induce fibroblast activity and promote new collagen synthesis.