• Title/Summary/Keyword: Matrix formation

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Secreotory Leukocyte Protease Inhibitor Regulates Bone Formation via RANKL, OPG, and Runx2 in Rat Periodontitis and MC3T3-E1 Preosteoblast

  • Seung-Yeon Lee;Soon-Jeong Jeong;Myoung-Hwa Lee;Se-Hyun Hwang;Do-Seon Lim;Moon-Jin Jeong
    • Journal of dental hygiene science
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    • v.23 no.4
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    • pp.282-295
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    • 2023
  • Background: Secretory leukocyte protease inhibitor (SLPI) protects tissues from proteases and promotes cell proliferation and healing. SLPI also reduces periodontal inflammation and alveolar bone resorption by inhibiting proinflammatory cytokine expression in rat periodontal tissues and osteoblasts. However, little is known of the role of SLPI in the expression of osteoclast regulatory factors from osteoblasts, which are crucial for the interaction between osteoblasts and osteoclasts. Therefore, we aimed to determine the effects of SLPI on the regulation of osteoclasts and osteoblasts in LPS-treated alveolar bone and osteoblasts. Methods: Periodontitis was induced in rats using LPS. After each LPS injection, SLPI was injected into the same area. Immunohistochemical analysis was performed with antibodies against SLPI, RANKL, OPG, and Runx2 in the periodontal tissue. RT-PCR and western blotting were performed to determine the expression levels of SLPI, RANKL, OPG, and Runx2 in LPS- and SLPI/LPS-treated MC3T3-E1 cells. SLPI/LPS-treated MC3T3-E1 cells were also stained with Alizarin Red S. Results: Immunohistochemical analysis showed that the expression levels of SLPI, OPG, and Runx2 were higher while that of RANKL was lower in the LPS/SLPI group relative to those in the LPS group. The mRNA and protein expression of SLPI, OPG, and Runx2 was higher in SLPI/LPS/MC3T3-E1 cells than in LPS/MC3T3-E1 cells, and RANKL expression was lower. During differentiation, OPG and Runx2 protein levels were higher whereas RANKL levels were lower in SLPI/LPS/MC3T3-E1 than in LPS/MC3T3-E1 cells on days 0, 4, 7, and 10. In addition, mineralization and matrix deposition were higher in SLPI/LPS/MC3T3-E1 than in LPS/MC3T3-E1 on days 7 and 10. SLPI decreased RANKL expression in LPS-treated alveolar bone and osteoblasts but increased the expression of OPG and Runx2. Conclusion: SLPI can be considered as a regulatory molecule that indirectly regulates osteoclast activation via osteoblasts and promotes osteoblast differentiation.

Research on Characterizing Urban Color Analysis based on Tourists-Shared Photos and Machine Learning - Focused on Dali City, China - (관광객 공유한 사진 및 머신 러닝을 활용한 도시 색채 특성 분석 연구 - 중국 대리시를 대상으로 -)

  • Yin, Xiaoyan;Jung, Taeyeol
    • Journal of the Korean Institute of Landscape Architecture
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    • v.52 no.2
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    • pp.39-50
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    • 2024
  • Color is an essential visual element that has a significant impact on the formation of a city's image and people's perceptions. Quantitative analysis of color in urban environments is a complex process that has been difficult to implement in the past. However, with recent rapid advances in Machine Learning, it has become possible to analyze city colors using photos shared by tourists. This study selected Dali City, a popular tourist destination in China, as a case study. Photos of Dali City shared by tourists were collected, and a method to measure large-scale city colors was explored by combining machine learning techniques. Specifically, the DeepLabv3+ model was first applied to perform a semantic segmentation of tourist sharing photos based on the ADE20k dataset, thereby separating artificial elements in the photos. Next, the K-means clustering algorithm was used to extract colors from the artificial elements in Dali City, and an adjacency matrix was constructed to analyze the correlations between the dominant colors. The research results indicate that the main color of the artificial elements in Dali City has the highest percentage of orange-grey. Furthermore, gray tones are often used in combination with other colors. The results indicated that local ethnic and Buddhist cultures influence the color characteristics of artificial elements in Dali City. This research provides a new method of color analysis, and the results not only help Dali City to shape an urban color image that meets the expectations of tourists but also provide reference materials for future urban color planning in Dali City.

Genetic Studies on the Sea Urchin Embryogenesis and Skeletogenesis (성게의 발생과 뼈대형성의 유전학적 연구)

  • Lee, Youn-Ho
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.6 no.4
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    • pp.265-273
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    • 2001
  • The sea urchin has been used as sea food in many countries. This species has also been an important organism of embryological studies for more than a century. In recent years, sea urchin embryos are being used as testing materials for toxicity of pollutants and toxins. Usefulness of sea urchin embryos as experimental models comes from the easiness in obtaining sea urchin samples and a lot of gametes, in rearing embryos in the laboratory, in observing the cellular movement and organ formation during the embryogenesis and in manipulating blastomeres and genetic maferials. The sea urchin in itself is a key organism for the understanding of deuterostome evolution from the protostomes and of indirect development of marine invertebrates which undergo the planktotrophic larval stage. A fertilized sea urchin egg goes through rapid cleavage and becomes a 60 cell embryo 7hr after fertilization. It then develops into a morula, a blastula, a gastrula and finally a pluteus larva approximately 70 hr after fertilization. At the 60 cell stage, the embryo comprises of five territories that express territory-speciflc genes and later form different organs. Micromeres at the vegetal pole ingress into the blastoceol and become the primary mesenchyme cells(PMCs). PMCs express genes involved in skeletogenesis such as SM30, SM37, SM50, PM27, msp130. Among the genes, SM37 and SM50 are considered to be members of a gene family which is characterized by early blastula expression, Glycine-Proline-Glutamine rich repeat structures and spicule matrix forming basic proteins. Genetic studies on the sea urchin embryos help understand the molecular basis of indirect development of marine invertebrates and also of the biomineralization common to the animal kingdom.

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Effect of ω3-Fatty Acid Desaturase Gene Expression on Invasion and Tumorigenicity in Human Tongue Squamous Cell Carcinoma Cells (인체 혀의 편평세포암 세포에서 ω3-fatty acid desaturase 유전자 발현이 침윤 및 종양형성에 미치는 영향)

  • Hong, Tae-Hwa;Shin, Soyeon;Han, Seung-Hyeon;Hwang, Byung-Doo;Lim, Kyu
    • Journal of Life Science
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    • v.28 no.8
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    • pp.945-954
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    • 2018
  • Omega-3 polyunsaturated fatty acids (${\omega}3$-fatty acid) have been found to possess anticancer properties in a variety of cancer cell lines and animal models, but their effects in human tongue squamous cell carcinomas (SCCs) remain unclear. This study was designed to examine the effect of ${\omega}3$-fatty acid desaturase (fat-1) gene expression on invasion and tumorigenicity in human tongue SCC cells and the molecular mechanism of its action. Docosahexaenoic acid (DHA) treatment inhibited in vitro invasion in a dose-dependent manner. In zymography, matrix metalloproteinase-9 (MMP-9) and Matrix metallopeptidase-2 (MMP-2) activities were reduced, and MMP-9 and MMP-2 promoter activities were inhibited by the DHA treatment. In addition, cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF) promoter reporter activities were inhibited in SCC-4 and SCC-9 cells after the DHA treatment. To investigate the effect of a high level of endogenous ${\omega}3$ fatty acids, a stable SCC-9 cell line expressing the ${\omega}3$-desaturase gene (fSCC-9sc) was generated. The growth rate and colony-forming capacity of fSCC-9sc were remarkably decreased as compared with those of fSCC-9cc. Likewise, the tumor size and volume of fSCC-9sc implanted into nude mice were significantly inhibited, with increases in the cell death index. Furthermore, a transwell chamber invasion assay showed a reduction in cell invasion of the fSCC-9sc lines when compared with that of the fSCC-9cc line. These findings suggested that fat-1 gene expression inhibited tumorigenicity, as well as invasion in human tongue SCC cells. Thus, utilization of ${\omega}3$ fatty acids may represent a promising therapeutic approach for chemoprevention and the treatment of human tongue SCCs.

EVALUATION OF OSTEOGENIC ACTIVITY AND MINERALIZATION OF CULTURED HUMAN PERIOSTEAL-DERIVED CELLS (배양된 인간 골막기원세포의 조골활성 및 골기질 형성의 평가)

  • Park, Bong-Wook;Byun, June-Ho;Lee, Sung-Gyoon;Hah, Young-Sool;Kim, Deok-Ryong;Cho, Yeong-Cheol;Sung, Iel-Yong;Kim, Jong-Ryoul
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.6
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    • pp.511-519
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    • 2006
  • Autogenous bone grafts have been considered the gold standard for maxillofacial bony defects. However, this procedure could entail a complicated surgical procedure as well as potential donor site morbidity. Possibly the best solution for bone-defect regeneration is a tissue engineering approach, i.e. the use of a combination of a suitable scaffold with osteogenic cells. A major source of osteogenic cells is the bone marrow. Bone marrow-derived mesenchymal stem cells are multipotent and have the ability to differentiate into osteoblastic, chondrocytic, and adipocytic lineage cells. However, the isolation of cells from bone marrow has someproblems when used in clinical setting. Bone marrow aspiration is sometimes potentially more invasive and painful procedure and carries of a risk of morbidity and infection. A minimally invasive, easily accessible alternative would be cells derived from periosteum. The periosteum also contains multipotent cells that have the potential to differentiate into osteoblasts and chondrocytes. In the present study, we evaluated the osteogenic activity and mineralization of cultured human periosteal-derived cells. Periosteal explants were harvested from mandibule during surgical extraction of lower impacted third molar. The periosteal cells were cultured in the osteogenic inductive medium consisting of DMEM supplemented with 10% fetal calf serum, 50g/ml L-ascorbic acid 2-phosphate, 10 nmol dexamethasone and 10 mM -glycerophosphate for 42 days. Periosteal-derived cells showed positive alkaline phosphatase (ALP) staining during 42 days of culture period. The formation of ALP stain showed its maximal manifestation at day 14 of culture period, then decreased in intensity during the culture period. ALP mRNA expression increased up to day 14 with a decrease thereafter. Osteocalcin mRNA expression appeared at day 7 in culture, after that its expression continuously increased in a time-dependent manner up to the entire duration of culture. Von Kossa-positive mineralization nodules were first present at day 14 in culture followed by an increased number of positive nodules during the entire duration of the culture period. In conclusion, our study showed that cultured human periosteal-derived cells differentiated into active osteoblastic cells that were involved in synthesis of bone matrix and the subsequent mineralization of the matrix. As the periosteal-derived cells, easily harvested from intraoral procedure such as surgical extraction of impacted third molar, has the excellent potential of osteogenic capacity, tissue-engineered bone using periosteal-derived cells could be the best choice in reconstruction of maxillofacial bony defects.

Chondrogenesis of Human Adipose Tissue Derived Mesenchymal Stem Cells (ATMSCs) Seeded in Gelatin-Chondroitin-Glucosamine Scaffold (Gelatin-Chondroitin-Glucosamine Scaffold에 접종한 인간지방조직-유래 중간엽 줄기세포의 연골형성)

  • Kim, Eung-Bae;Hong, Soon-Gab;Do, Byung-Rok;Kim, Hae-Kwon;Lee, Joon-Yeong
    • Development and Reproduction
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    • v.15 no.2
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    • pp.99-111
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    • 2011
  • The present experiment was performed to evaluate the chondrogenic differentiation potential of human adipose tissue-derived mesenchymal stem cells (ATMSCs) in the chondrogenic induction medium (CIM) with transforming growth factor-${\beta}1$ (TGF-${\beta}1$) and to evaluate the chondrogenic differentiation of ATMSCs seeded in gelatin-chondroitinglucosamine scaffold (GCG-scaffold). ATMSCs and mouse chondrocytes were cultured in the basic medium and CIM without TGF-${\beta}1$ (CIM1) or with TGF-${\beta}1$ (CIM2) for chondrogenic differentiation potential. The chondrogenic differentiation of ATMSCs was evaluated by glycosaminoglycan (GAG) synthesis and histochemical staining. In pellet culture, GAG synthesis of ATMSCs and chondrocyte was increased in culture on 14 days, but higher in CIM1 than basic medium, especially highest in CIM2. Cartilage matrix was observed in ATMSCs cultured in CIM2 on 14 days by Safranin O and trichrome staining. In well plate culture, proliferation of ATMSCs was continuously increased in culture on 10 days and higher in CIM than basic medium. The cell adhesion rate of ATMSCs seeded in flask or scaffolds was continuously increased during culture period, but higher in scaffold than flask. GAG synthesis of ATMSCs seeded in scaffolds showed no change in control group. In the CIM groups, GAG synthesis of ATMSCs was continuously increased than control group during culture period, especially very high in CIM2 and in the GCG-scaffold was slightly higher than the gelatin scaffold (G-scaffold). The present results demonstrated that ATMSCs showed an low chondrogenic differentiation potential, compared to mouse chondrocytes for 14 days of culture. TGF-${\beta}1$ is important factor in chondrogenic differentiation of ATMSCs. Gelatin scaffold was considered to increasing the effective chondrogenic differentiation environment. ATMSCs seeded in GCG-scaffold was more effective in chondrogenesis than in G-scaffold. Conclusively, the present results demonstrated that the treatment of chondroitin and glucosamine in the scaffold was more effective to promote the cartilage matrix formation.

Effects of the Draronis sanguis on Antioxidation and MMP-1 Expression in Human Dermal Fibroblast (혈갈(Draconis Sanguis)의 항산화와 사람섬유아세포에서 MMP-1 발현저해 효과)

  • Sim Gwan Sub;Kim Jin Hui;Kim Jin Hwa;Lee Dong Hawn;Park Sung Min;Lee Bum Chun;Pyo Hyeong Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.4 s.48
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    • pp.439-444
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    • 2004
  • UV irradiation produces free radicals and related reactive oxygen species (ROS), and these are injury to all most of organisms of skin cells and extracellular matrix (ECM). In addition, free radicals and ROS stimulate the overexpression of matrix metalloproteinases (MMPs) that can degrade most components of ECM such as collagen. Since collagen constitutes almost of skin connective tissue, their disarrangement causes wrinkle formation and droop of skin. Therefore, scavenging activity on free radicals, ROS and suppression of MMP-1 is expected to prevent skin photoaging. In this study, to investigate the relationship between photoaging and Draconis sanguis, we examined the effects of antioxidant, in vitro MMP inhibition and expression of UVA-induced MMP-1 in human dermal fibroblasts. Draconis sanguis was found to show scavenging activities of radicals and ROS with the $IC_{50}$ values of $183{\;}{\mu}g/mL$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and $30{\;}{\mu}g/mL$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. Draconis sunguis inhibited the activities of MMP-1 in a does-dependent manner and the $IC_{50}$ value calculated from semi-log plots was $200{\;}{\mu}g/mL$. Also, UVA induced MMP expression was reduced $74\%$ by treatment with Draconis sanguis, and MMP-1 mRNA expression was reduced in a dose-dependent manner. Therefore Draconis sanguis was able to significantly inhibit MMP expression in protein and mRNA level. All these results suggested that Draconis sanguis may act as an anti-photoaging agent by antioxidation and reducing UVA-induced MMP-1 production.

The Occurrence and Origin of a Syn-collisional Mélange in Timor (티모르섬 충돌 동시성 멜란지의 산상 및 기원)

  • Park, Seung-Ik;Koh, Hee Jae;Kim, Sung Won;Kihm, You Hong
    • Economic and Environmental Geology
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    • v.47 no.1
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    • pp.1-15
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    • 2014
  • The Bobonaro m$\acute{e}$lange is one of the youngest syn-collisional m$\acute{e}$langes, located between the Indo-Australian and Eurasian plates. The m$\acute{e}$lange has formed in association with a collision between the Australian continental margin and the Banda arc initiated in Neogene. The Suai area at the southern part of Timor is a good place to examine the genetic relationship between the m$\acute{e}$lange and other rock sequences because various tectonostratigraphic units coexist in the area. In this study, we present the structural characteristics and spatial distribution of the Bobonaro m$\acute{e}$lange investigated as a part of 1:25K scale geologic mapping in the area, and discuss on the origin of the m$\acute{e}$lange. The Bobonaro m$\acute{e}$lange in the Suai area is composed of unmetamorphosed clay matrix and blocks of various lithologies. The clay matrix mainly is reddish brown or greenish gray in colour, and has scaly texture. Most blocks are allochthonous, but mostly derived from nearby formations. Based on the internal structure and relationship with surrounding rocks, the Bobonaro m$\acute{e}$lange is genetically classified into 1) diapiric m$\acute{e}$lange; 2) tectonic m$\acute{e}$lange; and 3) broken formation. The spatial distribution of the Bobonaro m$\acute{e}$lange indicates that it intruded all pre-collisional units including the lower Australian continental margin unit(Gondwana megasequence) and the Banda arc unit. Taking the field evidences and previous genetic models into consideration, the Bobonaro m$\acute{e}$lange is interpreted to be mainly formed as a diapiric m$\acute{e}$lange originated from Gondwana megasequence, consistently effected by faulting events. This study reflects that diapiric m$\acute{e}$lange is a significant component in recent accretionay-collision belts. It suggests that diapiric process should be considered as a main genetic factor even in ancient m$\acute{e}$lange.

AN EXPERIMENTAL STUDY ON THE EFFECT OF Ca(OH)2 UPON THE HEALING PROCESS OF THE PULP AND PERIAPICAL TISSUE IN THE DOGS' TEETH (수산화칼슘이 손상치수조직 및 치근조직의 치유에 미치는 영향에 관한 연구)

  • Lim, S.S.;Yoon, S.H.;Lee, C.S.;Lee, M.J.;Kim, Y.H.;Kwon, H.C.;Um, C.M.
    • Restorative Dentistry and Endodontics
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    • v.8 no.1
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    • pp.123-131
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    • 1982
  • The purpose of this study was to observe the responses of the remaining pulp tissue after pulpotomy upon the several kinds of $Ca(OH)_2$ products and the responses of periapical tissue upon some root canal filling materials after extirpation. For pulpotomy, the class V cavities were prepared on the premolars, molars and upper canines, and the pulp was amputated. Each drug was placed over the amputated tissue and cavity was sealed with zinc oxide eugenol cement. The drugs which were used for the study were Dycal (Caulk Co. U.S.A.), Cavitec (Kerr Co. U.S.A.), Calvital, Nobudyne and Neodyne (Neo Dental Chemical Products). For extirpation, the endodontic cavities were prepared on the lingual surfaces of anterior teeth, and the pulp tissues were extirpated as routine method. After enlarging, irrigation, and measuring of root length by taking X-ray, each root canal filling material was filled in the canal with gutta percha cone, and endodontic cavity was sealed with zinc oxide eugenol cement. Zinc oxide eugenol, $Ca(OH)_2$ (Eli Lilly Co. U.S.A.) and Vitapex (Neo Dental Chemical Products) were used as root canal filling materials. Animals were sacrificed after 1, 3 and 6 weeks following the operation. The teeth were decalcified in formic acid, sectioned and stained with hematoxylin eosin. Microscopic examination revealed as follows. 1. Dycal: The dentin bridge formation was observed at the 3rd week after pulpotomy. Inflammatory conditions which were infiltration of inflammatory cells and dilatation of blood vessels were kept in remaining pulp tissue at the 6th week. 2. Calvital: The dentin bridge was observed at the 1st week after pulpotomy. As the time clasped, the pulp tended to be the fibrous degeneration. 3. Cavitec, Nobudyne and Neodyne: In the case of Cavitec and Nobudyne, the incompleted and irregular dentin bridge was observed at the 6th week, and in Neodyne, was observed at the 3rd week. The severe inflammatory changes were seen in the remaining pulp tissue. As the time clasped, the fibrous degeneration tended to spread in the remaining pulp tissue. 4. $Ca(OH)_2$: Osteocementum was formed at the 3rd week, the matrix of cementum and dentin were resorted, and infiltration of lymphocytes was seen in periapical tissue when $Ca(OH)_2$ was used as canal-filling materials. S. ZOE and Vitapex The cementum like substance was seen in periapical portion at the 1st week, when ZOE and Vitapex were used as root canal filling materials. As the time elapsed, the matrix of cementum and dentin tended to be resorted. At the 6th week, the inflammatory condition of periapical tissue was continued in the case of ZOE, but was reduced in the case of Vitapex.

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Therapeutic Effects of Curdrania tricuspidata Leaf Extract on Osteoarthritis (골관절염 실험모델에서 꾸지뽕나무 추출물의 골관절염 억제효과 연구)

  • Nam, Da-Eun;Kim, Ok Kyung;Lee, Jeongmin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.5
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    • pp.697-704
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    • 2013
  • The inhibitory effect of ethanol extracts from Curdrania tricuspidata leaves (CTL) on osteoarthritis was investigated in primary cultured rat cartilage cells and a monosodium-iodoacetate (MIA)-induced arthritis rat model. To identify the effects of CTL 80% ethanol extracts (CTL80) and CTL 10% ethanol extracts (CTL10) against $H_2O_2$ treatment in vitro, cell survival was measured by the MTT assay. Cell survival after $H_2O_2$ treatment increased with CTL80 and CTL10 close to normal up to $300{\mu}g/mL\;H_2O_2$. The mRNA expression of matrix metalloproteinases (MMPs) was determined MMP-7 and MMP-13 (known catabolic factors), were significantly inhibited by CTL 80 and CTL10; a $200{\mu}g/mL$ dose of CTL80 especially decreased MMP-13 expression. In vivo, osteoarthritis was induced by an intra-articular injection of MIA into the knee joints of rats, then CTL80 and CTL10 orally administered daily for 35 days. After the animals were sacrificed, histological evaluations of their knee joints revealed a reduction in polymorphonuclear cell infiltration and smooth synovial lining in the CTL80-500 group. Micro-CT analysis of hind paws from CTL80-500 and CTL10 showed a protection against osteophyte formation, soft tissue swelling, and bone resorption. In conclusion, CTL ethanol extracts are effective in ameliorating joint destruction and cartilage erosion in MIA-induced rats. CTL decreases and normalizes articular cartilage through preventing extracellular matrix degradation and chondrocyte injury, and could potentially serve as a therapeutic treatment for humans.