• 대한의생명과학회지
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• 제14권3호
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• pp.179-186
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• 2008

Matrix metalloproteinases (MMPs), also designated matrixins, hydrolyze components of the extracellular matrix. These proteinases playa central role in many biological processes, such as embryogenesis, normal tissue remodeling, wound healing, and angiogenesis, and in diseases such as atheroma, arthritis, cancer, and tissue ulceration. In previous data, disruption of the actin cytoskeleton by cytochalasin D (CD) inhibited NO-induced apoptosis, dedifferentiation, cyclooxygenase (COX)-2 expression, and prostaglandin $E_2$ production in chondrocytes cultured on plastic or during cartilage explants culture. In this study, we investigated the effects of the actin cytoskeleton architecture on MMP-2 expression and dedifferentiation by CD in rabbit articular chondrocytes. Rabbit articular chondrocytes were prepared from cartilage slices of 2-weeks-old New Zealand white rabbits by enzymatic digestion. CD was used as a disruptor of actin cytoskeleton. In this experiments measuring CD dose response, primary chondrocytes were treated with various concentrations of CD for 24h. The actin disruption was determined by immunostaining. MMP-2 expression levels were determined by immunoblot analysis and Reverse transcriptase-Polymerase chain reaction (RT-PCR) and MMP-2 activity was determined by gelatin zymography. We found that cell morphological change and up-regulation of MMP-2 expression by CD as determined via immunostaining, gelatin zymography and immunoblotting. Moreover, CD induced MMP-2 transcription was detected by RT-PCR. Also, CD-induced type II collagen expression was inhibited by MMP-2 inhibitor I treatment. Our results indicate that CD up-regulated MMP-2 activation causes dedifferentiation of articular chondrocyte.

• International Journal of Oral Biology
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• 제49권1호
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• pp.18-25
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• 2024

Despite advancements in therapeutic approaches, radiotherapy and cisplatin-based chemotherapy remain primary noninvasive treatments for patients with oral squamous cell carcinoma (OSCC). Moreover, the 5-year survival rate for patients with OSCC has remained almost unchanged for several decades, and many side effects of chemotherapy still exist. In this study, three-dimensional (3D) models of OSCC were established using fibroblasts, and the efficacy of various biological inhibitors was evaluated. A culture of epithelial cells with two types of fibroblasts (hTERT-hNOFs and cancer-associated fibroblasts) within a type I collagen matrix resulted in the formation of a continuous layer of tightly packed cells compared to models without fibroblasts. Furthermore, the effects of biological chemicals, including Y27632, latrunculin A, and verteporfin, on these models were investigated. The stratified formation of the epithelial layer and invasion in OSCC 3D-culture models were effectively inhibited by verteporfin, whereas invasion was weakly inhibited by Y27632 and latrunculin. Collectively, the developed OSCC 3D-culture models established with fibroblasts demonstrated the potential for drug screening, with verteporfin showing promising efficacy.

• Journal of Chest Surgery
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• 제40권5호
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• pp.329-340
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• 2007

배경: Matrix metalloproteinase (MMP) 차단은 심근경색 후 좌심실 확장에 대한 가능한 치료 전략으로 대두되고 있다. 선택적 MMP 차단제의 투여가 심근경색 후 초기 단계에 MMP가 대량으로 분비되는 짧은 기간을 차단하는 것이 좋을 것인지, 초기 전체 기간 동안 차단하여야 할 것인지를 알아보고자 하였다. 대상 및 방법: 토끼를 이용하여 기관 삽관 하에 전신 마취를 하고 흉골 정중절개한 다음 좌전 하행지 관상동맥을 결찰하여 심근경색을 만들었다. 실험군은 3군으로 나누었다. 심근경색 단독(MI only 군)군은 7예, MMP 차단제 5일 투여군(MMPI 5d 군)은 6예, MMP 차단제 9일 투여군(MMPI 9d 군)은 5예이었다. MMP 차단제로는 MMP-2와 MMP-9에 대한 선택적 차단제인 CG2300을 사용하였다. 각 군은 심장초음파도 검사를 4회 시행하였는데, 술 전, 술 후 1주, 2주 및 3주에 하였다. 검사는 2D 심초음파도를 사용하여 EDD, ESD 및 EF를 측정하였다. 술 후 4주에 희생한 토끼의 심장을 western blotting과 zymography를 하여 MMP-2와 MMP-9의 단백질과 활성의 변화를 조사하였고, 경색부위를 병리학적 조직검사를 하였다. 결과: 심초음파도 검사상, MI only군에서는 대체로 술 전에 비하여 술 후 EDD와 ESD가 증가한 추세로 좌심실이 확장하였음을 알 수 있었다. MMP 차단제 9일 투여군에서는 심근경색 단독군과 MMP 차단제 5일 투여군에 비해 좌심실의 확장이 감소한 경향을 보였다. EF는 MMP 차단제 9일 투여군에서 술 후에 술 전과 큰 변동이 없었으며, 다른 군들보다 높은 경향이었다. MMP 단백질의 발현과 활성 변화를 보면, 심근경색 단독군, MMP 차단제 5일 및 9일 투여군 등 3군을 정상 심장군과 비교하였을 때 MMP-2 단백질 발현과 활성 변화는 일어나지 않았다. 그리고 MMP-9의 단백 발현 및 활성은 검출되지 않았다. 병리학적 조직 소견을 보면 심근경색 단독군에서 심한 교원질 침착이 있었다. MMP 차단제 5일 투여군과 9일 투여군에서는 교원질 축적이 감소된 경향을 보였다. MMP 9일 투여군에서는 모세혈관의 수가 증가한 것을 볼 수 있었다. 결론: 관상동맥을 결찰하여 심근경색을 유도하면 술 후 빠른 시간 내에 심실이 확장되며 MMP 차단제를 투여할 경우 심실의 확장이 완화됨을 알 수 있었다. MMP 차단제의 효과는 초기의 대부분 기간을 차단하는 것이 좋다고 생각된다. MMP 차단제가 혈관신생을 증가시켜 심실 재형성을 완화할 수 있는 것으로 분석된다.

• 대한화장품학회지
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• 제23권3호
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• pp.24-38
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• 1997

Using the all-trans-retinol which is double-capsulated with matrix, we investigated its stability and the change of the epidermal thickness. The proprietary MDC comprise two steps of capsulation of retinol, i.e., primary microcapsulation with collagen and then secondary capsulation with gellan gum. We compared the activity of all-trans-retinol in various forms such as (1) simply in O/W, (2) in W/O emulsion, (3) in primary capsulted form in O/W emulsion, or (4) in MDC in O/W emulsion. After storage at 45$^{\circ}C$ for 4 weeks, retinol in MDC in O/W emulsion retained 92% of the activity compared to the standard material upon HPLC analysis, whereas the primary capsule gave 70%, the O/W emulsion form 47% and the W/O emulsion 78%. The retinol in MDC in O/W induced the siginificant increase in epidermal thickness compared to the vehicle.

• Journal of Periodontal and Implant Science
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• 제48권1호
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• pp.34-46
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• 2018

Purpose: The purpose of this study was to evaluate the effects of 1,25-dihydroxyvitamin $D_3$ on the proliferation, differentiation, and matrix mineralization of MC3T3-E1 osteoblast-like cells in vitro. Methods: MC3T3-E1 osteoblastic cells and 1,25-dihydroxyvitamin $D_3$ were prepared. Cytotoxic effects and osteogenic differentiation were evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, alkaline phosphatase (ALP) activity assay, ALP staining, alizarin red S staining, and reverse transcription-polymerase chain reaction (RT-PCR) for osteogenic differentiation markers such as ALP, collagen type I (Col-I), osteocalcin (OCN), vitamin D receptor (VDR), and glyceraldehyde 3-phosphate dehydrogenase. Results: The MTT assay showed that 1,25-dihydroxyvitamin $D_3$ did not inhibit cell growth and that the rate of cell proliferation was higher than in the positive control group at all concentrations. ALP activity was also higher than in the positive control group at low concentrations of 1,25-dihydroxyvitamin $D_3$ ($10^{-10}$, $10^{-12}$, and $10^{-14}M$). RT-PCR showed that the gene expression levels of ALP, Col-I, OCN, and vitamin D receptor (VDR) were higher at a low concentration of 1,25-dihydroxyvitamin $D_3$ ($10^{-12}M$). Alizarin red S staining after treatment with 1,25-dihydroxyvitamin $D_3$ ($10^{-12}M$) showed no significant differences in the overall degree of calcification. In contrast to the positive control group, formation of bone nodules was induced in the early stages of cell differentiation. Conclusions: We suggest that 1,25-dihydroxyvitamin $D_3$ positively affects cell differentiation and matrix mineralization. Therefore, it may function as a stimulating factor in osteoblastic bone formation and can be used as an additive in bone regeneration treatment.

• Journal of Periodontal and Implant Science
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• 제26권2호
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• pp.429-447
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• 1996

Transforming growth factor $-{\beta}$ is one of the polypeptide growth factors that mediate the activity of mesenchymal cells and regulate wound healing process via cell proliferation, migration and extracellular matrix formation. The purposes of this study is to evaluate the effects of transforming growth factor $-{\beta}$ on the protein synthetic activity of human periodontal ligament cells and human gingival fibroblasts. The cells which were prepared were primary cultured gingival fibroblasts and periodontal ligament cells from humans, and the fourth or sixth subpassage were used in the experiments. Cells were seeded and at a confluent state, 0, 0.5, I, 2.5, 5, 10 ng/ml $TGF-{\beta}$ and $2{\mu]Ci/ml\;[^3H]$ proline were added to the cells and cultured for 24 hours. Then, 1 and 5 ng/ml concentrations were selected and added to confluent cells and cultured for 24 and 48 hours. They were labeled with $2{\mu}Ci/ml\;[^3H]$ proline for 24 hours and a collagen assay was done by the Peterkofsky and Diegelman method. The results were presented as the mean disintegration per minute (dpm) per well and S.D. of four determinations, The results were as follows. : The total protein, collagen and noncollagenous protein synthesis in periodontal ligament cells and gingival fibroblasts were increased dose- dependently by transforming growth factor-p to 2.5-5 ng/ml concentration and decreased at 10 ng/ml concentration. The percent of collagen was slightly changed according to the concentration of transforming growth factor-po The effect of transforming growth $factor-{\beta}$ was not specific for collagen synthesis since it increased the total, noncollagenous and collagenous protein, simultaneously. In the comparison of protein synthetic activity between the human periodontal ligament cells and human gingival fibroblasts, the human gingival fibroblasts had higher activities than the human periodontal ligament cells at all times and concentrations of $TGF-{\beta}$. In the comparison of protein synthetic activity between the 24 hour effect and the 48 hour effect of $TGF-{\beta}$, the 48 hour cultured cells' synthetic activity decreased more than the 24 hour cultured cells at human periodontal ligament cells and human gingival fibroblasts. In conclusion, $TGF-{\beta}$ has important roles in the stimulation of protein synthesis in human periodontal ligament cells and human gingival fibroblasts. Thus, it may be useful for clinical application in periodontal regenerative procedures.

• 대한화장품학회지
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• 제44권4호
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• pp.389-397
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• 2018

별불가사리(Asterina pectinifera Muller and Troschel)는 우리나라 전국 연안에서 흔히 볼 수 있는 토속종으로, 패류 양식장에 피해를 주는 불가사리류 중의 하나이다. 별불가사리 퇴치를 위해 대부분 건조하여 비료로 활용하고 있으며, 고부가가치 창출을 위한 다양한 연구가 진행되었으나 실제 활용은 미미한 실정이다. 따라서, 별불가사리로부터 피부 유용성분을 밝혀 새로운 활용 방안을 모색하고자 하였다. 성분연구를 통해 별불가사리로 부터 2종의 polyhydroxysteroid와 1종의 saponin을 분리하였으며, 이의 구조를 각각 $5{\alpha}$-cholestane-$3{\beta},6{\alpha},7{\alpha},8,15{\alpha},16{\beta},26$-heptol, $5{\alpha}$-cholestane-$3{\beta},4{\beta},6{\alpha},7{\alpha},8,15{\beta},16{\beta},2$6-octol 및 asterosaponin $P_1$으로 동정하였다. 이 물질들의 피부 효능을 확인한 결과 asterosaponin $P_1$이 표피 줄기세포의 증식을 촉진시키고, 각질형성세포에서 히알루론산을 합성하는 효소인 hyaluronan synthase-2와 hyaluronan synthase-3 유전자의 발현을 증가시킴을 확인하였다. 또한, asterosaponin $P_1$은 섬유아세포에서 진피의 주요 콜라겐인 type 1 콜라겐의 생합성을 촉진하는 효능을 보였다. 이상의 결과들로부터, 별불가사리로부터 분리한 asterosaponin $P_1$은 노화에 동반되는 피부 증상을 개선하는 항노화 화장품 소재로 활용될 수 있을 것으로 판단된다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제45권3호
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• pp.123-128
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• 2019

Demineralized dentin matrix (DDM) has been used as a recombinant human bone morphogenetic protein-2 (rhBMP-2) carrier in many clinical trials. To optimize the clinical safety and efficacy of rhBMP-2 with DDM, efforts have been made to improve the delivery of rhBMP-2 by 1) lowering the administered dose, 2) localizing the protein, and 3) prolonging its retention time at the action site as well as the bone forming capacity of the carrier itself. The release profile of rhBMP-2 that is associated with endogenous BMP in dentin has been postulated according to the type of incorporation, which is attributed to the loosened interfibrillar space and nanoporous dentinal tubule pores. Physically adsorbed and modified, physically entrapped rhBMP-2 is sequentially released from the DDM surface during the early stage of implantation. As DDM degradation progresses, the loosened interfibrillar space and enlarged dentinal tubules release the entrapped rhBMP-2. Finally, the endogenous BMP in dentin is released with osteoclastic dentin resorption. According to the postulated release profile, DDM can therefore be used in a controlled manner as a sequential delivery scaffold for rhBMP-2, thus sustaining the rhBMP-2 concentration for a prolonged period due to localization. In addition, we attempted to determine how to lower the rhBMP-2 concentration to 0.2 mg/mL, which is lower than the approved 1.5 mg/mL.

• Journal of Periodontal and Implant Science
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• 제33권3호
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• pp.359-372
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• 2003

Among objectives of periodontal therapy. the principal one is the morphological and functional reconstruction of lost periodontal supporting tissues. This includes de novo formation of connective tissue attachment and the regrowth of alveolar bone. The use of enamel matrix derivative(EMD) may be a suitable means of regeneration new periodontal attachment in the infrabony defects. Implant used to replace lost tooth but, implantitis occurred after installation. The purpose of this study was to investigate the effects of EMD on differentiation and growth of osteoblast in titanium disc. Twentyfive millimeter diameter and 1mm thick Ti disc which was coated 25, 50, 100, 200${\mu}g$/ml of EMD(Emdogain(R)) used as experimental group, 25, 50, 100, 200ng/d of rhBMP-2 as positive control group, and no coat as negative control group. A human osteosarcoma cell line Saos-2 was cultured in Ti disc and cell proliferation and Alkaline phosphatase (ALP) activity were measured at 1 and 6 days. PCR was performed at 2 and 8 hours. Semi-quantitative RT-PCR for mRNA expressions of various osteoblastic differentiation markers -type I collagen, ALP, osteopontin, and bone sialoprotein - were performed at appropriate concentrations based upon the results of MTT and ALP assay. Cultured cell-disc complexes were prepared for scanning electron microscopy (SEM) at 2 hour. Data were analyzed using Mann-Whitney and repeated- measures 1-way analysis of variance(SPSS software version 10,SPSS. Chicago. IL). After culture, there was more osteoblast in EMD100${\mu}g$/ml than in EMD50, 200${\mu}g$/ml on day 6. There was significant difference in experimental and positive control group compared control group, as times go by(1 and 6 days). Alkaline phosphatase activity was different significantly in EMD100, 200${\mu}g$/ml and BMP100, 200${\mu}g$/ml on day 6. The results of reverse transcriptase-polymerase chain reaction (RT-PCR) showed that expression of mRNA for ALPase, collagen type I, osteopontin. hone sialoprotein and BMP-2 was detected at 2 hour and 8 hour in EMI 200${\mu}g$/ml subgroup and BMP100ng/ml subgroup. The results of this study suggest that application of enamel matrix derivative on osteoblast attached to titanium surface facilitate the expression of bone specific protein and the differentiation and growth of osteoblast.

• Journal of Nutrition and Health
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• 제56권3호
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• pp.231-246
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• 2023

본 연구에서는 MIA로 골관절염을 유도한 SD 흰쥐에서 인도산 보스웰리아 검레진을 주정 추출 후, 헥산으로 지방 제거 공정을 추가하여 제조한 보스웰리아 검레진 추출물인 FJH-UBS의 항골관절염 효능을 평가하기 위해 실시하였다. FJH-UBS는 40 또는 80 mg/kg BW/day 용량으로 5주간 경구투여하였고, FJH-UBS를 2주간 투여 후 MIA (3 mg/50 µL/rat)를 무릎 관절강 내에 주사하여 골관절염을 유도하였다. MIA 유도 골관절염 동물모델에서 FJH-UBS는 무릎 관절의 부종을 감소시키고 연골의 분해를 억제하였으며, 연골 내 type II collagen과 aggrecan 발현을 증가시켰다. FJH-UBS (80 mg/kg BW/day)는 혈청 내 PGE2, LTB4, IL-1β, 및 IL-6 함량을 감소시켰고, MMP-13 함량을 감소시켰다. FJH-UBS (80 mg/kg BW/day)는 연골 활막 내 iNOS, COX-2, 5-LOX, IL-1β, IL-6 및 TNF-α 발현을 감소시켰고, MMP-2, MMP-9 및 MMP-13 발현을 감소시켰다. 이 결과는 FJH-UBS가 염증매개물질과 염증성 cytokine의 발현감소를 통해 염증 반응을 억제하고, MMPs의 발현을 억제하여 연골 기질의 분해를 억제함으로서 항골관절염 효능을 나타냄을 의미하며 이는 관절 및 연골 건강 개선 기능성 원료로 FJH-UBS의 활용 가능성을 제시한다.