• The korean journal of orthodontics
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• v.26 no.2 s.55
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• pp.187-196
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• 1996

Underlying malocclusions and dentofacial deformities are often related to variations in the craniofacial development. Type I and type II collagens are considered the major collagens of bone and cartilage respectively. Monitoring the patterns of those protein expressions during development will Provide a basis for the understanding of normal and abnormal growths. This study was undertaken to investigate the morphogenetic changes and the expression patterns of type I and II collagen proteins involved in the developing mandible of human embryos and fetuses. 50 embryos and fetuses were studied with Hematoxylin and Eosin, Alcian, blue-PAS, Masson Trichrome, md Immunohistochemical stains. The results were as follows : 1. A 13.5 mm embryo showed the stomatodeum with dental lamina, maxillary and mandibular processes. Meckel's cartilage appeared in the mandibular arch of a 20.5 mm embryo. New bone formation was bilaterally initiated at the outer side of middle portion of Meckel's cartilage of 22-38 mm embryos. 2. Meckel'cartilage was resorbed at the 15th week fetus. The endochondral ossification was observed where there was direct replacement of cartilage by bone. Meckel'cartilage disappeared and membraneous ossification were observed at the 25th week. 3. Before the appearance of Meckel's cartilage, the expression of type I collagen was moderate at the odontogenic epithelium of maxillary & mandibular process, but mild for the expression of type II collagen. 4. During the appearance of Meckel's cartilage and new bone formation, the immunoactivity of type II collagen was more expressed than type I collagen at the Meckel's cartilage and new bone. 5. During intrarmembranous bone formation, the expression of type II collagen was rare in the bony trabeculae. There was a switch for the expression of collagens from type II to type I during the appearance of Meckel's cartilage.

• The korean journal of orthodontics
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• v.26 no.4
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• pp.455-467
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• 1996

This study was designed to evaluate the expression of type I collagen in periodontal tissue during the experimental movement of rat incisors. Twenty-one Sprague-Dawley rats were divided into a control group(3 rats), and experimental groups(18 rats) where a force(75g) from helical springs across the maxillary incisors was applied. Experimental groups were sacrificed at 12 hours, 1, 4, 7, 14 and 28 days after force application, respectively. And tissue slides of control and experimental groups were studied histologically and immunohistochemically by LSAB(Labelled streptavidine Biotin) immunohistochemical staining for type I collagen. The results were as follows: 1. Until 28-day after force application, periodontal fibers were strectched on the tension side, and compressed in pressure side, and the arrangement of periodontal fibers was not recovered by that time. 2. The degree of type I collagen expression in control group was rare in the oral epithelium, predentin, pulp and periodontal ligament, but was mildly positive in osteoblasts, acellular cementum, cementoblasts, intermaxillary suture. 3. At acellular cementum of experimental group, the expression of type I collagen was moderate in 1-day and severe in 7-day, which was maintained until 28-day. 4. Type I collagen was observed in the newly formed fibrous connective tissue and osteoblasts at intermaxillary suture, moderately in 1-day, and severely in 14-day. 5. The tension side of periodontal ligament showed a more positive expression of type I collagen than the pressure side in 4-day. The degree was highest in 7-day and was not differentiated between sides in 14-day. 6. In the side wall of bone matrix on which osteoblasts were attached, type I collagen was expressed severely, especially in 7-day. From the above findings, we could suggest that bone remodeling in tooth movement be intimately related to the cell differentiation and the resulting formation of type I collagen.

• Journal of Periodontal and Implant Science
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• v.35 no.1
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• pp.133-152
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• 2005

1. 목적 Prostaglandin은 치주질환과 관련된 국소적 골 대사에 중요한 역할을 한다. ${\Delta}^{12}PGJ_2$는 생체 내에서 혈장의 존재 하에 형성되는 천연 $PGD_2$ 대사산물이며 peroxisome- proliferator에 의해 활성화되는 감마 수용체 (PPAR ${\Gamma}$)에 대해 높은 친화성을 갖는 리간드로서 핵 수용체군에 속하는 전사조절인자이다. 이 연구의 목적은 골화 과정에서 ${\Delta}^{12}PGJ_2$의 역할을 규명하기 위해, 조골세포주의 증식과 분화에 미치는 영향과 그에 관련된 세포기전을 조사하는 데에 있다. 2. 방법 인간 골육종세포주인 Saos-2 (ATCC.HTB 85)와 쥐의 조골세포주 (MC3T3-E1)를 배양한 후 실험군에 농도가 각각 $10^{-5}$, $10^{-6}$, $10^{-7}$, $10^{-8}$, $10^{-9}$ 몰인 ${\Delta}^{12}PGJ_2$와 ciglitazone (합성 PPAR 감마 길항체)를 첨가하였다. 조골세포에서 PPAR 감마의 발현을 관찰하기 위해 역전사효소-중합효소연쇄반응(RT-PCR)을 특정한 primer를 이용하여 시행하였다. 세포 증식은 1일, 2일, 3 일째에 MIT 분석법으로 측정하였고, 2 일째에 알칼리성 인산효소 (ALPase) 생산을 측정하였다. 위의 결과에서 얻은 적정한 농도에서 다양한 조골세포 분화의 표지자들-제 1 형 교원질, 알칼리성 인산효소, osteopontin 및 bone sialoprotein-에 대한 간이 정량적 역전사효소-중합효소연쇄반응 (semiquantitative RT-PCR)을 실시하였으며 골결절 형성에 대한 효과를 알아보고자 석회화 분석도 시행하였다. 3. 결과 ${\Delta}^{12}PGJ_2$와 ciglitazone 모두 Saos-2 세포주의 증식을 촉진시켰다 .$10^{-8}$ 몰의 ${\Delta}^{12}PGJ_2$와 $10^{-6}$몰의 ciglitazone을 첨가한 실험군을 대조군과 비교했을 때, 시간에 비례하여 세포 증식률이 증가되었다. 알칼리성 인산효소의 활성화 검사에서도 증식률에서와 유사한 결과를 보여주었다. 간이 정량적 RT-PCR에서는 ${\Delta}^{12}PGJ_2$로 처리한 군의 경우 제 1 형 교원질, 알칼리성 인산효소, osteopontin, 그리고 bone sialoprotein의 상대적 mRNA 수준이 유의하게 높았다. 석회화 분석에서는 MC3T3-E1 세포를 $10^{-6}$ 몰의 ${\Delta}^{12}PGJ_2$로 처리한 군과 $10^{-5}$ 몰의 ciglitazone으로 처리한 군에서 현저한 골결절 형성을 보였다. 이러한 결과들은 ${\Delta}^{12}PGJ_2$가 유용한 골 유도물질이 될 수 있으며 또한 그 작용기전이 PPAR 감마-의존형 경로와 연관되어 있음을 보여준다.

• Journal of the Korean Arthroscopy Society
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• v.3 no.2
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• pp.85-90
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• 1999

Purpose : The purpose of this study is to evaluate the clinical and histological results of the osteoarthritic patients who had second look arthroscopy after microfracture surgery. Materials and Methods : From Oct. 1997 to Dec. 1998, 46 patients, 48 knees were treated by microfracture technique. In the 22 patients, 24 knees, 'second-look' arthroscopies and biopsies were performed at 6 months following microfracture. Three patients were men and 19 patients were women. Average age of the patients were 58 years (range, 40-75 years). The average follow up period was 12 months(7-20 months). We analysed clinical results according to the nine-point scale. Also we observed type II collagen formation with immunohistochemical staining. Results : Clinical results were excellent in 83% and good in 17%. Among the 24 knees, more than 80% areas of chondral defect were covered with regenerated cartilage in 21 knees. Histologically, the regenerated tissue appears to be a hybrid of hyaline cartilage and fibrocartilage. Regenerated cartilage contains variable amount of type II collagen with immunohistochemical staining. Conclusion : Most of the patients had significant improvement clinically. 'Second-look' showed that the chondral defect areas were covered with newly grown grayish white tissue. Microfracture in the full thickness chondral defect provides and enriched environment for cartilaginous tissue regeneration.

• Journal of Periodontal and Implant Science
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• v.36 no.2
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• pp.397-408
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• 2006

치주질환은 치아 지지조직의 파괴로 튿정 지어지는 감염성 질환으로서 이것은 주로 조직의 교원질 성분을 분해시키는 MMP(matrix metalloproteinase)에 의해 이루어진다. 한편, 여러 연구에서 당뇨병과 치주질환의 심도와의 관계에 대한 논의가 있어왔으며 당뇨병이 치주염을 포함한 구강 감염에 대한 감수성을 증가시키며 역으로 만성 치주염의 염증성 매개물질에 의해 인슐린 작용이 방해받을 수 있음을 보고하였다. 본 연구의 목적은 만성 치주염 환자와 제 2형 당뇨병을 동반한 만성 치주염 환자의 치은조직에서 MMP-13과 elastase의 발현 양상을 비교하여 병리 기전의 차이점을 규명하고, 두 단백질간의 상호관계를 알아보기 위한 것이다. 경북대 병원 치주과에 내원한 환자의 비당뇨 환자의 정상 치은 부위, 비당뇨 환자의 만성 치주염 부위, 제 2형 당뇨병 환자의 만성 치주염 부위에서 각각 8개의 변연 치은을 채득하여 액화 질소에 급속 동결시켰다. 모든 조직 샘플에서 동량의 단백질을 western blotting을 통해 분석하여였고, densitometer를 이용하여 정량한 후 ANOVA 분석으로 통계처리 하였다. 결과분석에서 MMP-13은 제 2형 당뇨를 가진 환자의 염증성 조직에서 가장 높게 발현되었고 전신적으로 건강한 환자의 염증성 조직과 정상 조직에서는 유사한 양상을 보였으며 그 차이는 통계적으로 유의하였다. 또한 elastas는 그룹간에 유의한 차이 없이 유사한 양상으로 발현되었고, 염증성 조직에서도 당뇨병의 유무에 관계없이 유사하게 나타났다. 한편 조직내 MMP-13과 elastase의 발현 양상간에 유의한 상관관계는 보이지 않았다. 결론적으로 MMP-13은 당뇨병을 동반한 환자의 치은 염증초직에서 유의하게 증가되어 나타났으며, 전신적으로 건강한 환자의 염증조직과 당뇨를 동반한 환자의 염증조직에서 MMP-13과 elastase의 발현양상은 큰 상관관계가 발견되지 않았다.

• Journal of Korean Orthopaedic Sports Medicine
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• v.4 no.1
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• pp.18-28
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• 2005

Purpose: The purpose of this study is to evaluate the clinical, radiological and histopathological results after microfracture surgery for degenerative arthritis of the knee. Materials and Methods: From Oct. 1997 to Dec. 1998, 48 knees in 46 patients were treated by microfracture technique. Their mean age at the time of operation was 56 years(range, 40-75 years) and mean period of follow-up study was one year(range, 7-20 months). For 24 knees in 22 patients, 'second-look' arthroscopies and biopsies were performed at 6 months following microfracture. At the last follow up clinical results were evaluated with Baumgaertner's scale. The specimens of 24 cases were stained with H-E, Safranin-O, and Masson's trichrome. Eighteen of 24 cases were stained immunohistochemically and the Western blotting test was performed on 12 cases for type II collagen. We analyzed the relationship of the Western blotting for type II collagen with clinical score, preoperative varus deformity, joint space widening in radiological result, extent of repaired articular cartilage in '2nd-look' arthroscopic findings, patient's age and weight. Results: Clinical results were excellent in 90% and good in 10%. Among the 24 knees, more than 80% of areas of chondral defect were covered with regenerated cartilage in 21 knees Histologically, the repaired tissue appears to be a hybrid of hyaline cartilage and fibrocartilage. Repaired cartilage contains variable amounts of type II collagen with immunohistochemical staining. The results of the Western blotting test were similar. The amounts of type II collagen formation had positive correlation with the extent of repaired cartilage and preoperative varus deformity. Conclusion: 'Second-look' showed that the chondral defect areas were covered with newly grown grayish white tissue. Articular cartilage repair was confirmed with histological and immunohisto-chemical study qualitatively, and the amount of type II collagen was calculated with the Western blotting test quantitatively. The exact nature and fate of repaired cartilagenous tissues need further long term follow-up study. The results of this study provide the rationale to select osteoarthritic patients indicated for microfracture surgery.

• The korean journal of orthodontics
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• v.23 no.3 s.42
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• pp.415-425
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• 1993

Fluoride is one of the most potent stimulators of bone formation in vivo. But its direct effects on osteoblast is not yet clear This study was to investigate the effects of Sodium fluoride on alkaline phosphatase(ALP) activity, cAMP formation responsive to parathormone(PTH) and type I $\alpha$ 2 collagen ribonucleic acid (mRNA) level in Murin osteoblast-like (MC3T3-E1) cells. The cells were cultured in $\alpha-Minimal$ essential medium $(\alpha-MEM)$ supplemente with $10\%$ fetal bovine serum (FBS) and then changed to $0.1\%$ FBS with various concentration of Sodium fluoride. The ALP activity was assayed by the method of Lowry with disodium phenyl phosphated as substrate. cAMP formation was measured by Radioimmuno Assay(RIA). Type I $\alpha$ 2 collagen ribonucleic acid(mRNA) expression was studied by Nothern blot analysis. The results were as follows: 1. cAMP level was increased by PTH in MC3T3-E1 cells. 2. Sodium fluoride showed the tendency of inhibitory effects on cAMP responsiveness to PTH in MC3T3-E1 cells. 3. Sodium fluoride increased ALP activity at cocentration of $2{\mu}M,\;4{\mu}M,\;and\;10{\mu}M$ significantly different from control at the 0.001 level. ALP activity revealed maximum value at $10{\mu}M$ in this study. 4. Nothern blot analysis of Sodium fluoride treated cells, using Type I $\alpha$ 2 collagen prove, revealed significant increase at $10{\mu}M$ in MC3T3-E1 cells.

• Korean Journal of Head & Neck Oncology
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• v.13 no.2
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• pp.180-186
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• 1997

Objectives: Salivary gland tumors pose considerable difficulty in diagnostic and prognostic assessment based on the histopathologic features alone. We studied the expression of type IV collagen and fibronectin in salivary gland tumors with special emphasis on the differential diagnostic significance. Materials and Methods: We did immunohistochemical stain on paraffin embedded tissues of 33 benign and 24 malignant salivary gland tumors using monoclonal antibody for type IV collagen and polyclonal antibody for fibronectin. Results: 1) Well preserved linear basement membrane-like staining of type IV collagen was detected in duct-cell-derived benign salivary gland tumors. But pleomorphic adenoma exhibited a heterogeneous pattern as focal augmentation and interruption. 2) In malignant tumors, type IV collagen was distributed in an irregular, interrupted manner or completely absent. Adenoid cystic carcinomas displayed a marked staining of the basal membrane associated substances in the pseudocysts. 3) The staining pattern of fibronectin was similar to that of type IV collagen execpt more dense in the stroma. 4) Salivary gland tumors which have a prominent myoepithelial cell component revealed a particular deposition of basement membrane materials adjacent to the myoepithelial cells. Conclusion: The study of the basal membrane substances may be helpful for differential diagnosis of benign and malignant salivary gland tumors and identifying special features of salivary gland tumors such as pseudocystic pattern of adenoid cystic carcinoma. Also we think that the myoepithelial cells contribute to the formation of basement membrane materials.

• Journal of Yeungnam Medical Science
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• v.23 no.2
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• pp.182-192
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• 2006

Background: Atherosclerosis has emerged as the leading cause of death in developed countries. At present, human umbilical vein endothelial cells (HUVEC) are most commonly used for the investigation of Endothelial cells (EC). However, HUVEC are not found in arteries but only in veins. Currently there are many reports on methods used to isolate EC;, most of these methods require special equipment to remove contaminating smooth muscle cells (SMC). Materials and Methods: The method described here may be used to isolate not only ECs but also SMCs;,the approach presented here did not require special equipment. Rat aorta was treated with 2 mg/ml of type II collagenase solution for 45 minutes. The isolated cells from the aorta were incubated in medium G for a week;, only ECs could be separated. After the collagenase treatment, the rest of aorta was cut lengthwise, and left undisturbed to obtain SMCs in the culture dish for 10 days. To verify the purity of the isolated cells, we performed immunofluorescence and evaluated the results with transmission electron microscopy analysis. Results: The immunofluorescence study demonstrated specific expression of CD31 and ${\alpha}$-smooth muscle actin in the isolated ECs and SMCs, respectively. Cultured ECs and SMCs showed their own fine structure characteristics. Conclusion: These results suggest that this method for isolating ECs and SMCs may be especially useful for the study of atherosclerosis.

• YAKHAK HOEJI
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• v.56 no.1
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• pp.9-13
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• 2012

We previously reported that the extract of Taraxacum platycarpum (AF-343) had several biological properties such as skin hydration and anti-inflammatory effects, thereby AF-343 be a promising anti-atopic dermatitis agent. However, few studies have been conducted to evaluate its effect on modulation of extracellular matrix proteins in human skin fibroblasts. The purpose of this study was to investigate the expressions of type I collagen, MMP-1, Smad2/3, and TIMP-1 proteins in AF-343-treated human skin fibroblasts. Human skin fibroblasts were treated by various concentrations of AF-343 (0~2 mg/ml). The expressions of type I collagen, matrix metalloproteinase-1 (MMP-1), Smad2/3, and TIMP-1 proteins were analyzed by Western blot analysis. In addition, level of type I collagen mRNA was analyzed by CAT assay. Expression of type I collagen protein was increased in AF-343-treated human skin fibroblasts by dose and time-dependent manners. Consistent with this result, the expressions of phospho-Smad2/3 in skin fibroblasts were increased and MMP-1 expression was decreased by AF-343 treatment. TIMP-1 expression was not significantly changed in AF-343 treated skin fibroblasts. Extract of Taraxacum platycarpum (AF-343)-induced up-regulation of type I collagen expression was through increased expression of phospho-Smad2/3. These results were occurred combined with down-regulation of MMP-1 in skin fibroblasts. Taken together, this study indicated that AF-343 has property of the modulation of ECM in tissue as well as skin hydration and anti-inflammation.