• Maxillofacial Plastic and Reconstructive Surgery
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• 제30권2호
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• pp.133-141
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• 2008

Stem cells have self-renewal capacity, long-term viability, and multiline age potential. Adult bone marrow contains mesenchymal stem cells. Bone marrow-derived mesenchymal stem cells (BMSCs) are progenitors of skeletal tissue components and can differentiate into adipocytes, chondrocytes, osteoblasts, and myoblasts in vitro and undergo differentiation in vivo. However, the clinical use of BMSCs has presented problems, including pain, morbidity, and low cell number upon harvest. Recent studies have identified a putative stem cell population within the adipose tissue. Human adipose tissue contains pluripotent stem cells simillar to bone marrow-derived stem cells that can differentiate toward the osteogenic, adipogenic, myogenic, and chondrogenic lineages. Human adipose tissue-derived stem cells (ATSCs) could be proposed as an alternative source of adult bone marrow stem cells, and could be obtained in large quantities, under local anesthesia, with minimal discomfort. Human adipose tissue obtained by liposuction was processed to obtain ATSCs. In this study, we compared the osteogenic differentiation of ATSCs in a specific osteogenic induction medium with that in a non-osteogenic medium. ATSCs were incubated in an osteogenic medium for 28 days to induce osteogenesis respectively. Osteogenic differentiation was assessed by von Kossa and alkaline phosphatase staining. Expression of osteocyte specific bone sialoprotein, osteocalcin, collagen type I and alkaline phosphatase, bone morphogenic protein 2, bone morphogenic protein 6 was confirmed by RT-PCR. ATSCs incubated in the osteogenic medium were stained positively for von Kossa and alkaline phosphatase staining. Expression of osteocyte specific genes was also detected. Since this cell population can be easily identified through fluorescence microscopy, it may be an ideal source of ATSCs for further experiments on stem cell biology and tissue engineering. The present results show that ADSCs have an ability to differentiate into osteoblasts. In the present study, we extend this approach to characterize adipose tissue-derived stem cells.

• 대한의생명과학회지
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• 제23권3호
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• pp.230-237
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• 2017

Changes in the activity of alkaline phosphatase (ALP) isoenzymes and isoforms in human serum have a major diagnostic value, therefore the regulation of ALP activities is a valuable target for therapeutic interventions. To assess the pharmacological activity of retinoids, i.e., all-trans retinoic acid and 13-cis retinoic acid, their tissue-specific inhibitory effect on human serum ALP activity was elucidated by chemical inhibition methods, heat-sensitive inactivation, and wheat-germ lectin precipitation test. Retinoids showed significant inhibition of the total ALP activity in human serum at a concentration of 5 mM. All-trans retinoic acid (5 mM) and 13-cis retinoic acid (5 mM) inhibited ALP activities by up to 12% and 15%, respectively, compared to that by guanidine hydrochloride (200 mM). L-phenylalanine (100 mM) and urea (30 mM) had no further inhibitory effect on ALP activities in human serum pretreated with retinoids (5 mM). Retinoids significantly inhibited ALP activities by up to 20% compared with that of tetramisole (30 mM). The ALP activities in retinoid-pretreated serum remained unchanged after the heat inactivation process. These results suggest that retinoids are inhibitors of the intestinal ALP isoenzyme. Remarkably, retinoids revealed potent inhibitory activities against ALP in wheat-germ lectin precipitant serum, indicating that they also function as inhibitors of the bone ALP isoform. The results show that retinoids inhibit the specific tissue-derived human serum ALP activities, moreover, the inhibitory effect of retinoids against bone ALP activity suggests their clinical utility as monitoring and prevention of metastasis of bone cancer.

• 대한치과교정학회지
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• 제24권1호
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• pp.105-114
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• 1994

The movement of teeth during orthodontic treatment requires bone remodeling process of bone formation and bone resolution. To find out the changes occuring in the cell itself, mechanical stress was applied to the cell populations involved in the bone metabolism. Bone tissue cell populations were isolated from fetal rat calvaria and divided into OC and OB groups. Following results were obtained from measuring the changes in acid & alkaline phosphatease activity, cyclic AMP and $PGE_2$ production in time lapse after the application of mechanical stress. 1. In case of the marker enzyme of specific bone tissue cell, acid phosphatase activity was high in OC group and alkaline phosphatase activity was high in OB group. 2. After the mechanical stress was applied, acid phosphatase activity was decreased in both OC and OB groups and alkaline phosphatase activity was increase in OB group. 3. When the mechanical stress was applied for 15, 30 and 60 minutes, the production of $PGE_2$ increased in both OC and OB groups, as the time span increased. 4. When the mechanical stress was applied for 20 and 40 minutes, the production of $PGE_2$ increased in both OC and OB groups, as the time span increased.

• 대한골관절종양학회지
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• 제7권2호
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• pp.41-50
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• 2001

목적 : 골육종 환자의 조직과 혈청에서 DNase, RNase, 5'-nucleotidase, alkaline phosphatase 및 amylase 활성도를 측정하여 그 활성도의 변동을 보았고 이들 효소가 골육종의 표지자로 이용될 수 있는지 알아보았다. 대상 및 연구방법 : 골육종 환자의 혈청과 종양조직은 1 2예로부터 얻었으며, 이에 대한 대조조직은 동일 환자의 정상부위에서 채취하였다. 조직 추출액을 얻어서 핵산, 단백함량, 그리고 효소 활성도의 측정에 사용하였다. DEAE-cellulose chromatography에 의해 골육종 조직의 acid DNase, neutral RNase, RNase inhibitor와 단백을 분리하고 그활성도와 단백 함량을 측정하였다. 결과 : 골육종 조직의 acid DNase, RNase, 5'-nucleotidase, alkaline phosphatase 활성도는 유의하게 상승하였고, 효소활성도의 양성율도 높게 나타났다. neutral RNase의 활성도가 특히 높았으며, RNase inhibitor는 neutral RNase와 복합을 이루면서 그 활성도가 유의하게 증가하였다. 혈청 neutral RNase 활성도가 유의하게 상승하였다. DEAE-cellulose column chromatography에 의해 acid DNase는 단일효소로서, 그리고 neutral RNase는 5개의 isozyme으로 분리되었다. 결론 : 이들 효소의 병용이 골육종의 생화학적 표지자로서 이용될 수 있을 가능성을 시사하였다. 또한 혈청 neutral RNase활성도의 측정이 골육종의 생화학적 표지자로서의 역할을 시사하였다. acid DNase와 neutral RNase가 골육종의 발생과 억제과정에 작용하고 있을 가능성을 시사하였다.

• 생명과학회지
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• 제16권7호
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• pp.1207-1213
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• 2006

Tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$는 염증성 골질환에서의 골조직의 손실과 밀접한 관련이 있다. 본 연구에서는 인체 골수 유래 중간엽 줄기세포의 골세포로의 분화과정에 대한 $TNF-{\alpha}$의 영향을 조사하였다. $TNF-{\alpha}$는 골수 유래 중간엽 줄기세포의 골세포로의 분화를 나타내는 표시인 세포외 무기질 축적과 alkaline phosphatase의 발현의 증가를 일으켰으며 2ng/ml의 농도에서 최대의 증가를 나타내었다. $TNF-{\alpha}$에 의한 골세포로의 분화는 $NF_kB$의 저해제에 의해서는 영향받지 않았으나 JNK 특이 저해제인 SP600125에 의해 완벽하게 억제되었다. 이는 $TNF-{\alpha}$에 의한 골수 유래 중간엽 줄기세포의 골세포로의 분화과정에 JNK가 중요한 역할을 한다는 것을 제시한다.

• Journal of Animal Science and Technology
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• 제63권1호
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• pp.114-124
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• 2021

The objective of this study was to characterize the enzymatic hydrolysis of lipopolysaccharide (LPS) by wheat phytase and to investigate the effects of wheat phytase-treated LPS on in vitro toxicity, cell viability and release of a pro-inflammatory cytokine, interleukin (IL)-8 by target cells compared with the intact LPS. The phosphatase activity of wheat phytase towards LPS was investigated in the presence or absence of inhibitors such as L-phenylalanine and L-homoarginine. In vitro toxicity of LPS hydrolyzed with wheat phytase in comparison to intact LPS was assessed. Cell viability in human aortic endothelial (HAE) cells exposed to LPS treated with wheat phytase in comparison to intact LPS was measured. The release of IL-8 in human intestinal epithelial cell line, HT-29 cells applied to LPS treated with wheat phytase in comparison to intact LPS was assayed. Wheat phytase hydrolyzed LPS, resulting in a significant release of inorganic phosphate for 1 h (p < 0.05). Furthermore, the degradation of LPS by wheat phytase was nearly unaffected by the addition of L-phenylalanine, the inhibitor of tissue-specific alkaline phosphatase or L-homoarginine, the inhibitor of tissue-non-specific alkaline phosphatase. Wheat phytase effectively reduced the in vitro toxicity of LPS, resulting in a retention of 63% and 54% of its initial toxicity after 1-3 h of the enzyme reaction, respectively (p < 0.05). Intact LPS decreased the cell viability of HAE cells. However, LPS dephosphorylated by wheat phytase counteracted the inhibitory effect on cell viability. LPS treated with wheat phytase decreased IL-8 secretion from intestinal epithelial cell line, HT-29 cell to 14% (p < 0.05) when compared with intact LPS. In conclusion, wheat phytase is a potential therapeutic candidate and prophylactic agent for control of infections induced by pathogenic Gram-negative bacteria and associated LPS-mediated inflammatory diseases in animal husbandry.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제32권2호
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• pp.97-106
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• 2010

Aim of the study: An alternative source of adult stem cells that could be obtained in large quantities, under local anesthesia, with minimal discomfort would be advantageous. Adipose tissue could be processed to obtain a fibroblast-like population of cells or adipose tissue-derived stromal cells (ATSCs). This study was performed to confirm the availability of ATSCs in bone tissue engineering. Materials amp; Methods: In this study, adipose tissue-derived mesenchymal stem cell was extracted from the liposuctioned abdominal fat of 24-old human and cultivated, and the stem cell surface markers of CD 105 and SCF-R were confirmed by immunofluorescent staining. The proliferation of bone marrow mesenchymal stem cell and ATSCs were compared, and evaluated the osteogenic differentiation of ATSCs in a specific osteogenic induction medium. Osteogenic differentiation was assessed by von Kossa and alkaline phosphatase staining. Expression of osteocyte specific BMP-2, ALP, Cbfa-1, Osteopontin and osteocalcin were confirmed by RT-PCR. With differentiation of ATSCs, calcium concentration was assayed, and osteocalcin was evaluated by ELISA (Enzyme-linked immunosorbant assay). The bone formation by 5-week implantation of HA/TCP block loaded with bone marrow mesenchymal stem cells and ATSCs in the subcutaneous pocket of nude mouse was evaluated by histologic analysis. Results: ATSCs incubated in the osteogenic medium were stained positively for von Kossa and alkaline phosphatase staining. Expression of osteocyte specific genes was also detected. ATSCs could be easily identified through fluorescence microscopy, and bone formation in vivo was confirmed by using ATSC-loaded HA/TCP scaffold. Conclusions: The present results show that ATSCs have an ability to differentiate into osteoblasts and formed bone in vitro and in vivo. So ATSCs may be an ideal source for further experiments on stem cell biology and bone tissue engineering.

• 한국수의병리학회지
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• 제3권1호
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• pp.1-5
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• 1999

This experiment was carried out to establish the immunohistochemical diagnostic method for infectious pancreatic necrosis in the monolayers of CHSE-214 cell cultures and paraffin-embedded tissue sections from rainbow trout infected with infectious pancreatic necrosis virus(IPNV). Specific identification of IPNV antigens was often demonstrated in the pancreatic exocrine cells, and less in the intestinal mucous epithelia and the renal hemopoietic tissues by the use of monoclonal antibodies against capsid protein VP2. The specific reaction was seen as a distinct red cytoplasmic color, often as small granules of various sizes. The result showed that streptavidin alkaline phosphatase immunohistochemisry specifically identified IPNV antigens in both infected cell cultures and tissue sections.

• Asian Pacific Journal of Cancer Prevention
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• 제14권8호
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• pp.4627-4634
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• 2013

Chemoprotection refers to the use of specific natural or synthetic chemical agents to suppress or prevent the progression to cancer. The purpose of this study is to assess the protective effect of aspirin, vitamin C or zinc in a dimethyl hydrazine (DMH) colon carcinoma model in rats and to investigate the effect of these supplements on changes associated with colonic zinc status. Rats were randomly divided into three groups, group 1 (aspirin), group 2 (vitamin C) and group 3 (zinc), each being subdivided into two groups and given subcutaneous injection of DMH (30 mg/kg body wt) twice a week for 3 months and sacrificed at 4 months (A-precancer model) and 6 months (B-cancer model). Groups 1, 2, 3 were simultaneously given aspirin, vitamin C, or zinc supplement respectively from the beginning till the end of the study. It was observed that 87.5% of rats co-treated with aspirin or vitamin C showed normal colonic histology, along with a significant decrease in colonic tissue zinc at both time points. Rats co-treated with zinc showed 100% reduction in tumor incidence with no significant change in colonic tissue zinc. Plasma zinc, colonic CuZnSOD (copper-zinc superoxide dismutase) and alkaline phosphatase activity showed no significant changes in all 3 cotreated groups. These results suggest that aspirin, vitamin C or zinc given separately, exert a chemoprotective effect against chemically induced DMH colonic preneoplastic progression and colonic carcinogenesis in rats. The inhibitory effects are associated with maintaining the colonic tissue zinc levels and zinc enzymes at near normal without significant changes.

• 한국식품영양과학회지
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• 제21권3호
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• pp.241-246
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• 1992

L-Ascorbic acid (AsA) 생합성이 불가능한 guinea pig을 실험 동물로 하여 collagne 함량이 높은 조직인 폐 및 피부중의 AsA함량과 동일조직중 collagne 함량이 높은 조직인 폐 및 피부중의 AsA 함량과 동일조직중 collagen의 proline잔기의 수산화율을 조사하여 collagne 생합성에 대한 조직중 AsA의 영향에 대해서 알아보았다. Guinea pig(체중 약 250g)를 AsA 무투여군(A), 투여군(B), 300mg/day 투여군(C)으로 나눠 14일간 사육한 후, 마취하에서 개복하여 복부 대동맥으로부터 채혈함과 동시에 간장과 폐를 적출하였으며, 등부위의 피부를 채취하여 분석용 시료로 하였다. 이들 시료로부터 혈청중 alkaline phosphatase (ALP)활성과 각조직중의 AsA 함량, proline 함량 및 그 수산화율, (1-$^{14}$ C) proline 의 incorporation 양을 측정하였다. 그 결과, AsA 토여군인 B, C군의 경우 순조로운 체중증가와 함께 혈청중 ALP활성도 정상값을 나타냈으며 현저한 ALP활성 저하가 관찰되었다. 한편, AsA 함량이 높을수록 (1-14C) proline 의 incorporation 양이 많고 collagne 중의 hydroxyproline 함량도 증가하는 것으로 나타나, 조직중의 collagen합성량과 AsA함량과의 사이에는 높은 상관관계가 존재함이 확인되었다.