• Maxillofacial Plastic and Reconstructive Surgery
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• v.30 no.2
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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.

• Biomedical Science Letters
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• v.23 no.3
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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.

• The korean journal of orthodontics
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• v.24 no.1 s.44
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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.

• The Journal of the Korean bone and joint tumor society
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• v.7 no.2
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• pp.41-50
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• 2001

Purpose : To investigate biochemical markers for osteosarcoma, activities of deoxyribocuclease(DNase), ribonuclease(RNase), 5'-nucleotidase, alkaline phosphatase and amylase were determined in the osteosarcoma tissue and serum of patients with osteosarcoma. Also studied were DNase, RNase in osteosarcoma tissue, isolating the enzymes from the sarcoma tissue and investigating the sarcoma specific enzymes. Materials and Methods : The experimental tissue and serum were obtained from twelve patients with osteosarcoma. The control group were obtained from the normal healthy tissue of the same patients. The tissue were centrifugalized to obtain extracts. The extracts were analized for the estimation of nucleic acid, protein contents and enzyme activities. And then each enzymes were isolated and analized by DEAE-cellulose chromatography and estimated for activities. Result : Activities of acid DNase, RNase, 5'-nucleotidase and alkaline phosphatase were significantly increased in osteosarcoma tissue. Neutral RNase in osteosarcoma tissue was shown to bo highly active, exhibiting secretory form of RNase inhibitor associated with the RNase was also increased. In the serum of patients with osteosarcoma, RNase activity was significantly increased. DEAE-cellulose column chromatographical analysis revealed that acid DNase was isolated as a single enzyme and neutral RNase as five isozymes in osteosarcoma tissue. Conclusion : The results indicated that combination of these enzymes could be used as markers for osteosarcoma. The results indicated that acid DNase and neutral RNase might play a role in genesis of sarcoma and suppression of sarcoma.

• Journal of Life Science
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• v.16 no.7 s.80
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• pp.1207-1213
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• 2006

Tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ has been implicated in skeletal diseases by promoting bone loss in inflammatory bone diseases. In the present study, we examined the effects of $TNF-{\alpha}$ on osteoblastic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs). $TNF-{\alpha}$ dose-dependently promoted matrix mineralization of hBMSCs with a maximal stimulation at 2ng/ml. $TNF-{\alpha}$ increased expression of alkaline phosphatase, which plays a crucial role for the matrix deposition. The $TNF-{\alpha}-stimulated$ osteoblastic differentiation was not affected by $NF_kB$ inhibitors, BAY and SN50. However, a JNK-specific inhibitor, SP600125 completely abolished the $TNF-{\alpha}-stimulated$ matrix mineralization and expression of alkaline phosphatase. These results suggest that $TNF-{\alpha}$ enhances osteoblastic differentiation of hBMSCs through JNK-dependent pathway.

• Journal of Animal Science and Technology
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• v.63 no.1
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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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• v.32 no.2
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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.

• Korean Journal of Veterinary Pathology
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• v.3 no.1
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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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• v.14 no.8
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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.

• Journal of the Korean Society of Food Science and Nutrition
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• v.21 no.3
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• pp.241-246
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• 1992

To clarify the requirement of L-ascorbic acid (AsA) in collagen synthesis, the incorporation of 1-$^{14}$ C-proline into the tissues of guinea pigs and the specific radioactivity ratio (proline/hydroxyproline) in collagen were investigated. Male guinea pigs maintained on the AsA-deficient diet were divided into three groups ; group A (AsA-deficient animals) : group B (control animals) supplemented with 5mg AsA/day ; group C (high dose animals) with 300mg AsA/day, and orally supplemented with or with-out AsA for 14 days. Collagen synthesis was estimated by measuring the incorporation of labeled pro-line into collagen in lung and dorsal skin, and the hydroxyproline contents in lung and skin. The AsA contents in the tissues were determined by high-peforrnance liquid chromatography (HPLC), and serum alkaline phosphatase activity was also measured. The serum alkaline phosphatase activity of AsA deficient group was very low as compared with those of AsA supplemented group. Incorporation of labelled proline into collagen and its specific radioactivity ratio in collagen increased with increasing levels of AsA in the tissues. There was a significantly positive relationship between the levels of AsA and hydroxyproline in the tissues.