• The korean journal of orthodontics
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• v.31 no.4 s.87
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• pp.447-458
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• 2001

The purpose of this study was to evaluate 1) in vivo, the expression of chondroitin 4-sulfate (CH-4S), a structural element of glycosaminoglycans(GAGs), in periodontal tissue during the experimental movement of rat incisors, by labelled streptavidine biotin immunohistochemical staining for CH-4S, 2) In vitro, the expression of CH-4S in cultured human periodontal ligament(PDL) cells supplemented with 10ng/ml of $TGF-{\beta}_1$, 20ng/ml of PDGF-BB, 1ng/ml $TNF-\alpha$, or $1{\mu}g/ml$ LPS by western blot analysis. The results of this study were as follows ; 1. The expression of CH-4S was stronger in pulp, PDL, osteoblasts, osteoclasts and osteocytes in experimental group than in control group, but was rare in dentin, and cementum of experimental groups, regardless of the duration of force application, which was not different from that of control group. 2. In experimental group, the expression of CH-4S in pulp began to increase at 1 day after force application and got to the highest degree at 7 days. After 14 days, the expression in CH-4S immunoreactivity was decreased, and became similar to that of control group at 28 days. 3. The expression of CH-4S in PDL was noted in adjacent to alveolar bone. PDL showed higher intensity of immunolabelling after 1 day of orthodontic tooth movement. And the expression was more stronger in the tension side than that of pressure side of PDL at 1 day, but more stronger in the pressure side than that of tension side of PDL at 4 days. After 7 days, a decrease in CH-4S expression was observed. 4. The expression of CH-4S in alveolar bone got to the highest degree at 4 days, and At 7 days, a decrease in CH-4S expression was observed. 5. PDGF-BB notably raised the expression of CH-4S in the PDL cells at 3 days of cultivation 6. The expression of CH-4S of PDL cells was decreased with the application of $TNF-\alpha$ at 1 day. 7. Admixture of $TGF-{\beta}_1$ and PDGF-BB got more expression of CH-4S in PDL as compared to only $TGF-{\beta}1$ or PDGF-BB. A similar decrease of the expression of CH-4S was observed in the case of application of LPS or $TNF-\alpha$.

• Journal of Embryo Transfer
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• v.28 no.3
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• pp.281-287
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• 2013

A major barrier to progress in pig to primate organ transplantation or cell therapy is the presence of terminal ${\alpha}$-1,3-galactosyl epitopes on the surface of pig cells. Therefore, the purpose of this experiment was to establish and cha- racterize mesenchymal stromal/stem cells (MSCs) derived from ${\alpha}$-1,3-galactosyltransferase (GalT) knock out (GalT KO) pig to confirm their potential for cell therapy. Bone marrow (BM)-MSCs from GalT KO pig of 1 month old were isolated by Ficoll-Paque PLUS gradient and cultured with A-DMEM + 10% FBS on plastic dishes in 5% $CO_2$ incubator at 38.5. GalT KO BM-MSCs were analyzed for the expression of CD markers ($CD45^-$, $29^+$, $90^+$ and $105^+$) and in vitro differentiation ability (adiopogenesis and osteogenesis). Further, cell proliferation capacity and cell aging of GalT KO BM-MSCs were compared to Wild BM-MSCs by BrdU incorporation assay (Roche, Germany) using ELISA at intervals of two days for 7 days. Finally, the cell size was also evaluated in GalT KO and Wild BM-MSCs. Statistical analysis was performed by T-test (P<0.05). GalT KO BM-MSCs showed fibroblast-like cell morphology on plastic culture dish at passage 1 and exhibited $CD45^-$, $29^+$, $90^+$ and $105^+$ expression profile. Follow in ginduction in StemPro adipogenesis and osteogenesis media for 3 weeks, GalT KO BM-MSCs were differentiated into adipocytes, as demonstrated by Oilred Ostaining of lipid vacuoles and osteocytes, as confirmed by Alizarinred Sstaining of mineral dispositions, respectively. BrdU incorporation assay showed a significant decrease in cell proliferation capacity of GalT KO BM-MSCs compared to Wild BM-MSCs from 3 day, when they were seeded at $1{\times}10^3$ cells/well in 96-well plate. Passage 3 GalT KO and Wild BM-MSCs at 80% confluence in culture dish were allowed to form single cells to calculate cell size. The results showed that GalT KO BM-MSCs($15.0{\pm}0.4{\mu}m$) had a little larger cell size than Wild BM-MSCs ($13.5{\pm}0.3{\mu}m$). From the above findings, it is summarized that GalT KO BM-MSCs possessed similar biological properties with Wild BM-MSCs, but exhibited a weak cell proliferation ability and resistance to cell aging. Therefore, GalT KO BM-MSCs might form a good source for cell therapy after due consideration to low proliferation potency in vitro.

• Development and Reproduction
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• v.11 no.3
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• pp.167-177
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• 2007

In the present study, we isolated three human adult stem cells including adipose tissue-derived stem cells(HAD), umbilical cord-derived stem cells(HUC), and amnion-derived stem cells(HAM) and analysed their characteristics. And we examined whether HAD could be used as therapeutical cells for the heart diseases. Both HAM and HUC appeared very similar morphology but HAD was different. Doubling time of HUC was most fast, but total doubling numbers of HUC was same with HAM. Total doubling numbers of HAD was much more than others. Expression patterns of genes and proteins of three human adult stem cells were very similar. Also they were differentiated into adipocytes, osteocytes, and chondrocytes. In addition, they expressed many cardiomyocyte-related genes. But expression pattern of genes is a little different. When HAD were cultivated in the presence or absence of various combinations of BMP and FGF after 5-azacytidine expose for 24 h, expression of Cmlc-1, and ${\alpha}1c$ genes was significantly increased. However, expression of troponin T, troponin I and Kv4.3 genes was not changed. Based on these observations, it is suggested that HAD, HUC, and HAM might be used as potentially therapeutical cells for clinical application.

• The korean journal of orthodontics
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• v.31 no.1 s.84
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• pp.107-120
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• 2001

Nitric oxide(NO) has been reported to be one of the mediators relating to bone remodelling. Nitric oxide is synthesized from L-arguinine by nitric oxide synthetase(NOS), which is largely divided Into two groups. One group which is composed of $NOS_1\;and\;NOS_3$, is dependent of calcium or calmodulin. The other consisted of $NOS_2$, which is independent of calcium or calmodulin. NOS is thought to be a possible intermediate affecting in the course of tooth movement. This study was designed to evaluate the expression of nitrous oxide synthetase(NOS) in periodontal tissue during the experimental movement of rat incisors, by LSAB(labelled streptavidine biotin) immunohistochemical staining for $NOS_2\;and\;NOS_3$. Twenty seven Sprague-Dawley rats were divided into a control group(3 rats), and 6 experimental groups(24 rats), to which 75g of force was applied, with helical springs across the maxillary incisors. Rats of experimental groups were sacrificed at 12 hours, 1, 4, 7, 14 and 28 days after force application, respectively. After that, the tissues of the control group and experimental groups were studied immunohistochemically. The results were as follows: 1. In control group, the expression of $NOS_3$ was rare in gingiva, dentin, periodontal ligament and alveolar bone, and was mild in the capillaries of pulp and intermaxillary suture. And the expression of $NOS_2$ showed similar pattern to that of $NOS_3$. 2. There were no differences in the expression of $NOS_2\;or\;NOS_3$ in dentin, gingiva, cementum, cementoblast and odontoblast, between control and experimental groups, regardless of the duration of the force application. 3. The expression of $NOS_3$ began to increase at 4 days and showed to the highest degree at 7 days after force application, in the apical region of pressure side of periodontal ligament in experimental groups. 4. The expression of $NOS_3$ in alveolar bone was rare until 7 days, after which it increased to mild degree at 14 days through 28 days in experimental group. But there was no difference between pressure and tension side of periodontal ligament. 5. The expression of $NOS_2$ in periodontal ligament was mild from 7 days after force application, regardless of the side of periodontium, which was generally more evident than that of $NOS_3$. 6. The expression of $NOS_2$ in alveolar bone increased to mild degree at 14 days after force application, and it was evident in osteoblasts, osteoclasts and osteocytes. And the expression of $NOS_2$ was little more stronger in the tension side than that of pressure side of alveolar bone.

• The korean journal of orthodontics
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• v.31 no.5 s.88
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• pp.525-534
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• 2001

Bone remodeling in response to force requires coordinated actions of osteoblasts, osteoclasts, osteocytes, and periodontal ligament cells. Coordination among these cells may be mediated, in part, by cell-to-cell communication via gap junctions. This study was designed to evaluate the expression of gap junction, connection 43 In periodontal tissue during the experimental movement of rat's incisors, by LSAB(labelled streptavidine biotin) immunohistochemical staining for connexin 43. Twenty seven Sprague-Dawley rats were divided into a control group(3 rats), and 6 experimental groups(24 rats) where 75g of force was applied from helical springs across the maxillary incisors. Rats of experimental groups were sacrificed at 12 hours, 1, 4, 7, 14 and 28 days after force application, respectively. And the tissues of a control group and experimental groups were studied immunohistochemically. The results were as follows : 1. In control group, the expression of connexin 43 was rare in gingiva, dentin, cementum, periodontal ligament, and bone cells. 2. In experimental group, the expression of connexin 43 was increased in pulp, periodontal ligament, osteoblasts, and osteoclasts, comparing to that in control. And it was rare in gingiva, dentin, and odontoblasts regardless of the duration of force application, which was not different from that of control group. 3. The expression of connexin 43 in pulp of experimental group began to increase in 4-day after force application and got to the highest degree at 7-day. And it decreased after 14-day to be similar to that of control group at 28-day. 4. The expression of connexin 43 in periodontal ligament was noted in small capillaries adjacent to alveolar bone, showing higher intensity of immunolabelling after 4-day And it was stronger in the pressure side than in tension side of periodontal ligament. After 7-day, decrease in connexin 43 expression was observed. 5. The expression of connexin 43 in alveolar bone began to increase 1-day, reached to the highest degree at 4-day, and decreased at 7-day. And the expression in osteoclasts was more than that in osteoblasts or osteocyte at 7-day.