• Title/Summary/Keyword: Bone matrix

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Expression of Morphogenic Protein Genes in Juvenile Red Spotted Grouper (Epinephelus akaara) with Deformity (붉바리(Epinephelus akaara) 기형 발생 치어의 형태형성 유전자 발현)

  • You, Jin Ho;Mun, Seong Hee;Oh, Hyeon Ji;Baek, Hea Ja;Lee, Young-Don;Lee, Chi Hoon;Kwon, Joon Yeong
    • Journal of Marine Life Science
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    • v.4 no.1
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    • pp.38-43
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    • 2019
  • The deformity occurring at the early developmental stage of red spotted grouper (Epinephelus akaara) causes detrimental effects on the process of juvenile production. In this study, we have compared the expressions of several key genes (insulin like growth factor 1: IGF-1, bone morphogenic protein 4: BMP4, peroxisome proliferator-activated receptors γ: PPARγ, matrix Gla protein: MGP) for morphogenesis between normal and 2 types (cephalic and jaw) of deformed juvenile fish. Expression of these genes were investigated in the brain, liver and muscle of each group of fish (n=20) by real-time PCR. Expression of IGF-1 and BMP4 mRNA in the brain and liver showed significant difference between normal and deformed fish (p<0.05). However, no difference was observed in the expression of PPARγ and MGP mRNA between normal and deformed fish in any tissues. It seems certain that IGF-1 and BMP4 are associated with the state of deformity in juvenile red spotted grouper.

In Vivo Experiment of Tissue-Engineered Artificial Vessel (조직공학적 인조혈관의 생체 내 이식 실험)

  • 임상현;조승우;홍유선;김병수;유경종;장병철;최차용
    • Journal of Chest Surgery
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    • v.37 no.3
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    • pp.220-227
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    • 2004
  • The number of patients with coronary artery disease and peripheral vascular disease are increasing, and the need of small diameter vessel is also increasing. We developed small diameter artificial vessel and experimented in vivo. We got allogenic valve from mongrel dogs, and removed all cells from the allogenic valve. Then, we seeded autologous bone marrow cells onto the decellularized scaffold. After implantation of artificial vessel into the canine carotid artery, we performed angiography regularly. In case of vessel occlusion or at 8 weeks after operation, we euthanized dogs, and retrieved the implanted artificial vessels. Control vessels were all occluded except one (which developed aneurysmal dilatation). But autologous cell seeded vascular graft were patent by 4 weeks in one, by 6 in one and by 8 weeks in two. Histologic examination of patent vessel revealed similar structure to native artery. Tissue-engineered vascular graft manufactured with decellularized allogenic matrix and autologous bone marrow cells showed that tissue engineered graft had similar structure to native artery.

Heat or radiofrequency plasma glow discharge treatment of a titanium alloy stimulates osteoblast gene expression in the MC3T3 osteoprogenitor cell line

  • Rapuano, Bruce E.;Hackshaw, Kyle;Macdonald, Daniel E.
    • Journal of Periodontal and Implant Science
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    • v.42 no.3
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    • pp.95-104
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    • 2012
  • Purpose: The purpose of this study was to determine whether increasing the Ti6Al4V surface oxide negative charge through heat ($600^{\circ}C$) or radiofrequency plasma glow discharge (RFGD) pretreatment, with or without a subsequent coating with fibronectin, stimulated osteoblast gene marker expression in the MC3T3 osteoprogenitor cell line. Methods: Quantitative real-time polymerase chain reaction was used to measure changes over time in the mRNA levels for osteoblast gene markers, including alkaline phosphatase, bone sialoprotein, collagen type I (${\alpha}1$), osteocalcin, osteopontin and parathyroid hormone-related peptide (PTH-rP), and the osteoblast precursor genes Runx2 and osterix. Results: Osteoprogenitors began to differentiate earlier on disks that were pretreated with heat or RFGD. The pretreatments increased gene marker expression in the absence of a fibronectin coating. However, pretreatments increased osteoblast gene expression for fibronectin-coated disks more than uncoated disks, suggesting a surface oxide-mediated specific enhancement of fibronectin's bioactivity. Heat pretreatment had greater effects on the mRNA expression of genes for PTH-rP, alkaline phosphatase and osteocalcin while RFGD pretreatment had greater effects on osteopontin and bone sialoprotein gene expression. Conclusions: The results suggest that heat and RFGD pretreatments of the Ti6Al4V surface oxide stimulated osteoblast differentiation through an enhancement of (a) coated fibronectin's bioactivity and (b) the bioactivities of other serum or matrix proteins. The quantitative differences in the effects of the two pretreatments on osteoblast gene marker expression may have arisen from the unique physico-chemical characteristics of each resultant oxide surface. Therefore, engineering the Ti6Al4V surface oxide to become more negatively charged can be used to accelerate osteoblast differentiation through fibronectin-dependent and independent mechanisms.

Differentiation of Osteoblast Progenitor Cells from Human Umbilical Cord Blood (제대혈액에서 골조직 특이세포로의 분화)

  • Hong, Seung-Jin;Lee, Eun-A;Chae, Gue-Tae;Han, Hoon
    • IMMUNE NETWORK
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    • v.2 no.3
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    • pp.166-174
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    • 2002
  • Background: Human umbilical cord bloods, which could be taken during the delivery are utilized as a source of hematopoietic stem cells. Also in cord blood, there are several kinds of stem cells such as endothelial and mesenchymal stem cells. Methods: We isolated the mesenchymal stem cells from human umbilical cord bloods and confirmed the differentiation of these cells into osteoblast progenitor cells. The mesenchymal stem cells derived from umbilical cord blood have the ability to differentiate into specific tissue cells, which is one of characteristics of stem cells. These cells were originated from the multipolar shaped cells out of adherent cells of the umbilical cord blood mononuclear cell culture. Results: The mesenchymal stem cells expressed cell surface antigen CD13, CD90, CD102, CD105, ${\alpha}$-smooth muscle actin and cytoplasmic antigen vimentine. Having cultrued these cells in bone formation media, we observed the formation of extracellular matrix and the expression of alkaline phosphatase and of mRNA of cbfa-1, ostoecalcin and type I collagen. Conclusion: From these results we concluded that the cells isolated from the umbilical cord blood were mesenchymal stem cells, which we could differentiate into osteoblast when cultured in bone formation media. In short, it is suggested that these cells could be used as a new source of stem cells, which has the probability to alternate the embryonic stem cells.

GENE-EXPRESSION PROFILING OF TITANIUM-CELL INTERACTION

  • Kim, Chang-Su;Hwang, Jung-Won;Ryu, Jae-Jun;Shin, Sang-Wan;Sohn, Sung-Hwa;Kim, Ki-Nam;Kim, Meyoung-Kon
    • The Journal of Korean Academy of Prosthodontics
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    • v.43 no.3
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    • pp.393-408
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    • 2005
  • Statement of problem. In the process of bone formation, titanium (Ti) surface roughness is an important factor modulating osteoblastic function. Purpose. This study was carried out to determine the effect of different Ti surface on biologic responses of a human osteoblast-like cell line (MG63). Materials and methods. MG63 cells were cultured on S (smooth), SLA (sandblasted largegrit & acid etching), HA (hydroxyapatite) Ti. The morphology and attachment of the cells were examined by SEM. The cDNAs prepared from total RNAs of MG63 were hybridized to a human cDNA microarray (1,152 elements). Results. The appearances of the surfaces observed with SEM were different in the three types of dental substrates. The surface of SLA and HA were shown to be rougher than S. MG63 cells cultured on SLA and HA were cell-matrix interaction. In the expression of genes involved in osseointegration, upregulated genes were bone morphogenetic protein, Villin, Integrin, Insulin-like growth factors in different surfaces. Downregulated genes were fibroblast growth factor receptor 4, Bcl 2-related protein, collagen, CD4 in different surfaces. Conclusion. The attachment and expression of key osteogenic regulatory genes were enhanced by surface roughness of the dental materials.

THE ROLE OF BONE MORPHOGENETIC PROTEIN IN THE TOOTH CULTURE (치아 기관배양시 골형성단백의 역할에 관한 연구)

  • Chung, Il-Hyuk;Chung, Jong-Hoon;Choung, Pill-Hoon
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.30 no.5
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    • pp.438-443
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    • 2004
  • Objectives : The proper development of the facial structures relies upon a sequence of tightly regulated signaling interactions between the ectoderm and mesoderm involving the participation of several families of signaling molecules. Among these, bone morphogenetic proteins (BMPs) have been suggested to be a key signal that regulates the development of the mandible and the initiation and morphogenesis of the teeth. The aim of this study was to examine the artificial development of the mandibular structures and to examine the role of BMPs on tooth morphogenesis and differentiation using an organ culture system. Materials and Methods : The tooth germs from Ed 11.5, 13.5 mice were dissected, and transplanted into the diastema of the mandible primordia. The mandibles containing the transplanted tooth germs were cultured in vitro. During this period, beads soaked with BMP4 were implanted around the transplanted tooth germs. In addition, a diastema block containing the transplanted tooth germ was dissected, then transferred to an adult mouse kidney. After the organ culture, the developing mandibular explant was removed from the kidney and prepared for the tissue specimens. Odontogeneis of the transplanted tooth germs was examined after Hematoxylin-eosin, Masson-trichrome staining. Results : Proliferation and differentiation of the tooth germs cultured in the diastema was observed. In the BMP4-treated tooth germs, the formation of the first and second molars was noted. The crown of the developing tooth showed the formation of a mature cusp with the deposition of enamel and dentin matrix. In conclusion, it was confirmed that BMP4 is involved in the formation of a dental crown and the differentiation of ameloblasts and odontoblasts of the molar tooth during the development of the transplanted tooth germs.

Osteocalcin Expression and Mineralization in Developing Tooth of Xenopus laevis

  • Park, Jung Hoe;Kwon, Ki-Tak;Park, Byung Keon;Lee, Young-Hoon
    • International Journal of Oral Biology
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    • v.40 no.1
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    • pp.1-9
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    • 2015
  • Osteocalcin (OC) is the most abundant noncollagenous protein of extracellular matrix in the bone. In an OC deficient mouse, bone formation rates are increased in cancellous and cortical bones. OC is known as a negative regulator of mineral apposition. OC is also expressed in the tooth of the rat, bovine, and human. However, little is known about OC during tooth development in Xenopus. The purpose of this study is to compare the expression of OC with mineralization in the developing tooth of Xenopus, by using von Kossa staining and in situ hybridization. At stage 56, the developmental stage of tooth germ corresponds to the cap stage, and an acellular zone was apparent between the dental papilla and the enamel organ. From stage 57, calcium deposition was revealed by von Kossa staining prior to OC expression, and the differentiated odontoblasts forming predentin were located at adjoining predentin. At stage 58, OC transcripts were detected in the differentiated odontoblasts. At stage 66, OC mRNA was expressed in the odontoblasts, which was aligned in a single layer at the periphery of the pulp. These findings suggest that OC may play a role in mineralization and odontogenesis of tooth development in Xenopus.

Topical application of herbal formula for the treatment of ligature-induced periodontitis

  • Kim, Mi Hye;Choi, You Yeon;Lee, Hye Ji;Lee, Haesu;Park, Jung-Chul;Yang, Woong Mo
    • Journal of Periodontal and Implant Science
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    • v.45 no.4
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    • pp.145-151
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    • 2015
  • Purpose: The aim of this study was to investigate the therapeutic effects of a herbal formula, PerioH-035, containing Angelica sinensis, steamed Rehmannia glutinosa, Angelica dahurica, Cimicifuga heracleifolia, and Zanthoxylum piperitum on the periodontal breakdown in a well-established ligature-induced periodontitis model in rats. Methods: Sprague-Dawley rats were randomly assigned to 1 of 4 groups: NL (non-ligatured), L (ligatured), P1 (ligatured and treated with 1 mg/mL PerioH-035), P100 (ligatured and treated with 100 mg/mL PerioH-035). Periodontitis was induced by placing a ligature around the mandibular first molars. PerioH-035 was topically applied to both sides of the first molar for 2 weeks. The right side of the mandibles was retrieved for micro-computed tomography (CT) and methylene blue staining to analyze alveolar bone loss. The left side of the mandibles was histologically analyzed by TRAP and H&E staining. The MMP-9 mRNA level in gingival tissue was investigated by RT-PCR. Results: Alveolar bone resorption was significantly reduced in the PerioH-035-treated groups. The number of dense multi-nucleated cells found to be TRAP-positive by staining in the ligatured rats was markedly decreased by PerioH-035 application. In addition, periodontal tissue destruction, especially cementum demineralization, was ameliorated in the P1 and P100 groups. Moreover, gingival tissue from the PerioH-035-treated group showed a decrease in the MMP-9 mRNA level, resulting in recovery of collagen degradation. Conclusions: These results suggest that PerioH-035 has therapeutic effects on periodontitis, and thus, PerioH-035 shows promise as a treatment for periodontitis.

Effects of Microcurrent and High Voltage Pulsed Galvanic Current Stimulation on Fibular Fracture Healing of the Rabbits (미세전류자극과 고전압 맥동직류 통전이 토끼의 비골 골절치유에 미치는 효과)

  • Ko, Seung-Hyun;Yoon, Bum-Chul;Kim, Ji-Sung;Min, Kyung-Ok
    • The Journal of the Korea Contents Association
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    • v.11 no.10
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    • pp.286-292
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    • 2011
  • This study investigated the degree of fracture healing using cathode stimulation of microcurrent, cathode and anode stimulation of High Voltage Pulsed Galvanic Current (HVPGC). Measures were performed by X-ray test and Hematoxylin-Eosin stain and Masson's trichrome stain and osteocalcin-positive immunoreactivity. In the measure of X-ray, microcurrent stimulation group revealed more rapid recovery than the groups of HVPGC's cathode and anode stimulation in bone union degrees. Microcurrent group showed significant difference statistically (p<0.05). However, the groups of HVPGC's cathode and anode stimulation didn't show significant difference statistically(p>0.05). In the histologic examination with Hematoxylin-Eosin and Masson's trichrome, microcurrent stimulation group was observed more proliferation of irregular woven bones than the groups of HVPGC's cathode and anode stimulation. Osteocalcin-positive immunoreactivity was observed more osteoblast, osteocyte, osteoclast, bone matrix than the groups of HVPGC's cathode and anode stimulation. Microcurrent stimulation can be considered an effective way during healing of fresh fracture and it can show more effective method than HVPGC's cathode and anode stimulation in the fracture healing.

Osteopontin Potentiates Pulmonary Inflammation and Fibrosis by Modulating IL-17/IFN-γ-secreting T-cell Ratios in Bleomycin-treated Mice

  • Oh, Keunhee;Seo, Myung Won;Kim, Young Whan;Lee, Dong-Sup
    • IMMUNE NETWORK
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    • v.15 no.3
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    • pp.142-149
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    • 2015
  • Lung fibrosis is a life-threatening disease caused by overt or insidious inflammatory responses. However, the mechanism of tissue injury-induced inflammation and subsequent fibrogenesis remains unclear. Recently, we and other groups reported that Th17 responses play a role in amplification of the inflammatory phase in a murine model induced by bleomycin (BLM). Osteopontin (OPN) is a cytokine and extracellular-matrix-associated signaling molecule. However, whether tissue injury causes inflammation and consequent fibrosis through OPN should be determined. In this study, we observed that BLM-induced lung inflammation and subsequent fibrosis was ameliorated in OPNdeficient mice. OPN was expressed ubiquitously in the lung parenchymal and bone-marrow-derived components and OPN from both components contributed to pathogenesis following BLM intratracheal instillation. Th17 differentiation of $CD4^+$ ${\alpha}{\beta}$ T cells and IL-17-producing ${\gamma}{\delta}$ T cells was significantly reduced in OPN-deficient mice compared to WT mice. In addition, Th1 differentiation of $CD4^+$ ${\alpha}{\beta}$ T cells and the percentage of IFN-$\gamma$-producing ${\gamma}{\delta}$ T cells increased. T helper cell differentiation in vitro revealed that OPN was preferentially upregulated in $CD4^+$ T cells under Th17 differentiation conditions. OPN expressed in both parenchymal and bone marrow cell components and contributed to BLM-induced lung inflammation and fibrosis by affecting the ratio of pathogenic IL-17/protective IFN-$\gamma$ T cells.