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Secreotory Leukocyte Protease Inhibitor Regulates Bone Formation via RANKL, OPG, and Runx2 in Rat Periodontitis and MC3T3-E1 Preosteoblast

  • Seung-Yeon Lee;Soon-Jeong Jeong;Myoung-Hwa Lee;Se-Hyun Hwang;Do-Seon Lim;Moon-Jin Jeong
    • Journal of dental hygiene science
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    • v.23 no.4
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    • pp.282-295
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    • 2023
  • Background: Secretory leukocyte protease inhibitor (SLPI) protects tissues from proteases and promotes cell proliferation and healing. SLPI also reduces periodontal inflammation and alveolar bone resorption by inhibiting proinflammatory cytokine expression in rat periodontal tissues and osteoblasts. However, little is known of the role of SLPI in the expression of osteoclast regulatory factors from osteoblasts, which are crucial for the interaction between osteoblasts and osteoclasts. Therefore, we aimed to determine the effects of SLPI on the regulation of osteoclasts and osteoblasts in LPS-treated alveolar bone and osteoblasts. Methods: Periodontitis was induced in rats using LPS. After each LPS injection, SLPI was injected into the same area. Immunohistochemical analysis was performed with antibodies against SLPI, RANKL, OPG, and Runx2 in the periodontal tissue. RT-PCR and western blotting were performed to determine the expression levels of SLPI, RANKL, OPG, and Runx2 in LPS- and SLPI/LPS-treated MC3T3-E1 cells. SLPI/LPS-treated MC3T3-E1 cells were also stained with Alizarin Red S. Results: Immunohistochemical analysis showed that the expression levels of SLPI, OPG, and Runx2 were higher while that of RANKL was lower in the LPS/SLPI group relative to those in the LPS group. The mRNA and protein expression of SLPI, OPG, and Runx2 was higher in SLPI/LPS/MC3T3-E1 cells than in LPS/MC3T3-E1 cells, and RANKL expression was lower. During differentiation, OPG and Runx2 protein levels were higher whereas RANKL levels were lower in SLPI/LPS/MC3T3-E1 than in LPS/MC3T3-E1 cells on days 0, 4, 7, and 10. In addition, mineralization and matrix deposition were higher in SLPI/LPS/MC3T3-E1 than in LPS/MC3T3-E1 on days 7 and 10. SLPI decreased RANKL expression in LPS-treated alveolar bone and osteoblasts but increased the expression of OPG and Runx2. Conclusion: SLPI can be considered as a regulatory molecule that indirectly regulates osteoclast activation via osteoblasts and promotes osteoblast differentiation.

MC21/CTF and VERA multiphysics solutions to VERA core physics benchmark progression problems 6 and 7

  • Kelly, Daniel J. III;Kelly, Ann E.;Aviles, Brian N.;Godfrey, Andrew T.;Salko, Robert K.;Collins, Benjamin S.
    • Nuclear Engineering and Technology
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    • v.49 no.6
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    • pp.1326-1338
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    • 2017
  • The continuous energy Monte Carlo neutron transport code, MC21, was coupled to the CTF subchannel thermal-hydraulics code using a combination of Consortium for Advanced Simulation of Light Water Reactors (CASL) tools and in-house Python scripts. An MC21/CTF solution for VERA Core Physics Benchmark Progression Problem 6 demonstrated good agreement with MC21/COBRA-IE and VERA solutions. The MC21/CTF solution for VERA Core Physics Benchmark Progression Problem 7, Watts Bar Unit 1 at beginning of cycle hot full power equilibrium xenon conditions, is the first published coupled Monte Carlo neutronics/subchannel T-H solution for this problem. MC21/CTF predicted a critical boron concentration of 854.5 ppm, yielding a critical eigenvalue of $0.99994{\pm}6.8E-6$ (95% confidence interval). Excellent agreement with a VERA solution of Problem 7 was also demonstrated for integral and local power and temperature parameters.

Comparison of the Effectiveness of Sling Exercise and McKenzie Exercise in Patients with Acute Cervical Whiplash Associated Disorder Following Rear-end Collision

  • Jeong, Mo-Beom;Kim, Jae-Yun;Lee, Dong-Woo
    • The Journal of Korean Physical Therapy
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    • v.28 no.5
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    • pp.314-320
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    • 2016
  • Purpose: This study compared the effectiveness of sling exercise and McKenzie exercise in patients with acute cervical whiplash associated disorder (WAD) caused by rear-end collision. Methods: Thirty WAD patients were randomly assigned to one of three groups; a sling exercise (SE) group, a McKenzie exercise (McE) group, and a control group. Members of the SE and McE exercised three times a week for four weeks under the researcher's guidance. Three groups performed TENS treatment three times a week for four weeks. Diagnostic radiological equipment was used to measure whiplash injury. Visual analog scale (VAS), neck disability index (NDI), and range of motion (ROM) were used in this study. Results: The three groups showed a significant pre-post treatment difference in measures of VAS and NDI (p<0.05). The SE group showed a significant pre-post treatment difference in measures of flexion and extension changes compared to the McE and control group (p<0.05). The SE group also showed significantly greater improvement in the VAS, NDI, and ROM changes than the control group (p<0.05). Conclusion: These findings indicate that sling exercise is a stronger and more aggressive intervention for treatment of acute WAD patients.

THE EFFECTS OF LOW INTENSITY-ULTRASOUND ON THE ACTIVITY OF MC3T3-E1 CELL IN VITRO (저강도 초음파 조사시 MC3T3-E1 세포의 활성에 미치는 영향)

  • Choi, You-Sung;Lee, Sang-Chull
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.27 no.2
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    • pp.103-110
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    • 2001
  • This study was performed to observe the effect of ultrasound(1.0MHz, $0.75W/cm^2\;and\;1.0W/cm^2$) irradiation on cultured MC3T3-E1 cell, osteoblastic like cell with respect to the proliferation, protein synthesis, and alkaline phosphatase activity of the cells. The results were as follows: 1. The proliferation of MC3T3-E1 cells was increased on ultrasound irradiated group compared with control group. 2. The protein synthesis was not apparently increased on ultrasound irradiated group compared with control group. 3. The alkaline phosphatase activity level was not apparently increased on ultrasound irradiated group compared with control group. From the above results and other literatures, we could suggest that the ultrasound with the appropriate intensity and frequency may have important roles in stimulation of cell proliferation. Therefore the ultrasound may be used in the acceleration of the bone regeneration and bone fracture healing.

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Cottonseed Extract Improves the Function of Osteoblastic MC3T3-E1 Cells

  • Choi, Eun-Mi;Lee, Kyung-Hee
    • Food Quality and Culture
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    • v.2 no.2
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    • pp.85-88
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    • 2008
  • We have investigated the effects of cottonseed extract on the proliferation, differentiation and lipopolysaccharide (LPS)-induced production of local factors in murine clonal osteoblastic MC3T3-E1 cells. Ethanol extract of cotton seed ($4{\sim}63{\mu}g/mL$) significantly increased the proliferatin of MC3T3-E1 cells (p<0.05). Moreover, cottonseed extract ($10{\sim}50{\mu}g/mL$) caused a significant elevation of alkaline phosphatase (ALP) activity and collagen content in the cells. Lipopolysaccharide (LPS) is a potent stimulator of bone resorption in inflammatory diseases. We examined the effect of cottonseed extract on the LPS-induced production of tumor necrosis factor a (TNF-$\alpha$) and nitric oxide (NO) in MC3T3-E1 cells. Treatment with cottonseed extract ($10{\sim}50{\mu}g/mL$) decreased the $5{\mu}g/mL$ LPS-induced production of TNF-$\alpha$ and NO in osteoblasts, suggesting that the antiresorptive action of cottonseed extract may be mediated by decrease in these local factors. This study suggests that cottenseed may contribute to antiresorptive action against osteoblastic cells, resulting in a beneficial effect in promoting the function of osteoblastic cells.

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Relation of Expression Levels of Melanin Synthesis Genes according to the MC1R Genotypes with the Coat Color Patterns in Hanwoo, Jeju Black Cattle and Holstein (한우와 제주흑우, 홀스타인에서 MC1R 유전자형에 따른 melanin 생합성 유전자들의 발현수준과 모색 출현양상의 관계)

  • Lee, Sung-Soo;Yang, Young-Hoon;Cho, In-Cheol;Kim, Nam-Young;Ko, Moon-Suck;Jung, Ha-Yeon;Han, Sang-Hyun
    • Journal of Life Science
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    • v.19 no.3
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    • pp.384-389
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    • 2009
  • This study was carried out to elucidate the relation between expression levels of three melanin synthesis genes (Tyrosinase, Tyrosinase-related protein 1 and Dopachrome tautomerase) according to the Melanocortin-1 receptor genotypes with coat color patterns in Hanwoo cattle, Jeju black cattle and Holsteins. Using real-time semiquantitative reverse transcription-PCR assay (RT-PCR), the expression levels of these three genes were analyzed in skin tissues from four representative coat colored areas: yellowish-brown from MC1R e/e Hanwoo, wild type black from $E^+/E^+$ Jeju black cattle (JBC), and dominant black and white pied regions from $E^D/E^D$ Holstein. The TYR, TYRP1 and DCT genes showed higher expression levels of 4.5, 2.3 and 2.5 times higher in the black skin area of Holsteins than those of from JBC, respectively (p<0.001). In addition, the expression levels of these three genes from JBC were significantly higher than those from Hanwoo cattle (p<0.001). These results show that coat color phenotypes in Hanwoo cattle, JBC and Holsteins is directly correlated with TRY, TYRP1 and DCT transcription levels, which probably reflected involvement with MC1R genotypes; e/e in Hanwoo, $E^+/E^+$ in JBC and $E^D/E^D$ in Holsteins. Consequently, this study suggested that the status of MC1R protein may not only induce the transcription activities of a series of TYR and its related genes responsible for melanin synthesis, but also determine the levels of total melanin contents in bovine skin.

Effect of complex mechanical stimuli for MC3T3-E1 cells (MC3T3-E1 세포에 대한 복합 기계적 자극의 영향)

  • Kang, Kyung-Shin;Lee, Seung-Jae;Cho, Dong-Woo
    • Proceedings of the KSME Conference
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    • 2008.11a
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    • pp.1700-1703
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    • 2008
  • The purpose of this study was to examine the effects of various mechanical stimuli for MC3T3-E1 cells. Among the several mechanical stimulations, we focused on compressive stain and ultrasound. In this study, we developed a bioreactor capable of applying controlled stimuli to scaffolds. PLLA/PCL scaffold was fabricated by using salt-leaching method. We performed dynamic cell culture using preosteoblasts MC3T3-E1 cells with 1MHz, 30mW/cm2 ultrasound and 10% of compressive strain. Result of CCK-8 analysis at 1, 4, 7, 10 days showed that mechanical stimuli had no significant effect for cell proliferation. However, those stimuli influenced ALP(Alkaline phopatase) activity, which is one of differentiation marker.

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A New Record of the Genus Ectopsocus McLachlan, 1899 (Psocodea: Ectopsocidae) from South Korea (한국의 미기록속 Ectopsocus McLachlan, 1899 (다듬이목: 외다듬이벌레과)의 보고 )

  • Jaeyun Kim;Ki-Jeong Hong;Wonhoon Lee
    • Korean journal of applied entomology
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    • v.62 no.2
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    • pp.109-112
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    • 2023
  • The genus Ecopsocus is recorded from Korea for the first time with E. briggsi McLachlen, 1899. This species was collected on southern regions, Geoje-si, Jinju-si, Ulsan-si, and Jeju-do, in Korea. In this study, illustration of diagnostic characters of E. briggsi is provided.

Commercial Application of Porcine MC1R Gene Polymorphisms to Korean Pork Industry (돼지 MC1R 유전자변이의 양돈산업 적용)

  • Ha, You-Kyoung;Choi, Jung-Suk;Kim, Sang-Wook;Choi, Yang-Il;Lee, Seug-Soo;Choi, Jae-Won;Jeon, Soon-Hong;Kim, Kwan-Suk
    • Journal of Animal Science and Technology
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    • v.51 no.3
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    • pp.193-200
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    • 2009
  • The pork from black-coated pigs is famous among-consumers for better eating quality. The loci affecting black coat color was identified in pig chromosome 6 in which several genetic effects on pork quality have been reported. The melanocortin 1 receptor (MC1R) gene is a major gene which plays a key role in regulation of eumelanin (black/brown) and phaeomelanin (red/yellow). In this study, the MC1R gene polymorphism was analyzed for pig breed determination and genetic association with pork quality traits. MC1R Ala243Thr variation was analyzed to determine a specific genotype for four commercial pig breeds (Landrace, Yorkshire, Berkshire and, Duroc) and a Korean native pigs (KNP). Then we developed original KNP-specific DNA markers to determine the pork from black-coated pigs using MC1R DNA sequences. The total length of the MC1R coding sequence ranged 1451bp in KNP. KNP had the 0201 allele pertaining to $E^{D1}$ but some of the KNP had the $E^P$ allele, probably reflecting the geneticintrogression of $E^P$ allele into KNP. Furthermore, a relationship between Leu102Pro single nucleotide polymorphism (SNP) genotype and pork quality phenotype were analyzed in F2 reciprocal-crossbred population between KNP and Yorkshire. Association analysis indicated that the allele of the MC1R gene has no effect on pork quality. These results suggest that black coat-color is not directly associated with preferred pork quality, but the black-coat color pig breed may have other genetic components for superior pork quality.

Effects of Astragalus Membranaceus, Innamomum Cassia, Phellodendron Amurensis(BHH10) on MC3T3-E1 Cells Proliferation, Differntiation and Bone Mineralized Formation (MC3T3-E1 세포주에서 황기.계지.황백 처방(BHH10)의 골형성 촉진 효능 연구)

  • Lee, Mi Lim;Huh, Jeong Eun;Nam, Dong Woo;Seon, Jong In;Kang, Jung Won;Kim, Sung Hoon;Choi, Do Young;Lee, Jae Dong
    • Journal of Acupuncture Research
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    • v.29 no.6
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    • pp.11-21
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    • 2012
  • Objectives : BHH10 is traditional medicine herb used for enhancing body resistance against various diseases. The aim of this study was to identify BHH10 extract induces osteogenic activity in human osteoblast-like MC3T3-E1 cells. Methods : MC3T3-E1, pre-osteoblast cell line, were treated with BHH10 of various concentrations($0.1{\mu}g/mL$, $1{\mu}g/mL$, $10{\mu}g/mL$). And then, the effect of BHH10 on osteoblast differentiation was examined by alkaline phosphatase(ALP) activity, von Kossa staining and RT-PCR for osteoblast differentiation markers such as osteocalcin(OCN), osteopontin(OPN). Results : BHH10 had dose-dependent effect on the viability of osteoblastic cells, and dose-dependently increased alkaline phosphatase(ALP) activity. BHH10 markedly increased mRNA expression for OCN, OPN in MC3T3-E1 cells. Also, BHH10 significantly induced mineralization in the culture of MC3T3-E1 cells. Conclusions : In conclusion, these results propose that BHH10 can play an important role in osteoblastic bone formation, osteogenesis, and may possibly lead to the development of bone-forming drugs.