• Title/Summary/Keyword: Alizarin red S

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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.

Identification of Matrix Mineralization-Related Genes in Human Periodontal Ligament Cells Using cDNA Microarray (cDNA microarray에 의한 치주인대세포의 광물화 결절형성에 관여하는 유전자들의 분석)

  • Shin, Jae-Hee;Park, Jin-Woo;Yeo, Shin-Il;Noh, Woo-Chang;Kim, Moon-Kyu;Kim, Jung-Chul;Suh, Jo-Young
    • Journal of Periodontal and Implant Science
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    • v.37 no.sup2
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    • pp.447-463
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    • 2007
  • Periodontal ligament (PDL) cells have been known as multipotential cells, and as playing an important rolesin periodontal regeneration. The PDL cells are composed of heterogeneous cell populations which have the capacity to differentiate into either cementoblasts or osteoblasts, depending on needs and conditions. Therefore, PDL cells have the capacity to produce mineralized nodules in vitro in mineralization medium which include ascorbic acid, ${\beta}$-glycerophosphate and dexamethasone. In spite of these well-known osteoblast like properties of PDL cells, very little is known about the molecules involved in the formation of the mineralized nodules in the PDL cells. In the present study, we analysed gene-expression profiles during the mineralization process of cultured PDL cells by means of a cDNA microarray consisting of 3063 genes. Nodules of mineralized matrix were strongly stained with alizarin red S on the PDL cells cultured in the media with mineralization supplements. Among 3,063 genes analyzed, 35 were up-regulated more than two-fold at one or more time points in cells that developed matrix mineralization nodules, and 38 were down-regulated to less than half their normal level of expression. In accord with the morphological change we observed, several genes related to calcium-related or mineral metabolism were induced in PDL cells during osteogenesis, such as IGF-II and IGFBP-2. Proteogycan 1, fibulin-5, keratin 5, ,${\beta}$-actin, ${\alpha}$-smooth muscle actin and capping protein, and cytoskeleton and extracellular matrix proteins were up-regulated during mineralization. Several genes encoding proteins related to apoptosis weredifferentially expressed in PDL cells cultured in the medium containing mineralization supplements. Dkk-I and Nip3, which are apoptosis-inducing agents, were up-regulated, and Btf and TAXlBP1, which have an anti-apoptosis activity, were down-regulated during mineralization. Also periostin and S100 calciumbinding protein A4 were down-regulated during mineralization.

Osteogenesis of Human Adipose Tissue Derived Mesenchymal Stem Cells (ATMSCs) Seeded in Bioceramic-Poly D,L-Lactic-co-Glycolic Acid (PLGA) Scaffold (Bioceramic-Poly D,L-Lactic-co-Glycolic Acid(PLGA) Scaffold에 접종한 인간지방조직-유래 중간엽 줄기세포의 골 형성)

  • Kang, Yu-Mi;Hong, Soon-Gab;Do, Byung-Rok;Kim, Hae-Kwon;Lee, Joon-Yeong
    • Development and Reproduction
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    • v.15 no.2
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    • pp.87-98
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    • 2011
  • The present experiment was performed to evaluate the osteogenic differentiation of human adipose tissue derived mesenchymal stem cells (ATMSCs) seeded in bioceramic-poly D,L-latic-co-glycolic acid (PLGA) scaffold. Osteogenic differentiation of ATMSCs were induced using the osteogenic induction (OI) medium. ATMSCs were cultured with OI medium during 28 days in well plate. The proliferation of ATMSCs in OI medium group was significantly increased for 14 days of plate culture but slowed after 21 days. On the other hand, proliferation in the control group showed constant increase for 28 days of culturing. The alkaline phosphatase (ALP) activity of ATMSCs in OI medium group increased during the 21 days of culture but decreased on 28 days. However, in control group ALP activity of ATMSCs was continuously decreased as time goes. Nodule was observed at 21 days of culture in OI medium group and confirmed accumulation of calcium in cell by alizarin red staining. ATMSCs were seeded in PLGA scaffold or in Bioceramic-PLGA scaffold, and cultured with OI medium. ALP activity of ATMSCs by osteoblast differentiation in each scaffold increased on 21 days of culture and decreased rapidly on 28 days. ALP activity of ATMSCs was increased highly in Bioceramic-PLGA scaffold compared to PLGA scaffold on 21 days of culturing. SEM-EDS analysis demonstrated that calcium and phosphate content and Ca/P ratio in Bioceramic-PLGA scaffold increased higher than in PLGA scaffold. Biodegradability of scaffold at 56 days after implantation showed that Bioceramic-PLGA scaffold was more biodegradable than PLGA scaffold. The results demonstrated that the differentiation of ATMSCs to osteoblast were more effective in scaffold culture than well plate culture. Bioceramic increased cell adhesion rate on scaffold and ALP activity by osteoblast differentiation. Also, bioceramic was considered to increase the calcium and phosphate in scaffold when ATMSCs was mineralized by osteogenic differentiation. Bioceramic-PLGA scaffold enhanced the osteogenesis of seeded ATMSCs compared to PLGA scaffold.

The influence of magnet on tissue healing after immediate implantation in fresh extraction sites in dogs (성견에서 발치 후 즉시 식립 임플란트에 설치한 자석이 주위 조직에 미치는 영향)

  • Yu, Seok-Min;Cho, In-Ho;Shin, Soo-Yeon
    • The Journal of Korean Academy of Prosthodontics
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    • v.47 no.4
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    • pp.435-444
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    • 2009
  • Statement of problem: The clinical use of electric and electomagnetic fields for fracture healing applications began in the early 1970s. Since then, several technologies have been developed and shown to promote healing of fractures. Developments of these devices have been aided in recent years by basic research and several well controlled clinical trials not only in the medical field but in dentistry. Purpose: The purpose of this study was to compare alveolar bone reduction following immediate implantation using implants onto which magnets were attached in fresh extracted sockets. Material and methods: Four mongrel dogs were involved. Full buccal and lingual mucoperiosteal flaps were elevated and third and fourth premolars of the mandible were removed. Implants with magnets and implants without magnets were installed in the fresh extracted sockets and after 3 months of healing the animals were sacrificed. The mandibles were dissected and each implant sites were sampled and processed for histological examination. Results: The marginal gaps that were present between the implant and walls of the sockets at the implantation stage disappeared in both groups as a result of bone fill and resorption of the bone crest. The buccal bone crests were located apical of its lingual counterparts. At the 12 week interval the mean of marginal bone resorption in the control group was significantly higher than that of the magnet group. The majority of specimens in magnet group presented early bone formation and less resorption of the buccal marginal bone compared to the control group. Conclusion: Within the limitations of this study, it could be concluded that implants with magnets attached in the early stages of implantation may provide more favorable conditions for early bone formation and reduce resorption and remodeling of marginal bone.

Effects of Cultivated Wild Panax ginseng Extract on the Proliferation, Differentiation and Mineralization of Osteoblastic MC3T3-E1 Cells (산양삼(cultivated wild Panax ginseng) 추출물이 조골세포 활성에 미치는 영향)

  • Seo, Hyun-Ju;Eo, Hyun Ji;Kim, Hyun Jun;Jeon, Kwon Seok;Park, Gwang Hun;Hong, Se Chul;Jeong, Jin Boo
    • Korean Journal of Plant Resources
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    • v.33 no.4
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    • pp.227-236
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    • 2020
  • Panax ginseng C.A. Meyer (P. ginseng) is known to exert a wide range of pharmacological effects both in vitro and in vivo. Although studies on ginsenoside, antioxidant activity, and anticancer effect of the cultivated wild Panax ginseng (CWP) have been conducted, there is little research on the effect of CWP extract on bone metabolism. In this study, we investigated the potential anti-osteoporotic properties of CWP on the growth and differentiation of MC3T3-E1 cells. CWP significantly increased the viability and proliferation of MC3T3-E1 cells. CWP activated intracellular alkaline phosphatase (ALP) activity in MC3T3-E1 cells. In addition, CWP increased the mineralized nodules in MC3T3-E1 cells. Furthermore, CWP increased the expression of genes such as Runx2, ALP, OPN and OCN associated with osteoblast growth and differentiation in a dose-dependent manner.