• Title/Summary/Keyword: Pre-osteoblast

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Changes in pre-osteoblast cells associated with non-precious metal cores with dental implants: Pilot test (치과용 임플란트 적용 비귀금속 코어와 관련된 전조골세포의 변화)

  • Park, Jung-Hyun;Kang, Seen-Young;Kim, Jong-Woo;Kim, Jang-Ju;Kim, Woong-Chul;Kim, Ji-Hwan
    • Journal of Technologic Dentistry
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    • v.40 no.2
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    • pp.63-69
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    • 2018
  • Purpose: The purpose of this study is to investigate the non-precious metal core materials used in the dental laboratory to fabricate the implant superstructure by CAD / CAM method. And to observe and compare the morphology and distribution of the osteoblasts in relation to implant osseointegration. Methods: In this study, the mandibular right first molar tooth model was selected as an international standard to produce a single core. Using this model, the impression was made with the silicone rubber, the tooth model was scanned, and a single core was designed and 5-axis milling was performed. The materials used were Cobalt-Chromium and Nickel-Chromium, and the cores for dental implant top structures were fabricated according to the procedures of the dental labs. After the fabrication, the marginal area of the core was separated and cell culture experiment was performed. The osteoblast cells used MC3T3-E1, which is currently widely used. For morphological analysis of osteoblasts, cells were posttreated and observed using CLSM (Confocal Laser Scanning Microscope) and compared. Results: The cell adhesion behavior of the specimen surface measured by CLSM was uniformly distributed in specimen A (Cobalt-Chromium) than in specimen B (Nickel-Chromium). The distribution and changes of the cells were different in the two specimens. Conclusion : It is possible to confirm that specimen A (Cobalt-Chromium) is suitable for the living body through adhesion and proliferation of osteoblasts related to implant osseointegration in the non-precious metal superstructure used after implantation. It is considered that it is preferable to use Co-Cr when fabricating the superstructure.

Osteoporotic bone phenotype in Mats1/2 double-mutant mice (Mats1과 Mats2 이중결손 유전자 돌연변이에 의한 골감소증 기전에 대한 연구)

  • Oh, Juhwan;Choi, YunJeong;Ryu, Mi Heon;Bae, Moon-Kyoung;Kim, Hyung Joon
    • The Korean Journal of Oral and Maxillofacial Pathology
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    • v.42 no.6
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    • pp.159-165
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    • 2018
  • The Hippo pathway was originally discovered in Drosophila by genetic screening and it has been shown to be conserved in various organisms including human. Until now, the essential roles of Hippo pathway in regulating cell proliferation, apoptosis, tumorigenesis, and organ size control is extensively studied. Currently, Mats1/2 (Mob1a/1b), one of the important components in Hippo pathway, mutant mice were generated which has abnormal phenotype such as resistance to apoptosis and spontaneous tumorigenesis. Of note, Mats1/2 mutant mice also showed dental malocclusion. Therefore, in this study, we have evaluated the bone phenotype of Mats1/2 mutant mice. Although the mRNA expressions of Mats1 or Mats2 were observed in both osteoclastogenesis and osteoblastogenesis, the increase of Mats1 level was most prominent during osteoblastogenesis. The RANKL-induced osteoclast differentiation from bone marrow-derived macrophages (BMMs) was unaltered upon Mats1/2 mutation; however, the osteoblast differentiation using calvarial pre-osteoblasts was significantly reduced in Mats1/2 mutant mice compare to that of wild type mice. In accordance with in vitro results, Mats1/2 mutant mice showed decreased bone volume as well as increased trabecular separation in ${\mu}CT$ analyses. These results may provide novel prospect of the probable linkage between Hippo pathway and bone homeostasis.

Biocompatibility of Multilayer Poly Methyl Methacrylate (PMMA)/Poly Vinyl Alcohol (PVA) Bone Plate by Electrospinning Method (전기방사로 제조된 다층 Poly Methyl Methacrylate (PMMA)/Poly Vinyl Alcohol (PVA) Bone Plate의 생체적합성 평가)

  • Kwak, Kyung-A;Kim, Young-Hee;Thai, Van Viet;Lee, Byong-Taek;Song, Ho-Yeon
    • Korean Journal of Materials Research
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    • v.20 no.6
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    • pp.312-318
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    • 2010
  • Multilayer Poly methyl methacrylate (PMMA)/ Poly vinyl alcohol (PVA) bone plates were fabricated using electrospinning and in vitro investigations were carried out for pre-clinical biocompatibility studies. The initial cellular cytotoxicity of the methacrylate (PMMA)/ Poly vinyl alcohol (PVA) bone plates was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay using fibroblast-like L-929 cells. Cellular adhesion and differentiation studies were carried out using osteoblast-like MG-63 cells. As simulated body fluid (SBF) contains the same ionic concentration of body fluid and any bioactive material tends to deposit bone-like apatite on the samples surfaces into the SBF, in vitro bioactivity of the multilayer bone plates were investigated using SBF. We also studied the internal organization and tensile strength of the multilayer PMMA/PVA bone plates using micro-computed topography (${\mu}$-CT) and universal testing instrument (UTI, Korea) respectively. The cellular cytotoxicity study with MTT confirmed that the cellular viability was 78 to 90% which indicates good cyto-compatibility. Scanning electron microscopic findings revealed a good attachment and adhesion phenomenon of MG-63 cells onto the surfaces of the samples. Cellular differentiation studies also showed that osteogenic differentiation was switched on in a timely manner and affirmed along with that of the control group. Bone-like apatite formation on the surfaces was confirmed within 14 days of SBF incubation. Initial organizations of the multilayer PMMA/PVA bone plates were characterized as dense and uniform. The tensile strength of the post-pressing electronspun mat was higher than that of the pre-electronspun mat. These results suggest that a multilayer PMMA/PVA bone plate system is biocompatible, bioactive and a very good alternative bone plate system.

The Effect of Alkali- and Heat-Treated Titanium Surfaces on Differentiation of Osteoblast (티타니움 표면의 알칼리-열처리가 골모세포의 분화에 미치는 영향)

  • Kang, Choong Hee;Vang, Mong-Sook;Yang, Hong-so;Park, Sang-Won;Lim, Hyun-Pil
    • Journal of Dental Rehabilitation and Applied Science
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    • v.25 no.3
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    • pp.293-306
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    • 2009
  • In this study, the biological response of fetal rat calvarial cells on alkali- and heat-treated titanium was assessed. The results were as follows; Cell proliferation on alkali- and heat-treated surfaces showed significantly higher level than on the titanium-6aluminum-4vanadium (weight percentage: 6 % aluminum, 4 % vanadium, Ti-6Al-4V) surface (p<0.01). In ELISA analysis, concentration of $IL-1{\beta}$ and IL-6 were raised when the cells were grown to day 7. Pre-treatment with herbimycin, a known tyrosine kinase inhibitor, suppressed the production of IL-6 (p<0.01). In comparison to commercially pure titanium (grade II, cp-Ti) and Ti-6Al-4V alloy, alkali- and heat-treated titanium enhanced alkaline phosphatase activity (p<0.001). In RT-PCR analysis, alkaline phosphatase, bone sialoprotein, receptor activated nuclear factor ligand mRNA expression was increased alkali- and heat-treated titanium. Herbimycin and SB203580, p38 MAPK inhibitor, were repressed of $IL-1{\beta}-induced$ IL-6 mRNA expression. These results suggest that alkali- and heat-treated titanium stimulate osteoblasts differentiation and facilitate bone remodeling.

In Vitro and In Vivo Evaluation of Composite Scaffold of BCP, Bioglass and Gelatin for Bone Tissue Engineering

  • Kim, Woo Seok;Nath, Subrata Deb;Bae, Jun Sang;Padalhin, Andrew;Kim, Boram;Song, Myeong Jin;Min, Young Ki
    • Korean Journal of Materials Research
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    • v.24 no.6
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    • pp.310-318
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    • 2014
  • In this experiment, a highly porous scaffold of biphasic calcium phosphate (BCP) was prepared using the spongereplica method. The BCP scaffold was coated with 58S bioactive glass (BG) and sintered for a second time. The resulting scaffold was coated with gelatin (Gel) and cross-linked with [3-(3-dimethyl aminopropyl) carbodiimide] and N-Hydroxysuccinamide (EDC-NHS). The initial average pore size of the scaffold ranged from 300 to $700{\mu}m$, with more than 85 % porosity. The coating of BG and Gel had a significant effect on the scaffold-pore size, decreasing scaffold porosity while increasing mechanical strength. The material and surface properties were evaluated by means of several experiments involving scanning electron microscopy (SEM), energy-dispersive X-ray (EDX) and X-ray diffraction (XRD). Cytotoxicity was evaluated using MTT assay and confocal imaging of MC3T3-E1 pre-osteoblast cells cultured in vitro. Three types of scaffold (BCP, BCP-BG and BCP-BG-Gel) were implanted in a rat skull for in vivo evaluation. After 8 weeks of implantation, bone regeneration occurred in all three types of sample. Interestingly, regeneration was found to be greater (geometrically and physiologically) for neat BCP scaffolds than for two other kinds of composite scaffolds. However, the other two types of scaffolds were still better than the control (i.e., defect without treatment).

Anti-adipogenic and Pro-osteoblastogenic Activities of Spergularia marina Extract

  • Karadeniz, Fatih;Kim, Jung-Ae;Ahn, Byul-Nim;Kim, Mihyang;Kong, Chang-Suk
    • Preventive Nutrition and Food Science
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    • v.19 no.3
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    • pp.187-193
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    • 2014
  • This is an Open Access article distributed under the terms of the Creative Commons Attribution For decades, Spergularia marina, a local food that is popular in South Korea, has been regarded as a nutritious source of amino acids, vitamins, and minerals. While several halophytes are reported to possess distinct bioactivities, S. marina has yet to be promoted as a natural source of bioactives. In this study, the effects of S. marina on the adipogenic differentiation of 3T3-L1 fibroblasts and the osteoblastic differentiation of MC3T3-E1 pre-osteoblasts and C2C12 myoblast cells were evaluated. The anti-adipogenic effect of S. marina was assessed by measuring lipid accumulation and adipogenic differentiation marker expression. S. marina treatment significantly reduced lipid accumulation and notably decreased the gene levels of peroxisome proliferator-activated receptor ${\gamma}$, CCAAT/enhancer-binding protein ${\alpha}$, and sterol regulatory element binding protein 1c. In addition, S. marina enhanced osteoblast differentiation, as indicated by increased alkaline phosphatase activity and increased levels of osteoblastogenesis indicators, namely bone morphogenetic protein-2, osteocalcin, and type I collagen. In conclusion, S. marina could be a source of functional food ingredients that improve osteoporosis and obesity. Further studies, including activity-based fractionation, will elucidate the mechanism of action and active ingredients of S. marina, which would provide researchers with a better understanding of the nutraceutical and therapeutic applications of S. marina.

Inhibition of mRANKL Expression by Doxycycline in Rat Periodontal Ligament Cells (백서 치주인대세포에서 Doxycycline에 의한 mRANKL 발현 억제)

  • Cho, Kwan-Pyo;Cui, De-Zhe;Kim, Young-Joon
    • Journal of Periodontal and Implant Science
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    • v.36 no.2
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    • pp.335-344
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    • 2006
  • Osteoblast or bone marrow stromal cell-derived RANKL is the major effector molecule essential for osteoclastogenesis. Previous studies have shown that tetracyclines have beneficial therapeutic effects in the prevention and treatment of inflammatory bone disease including periodontal disease. Periodontal ligament cells are thought not only to play an important role in the progression of periodontal disease, but to play an important role in alveolar bone remodeling. Previous studies indicated that receptor activation of nuclear factor $\kappa\;B$ ligand (RANKL) and osteoprotegerin (OPG) are expressed in periodontal ligament cells by pro-inflammatory cytokine, such as $IL-1{\beta}$ and $TNF-{\alpha}$. This study was designed to investigate the inhibitory effect of doxycycline on RANKL and OPG mRNA in rat periodontal ligament cells induced by $IL-1{\beta}$ (1 ng/ml). The results are as follows; 1. MTT assay showed that doxycycline at the concentration of $1-50\;{\mu}g/m{\ell}$ didn't result in statistically significant cell death at day 1 and 3. 2. RANKL mRNA expression was increased to 2.6 folds by $IL-1{\beta}$. When cells were treated with doxycycline ($50{\mu}g/m{\ell}$), $IL-1{\beta}$ -induced mRANKL expression was reduced by 33%. In contrast to RANKL, OPG mRNA expression was not inhibited by pre-treatment with doxycycline. These results suggest that doxycycline decrease the expression of mRANKL resulting in regulation of osteoclastogenesisp in rat periodontal ligament cells.

Inhibition of MMP-13 mRNA expression by ginseng saponin in fetal rat calvarial cells (백서 태자 두개관세포에서 인삼 사포닌에 의한 MMP-13 mRNA 발현 억제)

  • Kim, Yang-Yi;Ciu, De-Zhe;Kim, Young-Joon
    • Journal of Periodontal and Implant Science
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    • v.35 no.2
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    • pp.277-288
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    • 2005
  • There is a potential role of collagenase-3 in alveolar bone loss and periodontal disease progression, we need to develope or find chemotherapeutic drugs or herbal agents which may regulate the expression of MMP-13. Ginseng saponin, one of the major components of Korea ginseng(panax ginseng) root, has many various biologic effects, such as cytotoxic effect, tumoricidal effects, cytokine regulations, and protein biosynthesis effect. The purpose of this study was to determine the effects of Korea red ginseng saponin on MMP-13 gene expression in osteoblasts. The experimental groups were cultured with ginseng saponin in concentration of 1.0, 10, 25, 50, 100, 250 and $500{\mu}g/ml$ for MTT assay. Primary rat calvarial cells were pre-treated for 1 hour with ginseng saponin(100 ${\mu}g/ml$) and then stimulated with $IL-1{\beta}(1.0ng/ml)$ and PTH(10 nM). MMP-13 gene expression was evaluated by RT-PCR. The results were as follows: Ginseng saponin was cytotoxic to osteoblast at concentration exceeding $250{\mu}g/ml$ for longer than 24 hours in tissue culture(p<0.01). In RT-PCR analysis, steady state MMP-13 mRNA levels were increased approximately 350% by $IL-1{\beta}$, and 400% by PTH when normalized to untreated control. $IL-1{\beta}-indued$ MMP-13 mRNA expression was reduced 50% by pretreatment with ginseng saponin. But ginseng saponin didn't inhibit MMP-13 expression from PTH stimulated cells. This results suggest that ginseng saponin Inhibit $IL-1{\beta}-indued$ MMP-13 mRNA expression.

Identification of novel susceptibility genes associated with bone density and osteoporosis in Korean women

  • Bo-Young Kim;Do-Wan Kim;Eunkuk Park;Jeonghyun Kim;Chang-Gun Lee;Hyun-Seok Jin;Seon-Yong Jeong
    • Journal of Genetic Medicine
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    • v.19 no.2
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    • pp.63-75
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    • 2022
  • Purpose: Osteoporosis is a common calcium and metabolic skeletal disease which is characterized by decreased bone mass, microarchitectural deterioration of bone tissue and impaired bone strength, thereby leading to enhanced risk of bone fragility. In this study, we aimed to identify novel genes for susceptibility to osteoporosis and/or bone density. Materials and Methods: To identify differentially expressed genes (DEGs) between control and osteoporosis-induced cells, annealing control primer-based differential display reverse-transcription polymerase chain reaction (RT-PCR) was carried out in pre-osteoblast MC3T3-E1 cells. Expression levels of the identified DEGs were evaluated by quantitative RT-PCR. Association studies for the quantitative bone density analysis and osteoporosis case-control analysis of single nucleotide polymorphism (SNPs) were performed in Korean women (3,570 subjects) from the Korean Association REsource (KARE) study cohort. Results: Comparison analysis of expression levels of the identified DEGs by quantitative RT-PCR found seven genes, Anxa6, Col5a1, Col6a2, Eno1, Myof, Nfib, and Scara5, that showed significantly different expression between the dexamethason-treated and untreated MC3T3-E1 cells and between the ovariectomized osteoporosis-induced mice and sham mice. Association studies revealed that there was a significant association between the SNPs in the five genes, ANXA6, COL5A1, ENO1, MYOF, and SCARA5, and bone density and/or osteoporosis. Conclusion: Using a whole-genome comparative expression analysis, gene expression evaluation analysis, and association analysis, we found five genes that were significantly associated with bone density and/or osteoporosis. Notably, the association P-values of the SNPs in the ANXA6 and COL5A1 genes were below the Bonferroni-corrected significance level.

Effects of Ulmus davidiana Planch(Ulmaceae) on mineralization, bone morphogenetic protein-2, alkaline phosphatase, type I collagen and collagennase-1 in bone cells (유근피가 골세포의 mineralization, bone morphogenetic protein-2, alkaline phosphatase, type I collagen 및 collagennase-1에 미치는 영향)

  • Byun, You-seok;Yoon, Jong-hwa;Hwang, Min-seob;Kim, Kap-sung;Jo, Hyun-seog
    • Journal of Acupuncture Research
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    • v.22 no.3
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    • pp.13-22
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    • 2005
  • By extracting the sample of Ulmus davidiana Planch(Ulmaceae), which was known to have the protection of damaged organ and the anti-inflammation action, it was experimented whether it is available for the application of treatment of osteoporosis. In the previous experiment, the extracts from Ulmus davidiana Planch(Ulmaceae) were confirmed to inhibit Cathepsin K through treating the cell of long bone, which contains osteoclast. Through this, it is suggested that Ulmus davidiana Planch(Ulmaceae) can play a role of prodrug as an inhibitor of absorbing bone ash in the treatment of osteoporosis. In the present experiment, a research in vitro Ulmus davidiana Planch(Ulmaceae) on the growth and sensibilization of osteoblast in a state that induced osteosis by using the cell tissue of MC3T3-El pre-osteoblastic was conducted. As a result, it could be confirmed that Ulmus davidiana Planch(Ulmaceae) has the strengthening function by enhancing the dosage and the activity of ALP depending on the time. The dosage was observed at the minimum of $50{\mu}g/m{\ell}$ and the maximum of $150{\mu}g/m{\ell}$. The enhancement in bone morphogenetic protein-2 at $100{\mu}g/m{\ell}$ UD could be observed, and it also increased the concentration of ALP mRNA within the cell of MC3T3-El. At $60{\mu}g/m{\ell}$ UD which indicated a little increase in Type I collagen mRNA for a long time of culture. However, it was shown to sharply inhibit the expression of gene in the culture between 15-20 days. These results suggest that Ulmus davidiana Planch(Ulmaceae) has an influence upon bone metabolism through thje sensibilization of osteoblast. Therefore, it could be known that utilized Ulmus davidiana Planch(Ulmaceae) can be positively applied for the general disease of bone metabolism through future studies.

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