• Title/Summary/Keyword: Cultured osteoblast

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DNA microarray analysis of gene expression of MC3T3-E1 osteoblast cell cultured on anodized- or machined titanium surface

  • Park, Ju-Mi;Jeon, Hye-Ran;Pang, Eun-Kyoung;Kim, Myung-Rae;Kang, Na-Ra
    • Journal of Periodontal and Implant Science
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    • v.38 no.sup2
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    • pp.299-308
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    • 2008
  • Purpose: The aim of this study was to evaluate adhesion and gene expression of the MC3T3-E1 cells cultured on machined titanium surface (MS) and anodized titanium surface (AS) using MTT test, Scanning electron micrograph and cDNA microarray. Materials and Methods: The MTT test assay was used for examining the proliferation of MC3T3-E1 cells, osteoblast like cells from Rat calvaria, on MS and AS for 24 hours and 48 hours. Cell cultures were incubated for 24 hours to evaluate the influence of the substrate geometry on both surfaces using a Scanning Electron Micrograph (SEM). The cDNA microarray Agilent Rat 22K chip was used to monitor expressions of genes. Results: After 24 hours of adhesion, the cell density on AS was higher than MS (p < 0.05). After 48 hours the cell density on both titanium surfaces were similar (p > 0.05). AS had the irregular, rough and porous surface texture. After 48 hours incubation of the MC3T3-E1 cells, connective tissue growth factor (CTGF) was up-regulated on AS than MS (more than 2 fold) and the insulin-like growth factor 1 receptor was down-regulated (more than 2 fold) on AS than MS. Conclusion: Microarray assay at 48 hours after culturing the cells on both surfaces revealed that osteoinductive molecules appeared more prominent on AS, whereas the adhesion molecules on the biomaterial were higher on MS than AS, which will affect the phenotype of the plated cells depending on the surface morphology.

Effect of NMDA receptor antagonist on Osteoblasts Damaged by Methylmercuric Chloride (메틸수은으로 손상된 골모세포에 대한 NMDA 수용체길항제의 영향)

  • Ha Dae Ho;Yang Hyun Woong;Lee Joung Hwa;Lee Kang Chag
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.412-415
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    • 2003
  • In order to elucidate the mechanism between cytotoxicity of methhylmercuric chloride(MMC) and oxygen radicals in cultured osteoblasts of neonatal mouse, cell viability was measured by MTT assay in osteoblasts treated with 1~50 μM MMC for 30 hours. And also, the protective effect of N-methyl D-aspartate(NMDA) receptor antagonist, D-2-amino-5-phosphovaleric acid(APV) was examined by cell viability in these cultures. Cell viability was significantly decreased in dose dependently after exposure of 30 μM MMC to cultured osteoblasts for 30 hours. Protective effect of APV against MMC-mediated toxicity was very effective in these cultures. From above the results, it suggests that MMC is toxic in cultured mouse osteoblasts and NMDA receptor antagonist such as APV is effective in blocking the osteotoxicity induced by MMC.

Effects of Irradiated Frozen Allogenic Bone and Musculoskeletal Transplant Foundation on Bone Formation in Human Fetal Osteoblasts (사람 태아 골모 세포에 대한 냉동 동종골과 근골격이식재의 골형성 유도에 관한 효과)

  • Yoon, Ho-Sang;Pi, Sung-Hee;Yun, Hyeong-Geun
    • Journal of Periodontal and Implant Science
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    • v.36 no.2
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    • pp.435-448
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    • 2006
  • The purpose of this study was to investigate the effects of ICB(Irradiated frozen allogenic bone, Rocky Mountain Tissue Bank, USA) and MTF(Decalcified freeze-dried bone allograft, Musculoskeletal Transplant Foundation, USA) on the cell proliferation and differentiation of human fetal osteoblasts. Human fetal osteoblasts (hFOB1) were cultured with $10\;ng/m{\ell}$of ICB and MTF. The negatvie control group was cultured with DMSO and positive control group was cultured with BMF ($2\;ng/m{\ell}$). MIT was performed to examine the viability of the cell, and alkaline phosphatase activity was analyzed to examine the mineralization. Calcium accumulation was also evaluated. ICB and MTF did not increase the rate of the cellular proliferation of hFOB1s while they enhanced ALP and calcium accumulation. The expression of osteocalcin (OC) and bone silaloprotein (BSP) increased in hFOB1 treated with ICB and MTF ($10\;ng/m{\ell}$). These results suggest that ICB and MTF stimulate osteoblastic activity of the hFOBl.

Study on the Effect of Flos Carthami on Cultured Osteoblasts Damaged by Methylmercuric Chloride (메틸수은으로 손상된 골모세포에 대한 홍화의 효과에 관한 연구)

  • Min Bu Ki;Hong Gi Youn;Oh Yeon Kyun;Shin Yong Il;Han Sun Hee;Lee Sang Bork;Shin Min Kyo;Jeon Byung Hoon;Song Ho Jun;Ryu Do Gon;Park Seung Taeck
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.4
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    • pp.764-767
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    • 2002
  • To evaluate the osteotoxic effect of methylmercuric chloride(MMC) on cultured mouse osteoblasts, cytotoxic effect was measured by MTT assay after cultured mouse osteoblasts were incubated with various concentrations of MMC for 20 hours. The protective effect of Flos Carthami(FC) against MMC-induced osteotoxicity was also examined in these cultures. MMC decreased cell viability of cultured mouse osteoblasts remarkably in a dose- and time-dependent manners. In protective effect of FC was remarkably effective in blocking the osteotoxicity induced by MMC. From aboved the results, it is suggested that MMC induce osteotoxicity, and the selective herba extract such as FC is very effective in blocking MMC-mediated neurotoxicity on cultured mouse osteoblasts.

Establishment of Validation Methods to Test the Biocompatibility of Titanium Dioxide

  • Kim, Mi-Ju;Lim, Hee-Joung;Lee, Byung Gun;Kim, Jong-Hoon;Choi, Jinsub;Kang, Hee-Gyoo
    • Bulletin of the Korean Chemical Society
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    • v.34 no.6
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    • pp.1857-1863
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    • 2013
  • Most of biomaterials come in direct contact with the body, making standardized methods of evaluation and validation of biocompatibility an important aspect to biomaterial development. However, biomaterial validation guidelines have not been fully established, until now. This study was to compare the in vitro behavior of osteoblasts cultured on nanomaterial $TiO_2$ surfaces to osteoblast behavior on culture plates. Comparisons were also made to cells grown in conditioned media (CM) that creates an environment similar to the in vivo environment. Comparisons were made between the different growth conditions for osteoblast adhesion, proliferation, differentiation, and functionality. We found that the in vivo-like system of growing cells in concentrated CM provided a good validation method for biomaterial development and in vivo implant therapy. The $TiO_2$ materials were biocompatible, showing similar behavior to that observed in vivo. This study provided valuable information that would aid in the creation of guidelines into standardization and evaluation of biocompatibility in $TiO_2$ biomaterials.

Graphene accelerates osteoblast attachment and biomineralization

  • Ren, Jia;Zhang, Xiaogang;Chen, Yao
    • Carbon letters
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    • v.22
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    • pp.42-47
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    • 2017
  • In this paper, the in vitro biocompatibility of graphene film (GF) with osteoblasts was evaluated through cell adhesion, viability, alkaline phosphatase activity, F-actin and vinculin expressions, versus graphite paper as a reference material. The results showed that MG-63 cells exhibited stronger cell adhesion, better proliferation and viability on GF, and osteoblasts cultured on GF exhibited vinculin expression throughout the cell body. The rougher and wrinkled surface morphology, higher elastic modulus and easy out-of-plane deformation associated with GF were considered to promote cell adhesion. Also, the biomineralization of GF was assessed by soaking in simulated body fluid, and the GF exhibited enhanced mineralization ability in terms of mineral deposition, which almost pervaded the entire GF surface. Our results suggest that graphene promotes cell adhesion, activity and the formation of bone-like apatite. This research is expected to facilitate a better understanding of graphene-cell interactions and potential applications of graphene as a promising toughening nanofiller in bioceramics used in load-bearing implants.

Adhesion of Human Osteoblasts Cell on TiN Thin Film Deposited by Cathodic Arc Plasma Deposition

  • Pham, Vuong Hung;Kim, Sun-Kyu;Le, Vinh Van;Kwon, Byoung-Se
    • Journal of the Korean institute of surface engineering
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    • v.41 no.6
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    • pp.264-268
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    • 2008
  • Interaction between human osteoblast and TiN films was conducted in vitro. TiN films were produced by cathodic arc plasma deposition. The surface was characterized by atomic force microscopy (AFM). TiN films, glass substrates and Ti films were cultured with human osteoblasts for 48 and 72 h hours. Actin stress fiber patterns and microtubules of osteoblasts were found slightly more organized and distributed on TiN films compared to those on the Ti films and the glass substrates. Human osteoblasts also showed slightly higher cell attachment, proliferation, and focal contact adhesion on TiN films compared to those on Ti films and glass substrates. Our results demonstrated that TiN films showed slightly better cellular adhesion of osteoblasts than Ti films and glass substrates in a short-time culture period.

The Effect of Intermittent Compressive Loading to Growth of Pre-osteoblast Cells (간헐적인 압축하중이 조골세포주 성장에 미치는 영향)

  • Choi, Sung-Kyu;Park, Jeong-Hun;Lee, Seung-Jae;Lee, In-Hwan;Kang, Sang-Sun
    • Journal of the Korean Society for Precision Engineering
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    • v.27 no.2
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    • pp.153-159
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    • 2010
  • Recently, it has been reported that mechanical stimulation takes a role in improving cell growth. Also, became generally known that skeletal system as bone or cartilage tissues take influence of compression loading. In this study, we fabricated a custom-made bioreactor and analyzed that conditions of compressive loading would influence cell growth. To compare the effect of intermittent compressive loading on cell-encapsulated agarose scaffold, we cultured preosteoblast cell (MC3T3-E1 cells) statically and dynamically. And dynamic culture conditions were produced by changing parameters such as the iteration time and interval delay time. Also, cellencapsulated agarose scaffold were subjected to 10 % dynamic compressive strain at 1㎐ frequency for 7 days. After cell culture, cell proliferation was assessed with PI stain assay for fluorescence images and flow cytometry (FACS).

Biocompatibility of Low Modulus Porous Titanium Implants Fabricated by Spark Plasma Sintering (방전플라즈마소결법에 의해 제조된 저탄성 타이타늄 다공질체의 생체적합성 평가)

  • Song, Ho-Yeon;Kim, Young-Hee;Chang, Se-Hun;Oh, Ik-Hyun
    • Korean Journal of Materials Research
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    • v.17 no.2
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    • pp.107-114
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    • 2007
  • Porous Ti compacts were fabricated by spark plasma sintering (SPS) method and their in vitro and in vivo biocompatibilities were investigated. Alkaline phosphatase (ALP) activity representing the activity of osteoblast was increased when osteoblast-like MG-63 cells were cultured on the Ti powder surface. Some genes related to cell growth were over-expressed through microarray analysis. The porous Ti compact with 32.2% of porosity was implanted in the subcutaneous tissue of rats to confirm in vivo cytotoxicity. 12 weeks post-operation, outer surface and inside the porous body was fully filled with fibrous tissue and the formation of new blood vessels were observed. No inflammatory response was confirmed. To investigate the osteoinduction, porous Ti compact was implanted in the femur of NZW rabbits for 4 months. Active in-growth of new bone from the surrounded compact bone was observed around the porous body. From the results, The porous Ti compacts fabricated by spark plasma sintering might be available for the application of the stem part of artificial hip joint.

The Effects of HAp and BCP Nano Powders Synthesized by Microwave-Assisted Synthesis on the Activation of Osteoblast and Osteoclast (마이크로웨이브법에 의해 제조된 HAp 및 BCP 분말이 뼈모세포 및 파골세포의 활성에 미치는 영향)

  • Song, Ho-Yeon;Min, Young-Ki;Yang, Hun-Mo;Mang, Joo-Yang
    • Korean Journal of Materials Research
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    • v.17 no.12
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    • pp.669-675
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    • 2007
  • Hydroxyapatite (HAp) and biphasic calcium phosphate (BCP) nano powders were synthesized using the microwave-assisted synthesis process dependent on pH and microwave irradiation time. The average size of a powder was less than 100 nm in diameter. Through in-vitro cytotoxicity tests by an extract dilution method, the HAp and BCP nano powders have shown to be cytocompatible for L-929 fibroblast cells, osteoblastlike MG-63 cells and osteoclast-like Raw 264.7 cells. The activation of osteoblast was estimated by alkaline phosphatase (ALP) activity. When the HAp and BCP were treated to MG-63 cells, alkaline phosphatase activities increased on day 3, compared with those of the untreated cells. Also, the collagen fibers increased when the HAp and BCP powders suspension were treated to MG-63 cells, compared to those of the untreated cells. Quantitative alizarin red S mineralization assays showed a trend toward increasing mineralization in osteoblast cultured with powder suspension. In conclusion, hydroxyapatite and biphasic calcium phosphate appeared to be a bone graft substitute material with optimal biocompatibility and could be further applied to clinical use as an artificial bone graft substitute.