• Title/Summary/Keyword: osteoblast adhesion

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EFFECT OF TITANIUM SURFACE ROUGHNESS ON CELL ADHESION OF HUMAN OSTEOBLAST-LIKE CELLS (MG63)

  • Yim Soon-Ho
    • The Journal of Korean Academy of Prosthodontics
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    • v.42 no.3
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    • pp.261-266
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    • 2004
  • Statement of problem. The effects of surface roughness have not or insufficiently been analyzed on earlier events such as cell adhesion though cell behavior most germane to implant performance is cell adhesion. Purpose. The purpose of this study was to evaluate cell adhesion of osteoblast-like cells (MG63) onto three types of titanium disks with varying roughness using the Elisa assay. Materials and methods. Representative disks from each group (SLA, HA, machined) were subjected to surface analysis and surface roughness was measured by the optical interferometer (Accura 2000, Intekplus Co., Seoul, Korea). Following this, MG63 cells were cultured on the titanium disks and released. Cell adhesion measurements using the Elisa assay were performed specifically at three points: after 24, 48, and 72 hours of culture. Results. Among the 3 types of surface analyzed, the SLA surface was the roughest with a Ra value of $1.114{\mu}m$ followed by HA coated surface and machined surface, consecutively. The optical density values for the SLA surface group was significantly higher than that of the machined and HA coated surface groups following 24 and 48 hours of culture. The cell culture on HA coated surface showed significantly higher values compared to the machined surface following 24, 48 and 72 hours of culture. Conclusion. The results suggest that surface treatment of titanium surfaces enhanced cell adhesion of human osteoblast-like cells (MG63).

Hydrothermal Synthesis of $TiO_2$ Nanowire Array for Osteoblast Adhesion

  • Yun, Young-Sik;Kang, Eun-Hye;Hong, Min-Eui;Yun, In-Sik;Kim, Yong-Oock;Yeo, Jong-Souk
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.275-275
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    • 2013
  • Osteoblast is one of cells related with osseointegration and many research have conducted the adhesion of osteoblast onto the surface of implant. In the osseointegration, biocompatibility of the implant and cell adhesion to the surface are important factors. The researches related to cell adhesion have a direction from micro-scaled surface roughness to nano-scaled surface roughness with advancing nanotechnology. A cell reacts and sense to stimuli from extracellular matrix (ECM) and topography of the ECM [1]. Thus, for better osseointegration, we should provide an environment similar to ECM. In this study, we synthesize TiO2 nanowires using hydrothermal reaction because TiO2 provides inertness to titanium on its surface and enables it used as an implant material for the orthopedic treatment such as fixation of the bone fracture [2]. Ti substrate is immersed into NaOH aqueous solution. The solution are heated at $140{\sim}200^{\circ}C$ for various time (10~720 minutes). After heat treatment, we take out the sample and immerse it into HCl aqueous solution for 1 hour. The acid treated sample is heated again at $500^{\circ}C$ for 3 hours [3]. Then, we culture osteoblast on the TiO2 nanowires. For investigating cell adhesion onto nanostructured surface, we conduct several tests such as MTT assay, ALP (Alkaline phosphatase) activity assay, measuring calcium expression, and so on. These preliminary results of the cell culture on the nanowires are foundation for investigating cell-material interaction especially with nanostructure interaction.

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Bioactive characteristics of an implant surface coated with a pH buffering agent: an in vitro study

  • Pae, Hyung-Chul;Kim, Su-Kyoung;Park, Jin-Young;Song, Young Woo;Cha, Jae-Kook;Paik, Jeong-Won;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.49 no.6
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    • pp.366-381
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    • 2019
  • Purpose: The purpose of this study was to evaluate the effectiveness of conventional sandblasted, large-grit, acid-etched (SLA) surface coated with a pH buffering solution based on surface wettability, blood protein adhesion, osteoblast affinity, and platelet adhesion and activation. Methods: Titanium discs and implants with conventional SLA surface (SA), SLA surface in an aqueous calcium chloride solution (CA), and SLA surface with a pH buffering agent (SOI) were prepared. The wetting velocity was measured by the number of threads wetted by blood over an interval of time. Serum albumin adsorption was tested using the bicinchoninic acid assay and by measuring fluorescence intensity. Osteoblast activity assays (osteoblast adhesion, proliferation, differentiation, mineralization, and migration) were also performed, and platelet adhesion and activation assays were conducted. Results: In both the wetting velocity test and the serum albumin adsorption assay, the SOI surface displayed a significantly higher wetting velocity than the SA surface (P=0.000 and P=0.000, respectively). In the osteoblast adhesion, proliferation, differentiation, and mineralization tests, the mean values for SOI were all higher than those for SA and CA. On the osteoblast migration, platelet adhesion, and activation tests, SOI also showed significantly higher values than SA (P=0.040, P=0.000, and P=0.000, respectively). Conclusions: SOI exhibited higher hydrophilicity and affinity for proteins, cells, and platelets than SA. Within the limits of this study, it may be concluded that coating an implant with a pH buffering agent can induce the attachment of platelets, proteins, and cells to the implant surface. Further studies should be conducted to directly compare SOI with other conventional surfaces with regard to its safety and effectiveness in clinical settings.

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

  • Pham, Vuong-Hung;Kim, Sun-Kyu
    • Journal of the Korean institute of surface engineering
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    • v.42 no.5
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    • pp.203-207
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    • 2009
  • Interaction between human osteoblast (hFOB 1.19) and CrN films was conducted in vitro. CrN films were produced by cathodic arc plasma deposition. The surface was characterized by atomic force microscopy (AFM). CrN films, glass substrates and TiN films were cultured with human osteoblasts for 48 and 72 hours. Actin stress fiber patterns and cell adhesion of osteoblasts were found less organized and weak on CrN films compared to those on the glass substrates and the TiN films. Human osteoblasts also showed less proliferation and less distributed microtubule on CrN films compared to those on glass substrates and TiN films. Focal contact adhesion was not observed in the cells cultured on CrN films, whereas focal contact adhesion was observed well in the cells cultured on glass substrates and TiN films. As a result, the CrN film is a potential candidate as a surface coating to be used for implantable devices which requires minimal cellular adhesion.

Thrombospondins Mediate the Adhesion of Osteoblast to Extracelluar Matrix

  • Lim, Dong-Jin;Bae, In-Ho;Jeong, Byung-Chul;Kim, Sun-Hun;Park, Bae-Keun;Kang, In-Chul;Lee, Shee-Eun;Song, Sang-Hun;Koh, Jeong-Tae
    • International Journal of Oral Biology
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    • v.33 no.3
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    • pp.105-111
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    • 2008
  • Thrombospondins (TSP-1, TSP-2) are secretory extracellular glycoproteins that are involved in a variety of physiological processes such as tumor cell adhesion, invasion, and metastasis. The present study was undertaken to elucidate the involvement of thrombospondins in the adhesion of osteoblast-like cells using the TSP-1 or TSP-2 antisense MG63 and MC3T3-E1 cell lines. For downregulation of TSPs expression, we prepared antisense constructs for TSP-1 and TSP-2 using the pREP4 an episomal mammalian expression vector, which be able to produce the specific antisense oligonucleotides around chromosome. MG63 and MC3T3-E1 osteoblast-like cells were transfected with the antisense constructs and nonliposomal Fugene 6, and then selected under hygromycin B (50 ${\mu}g/ml$) treatment for 2 weeks. Western blot analysis revealed that expression of the TSP proteins was downregulated in the antisense cell lines. The cell adhesion assay showed that adhesive properties of TSP-1 and TSP-2 antisense MG63 cells on the polystyrene culture plate were reduced to 17% and 21% of the control cells, respectively, and those of the TSP-1 and TSP-2 antisense MC3T3-E1 cells also decreased to 19% and 27% of control, respectively. Adhesion of TSP-1 and TSP-2 antisense MC3T3-E1 cells on Type I collagen-coated culture plate decreased to 27% and 76%, respectively. These results indicate that TSP-1 and TSP-2 proteins may have an important role in adhesion of osteoblast-like cells to extracellular matrix.

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.

CELLULAR ATTACHMENT AND GENE EXPRESSION OF OSTEOBLAST-LIKE CELLS ON ZIRCONIA CERAMIC SURFACES

  • Pae, Ah-Ran;Lee, Hee-Su;Kim, Hyeong-Seob; Baik, Jin;Woo, Yi-Hyung
    • The Journal of Korean Academy of Prosthodontics
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    • v.46 no.3
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    • pp.227-237
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    • 2008
  • STATEMENT OF PROBLEM: Zirconium oxide can be a substitute to titanium as implant materials to solve the esthetic problems of dark color in the gingival portion of implant restorations. PURPOSE: This study was performed to define attachment and growth behavior of osteoblast- like cells cultured on grooved surfaces of zirconium oxide and evaluate the genetic effect of zirconium oxide surfaces using the reverse transcriptase-polymerase chain reaction (RT-PCR). MATERIAL AND METHODS: MC3T3-E1 cells were cultured on (1) commercially pure titanium discs with smooth surface (T group), (2) yttrium-stabilized tetragonal zirconia polycrystal (Y-TZP) with machined surface (ZS group), and (3) Y-TZP with $100{\mu}m$ grooves (ZG group). Cell proliferation activity was evaluated through MTT assay and cell morphology was examined by SEM. The mRNA expression of Runx2, alkaline phosphatase, osteocalcin, TGF-${\beta}1$, IGF-1, G3PDH in E1 cells were evaluated by RT-PCR. RESULTS: From the MTT assay, after 48 hours of adhesion of MC3T3-E1 cells, the mean optical density value of T group and ZG group significantly increased compared to the ZS group. SEM images of osteoblast-like cells showed that significantly more cells were observed to attach to the grooves and appeared to follow the direction of the grooves. After 24 hours of cell adhesion, more spreading and flattening of cells with active filopodia formation occurred. Results of RT-PCR suggest that T group, ZS group, and ZG group showed comparable osteoblast-specific gene expression after 24 hours of cell incubation. CONCLUSION: Surface topography and material of implants can play an important role in expression of osteoblast phenotype markers. Zirconia ceramic showed comparable biological responses of osteoblast-like cells with titanium during a short-time cell culture period. Also, grooves influence cell spreading and guide the cells to be aligned within surface grooves.

Effect of Pore Structures of a Ti-49.5Ni (at%) Alloy on Bone Cell Adhesion (Ti-49.5Ni (at%)합금의 다공성 구조가 뼈 세포 흡착에 미치는 영향)

  • Im, Yeon-Min;Choi, Jung-Il;Khang, Dong-Woo;Nam, Tae-Hyun
    • Korean Journal of Materials Research
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    • v.22 no.2
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    • pp.66-70
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    • 2012
  • Ti-Ni alloys are widely used in numerous biomedical applications (e.g., orthodontics, cardiovascular science, orthopaedics) due to their distinctive thermomechanical and mechanical properties, such as the shape memory effect, superelasticity and low elastic modulus. In order to increase the biocompatibility of Ti-Ni alloys, many surface modification techniques, such as the sol-gel technique, plasma immersion ion implantation (PIII), laser surface melting, plasma spraying, and chemical vapor deposition, have been employed. In this study, a Ti-49.5Ni (at%) alloy was electrochemically etched in 1M $H_2SO_4$+ X (1.5, 2.0, 2.5) wt% HF electrolytes to modify the surface morphology. The morphology, element distribution, crystal structure, roughness and energy of the surface were investigated by scanning electron microscopy (SEM), energy-dispersive Xray spectrometry (EDS), X-ray diffractometry (XRD), atomic force microscopy (AFM) and contact angle analysis. Micro-sized pores were formed on the Ti-49.5Ni (at%) alloy surface by electrochemical etching with 1M $H_2SO_4$+ X (1.5, 2.0, 2.5) wt% HF. The volume fractions of the pores were increased by increasing the concentration of the HF electrolytes. Depending on the HF concentration, different pore sizes, heights, surface roughness levels, and surface energy levels were obtained. To investigate the osteoblast adhesion of the electrochemically etched Ti-49.5Ni (at%) alloy, a MTT test was performed. The degree of osteoblast adhesion was increased at a high concentration of HF-treated surface structures.

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.

Osteoblast adhesion and differentiation on magnesium titanate surface (마그네슘 티타네이트 표면의 조골세포 부착도와 분화)

  • Choi, Seung-Min;Lee, Jae-Kwan;Ko, Sung-Hee;Um, Heung-Sik;Chang, Beom-Seok
    • Journal of Periodontal and Implant Science
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    • v.35 no.4
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    • pp.851-861
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    • 2005
  • The nature of the implant surface can directly influence cellular response, ultimately affecting the rate and quality of new bone tissue formation. The aim of this in vitro study was to investigate if human osteoblast-like cells, Saos-2, would respond differently when plated on disks of magnesium titanate and machined titanium. Magnesium titanate disks were prepared using Micro Arc Oxidation(MAO) methods. Control samples were machined commercially pure titanium disks. The cell adhesion, proliferation and differentiation were evaluated by measuring cell number, and alkaline phosphatase(ALPase) activity at 1 day and 6 day after plating on the titanium disks. Measurement of cell number and ALPase activity in Saos-2 cells at 1 day did not demonstrate any difference between machined titanium and magnesium titanate. When compared to machined titanium disks, the number of cells was reduced on the magnesium titanate disks at 6 day, while ALPase activity was more pronounced on the magnesium titanate. Enhanced differentiation of cells grown on magnesium titanate samples was indicated by decreased cell proliferation and increased ALPase activity.