Calcification of MC3T3-E1 cells on titanium and zirconium

Umezawa, Takayuki; Chen, Peng; Tsutsumi, Yusuke; Doi, Hisashi; Ashida, Maki; Suzuki, Shigehiko; Moriyama, Keiji; Hanawa, Takao

doi:10.4012/dmj.2015-018

Cited by 6 publications

(9 citation statements)

References 30 publications

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“…Cell culture and induction of osteogenic differentiation As described previously 23,24) , a mouse preosteoblast cell line (MC3T3-E1, RIKEN BioResource Center, Ibaraki, Japan) was maintained in alpha modi ed Eagle s minimum essential medium (α-MEM; Gibco, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (Gibco) and an antibiotic/antimycotic (Gibco). Cells were seeded onto sterilized specimens at an approximate density of 5000 cells cm…”

Section: 3mentioning

confidence: 99%

Cytocompatibility of Ti–6Al–7Nb through High-Pressure Torsion Processing

et al. 2016

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In this study, we evaluated the cytocompatibility of high-pressure torsion (HPT)-processed Ti-6Al-7Nb alloys. Grain size of titanium (Ti) alloys decreased from 5 µm (without HPT processing, HPT-0) to 100 nm (at 2 GPa, HPT-2) or 70 nm (at 6 GPa, HPT-6) through HPT processing. Cell adhesion, proliferation, and differentiation were evaluated with mouse preosteoblast (MC3T3-E1). A locomotion trend was presented by cells cultured on HPT-2 to compare with other specimens, while, an immobilization trend was presented by cells cultured on HPT-6 to compare with other specimens. Akp2 (alkaline phosphatase 2) expression was higher in cells cultured on HPT-2 than that on HPT-0 and HPT-6, indicating enhanced osteogenic differentiation. Our results demonstrate that HPT-processed Ti-6Al-7Nb alloys offer good cytocompatibility.

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Section: 3mentioning

confidence: 99%

Cytocompatibility of Ti–6Al–7Nb through High-Pressure Torsion Processing

et al. 2016

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“…The proliferation of MC3T3‐E1 cells cultured on fsTi and mTi was evaluated by determining the number of attached cells with a Cell Counting Kit‐8 (Dojindo Laboratories, Kumamoto, Japan). The attached cells on each specimen were counted for five consecutive days according to the manufacturer's instructions . Furthermore, the absorbance of the sample at 450 nm was measured with a microplate reader (ChroMate® Microplate Reader; Awareness Technology, Inc., Palm City, FL) with a reference wavelength of 630 nm.…”

Section: Methodsmentioning

confidence: 99%

“…The attached cells on each specimen were counted for five consecutive days according to the manufacturer's instructions. 17,18 Furthermore, the absorbance of the sample at 450 nm was measured with a microplate reader (ChroMate V R Microplate Reader; Awareness Technology, Inc., Palm City, FL) with a reference wavelength of 630 nm.…”

Section: Cell Countingmentioning

confidence: 99%

“…The percentage of the total stained area of the specimen was measured automatically with the ImageJ software. [16][17][18] Cell calcification was quantified by extracting the alizarin red S from the stained specimens with 10% (vol/vol) acetic acid (Sigma-Aldrich) and measuring the absorbance at 405 nm with the ChroMate V R Microplate Reader (Awareness Technology, Inc.), according to a previously described procedure. 6,21 Statistical analysis All the reported results represent data from at least three independent determinations.…”

Section: Alizarin Red S Stainingmentioning

confidence: 99%

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Response of preosteoblasts to titanium with periodic micro/nanometer scale grooves produced by femtosecond laser irradiation

Chen

Miyake

Tsukamoto

et al. 2017

J Biomedical Materials Res

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To investigate the cellular response to designed topography in vitro, we studied the adhesion, proliferation, osteogenic differentiation, and calcification of mouse preosteoblasts (MC3T3-E1) cultured on titanium (Ti) surfaces with periodic micrometer scale grooves containing nanometer scale ripples in the vertical direction fabricated by single-shot, femtosecond laser irradiation (fsTi). The surface composition and chemical state of fsTi were almost the same as those of mirror-polished Ti without femtosecond laser irradiation (mTi). Cells cultured on fsTi were highly aligned, whereas the cell proliferation rate on fsTi was less than that on mTi. Higher gene expressions of Spp1 and Bglap1 were detected in cells cultured on fsTi than those on mTi, indicating that the periodic micro/nanometer scale grooves topography promoted osteogenic differentiation and calcification. This initial activation of osteoinduction on fsTi generated calcified deposits that were thicker and larger than those on mTi and hence, osteoconductivity was promoted on fsTi. Our findings indicate that femtosecond laser irradiation is a technique with potential for controlling biomaterial-cell interfaces and, in particular, the promotion of osseointegration of Ti. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 3456-3464, 2017.

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“…The ALP activity is very important for evaluating the osteogenic differentiation of cells 21) . After 6, 10, and 14 days of culture, the MC3T3-E1 cells cultured on all specimens were washed twice with ice-cold PBS and then distributed by sonication (UD100, Tomy Seiko, Japan) on ice.…”

Section: Osteogenic Differentiation Evaluationmentioning

confidence: 99%

Effect of strontium ions on calcification of preosteoblasts cultured on porous calcium- and phosphate-containing titanium oxide layers formed by micro-arc oxidation

et al. 2016

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INTRODUCTIONTitanium (Ti) and its alloys have been used in dentistry as implant materials because of their good mechanical properties, high corrosion resistance, and low cytotoxicity. However, their poor bone fusion properties limit their clinical applications 1) . Various techniques have been explored to modify the surfaces of Ti and its alloys to improve their osseointegration, including plasma spraying, sol-gel method, and electrochemical deposition [2][3][4][5][6][7][8][9][10][11][12] . Among these modification methods, micro-arc oxidation (MAO) has attracted special attention since it is a relatively convenient and effective technique for depositing oxide coatings on the surfaces of nonferrous metals [9][10][11][12] . More importantly, by using this technique, various desired elements can be introduced into the coated layers with porous structures 11) , such as calcium (Ca), phosphorus (P), magnesium (Mg) and strontium (Sr), which are trace elements in the human body playing a significant role in bone formation.Sr is considered a bioactive element that improves implant osseointegration. As was previously reported, Sr ions have been shown to enhance bone collagen synthesis and decrease bone resorption by inhibiting osteoclast resorbing activity [13][14][15][16][17][18][19][20] . Therefore, it is expected that a local release of Sr ions from implant surfaces would directly improve implant osseointegration.Since our previous studies have shown that MAO is a promising method to produce porous and firmly adherent bone formation concerned elements contained oxide deposition layers on metal substrates 11) , it is easy to optimize the Sr amounts introduced into the oxide deposition layers by altering MAO electrolyte compositions. Moreover, except for the first several days, a long-term stable release of metallic ions from the oxide deposition layers can be achieved by using the microporous structures formed during the MAO process 8) . In this study, Ti surfaces were modified by MAO using electrolytes with different Sr concentrations to promote calcification of preosteoblasts. Morphologies and elemental compositions of the surfaces as well as cross-sections of the Ti specimens obtained with and without MAO were characterized by scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS). A mouse preosteoblast MC3T3-E1 was utilized to study the calcification effect of Sr contents in the MAO-deposited layers on the Ti substrates in vitro. Osteogenic differentiation and calcified depositions of the MC3T3-E1 cells cultured on all specimens were evaluated using alkaline phosphatase (ALP) activity assay and alizarin red s staining, respectively. MATERIALS AND METHODS Ti substrate preparationCommercially pure Ti discs (grade 2, 8 mmφ×2.0 mm in thickness, Rare Metallic, Tokyo, Japan) were used in this study. Before the MAO treatment, Ti discs were mechanically polished with up to #800-grid SiC abrasive paper and then ultrasonically rinsed in acetone and ethanol. The obtained specimens (not subjected to...

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Calcification of MC3T3-E1 cells on titanium and zirconium

Cited by 6 publications

References 30 publications

Cytocompatibility of Ti–6Al–7Nb through High-Pressure Torsion Processing

Cytocompatibility of Ti–6Al–7Nb through High-Pressure Torsion Processing

Response of preosteoblasts to titanium with periodic micro/nanometer scale grooves produced by femtosecond laser irradiation

Effect of strontium ions on calcification of preosteoblasts cultured on porous calcium- and phosphate-containing titanium oxide layers formed by micro-arc oxidation

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