Culture human osteoblasts

Jevon M, Guo C, Ma B, et al Mechanisms of internalization of Staphylococcus aureus by cultured human osteoblasts. Infect Immun 1999 67 2677-2681. 

Torricelli P, Fini M, Giavaresi G, Giardino R, Gnudi S, Nicolini A, Carpi A. L-Arginine and L-lysine stimulation on cultured human osteoblasts. Biomed Pharma-cother 2002 56 492 97. 

Human osteoblast-like MG63 cells were cultured on the macroporous chitosan scaffolds reinforced with hydroxyapatite or calcium phosphate invert glass were fabricated using a thermally induced phase separation technique. 

Figure 22. Human embryonic kidney cells (A), rat vascular smooth muscle cells (B, C) and human osteoblast-like MG 63 cells (D) in cultures on micropattemed surfaces. A, B PTFE irradiated with UV light produced by a Xe2 -excimer lamp for 30 min in an ammonia atmosphere through a mask with holes 100 pm in diameter and center-to-center distance 300 pm C PE irradiated with Ar ions (energy 150 keV, ion dose lO ions/cm ) through a mask with holes 100 pm in diameter and center-to-center distance 200 pm fullerenes Qo deposited through a mask with rectangular holes with an average size of 128 3 pm per 98 8 pm on glass coverslips. Day 7 after seeding. A native cells in an inverted phase-contrast microscope B, C cells stained with hematoxylin and eosin, Olympus microscope IX 50 D cells stained with fluorescence-based LIVE/DEAD viability/cytotoxicity kit (Invitrogen), Olympus microscope IX 50. Bars 300 pm (A), 200 pm (B, D), Imm (C) [10,11].

Figure 27. Human osteoblast-like MG 63 cells in cultures on porous (A) or fibrous (B) poly(L-lactide-co-glycolide) scaffolds. A A summarizing picture of horizontal optical sections. The depth of cell ingrowth into the pores (average pore diameter of 400-600 mm) is indicated by spectral colors (blue 0-60 mm, green 80-160 mm, yellow 180-220 mm, orange 240-300 mm, red 320-400 mm, violet 420-480 mm). Day 14 after seeding, cells stained with propidium iodide. B cells grown for 4 days in static culture followed by 2 days in dynamic perfusion cell culture system. Cell membrane stained with Texas Red C2-maleimide and the nuclei counterstained with Hoechst 33342. Leica TCS SP2 confocal microscope, objective 5x (A) or lOx (B) [37].

Finally, the modified surfaces have been tested in cell culture with human osteoblasts. Although cell adhesion was found to be roughly equivalent on treated and raw surfaces, cell proliferation greatly improved after 10 days on the fluoridated surface [159]. In addition, the fluoridation treatment considerably reduced the degradation of the coating. 

Henriksen Z, Hiken JF, Steinberg TH, Jorgensen NR. 2006. The predominant mechanism of intercellular calcium wave propagation changes during long-term culture of human osteoblast-like cells. Cell Calcium 39 435 14. 

Schroder and co-workers (Schroder etal., 1999,2000) studied PolyP metabolism in bone tissues and osteblast cultures. They revealed that PolyP metabolism in human osteoblasts was modulated by stimulators of osteoblast proliferation and differentiation (Leyhausen et al., 1998). A combined treatment of the cells with dexamethasone, ft-glycerophosphate, epidermal growth factor (EGF), and ascorbic acid resulted in a dramatic decrease in PolyP content. This decrease is caused mainly by a decrease in the amount of soluble long-chain PolyPs. The amount of this PolyP fraction, but not the amount of insoluble long-chain PolyPs, further decreases after additional treatment of the cells with la, 25-dihydroxy vitamin D3. The decrease in PolyP content during treatment with dexamethasone, ft-glycerophosphate, EGF and ascorbic acid is accompanied by a decrease in exopolyphosphatase activity. However, additional treatment with la, 25-dihydroxyvitamin D3 results in a significant increase of the enzyme activity. Therefore, it is reasonable to assume that PolyP... 

Human osteoblasts like cells osteosarcoma SaOS-2 cells were cultured for 24 hours, after which varying doses of a water dispersible microemulsion preparation of lycopene or vehicle of the same dilution were added. The cells were further cultured for 24 to 144 h and the cell numbers were counted. Lycopene at 10 and 10 M had significant stimulatory effects on cell numbers, compared with the corresponding vehicle treatment, at all time points from 24 h to 144 h. The effects of lycopene on activity of the differentiation marker alkaline phosphatase activity in the absence or presence of dexamethasone were shown to be dependent on osteoblasts of human origin [79]. 

Tomas, H., Carvalho, G.S., Fernandes, M.H., Freire, A.P., and Abrantes, L.M. (1996) Effects of Co-Cr corrosion products and corresponding separate metal ions on human osteoblast-like cell cultures. J. 

Amaral IF et al (2007) Attachment, spreading and short-term proliferation of human osteoblastic cells cultured on chitosan films with different degrees of acetylation. J Biomater Sci Polym Ed 18(4) 469 85... 

Fuchs, S., Hofmann, A., Kirkpatrick, C.J. Microvessel-like structures from outgrowth endothelial cells from human peripheral blood in 2-dimensional and 3-dimensional co-cultures with osteoblastic lineage cells. Tissue Eng. 13, 2577-2588... 

The cytotoxicity tests were performed with the use of the direct contact method on the extracts obtained by an 8-d incubation of the polymer PSU and PSU/Ag composite samples, placed at the bottom of the well of a 24-well culture plate, in 2 ml of culture mediiun. The incubation was conducted at 37°C in air atmosphere with a 5% content of C02and 100% of relative air humidity. Next, 0.2 ml of ihe obtained extract and its fourfold dilution in a proper culture medium was dosed for the cultures of human osteoblasts (HTB-85 cell line, ATCC, USA) and human fibroblasts (CRL-7422 cell line, ATCC, USA) adhered to the bottom of the well of a 96-well culture plate. In the case of the control test, the extract of the examined samples was replaced by the proper volume of culture medium. The plates were incubated at 37°C in air atmosphere with a 5% content of CO and 100% of relative air humidity. The incubation time for two parallelly conducted experiments was 24 h and 48 h. The cytotoxicity of PSU and the PSU/Ag composites was measured by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium (MTT) assay [9, 10] and by lactate dehydrogenase (LDH) activity measured with the use of a commercial cytotoxicity assay kit (Roche Diagnostics GmbH, Mannheim, Germany), [11]. Each of the indications was repeated three times. 

TABLE 3 The lactate dehydrogenase (LDH) activity in supernatants of human osteoblasts and human fibroblasts cultures, expressed as cytotoxic effect (CT,%) after incubation with extracts of tested materials (PSU and PSU/Ag composites). 

Figure 10. Morphology of human osteoblast-like MG 63 cells in cultures on a polystyrene dish (A) a microscopic glass coverslip (B), thin continuous (C), thick continuous (D), thin micropattemed (E) and thick micropattemed (F) fullerene C60 layers. A-D native cultures E a culture stained with hematoxylin and eosin F a culture stained with LiVE/DEAD viability/cytotoxicity kit (Invitrogen). A-D day 5 after seeding, E-F day 7 after seeding. Olympus IX 50 microscope, DP 70 digital camera, obj. 20x, bar=200 pm except E, where bar=100 pm [62].

Figure 9.2 Oxygen-functionalized surfaces improve the attachment and culture of osteoblast-like cells in vitro. Scanning electron microscopy showing human primary osteoblast-like cells (isolated from femoral heads) cultured on ultrahigh molecular weight polyethylene (UHMWP) surfaces with increasing surface oxygen concentration and tissue culture polystyrene (TCPS). UV/ozone-treated UHMWP samples showed elevated cell densities of osteoblast-Mke cells and cell morphologies which paralleled those observed in vivo. Individual scale bars are shown on the SEM images of 10 and 20 pm, and for the optical microscope images (TCPS) the scale bar indicates 100 pm.

Hofmann A, Ritz U, Verrier S, Eglin D, Alini M, Fuchs S, et al. The effect of human osteoblasts on proliferation and neo-vessel formation of human umbUical vein endothelial cells in a long-term 3D co-culture on polyurethane scaffolds. Biomaterials November 2008 29(31) 4217-26. 

Alpar B, Giinay H, Geurtsen W, Leyhausen G (1999) Cytocompatibility of periodontal dressing materials in fibroblast and primary human osteoblast-like cultures. Clin Oral Investig 3 41 8... 

El-Amin, S.F., hu, H.H., Khan, Y., Burems, J., Mitchell, J., Tuan, R.S., and haurencin, C.T, Extracellular matrix production by human osteoblasts cultured on biodegradable polymers applicable for tissue engineering, BiomoterioZs 2003, 24(7), 1213-1221. 

The metaUic products released from the prosthesis because of wear, corrosion, and fretting may impair organs and local tissues. In vitro studies have indicated that particulate Co is toxic to human osteoblast-hke cell lines and inhibits synthesis of type-I collagen, osteocalcin and alkaline phosphatase in the culture medium. However, particulate Cr and CoCr alloy are well tolerated by cell lines with no significant toxicity. The toxicity of metal extracts in vitro have indicated that Co and Ni extracts at 50% concentration appear to be highly toxic since aU viability parameters were altered after 24 h. However, Cr extract seems to be less toxic than Ni and Co [Granchi et al, 1996]. 

Wiedmann-Al-Ahmad, M., R. Gutwald, G. Lauer, U. Hiibner, and R. Schmelzeisen. 2002. How to optimize seeding and culturing of human osteoblast-like cells on various biomaterials. Biomaterials 23 3319-3328. 

Hulbert, S. F. (1993). The use of alumina and zirconia in surgical implants. In L.L. Hench J. Wilson editors. Introduction to Bioceramics, (25-40) Publ. World Scientific. Josset, Y., Oum Hamed, Z., Dupont, C., et al. (2000). Examination of ziiconia, alumina ceramics and titanium interactions on human osteoblasts in culture. Key Eng Mat 192-1, 329-332. 

Moreover, the viability and proliferation of human osteoblast-like cells of the line MG63 (European Collection of Cell Cultures, Salisbury, UK) on nanocomposites were studied. After 1, 3 and 7 days cultured cells were harvested by trypsin and counted in a Biirker s chamber. 

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