Osteoblast cell culture

Tutak, W. et al. (2009) Toxicity induced enhanced extracellular matrix productionin osteoblastic cells cultured on single-walled carbon nanotube networks. Nanotechnology, 20 (25). 255101. 

Bradford PG, Maglich JM, Kirkwood KL. 2000. IL-1 (3 increases type 1 inositol trisphosphate receptor expression and IL-6 secretory capacity in osteoblastic cell cultures. Mol Cell Biol Res Commun 3 73-5. 

The application of tensile stresses to osteoblasts appears to cause induction of the BMP-4 gene in preosteoblastic cells and adjacent spindle-shaped fibroblasts suggesting that BMP-4 may play a pivotal role by acting as an autocrine and a paracrine factor for recruiting osteoblasts in tensile stress-induced osteogenesis. Intermittent hydrostatic pressure (IHC) up-regulates osteopontin (OPN) mRNA expression in osteoblast cell cultures. In both... 

Dos Santos et al. (2014) deposited titanium oxide coatings by PEO in a Ca-P-based electrolyte. The crystalline part of the sample consisted predominately of anatase with minor amounts of rutile, whereby Ca2+ and P043- ions were incorporated into the oxide phases. Cross-sectional images of the coating-substrate interface revealed the presence of voids elongated along the interface. Osteoblast cell cultured at the PEO coating verified the cytocompatibility of the anodised surface. The results of the cytotoxicity tests show satisfactory cell viability of the titanium dioxide films produced in this study. 

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... 

Figure 12-24. Fluorescence intensity of mouse osteoblast cells cultured on untreated and treated PCL...

Figure 7.14 Effect of support pore size (D=100, 60, 40 pm) on mineralized nodule formation of rat osteoblast cells cultured in vitro after 28 or 35 days using hydroxylapatite-coated (modified) or uncoated (unmodified) styrene Poly HI PE Polymer cell supports...

Synthetic hiomaterials, whether they are metallic, ceramic or polymeric, shall be thoroughly tested for their in vivo hiocompadbility. In vivo animal tests apparently place a burden on the shoulders of many materials-hased research groups since such animal tests require tedious histological examinations and careful interpretation. In vitro osteoblast cell culture tests, mainly reporting the live/dead cell numbers and the popular ALP (alkaline phosphatase) activity data, have thus been the most routine tests performed by the materials-based research groups which lack collaborative partnerships with the external veterinarians and skilled histologists. DMEM (or a-MEM) solutions are the media of choice in such in vitro cell culture tests. 

The spongious matrix of CG/ PB/ MA-MMA nanocomposite was in vitro tested on osteoblast cell cultures. In vitro test results are presented in Table 5. After 24 and 72 hours since hatching, the cells proliferate on matrix surface, the viability being of 97%, very dose to the reference value (control sample). These results are also supported by microscopy observation of oeUular density (Fig. 11). The collagen/layered silicate/MA-MMA ternary nanocomposite does not present any citotoxic response and the cells present normal phenotype. 

Figure 4.17 Proliferation kinetics of primary rat osteoblast cells cultured on PN-EA/EOB PN-EA/POB and TCPS. Initial seeding density was 50,000 cells/well. Reproduced with permission from L.S. Nair, D.A. Lee, J.D. Bender, E.W. Barrett, Y.E. Greish, P.W. Brown, H.R. Allcock and C.T. Laurencin, Journal of Biomedical Materials Research Part A, 2006, 76,1, 206. 2006, Wiley Periodicals, Inc. [14]...

Biocompatibile hydroxyapatite/coUagen bionanocomposites were also studied by Santos et al. [232] using osteoblast cell culture assay. Their results showed that, after 72 h, their bionanocomposites did adversely affect ceU morphology, and had similar cell viability and alkaline phosphatase activity related to the control. This demonstrated that these bionanocomposites can potentially be used in bone tissue replacement applications. 

Granet C, Laroche N, Vico L, Alexandre C, Lafage-Proust MH. 1998. Rotatmg-waU vessels, promising bioreactors for osteoblastic cell culture Comparison with other 3D conditions. Med Biol Eng Comput 36(4) 513-9. 

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].

Figure 29. Fiuman osteoblast-like MG 63 cells in cultures on material surfaces modified with carbon nanoparticles. A fullerene Cgo layers deposited on carbon fibre-reinforced carbon composites (CFRC), B fullerene C o layers deposited on microscopic glass coverslips, C terpolymer of polytetrafluoroethylene, polyvinyldifluoride and polypropylene, mixed with 4% of single-wall carbon nanohorns, D the same terpolymer with high crystalline electric arc multi-wall nanotubes, E diamond layer with hierarchically organized micro- and nanostmcture deposited on a Si substrate, F nanocrystalline diamond layer on a Si substrate. Standard control cell culture substrates were represented by a PS culture dish (G) and microscopic glass coverslip (FI). Immunofluorescence staining on day 2 (A) or 3 (B-Fl) after seeding, Olympus epifluorescence microscope IX 50, digital camera DP 70, obj. 20x, bar 100 pm (A, C, D, G,H)or 200 pm (B, E, F) [16].

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. 

Fig. 18.3. Raman spectral analysis of foetal osteoblast (FOB) differentiation. Unsupervised PCA of FOB cells cultured for 3 days in bioactive glass (BG) conditioned media (triangle) or control media (circle) (a). BG-treated cells formed a distinct cluster separate from control cells after 3 days culture. Least square (LS) analysis (which decomposes the cell spectra into the linear combination of Raman spectra obtained from the pure chemical constituents of the cell, e.g. nucleic acid, proteins, lipids, phospholipids and carbohydrates) of the relative RNA concentration of FOBs cultured for 1, 3 and 14 days in culture media (black) or BG condition media (grey), revealed a significantly reduced relative RNA concentration in FOBs culture in BG-conditioned media (b). FOBs cultured in BG-conditioned media appeared to accelerate FOB differentiation into mature adult osteoblast phenotypes (parallel gene and protein expression experiments confirmed this). Significant difference to control (p <0.05) [38]...

Differential 31P 1H CPMAS technique has been applied to study mineral crystals of calcium phosphate deposited during osteoblast calcification in cell culture.198 The samples harvested after 8 days of culture represent the minerals formed at the very early stage of calcification, in... 

Nitric oxide (NO) is considered to be a factor that participates significantly in bone remodeling, especially as a mediator of cytokines, and their activities in bone tissue. In cell cultures, the presence of NO leads to an increase of activity of alkaline phosphatase, and to an increase of the number of calcified nodules in the primary line of bone osteoblasts (C3). Low NO concentration, on the other hand, led to increased osteoclast formation. Damoulis and Hauschka (Dl) arrived at a similar conclusion the proinflammatory cytokines induce production of NO in various types of cells, including osteoblasts and osteoclasts. It is obvious that each of these cytokines can interfere in the process of bone remodeling independently of the permissive effect of NO. 

The non-pattemed CaCOj films could be observed to crystallize within 1 h by optical microscopy. However the patterned films stayed amorphous for 2-3 h under ambient conditions and were only completely crystalline after 24 h, which is probably due to the use of ethanol in the patterning procedure, as this is known to stabilize ACC (amorphous calcium carbonate). Subsequently cell culture experiments were performed and the results indicated that the CaCOj substrates support rat bone marrow stromal cell attachment, proliferation and differentiation into osteoblast and osteoclast-like cells. Moreover, mineral formation by the osteoblast-like cells was favored on the CaCOj films compared to the developed polymer films. Also, the osteoclast-like cells can degrade the CaC03 films. Therefore, these patterns of CaCOj films can be regarded as suitable 2D model substrates for bone cells. 

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