A serum substitute promotes osteoblast-like phenotypic expression in cultured cells from chick calvariae

Ramp, Warren K.; Dillaman, Richard M.; Lenz, Lois G.; Roer, Robert D.; Ballard, Timothy A.

doi:10.1016/0169-6009(91)90107-b

Cited by 19 publications

(10 citation statements)

References 25 publications

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“…This phenotypic expression is characteristic of avian osteoblasts and was previously investigated and documented. 18 The photomicrograph in Figure 1 and Ti alloy thin films were not significantly different (p ജ 0.05) from the polystyrene control group. Similarly, the results obtained regarding all variables for the thin-film coated dishes were not significantly different (p ജ 0.05) from the results obtained with the disks.…”

Section: Resultsmentioning

confidence: 81%

“…Previous studies with this system showed this to be an adequate sample size. 18,[21][22][23][24] A one-way analysis of variance, followed by the Tukey-Kramer (honestly significant difference) multiple comparison analysis, was performed. Individual group standard deviations were used to construct the 95% confidence intervals (CIs).…”

Section: Methodsmentioning

confidence: 99%

“…Further details of the isolation method and the osteoblastlike phenotypic expression of these chick embryo cells in culture were previously described. 18 The isolated cells were seeded (8.7 × 10 4 cells/cm 2 ) on uncoated 35-mm diamter polystyrene culture dishes (Corning Costar), on Ti alloy coated 35-mm diameter polystyrene culture dishes, or on machined and polished Ti alloy disks placed in uncoated 35-mm diameter plystyrene culture dishes. The medium volume was 2 mL/dish, and it was changed after the first 24 h and at 48-h intervals thereafter.…”

Section: Cell Culturementioning

confidence: 99%

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Comparison of growth and metabolism of avian osteoblasts on polished disks versus thin films of titanium alloy

Koontz¹,

Ramp²,

Peindl³

et al. 1998

J. Biomed. Mater. Res.

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The purpose of this study was to evaluate the efficacy of using high vacuum, thermal evaporation to deposit thin films of Ti-6Al-4V onto plates for subsequent cell culture investigations. Osteoblastic response to thin-film coated plates was compared to that of cells grown on Ti alloy disk inserts and uncoated culture plates. The Ti alloy disks were polished, cleaned, and passivated following a commercial protocol for orthopedic implants. Mean surface roughness was 262 nm for the Ti alloy disks and 4.756 nm for the coated culture plates. Osteoblasts isolated from 16-day chick embryo calvariae were cultured on polystyrene, thin films, and disks. At confluence, the cells were cultured an additional 48 h and were evaluated for cell number (DNA content), rate of glycolysis (lactate production), alkaline phosphatase activity (ALPase), and collagenous (3H-proline hydroxylation) and noncollagenous protein synthesis. Cell morphology was similar for the controls, disks, and thin-film groups. DNA, lactate, cell layer ALPase, 3H-hydroxyproline, and noncollagenous protein were not different (p > 0.05) among the control, thin-film, and disk groups. Medium ALPase was lower (p < 0.05) in the thin-film group compared to the control group. Although aluminum and vanadium percentages varied from nominal in the thin-film groups (11Al-2V as opposed to 6Al-4V), avian osteoblasts responded similarly to the Ti alloy thin films, disks, and uncoated culture plates for the smooth surfaces tested. The thin-film cell culture system used for elemental material studies appears to offer a promising method for the investigation of cellular response to alloyed biomaterials as well. Proper adjustments in alloy percentages before deposition, however, need to be made if thermal evaporation is utilized.

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Section: Resultsmentioning

confidence: 81%

Section: Methodsmentioning

confidence: 99%

Section: Cell Culturementioning

confidence: 99%

See 1 more Smart Citation

Comparison of growth and metabolism of avian osteoblasts on polished disks versus thin films of titanium alloy

Koontz¹,

Ramp²,

Peindl³

et al. 1998

J. Biomed. Mater. Res.

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“…Cells were harvested by centrifugation and rinsed 3 times in 25 mM HEPES-buffered Hanks' balanced salt solution (pH 7.4). The digestion step was repeated twice, and the 3 cell isolates were pooled in osteoblast growth medium consisting of Dulbecco's modified Eagle medium containing 25 mM HEPES, 10% Nu-serum IV (Collaborative Biomedical Products, Bedford, MA), 75 mg/mL glycine, 100 mg/mL ascorbic acid, 40 ng/mL vitamin B 12 , 2 mg/mL p-aminobenzoic acid, 200 ng/mL biotin, and 100 U/mL-100 mg/mL-0.25 mg/mL penicillin-streptomycin-Fungizone (pH 7.4) [35]. Cells were seeded in 6-well cluster plates and incubated at 37ЊC in a humidified incubator until they reached confluence (6-7 days).…”

Section: Methodsmentioning

confidence: 99%

Staphylococcus aureusInfection of Mouse or Human Osteoblasts Induces High Levels of Interleukin‐6 and Interleukin‐12 Production

et al. 1999

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Staphylococcus aureus is the principal causative agent of the inflammatory bone disease osteomyelitis. Unfortunately, the pathogenesis of this often chronic infection is poorly understood and is complicated by the recent observation that bone-forming osteoblasts can harbor S. aureus. Such an infection presents a significant challenge for the host immune response, because osteoblasts are not known to initiate protective cell-mediated immune responses. Cultured mouse and human osteoblasts infected with S. aureus were found to express high levels of interleukin (IL)-6 and IL-12p75, on the basis of complementary investigations demonstrating both S. aureus-induced up-regulation of expression of IL-6 and IL-12p40 mRNA and secretion of IL-6 and IL-12p75 by these cells. Additionally, a quantitative bioassay demonstrated that IL-12p75 secreted after infection was biologically active. These studies are the first to demonstrate induced IL-12p75 expression by osteoblasts and suggest a previously unrecognized role for osteoblasts in initiating immune responses after S. aureus infection.

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“…Cells were harvested by centrifugation and rinsed three times in 25 mM HEPES-buffered Hanks' balanced salt solution (pH 7.4; HBSS). The digestion step was repeated twice, and the three cell isolates were pooled in mouse osteoblast growth medium (OBGM), consisting of Dulbecco's modified Eagle's medium containing 25 mM HEPES, 10% fetal bovine serum (Sigma), 2 g of sodium bicarbonate per liter, 75 g of glycine/ml, 100 g of ascorbic acid/ml, 40 ng of vitamin B 12 /ml, 2 g of p-aminobenzoic acid/ml, 200 ng of biotin/ml, and penicillin (100 U/ml)-streptomycin (100 g/ml)-amphotericin B (Fungizone; 0.25 g) (pH 7.4) (27). Cells were seeded in six-well cluster plates and incubated at 37°C in a 5% CO 2 atmosphere until they reached confluence (6 to 7 days).…”

Section: Methodsmentioning

confidence: 99%

Staphylococcus aureusandSalmonella entericaSerovar Dublin Induce Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand Expression by Normal Mouse and Human Osteoblasts

et al. 2001

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Staphylococcus aureus and Salmonella enterica serovar Dublin invade osteoblasts and are causative agents of human bone disease. In the present study, we examined the ability of S. aureus and Salmonella serovar Dublin to induce the production of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) by normal osteoblasts. Normal mouse and human osteoblasts were cocultured with S. aureus or Salmonella serovar Dublin at different multiplicities of infection. Following initial incubation and examination of TRAIL expression, extracellular bacteria were killed by the addition of media containing the antibiotic gentamicin. Lysates and conditioned media from osteoblast cultures were then collected at various times following invasion and analyzed. The results demonstrated that S. aureus and Salmonella serovar Dublin are potent inducers of TRAIL expression by osteoblasts. Mouse and human TRAIL mRNA expression was induced by bacterial infection and demonstrated a dose-dependent response. Analysis of kinetics suggested that TRAIL mRNA was induced within 30 min after exposure to bacteria and that its level of expression remained relatively constant over the time period examined. mRNA molecules encoding TRAIL receptors were constitutively expressed by osteoblasts. Furthermore, TRAIL protein was detected as early as 45 min and up to 24 h following infection. The quantity of TRAIL protein produced also increased in a dose-dependent manner. Collectively, these findings suggest a mechanism whereby bacterial pathogens mediate bone destruction via osteoblast apoptosis.

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A serum substitute promotes osteoblast-like phenotypic expression in cultured cells from chick calvariae

Cited by 19 publications

References 25 publications

Comparison of growth and metabolism of avian osteoblasts on polished disks versus thin films of titanium alloy

Comparison of growth and metabolism of avian osteoblasts on polished disks versus thin films of titanium alloy

Staphylococcus aureusInfection of Mouse or Human Osteoblasts Induces High Levels of Interleukin‐6 and Interleukin‐12 Production

Staphylococcus aureusandSalmonella entericaSerovar Dublin Induce Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand Expression by Normal Mouse and Human Osteoblasts

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