Osteoblastic cells from rat long bone. I. Characterization of their differentiation in culture

Stringa, E.; Filanti, C.; Giunciuglio, Daniela; Albini, Adriana; Manduca, Paola

doi:10.1016/8756-3282(95)00100-r

Cited by 54 publications

(43 citation statements)

References 25 publications

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“…The isolated osteoblasts were cultured in monolayer at 373C and 5% CO in Dulbecco's Modi"ed Eagle Medium (Irvine Scienti"c, Santa Ana, CA) containing 10% of fetal bovine serum (Irvine Scienti"c,), 2% L-glutamin (BioWhittaker, Walkersville, MD) and 1% of PSF (100;, 10 000 U/ml Penicillin, 10 000 UG/ml strepzin, 25 UG/ml Fungizone; BioWhittaker). These cells were characterized as being of osteoblastic phenotype by testing for the formation of calcium phosphate salts using the von Kossa staining technique (data not shown) which is an indication of osteoblast mineralization [25] and by the presence of alkaline phosphatase [18]. Osteocalcin, another marker of osteoblast phenotype [26], was detected by EIA kit (Biomedical Technology, Stoughton, MA) when osteoblasts were cultured with vitamin D (CalBiochem, San Diego, CA) (data not shown).…”

Section: Cell Cultures 221 Osteoblastmentioning

confidence: 99%

The effects of calcium phosphate cement particles on osteoblast functions

et al. 2000

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Calcium phosphate cements (CPC) are increasingly used in the orthopedic "eld. This kind of cement has potential applications in bone defect replacements, osteosynthetic screw reinforcements or drug delivery. In vivo studies have demonstrated a good osteointegration of CPC. However, it was also observed that the resorption of CPC could create particles. It is known from orthopedic implant studies that particles can be responsible for the peri-implant osteolysis. Biocompatibility assessment of CPC should then be performed with particles. In this study, we quanti"ed the functions of osteoblasts in the presence of -TCP, brushite and cement particles. Two particle sizes were prepared. The "rst one corresponded to the critical diameter range 1}10 m and the second one had a diameter larger than 10 m. We found that CPC particles could adversely a!ect the osteoblast functions. A decrease in viability, proliferation and production of extracellular matrix was measured. A dose e!ect was also observed. A ratio of 50 CPC particles per osteoblast could be considered as the maximum number of particles supported by an osteoblast. The smaller particles had stronger negative e!ects on osteoblast functions than the larger ones. Future CPC development should minimize the generation of particles smaller than 10 m.

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Section: Cell Cultures 221 Osteoblastmentioning

confidence: 99%

The effects of calcium phosphate cement particles on osteoblast functions

et al. 2000

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“…Isolation and characterization of rat primary osteoblasts were prepared using techniques previously described (25,40). Briefly, femora collected from 3-wk-old female rats were dissected free of epiphysial cartilage, cleaned of marrow, and minced finely.…”

Section: Methodsmentioning

confidence: 99%

Temporal gradients in shear stimulate osteoblastic proliferation via ERK1/2 and retinoblastoma protein

Jiang

White

Stevens

et al. 2002

American Journal of Physiology-Endocrinology and Metabolism

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The effects of pulsatile flow (temporal gradients in fluid shear) on rat UMR106 cells and rat primary osteoblastic cells were studied. Pulsatile flow induced a 95% increase in S-phase UMR106 cells compared with static controls. In contrast, ramped steady flow stimulated only a 3% increase. Similar patterns of S-phase induction were also observed in rat primary osteoblastic cells. Pulsatile flow significantly increased relative UMR106 cell number by 37 and 62% at 1.5 and 24 h, respectively. Pulsatile flow also significantly increased extracellular signal-regulated kinase (ERK1/2) phosphorylation by 418%, whereas ramped steady flow reduced ERK1/2 activation to 17% of control. Correspondingly, retinoblastoma protein was significantly phosphorylated by pulsatile fluid flow. Inhibition of mitogen-activated protein (MAP)/ERK kinase (MEK)1/2 by U0126 (a specific MEK1/2 inhibitor) reduced shear-induced ERK1/2 phosphorylation and cell proliferation. These findings suggest that temporal gradients in fluid shear stress are potent stimuli of bone cell proliferation. shear stress; pulsatile fluid flow; osteoblasts (RE)MODELING OF BONE in the skeletal system is strongly influenced by mechanical loading and unloading. Exercise has been shown to stimulate increases in bone mass (24), whereas prolonged mechanical unloading results in bone atrophy (36,45). It has been hypothesized that exercise-induced (re)modeling of bone may result from stimulation of osteoblasts by interstitial fluid flow (IFF) through the porous structure of bone (10, 31). Interstitial fluid flows radially outward through the bone cortex and is driven by hydrostatic pressure gradients across the cortex and pressure gradients from mechanical loading (21,23,39,42). Theoretical models of IFF indicate significant fluid shear stresses in the canaliculi ranging between 6 and 30 dyn/cm 2 (44). During physical activity, rapid dynamic changes in mechanical loading cause transient changes in bone marrow pressure and IFF (30, 43). Rapid changes in IFF subject bone cells to large temporal gradients in interstitial fluid shear stress. Temporal shear stress gradients are defined as the rapid change in shear stress over a small period of time (Ͻ0.5 s) at any given location on the cell surface. Steady shear stress is devoid of temporal gradients and can be established if the onset and/or cessation of flow is slowly transitioned. In any given system of flow, if the change in flow is sudden, significant temporal gradients are generated (8,46). Given that similar temporal gradients in shear stress in other cell types have been shown to be promitogenic compared with steady fluid flow (2), repeated temporal gradients in shear stress through the bone matrix may also stimulate osteoblast proliferation (10, 43).A number of investigators have demonstrated that IFF stimulates elaborate mechanochemical signaling cascades in osteoblasts. Fluid shear has been shown to stimulate osteoblast production and release of cAMP, prostaglandin E 2 (PGE 2 ), inositol triphosphate, and nitric oxide ...

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“…The cells were then transferred by trypsinization to a 75 cm 2 tissue culture flask defining the cell passage number 2. These cells displayed typical phenotypes of osteoblasts as the polygonal morphology, the formation of calcium phosphate salts, the production of alkaline phosphatase, or the increase of osteocalcin production when these cells were cultured with 1,25(OH) 2 D 3 [29,30].…”

Section: Cellsmentioning

confidence: 99%

The influence of wear particles in the expression of osteoclastogenesis factors by osteoblasts

Pioletti

Kottelat

2004

Biomaterials

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Orthopedic implant failures are often associated with peri-implant osteolysis. Particles generated from the wear process have been suspected to play an important role in this situation. Indeed, the peri-implant osteolysis could be due to the presence of particles stimulating the osteoclastogenesis process. We hypothesize then that the presence of a low particle concentration positively influences osteoblasts to produce osteoclastogenesis factors. If true, this hypothesis would then support the idea that the particles could be at the origin of the process leading to implant loosening. To check the validity of this hypothesis, we quantified in vitro the production of different genes involved in the osteoclastogenesis process using primary isolated human osteoblasts treated or not with particles. Results showed that low concentrations of particles might have a stimulating effect on osteoblasts to produce osteoclastogenesis factors as demonstrated by the increase of RANKL and CSF-1 gene expression in the particle group. r

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Osteoblastic cells from rat long bone. I. Characterization of their differentiation in culture

Cited by 54 publications

References 25 publications

The effects of calcium phosphate cement particles on osteoblast functions

The effects of calcium phosphate cement particles on osteoblast functions

Temporal gradients in shear stimulate osteoblastic proliferation via ERK1/2 and retinoblastoma protein

The influence of wear particles in the expression of osteoclastogenesis factors by osteoblasts

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