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  • LONGITUDINAL IN-VIVO  (3)
  • differentiation  (3)
  • 1
    Keywords: ANGIOGENESIS ; PERFUSION ; metastases ; CONTRAST-ENHANCED MRI ; MINERAL DENSITY ; LONGITUDINAL IN-VIVO ; FAT-CONTENT
    Abstract: Background: Etiologic and pathophysiologic role of functional bone marrow processes is not fully understood especially in the case of osteoporosis. Purpose: To investigate the role of vascularization and diffusion in rat models of osteoporosis through a cross-correlation between non-invasive in-vivo imaging and invasive ex-vivo imaging of bone, bone marrow, and in particular of microcirculation. Material and Methods: Osteoporosis was induced in rats by combining ovariectomy (OVX) with calcium and Vitamin D3 deficiency, or with glucocorticoid (dexamethasone). For comparison, controls underwent a sham surgery. In in-vivo investigations, animals (n = 36) were examined by volumetric CT (VCT) and MRI at 1, 3, or 12 months post surgery. Using VCT, bone morphology was monitored and relative bone density r within pelvis was extracted. With DCE-MRI and DW-MRI, parameters A (amplitude), Kep (exchange rate constant), and ADC (apparent diffusion coefficient) were acquired for regions of lumbar vertebrae, pelvis, and femur. In ex-vivo investigations, selective histological sections of pelvis were either stained with hematoxylin and eosin (HE stain) for quantifying vessel size and density or immunostained for collagen IV and alpha-smooth muscle actin to assess vessel maturity (SMA/collagen IV ratio). Results: After 12 months, decrease in DCE-MRI parameter Kep was found in all locations of osteoporotic rats (strongest in femur and lumbar vertebrae) while no significant differences were seen for parameter A and DW-MRI parameter ADC. Furthermore, vessel rarefication and maturation were observed on the histological level in animals with osteoporotic phenotype. In particular in the pelvis, the osteoporotic individuals (irrespective of the osteoporosis inducers applied) exhibited decreased Kep, significantly reduced vessel density, significantly increased vessel maturity, as well as statistically unaltered A, ADC, and vessel diameter. Conclusion: Changes in microcirculation but not diffusion in the bone marrow of osteoporotic rats are detected by DCE-MRI and DW-MRI due to vessel rarefication and maturation.
    Type of Publication: Journal article published
    PubMed ID: 23319721
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  • 2
    Keywords: DISEASE ; metastases ; MUSCLE ; DCE-MRI ; MARROW ; LONGITUDINAL IN-VIVO ; FRACTURES ; VCT
    Abstract: Angiogenesis is pivotal for bone metabolism and bone defect healing. Yet the role of vascularization in osteoporosis and osteoporotic bone repair mechanisms is unclear. Here we investigated effects of osteoporotic phenotype on vascularization during bone defect healing in a rodent osteotomy model using volumetric computed tomography (VCT), dynamic contrast-enhanced VCT (DCE-VCT), dynamic contrast-enhanced MRI (DCE-MRI) and histology. In 16 rats, 8 with physiological bone status (SHAM) and 8 with osteoporotic bone status induced by ovariectomy (OVX) in combination with a vitamin D- and low calcium diet, wedge-shaped defects were created at the left distal femur and stabilized internally by T-shaped miniplate. MRI and VCT were performed in all animals 6 weeks after this procedure. By VCT, relative bone density in the defect was evaluated. Using DCE-VCT and DCE-MRI, parameters associated with regional blood volume were calculated in the bone defect, vicinity of the defect, surrounding muscles and bone marrow: Amplitude A and exchange rate constant Kep (DCE-MRI, respectively) as well as peak enhancement PE and area under the curve AUC (DCE-VCT, respectively). In animals of osteoporotic phenotype, bone density within the osseous defect was significantly reduced as compared to SHAM rats. Vascularization parameters determined by DCE-MRI and DCE-VCT in the defect were significantly elevated compared to the adjacent tissues for both SHAM and OVX groups. However, comparing SHAM and OVX rats, no statistically different values were found by DCE-MRI and DCE-VCT concerning any determined vascularization parameter within the bone defect. Furthermore, parameters of vascularization were increased for OVX as compared to SHAM rats within the bone marrow although significant difference was only found for A. In a rat osteotomy model we showed that at the reparative healing stage, osteoporotic phenotype did influence osteogenic but not angiogenic response within bone defect as imaged by DCE-MRI and DCE-VCT. This study provides insight into the relationship between angiogenesis and osteogenesis during osteoporosis-related compromised bone healing.
    Type of Publication: Journal article published
    PubMed ID: 23522092
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  • 3
    Keywords: radiation ; MRI ; metastases ; COMPUTED-TOMOGRAPHY ; MACHINE ; LONGITUDINAL IN-VIVO ; VCT ; VOLUMETRIC CT
    Abstract: Purpose Cone beam computed tomography (CBCT) has the disadvantage of providing non-quantitative results for bone density determination. The aim of this study is to calibrate CBCT results by using an internal reference (such as muscle) for quantitatively assessing bone density. Methods We developed a new method using the relative attenuation ratio between two nearby materials (such as bone and muscle) for systemic error correction in CBCT that depends on the relative object position in the image volume. Phantom calibration was performed to calculate the acquired attenuation ratio in Hounsfield units (HU), comparable to the results from clinical multislice spiral computed tomography (MSCT). In addition, a small animal study with an osteoporotic rat model was evaluated to show the feasibility of this presented method to quantitatively assess bone density using a CBCT system. Results The phantom study results showed that the calibration process successfully corrected the systemic inaccuracy from CBCT, and the calibrated HU values agreed with the values measured from MSCT. In the small animal study, the quantitative bone densities assessed from the calibrated CBCT results were consistent with the results from MSCT data. Conclusion A practical method to quantitatively estimate attenuation (HU) values for bone tissues from CBCT scans that are comparable to MSCT scans is proposed. The method may improve the quantification ability of CBCT scanning without any additional hardware requirements.
    Type of Publication: Journal article published
    PubMed ID: 23225074
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  • 4
    ISSN: 0021-9304
    Keywords: implant ; titanium ; osteoblasts ; prostaglandin ; indomethacin ; surface roughness ; 1α,25-(OH)2D3 ; differentiation ; Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine , Technology
    Notes: Surface roughness affects proliferation, differentiation (alkaline phosphatase and osteocalcin), local factor production [transforming growth factor (TGFβ) and prostaglandin E2 (PGE2)], and response to 1,25-(OH)2D3 (1,25) of MG63 osteoblast-like cells. In this study, we examined whether the effect of surface roughness on MG63 cells is mediated by prostaglandins produced by the cells. Unalloyed titanium (Ti) disks were pretreated with HF/HNO3 (PT) and then machined and acid-etched (MA). Disks were also coarse grit-sandblasted (SB), coarse grit-sandblasted and acid-etched (CA), or plasma-sprayed with Ti particles (PS). The surfaces, from smoothest to roughest, were PT, MA, CA, SB, and PS. MG63 cells were cultured to confluence on the Ti disks in the presence or absence of 10-7M indomethacin (Indo), a specific inhibitor of cyclooxygenase activity, resulting in decreased prostaglandin production. When the cells reached confluence, cell number, cell layer alkaline phosphatase specific activity (ALPase), and osteocalcin (OC) and latent TGFβ (LTGFβ) production were determined. In addition, confluent cultures which had been grown in the absence of Indo were exposed to 10-7M 1,25, 10-7M Indo, or a combination of the two for 24 h. On the rougher surfaces, cell number was decreased and ALPase, OC, and LTGFβ were increased. When indomethacin was present throughout the culture period, the effect of surface roughness on cell number, OC, and LTGFβ was abolished. ALPase was reduced, but surface roughness-dependent effects were still observed. Addition of indomethacin to confluent cultures for 24 h had no effect on any of the parameters examined, with one exception: Cells cultured on MA surfaces exhibited a more differentiated phenotype. 1,25 increased all parameters examined on SB, CA, and PS surfaces. When indomethacin was added with 1,25, the 1,25-dependent effects on cell number and OC and LTGFβ production were abolished; however, ALPase was unaffected. This indicates that bone cell response to systemic hormones may be modified by implant surface roughness. This effect may be mediated, at least in part, by prostaglandins produced by the same cells. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 41, 489-496, 1998.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1435-1803
    Keywords: Ischemic preconditioning ; rabbit ventricular myocytes ; HEK 293 cell line ; HIT-T15 cell line ; C2C12 cell line ; differentiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We investigated whether preconditioning could protect several cultured cell lines, to determine whether the protection is specific for cells derived from different myogenic and non-myogenic sources. Ischemia was simulated by centrifugation of cells into a pellet, and cell viability was determined by hypotonic trypan blue solution. Preconditioning was produced by brief exposures to either glucose-free solution or metabolic inhibition. Freshly isolated rabbit ventricular myocytes were studied to confirm that preconditioning occurs in this model. We then compared these results to those in several cultured cell lines, including HEK 293 cells derived from human embryonic kidney, HIT-T15 cells from Syrian hamster pancreatic islets, and C2C12 cells from mouse skeletal muscle. In the latter cell line, we also determined whether differentiation alters preconditioning. Preconditioning protected rabbit ventricular myocytes: the percentage of dead cells was decreased from 36.8±4.7% in the control group to 23.0±5.2% in the preconditioned group after 60 min and from 50.7±2.1% in the control group to 25.5±4.5% in the preconditioned group after 120 min ischemia (p〈0.02). In contrast, there was no protection from preconditioning in HEK 293 cells or HIT-T15 cells. Preconditioning did not protect C2C12 myoblasts either. Interestingly, after C2C12 myoblasts had differentiated into myotubes (induced by exposing the cells to low-serum medium), they could then be protected by preconditioning (46.3±3.6% in the control group vs 26.0±2.7% in the preconditioned group after 60 min and 67.4±3.6% in the control group vs 46.0±4.6% in the preconditioned group after 120 min ischemia; p〈0.05). In conclusion, protection from preconditioning is cell-type specific. The presence of endogenous KATP channels (which are plentiful in HIT-T15 cells) is insufficient to enable preconditioning of the cell. Among the various cell types studied, only differentiated muscle cells (rabbit ventricular myocytes and C2C12 myotubes) exhibited preconditioning.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 0021-9304
    Keywords: implant ; titanium ; osteoblasts ; surface roughness ; 1α,25- (OH)2D3 ; differentiation ; local factor ; Chemistry ; Polymer and Materials Science
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine , Technology
    Notes: Surface roughness has been shown to affect differentiation and local factor production of MG63 osteoblast-like cells. This study examined whether surface roughness alters cellular response to circulating hormones such as 1α,25-(OH)2D3. Unalloyed titanium (Ti) disks were pretreated with HF/HNO3 (PT) and then were machined and acid-etched (MA). Ti disks also were sandblasted (SB), sandblasted and acid etched (CA), or plasma sprayed with Ti particles (PS). The surfaces, from smoothest to roughest, were: PT, MA, CA, SB, and PS. MG63 cells were cultured to confluence on standard tissue culture polystyrene (plastic) or the Ti surfaces and then treated for 24 h with either 10-8M or 10-7M 1α,25-(OH)2D3 or vehicle (control). Cellular response was measured by assaying cell number, cell layer alkaline phosphatase specific-activity, and the production of osteocalcin, latent (L) TGFβ, and PGE2. Alkaline phosphatase activity was affected by surface roughness; as the surface became rougher, the cells showed a significant increase in alkaline phosphatase activity. Addition of 1α,25-(OH)2D3 to the cultures caused a dose-dependent stimulation of alkaline phosphatase activity that was synergistic with the effect caused by surface roughness alone. 1α,25-(OH)2D3 also caused a synergistic increase in osteocalcin production as well as local factor (LTGFβ and PGE2) production on the rougher CA, SB, and PS surfaces, but it had no effect on the production on smooth surfaces. The inhibitory effect of surface roughness on cell number was not affected by 1α,25-(OH)2D3 except on the SB surface. 1α,25-(OH)2D3 decreased cell number, increased alkaline phosphatase activity and osteocalcin production, and had no effect on LTGFβ or PGE2 production by MG63 cells grown on tissue culture polystyrene. These data suggest that bone cell response to systemic hormones is modified by surface roughness and that surface roughness increases the responsiveness of MG63 cells to 1α,25-(OH)2D3. They also suggest that the endocrine system is actively involved in normal bone healing around implants. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 39, 77-85, 1998.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
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