Factors influencing synthesis and mineralization of bone matrix from fetal bovine bone cells grown in vitro

2009 ◽  
Vol 7 (7) ◽  
pp. 727-741 ◽  
Author(s):  
S. William Whitson ◽  
Marcia A. Whitson ◽  
Daniel E. Bowers ◽  
Michael C. Falk
Keyword(s):  
Bone Cells ◽  
Bone Matrix ◽  
Bovine Bone ◽  
Factors Influencing ◽  
Fetal Bovine

scholarly journals Fetal bovine bone cells synthesize bone-specific matrix proteins.

1984 ◽  
Vol 99 (2) ◽  
pp. 607-614 ◽  
Author(s):  
S W Whitson ◽  
W Harrison ◽  
M K Dunlap ◽  
D E Bowers ◽  
L W Fisher ◽  
...  
Keyword(s):  
Type I Collagen ◽  
Bone Cells ◽  
Bone Sialoprotein ◽  
Cell System ◽  
Matrix Proteins ◽  
Type I ◽  
Bovine Bone ◽  
Isolated Cells ◽  
Noncollagenous Proteins

We isolated cells from both calvaria and the outer cortices of long bones from 3- to 5-mo bovine fetuses. The cells were identified as functional osteoblasts by indirect immunofluorescence using antibodies against three bone-specific, noncollagenous matrix proteins (osteonectin, the bone proteoglycan, and the bone sialoprotein) and against type 1 collagen. In separate experiments, confluent cultures of the cells were radiolabeled and shown to synthesize and secrete osteonectin, the bone proteoglycan and the bone sialoprotein by immunoprecipitation and fluorography of SDS polyacrylamide gels. Analysis of the radiolabeled collagens synthesized by the cultures showed that they produced predominantly (approximately 94%) type I collagen, with small amounts of types III and V collagens. In agreement with previous investigators who have employed the rodent bone cell system, we confirmed in bovine bone cells that (a) there was a typical cyclic AMP response to parathyroid hormone, (b) freshly isolated cells possessed high levels of alkaline phosphatase, which diminished during culture but returned to normal levels in mineralizing cultures, and (c) cells grown in the presence of ascorbic acid and beta-glycerophosphate rapidly produced and mineralized an extracellular matrix containing largely type I collagen. These results show that antibodies directed against bone-specific, noncollagenous proteins can be used to clearly identify bone cells in vitro.

scholarly journals Increased serum and bone matrix levels of transforming growth factor β1 in patients with GH deficiency in response to GH treatment

2011 ◽  
Vol 165 (3) ◽  
pp. 393-400 ◽  
Author(s):  
Thor Ueland ◽  
Tove Lekva ◽  
Kari Otterdal ◽  
Tuva B Dahl ◽  
Nicoleta Cristina Olarescu ◽  
...  
Keyword(s):  
Growth Factor ◽  
Cortical Bone ◽  
Transforming Growth Factor ◽  
Bone Cells ◽  
Gh Deficiency ◽  
Bone Matrix ◽  
Transforming Growth Factor Β1 ◽  
Gh Treatment

ObjectivePatients with adult onset GH deficiency (aoGHD) have secondary osteoporosis, which is reversed by long-term GH substitution. Transforming growth factor β1 (TGFβ1 or TGFB1) is abundant in bone tissue and could mediate some effects of GH/IGFs on bone. We investigated its regulation by GH/IGF1in vivoandin vitro.Design and methodsThe effects of GH substitution (9–12 months, placebo controlled) on circulating and cortical bone matrix contents of TGFβ1 were investigated in patients with aoGHD. The effects of GH/IGF1 on TGFβ1 secretion in osteoblasts (hFOB), adipocytes, and THP-1 macrophages as well as the effects on release from platelets were investigatedin vitro.ResultsIn vivoGH substitution increased TGFβ1 protein levels in cortical bone and serum.In vitro, GH/IGF1 stimulation induced a significant increase in TGFβ1 secretion in hFOB. In contrast, no major effect of GH/IGF1 on TGFβ1 was found in adipocytes and THP-1 macrophages. Finally, a minor modifying effect on SFLLRN-stimulated platelet release of TGFβ1 was observed in the presence of IGF1.ConclusionGH substitution increases TGFβ1in vivoandin vitro, and this effect could contribute to improved bone metabolism during such therapy, potentially reflecting direct effect of GH/IGF1 on bone cells.

scholarly journals Osteoblasts synthesize and respond to transforming growth factor-type beta (TGF-beta) in vitro.

1987 ◽  
Vol 105 (1) ◽  
pp. 457-463 ◽  
Author(s):  
P G Robey ◽  
M F Young ◽  
K C Flanders ◽  
N S Roche ◽  
P Kondaiah ◽  
...  
Keyword(s):  
Growth Factor ◽  
Transforming Growth Factor ◽  
Northern Analysis ◽  
Beta Activity ◽  
Bovine Bone ◽  
Osteosarcoma Cell ◽  
Tgf Beta ◽  
Fetal Bovine ◽  
Factor Type

Transforming growth factor-type beta (TGF-beta) has been identified as a constituent of bone matrix (Seyedin, S. M., A. Y. Thompson, H. Bentz, D. M. Rosen, J. M. McPherson, A. Conti, N. R. Siegel, G. R. Gallupi, and K. A. Piez, 1986, J. Biol. Chem. 261:5693-5695). We used both developing bone and bone-forming cells in vitro to demonstrate the cellular origin of this peptide. TGF-beta mRNA was detected by Northern analysis in both developing bone tissue and fetal bovine bone-forming cells using human cDNA probes. TGF-beta was shown to be synthesized and secreted by metabolically labeled bone cell cultures by immunoprecipitation from the medium. Further, TGF-beta activity was demonstrated in conditioned media from these cultures by competitive radioreceptor and growth promotion assays. Fetal bovine bone cells (FBBC) were found to have relatively few TGF-beta receptors (5,800/cell) with an extremely low Kd of 2.2 pM (high binding affinity). In contrast to its inhibitory effects on the growth of many cell types including osteosarcoma cell lines, TGF-beta stimulated the growth of subconfluent cultures of FBBC; it had little effect on the production of collagen by these cells. We conclude that bone-forming cells are a source for the TGF-beta that is found in bone, and that these cells may be modulated by this factor in an autocrine fashion.

scholarly journals Plasma Membrane Receptors Involved in the Binding and Response of Osteoclasts to Noncellular Components of the Bone

2021 ◽  
Vol 22 (18) ◽  
pp. 10097
Author(s):  
Divakar S. Karanth ◽  
Macey L. Martin ◽  
Lexie S. Holliday
Keyword(s):  
Bone Cells ◽  
Bone Matrix ◽  
Bone Diseases ◽  
Membrane Receptors ◽  
Membrane Structures ◽  
Regulatory Effects ◽  
Plasma Membrane Receptors ◽  
To Receive ◽  
New Strategies

Osteoclasts differentiate from hematopoietic cells and resorb the bone in response to various signals, some of which are received directly from noncellular elements of the bone. In vitro, adherence to the bone triggers the reduction of cell–cell fusion events between osteoclasts and the activation of osteoclasts to form unusual dynamic cytoskeletal and membrane structures that are required for degrading the bone. Integrins on the surface of osteoclasts are known to receive regulatory signals from the bone matrix. Regulation of the availability of these signals is accomplished by enzymatic alterations of the bone matrix by protease activity and phosphorylation/dephosphorylation events. Other membrane receptors are present in osteoclasts and may interact with as yet unidentified signals in the bone. Bone mineral has been shown to have regulatory effects on osteoclasts, and osteoclast activity is also directly modulated by mechanical stress. As understanding of how osteoclasts and other bone cells interact with the bone has emerged, increasingly sophisticated efforts have been made to create bone biomimetics that reproduce both the structural properties of the bone and the bone’s ability to regulate osteoclasts and other bone cells. A more complete understanding of the interactions between osteoclasts and the bone may lead to new strategies for the treatment of bone diseases and the production of bone biomimetics to repair defects.

Physiological Levels of Substrate Deformation Are Less Stimulatory to Bone Cells Compared to Fluid Flow

1999 ◽  
Author(s):  
Jun You ◽  
Clare E. Yellowley ◽  
Henry J. Donahue ◽  
Christopher R. Jacobs
Keyword(s):  
Fluid Flow ◽  
Matrix Protein ◽  
Bone Cells ◽  
Bone Matrix ◽  
Biological Response ◽  
Bone Cell ◽  
Osteoblastic Cells ◽  
Physiological Strain ◽  
Substrate Strain

Abstract It is believed that bone cells can sense mechanical loading and alter bone external shape and internal structure to efficiently support the load bearing demands placed upon it. However, the mechanism by which bone cells sense and respond to their mechanical environment is still poorly understood. In particular, the load-induced signals to which bone cells respond, e.g. fluid flow, substrate deformation, electrokinetic effects etc., are unclear. Furthermore, there are few studies focused on the effects of physiological strain (strain < 0.5%, Burr, 1996; Owan, 1997) on bone cells. The goal of this study was to investigate cytosolic Ca2+ mobilization (a very early signaling event) in response to different substrate strains (physiological or supra-physiological strains), and to distinguish the effects of substrate strain from those of fluid flow by applying precisely controlled strain without induced fluid flow. In addition, we quantified the effect of physiologically relevant fluid flow (Cowin, 1995) and substrate stretch on the expression of mRNA for the bone matrix protein osteopontin (OPN). A computer controlled stretch device was employed to apply different substrate strains, 0.1%, 1%, 5% and 10%. A parallel plate flow chamber was used to test cell responses to steady and oscillating flows (20dyn/cm2, 1Hz). Our data demonstrate that physiological strain (< 0.5%) does not induce [Ca2+]i responses in primary rat osteoblastic cells (ROB) in vitro. However, there was a significant (p < 0.05) increase in the number of responding cells at supra-physiological strains of 1, 5, and 10% suggesting that the cells were capable of a biological response. Similar results for human fetal osteoblastic cells (hFOB 1.19) and osteocyte-like cells (ML0-Y4) were obtained. Furthermore, compared to physiological substrate deformation, physiological fluid flow induced greater [Ca2+]i responses for hFOB cells, and these [Ca2+]i responses were quantitatively similar to those obtained for 10% substrate strain. Moreover we found no change in osteopontin mRNA expression after 0.5% strain stretch. Conversely, physiological oscillating flow (20dyn/cm2, 1Hz) caused a significant increase in osteopontin mRNA. These data suggest that, relative to fluid flow, substrate deformation may play less of a role in bone cell mechanotransduction.

OR 10 Effect of endodontic filling materials on fetal bovine bone formation in vitro

1999 ◽  
Vol 25 (4) ◽  
pp. 284
Author(s):  
S.J. Muraski ◽  
S.W. Whitson ◽  
C.J. Nielsen ◽  
D.E. Bowers ◽  
J.F. Hatton
Keyword(s):  
Bone Formation ◽  
Bovine Bone ◽  
Filling Materials ◽  
Fetal Bovine

scholarly journals In vitro induction of cartilage-specific macromolecules by a bone extract.

1983 ◽  
Vol 97 (6) ◽  
pp. 1950-1953 ◽  
Author(s):  
S M Seyedin ◽  
A Y Thompson ◽  
D M Rosen ◽  
K A Piez
Keyword(s):  
Morphological Changes ◽  
Bone Matrix ◽  
Type Ii Collagen ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bovine Bone ◽  
In Vitro System ◽  
In Vitro Induction ◽  
Inhibition Technique

An in vitro system has been developed to study the onset of chondrogenesis. Embryonic rat muscle mesenchymal cells, when treated in suspension culture with an extract of bovine bone matrix, synthesized cartilage-specific proteoglycan and type II collagen. The synthesis of these two macromolecules was assayed by the enzyme-linked immunosorbent assay inhibition technique. Further evidence of chondrogenesis was demonstrated by morphological changes of treated cells when cultured in firm agarose and stained for metachromatic matrix. Even with crude bone matrix extracts, the assay was sensitive at the microgram level and significant differences in cartilage macromolecules compared with controls were observed in 2-3 d. In vivo the same extract induced first cartilage and then bone.

scholarly journals Bone Matrix Proteins: Isolation and Characterization of a Novel Cell-binding Keratan Sulfate Proteoglycan (Osteoadherin) from Bovine Bone

1998 ◽  
Vol 141 (3) ◽  
pp. 839-847 ◽  
Author(s):  
Mikael Wendel ◽  
Yngve Sommarin ◽  
Dick Heinegård
Keyword(s):  
Guanidine Hydrochloride ◽  
Bone Matrix ◽  
Keratan Sulfate ◽  
Apparent Size ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bovine Bone ◽  
Metabolic Labeling ◽  
Cell Binding ◽  
Isolation And Characterization

A small cell-binding proteoglycan for which we propose the name osteoadherin was extracted from bovine bone with guanidine hydrochloride–containing EDTA. It was purified to homogeneity using a combination of ion-exchange chromatography, hydroxyapatite chromatography, and gel filtration. The Mr of the proteoglycan was 85,000 as determined by SDS-PAGE. The protein is rich in aspartic acid, glutamic acid, and leucine. Two internal octapeptides from the proteoglycan contained the sequences Glu-Ile-Asn-Leu-Ser-His-Asn-Lys and Arg-Asp-Leu-Tyr-Phe-Asn-Lys-Ile. These sequences are not previously described, and support the notion that osteoadherin belongs to the family of leucine-rich repeat proteins. A monospecific antiserum was raised in rabbits. An enzyme-linked immunosorbent assay was developed, and showed the osteoadherin content of bone extracts to be 0.4 mg/g of tissue wet weight, whereas none was found in extracts of various other bovine tissues. Metabolic labeling of primary bovine osteoblasts followed by immunoprecipitation showed the cells to synthesize and secrete the proteoglycan. Digesting the immunoprecipitated osteoadherin with N-glycosidase reduced its apparent size to 47 kD, thus showing the presence of several N-linked oligosaccharides. Digestion with keratanase indicated some of the oligosaccharides to be extended to keratan sulfate chains. In immunohistochemical studies of the bovine fetal rib growth plate, osteoadherin was exclusively identified in the primary bone spongiosa. Osteoadherin binds to hydroxyapatite. A potential function of this proteoglycan is to bind cells, since we showed it to be as efficient as fibronectin in promoting osteoblast attachment in vitro. The binding appears to be mediated by the integrin αvβ3, since this was the only integrin isolated by osteoadherin affinity chromatography of surface-iodinated osteoblast extracts.

scholarly journals Dynamics of reparative histogenesis of bone tissue in presence of some osteoplastic materials in vitro

2020 ◽  
Vol 4 (34) ◽  
pp. 46-50
Author(s):  
S. Yu. Ivanov ◽  
A. V. Volkov ◽  
D. A. De
Keyword(s):  
Bone Regeneration ◽  
Bone Tissue ◽  
Bone Matrix ◽  
Three Dimensional ◽  
Bone Defects ◽  
Control Group ◽  
Bovine Bone ◽  
Reparative Osteogenesis ◽  
The Impact

Currently, to solve the bone deficiency problem in the maxillofacial region, osteoplastic materials based on allogeneic and xenogenic collagen bone matrix are used, both in pure and in activated forms, by adding growth factors. It is impossible to determine the effectiveness and mechanisms of the osteoplastic materials effect on bone regeneration without a comprehensive study, including not only histological, but also morphometric studies of the structural components and cellular reactions in the impact area. Such studies provide reliable and objective information on the main processes taking place in bone regeneration.Purpose. To determine the spatial distribution of reparative osteogenesis in the presence of some osteoplastic materials in vitro.Materials and methods. Svetlogorsk breed pigs were used as a biomodel. Depending on the osteoplastic preparations used, the animals were divided into four groups of the two in each: 1st — a preparation based on a natural bovine bone graft was injected into bone defects. 2nd — a preparation based on collagenized porcine transplant was injected into bone defects. 3rd — a preparation consisting of 60 % hydroxyapatite (HA) and 40 % beta-tri-calcium phosphate; 4th — control group — the bone defect healed under a blood clot. Animals were removed from the experiment on the 45th day. We examined sections with a thickness of 20 μm using the method of light and fluorescence microscopy.Results. The results indicate different dynamics of the reparative osteogenesis in the presence of osteoplastic materials of different classes. In group 1, the filling of the defect with newly formed bone tissue is not uniform; in group 2, the filling of the defect with newly formed bone tissue is uniform; in group 3 the filling of the defect with non-formed bone tissue is uneven due to the pronounced hyperostosis; in the control group, the filling of the defect with newly formed bone tissue is not happening.Conclusion. Stimulation, the dynamics of reparative osteogenesis and the three-dimensional organization of bone regenerate depend on the osteoplastic material class, which requires further study of the dynamics and three-dimensional organization of bone regenerate to select the optimal bone-replacing agent.

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