scholarly journals Comparative analysis of local stimulation methods of reparative osteogenesis

Author(s):  
M. L. Mukhanov ◽  
A. N. Blazhenko ◽  
S. B. Bogdanov ◽  
A. S. Sotnichenko ◽  
T. V. Rusinova ◽  
...  

Objective To determine the ratio of the main growth factors when using various methods of local stimulation of reparative osteogenesis.Material and methods The study consisted of two parts: in the first part a comparative analysis of the content of growth factors by ELISA was carried out (PDGF – platelet derived growth factor, TGF – transforming growth factor, VEGF – vascular endothelial growth factor, IGF – insulin-like growth factor, BMP6 and BMP7 – morphogenetic proteins 6 and 7), capable of stimulating reparative osteogenesis in blood plasma, plateletrich plasma, red bone marrow and bone autoregenerate. The second part presented the results of approbation of the autoregenerate obtained according to the original method in the framework of an acute experiment on animals.Results The most important cytokines affecting the process of reparative osteogenesis are fibroblast growth factor – FGF1 and bone morphogenetic protein 7 – BMP7. Based on the results of a comparative enzymelinked immunosorbent assay, it has been established that the autoregenerate, obtained by the original method, and a bone marrow aspirate concentrate have the highest osteogenic potential.Conclusion Autoregenerate is an effective and promising means of local stimulation of reparative osteogenesis, and its transplantation is a simple and highly effective procedure.

1994 ◽  
Vol 303 (3) ◽  
pp. 713-721 ◽  
Author(s):  
G E Ysart ◽  
R M Mason

1. The effect of different batches of fetal bovine serum and of growth factors on [35S]sulphate incorporation into glycosaminoglycans and on UDP-sugar pools in explant cultures of bovine articular cartilage was investigated. 2. [35S]Sulphate incorporation was variably stimulated between 1.2- and 3.5-fold by four different batches of serum. The UDP-glucuronate pool size expanded 4.3-6.5-fold in the presence of serum, even in those cultures in which little stimulation of [35S]sulphate incorporation occurred. The UDP-N-acetylhexosamine and UDP-hexose pools expanded by about 1.5- and 2.0-fold respectively in the presence of serum. UDP-xylose was not detected. 3. Equilibrium-labelling and pulse-chase experiments with D-[1-3H]glucose indicated that the rate of flux through the UDP-sugar pools was unaffected by serum. UDP-hexose, UDP-N-acetylhexosamine and UDP-glucuronate have approximate half-lives (t1/2) of 7, 12 and 3-4 min respectively. At equilibrium, the 3H specific activities of UDP-hexose and UDP-N-acetylhexosamine were very similar but that for the UDP-glucuronate pool was much higher, especially in serum-supplemented cultures. The results suggest that UDP-glucuronate synthesis occurs via a pathway which is independent of the main UDP-hexose pathway. 4. Supplementing cultures with heat-treated serum had no effect on the serum-induced expansion of UDP-sugar pools but stimulation of [35S]sulphate incorporation into glycosaminoglycans was 50% lower than for native serum. Acid-treated serum promoted a 2-fold expansion of the UDP-glucuronate and UDP-N-acetylhexosamine pool over that obtained with native serum but was 20% less effective in stimulating [35S]sulphate incorporation than the latter. Prior dialysis of serum had no effect on its modulatory action on either [35S]sulphate incorporation or on the size of UDP-sugar pools. 5. Insulin-like growth factor 1 (IGF-1), transforming growth factor beta-1 (TGF beta-1), platelet-derived growth factor (PDGF) (BB homodimer) and epidermal growth factor (EGF) all stimulated [35S]sulphate incorporation into glycosaminoglycans as expected. The UDP-glucuronate pool expanded by 1.5- and 2.0-fold in the presence of IGF-1 and TGF beta-1 respectively, and by about 1.8-fold in the presence of PDGF or EGF. None of the factors investigated, or combinations of IGF-1 and TGF beta-1 or IGF-1 and EGF, stimulated expansion of the UDP-glucuronate pool to the same extent as native serum.(ABSTRACT TRUNCATED AT 400 WORDS)


Blood ◽  
1997 ◽  
Vol 90 (6) ◽  
pp. 2253-2259 ◽  
Author(s):  
Zhong-Ying Liu ◽  
Ramesh K. Ganju ◽  
Jian-Feng Wang ◽  
Karin Schweitzer ◽  
Babette Weksler ◽  
...  

Human bone marrow endothelial cells immortalized with the T antigen of SV40 (TrHBMEC) have previously been characterized by us with regard to their properties that are similar to primary marrow endothelial cells and their utility as a model system. We now report that TrHBMEC express a recently discovered signal transduction molecule termed RAFTK (related adhesion focal tyrosine kinase), also called Pyk2 or CAK-β. RAFTK, the second member of the focal adhesion kinase (FAK) family, is known to be activated in response to calcium flux in neuronal cells and integrin stimulation in megakaryocytes and B cells. We have studied the effects of cytokines on RAFTK activation in TrHBMEC. Treatment of TrHBMEC with the vascular endothelial growth factor (VEGF ), as well as the VEGF-related protein (VRP), the recently identified ligand for the FLT-4 receptor, resulted in enhanced tyrosine phosphorylation of RAFTK. Similar changes in RAFTK phosphorylation were observed upon stimulation of TrHBMEC with basic fibroblast growth factor (bFGF ) or oncostatin M (OSM). Stimulation of these cells with growth factors also resulted in an increase in RAFTK activity and the c-Jun NH2 -terminal kinase (JNK). RAFTK coimmunoprecipitated with the cytoskeletal protein paxillin through its C-terminal proline-rich domain in TrHBMEC. These results suggest that, in marrow endothelium, activation of RAFTK by VEGF, VRP, OSM, and bFGF represents a new element in the signal transduction pathways used by these growth factors and likely acts to coordinate signaling from their surface receptors to the cytoskeleton, thereby modulating cell growth and function.


1993 ◽  
Vol 137 (3) ◽  
pp. 369-374 ◽  
Author(s):  
S. C. Butterwith ◽  
C. D. Peddie ◽  
C. Goddard

ABSTRACT The development of adipose tissue is dependent on the growth and differentiation of fibroblast-like adipocyte precursor cells. Culture of adipocyte precursor cells in vitro has provided an ideal system for identifying potential regulators of proliferation and differentiation. We have demonstrated that both acidic fibroblast growth factor (aFGF) and basic fibroblast growth factor (bFGF) stimulate chicken adipocyte precursor DNA synthesis in a dose-dependent manner up to a concentration of 100 μg aFGF/l and 1 μg bFGF/l. The effect of bFGF was biphasic, so that in incubations with 25 μg bFGF/l, DNA synthesis was not significantly different from controls. In the presence of heparin, stimulation of DNA synthesis at 25 μg bFGF/l was 1·6-fold greater than at a concentration of 1 μg bFGF/l. Addition of heparin to incubations containing aFGF reduced the concentration required for maximum stimulation of DNA synthesis to 1 μg/l. Cells incubated with aFGF (1–100 μg/l) in combination with insulin-like growth factor-I (IGF-I), platelet-derived growth factor, transforming growth factor-α or transforming growth factor-β1 (TGF-β1) exhibited a marked synergistic increase in DNA synthesis. This was also the case when 1 μg bFGF/l was used, but at a concentration of 25 pg bFGF/l synergy was only seen with IGF-I and TGF-β1. These results suggest that both basic and acidic FGF are potentially important regulators of adipocyte hyperplasia and that their effect is modulated by constituents of the extracellular matrix and the presence of other growth factors. Journal of Endocrinology (1993) 137, 369–374


Blood ◽  
1997 ◽  
Vol 90 (6) ◽  
pp. 2253-2259 ◽  
Author(s):  
Zhong-Ying Liu ◽  
Ramesh K. Ganju ◽  
Jian-Feng Wang ◽  
Karin Schweitzer ◽  
Babette Weksler ◽  
...  

Abstract Human bone marrow endothelial cells immortalized with the T antigen of SV40 (TrHBMEC) have previously been characterized by us with regard to their properties that are similar to primary marrow endothelial cells and their utility as a model system. We now report that TrHBMEC express a recently discovered signal transduction molecule termed RAFTK (related adhesion focal tyrosine kinase), also called Pyk2 or CAK-β. RAFTK, the second member of the focal adhesion kinase (FAK) family, is known to be activated in response to calcium flux in neuronal cells and integrin stimulation in megakaryocytes and B cells. We have studied the effects of cytokines on RAFTK activation in TrHBMEC. Treatment of TrHBMEC with the vascular endothelial growth factor (VEGF ), as well as the VEGF-related protein (VRP), the recently identified ligand for the FLT-4 receptor, resulted in enhanced tyrosine phosphorylation of RAFTK. Similar changes in RAFTK phosphorylation were observed upon stimulation of TrHBMEC with basic fibroblast growth factor (bFGF ) or oncostatin M (OSM). Stimulation of these cells with growth factors also resulted in an increase in RAFTK activity and the c-Jun NH2 -terminal kinase (JNK). RAFTK coimmunoprecipitated with the cytoskeletal protein paxillin through its C-terminal proline-rich domain in TrHBMEC. These results suggest that, in marrow endothelium, activation of RAFTK by VEGF, VRP, OSM, and bFGF represents a new element in the signal transduction pathways used by these growth factors and likely acts to coordinate signaling from their surface receptors to the cytoskeleton, thereby modulating cell growth and function.


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