Effect of growth hormone on osteoblasts and demonstration of somatomedin-C/IGF I in bone organ culture

1984 ◽  
Vol 107 (1) ◽  
pp. 16-24 ◽  
Author(s):  
H. Stracke ◽  
A. Schulz ◽  
D. Moeller ◽  
S. Rossol ◽  
H. Schatz
Keyword(s):  
Growth Hormone ◽  
Alkaline Phosphatase ◽  
Organ Culture ◽  
Culture Medium ◽  
Bone Cells ◽  
Electron Microscopic ◽  
Continuous Increase ◽  
Somatomedin C ◽  
Igf I ◽  
Bone Organ Culture

Abstract. Bone organ culture makes it possible to observe the direct influence of hormones on bone cells. We studied the effect of growth hormone in vitro on embryonal rat tibiae during culture for 7 days, functionally by measuring the levels of alkaline phosphatase in the culture medium, and morphologically by means of semithin sections and electron microscopic examination. Since growth hormone (GH) is supposed to exert an indirect effect on bone cells, somatomedin-C/insulin-like growth factor I (SM-C/IGF I) as a possible mediator was also measured radioimmunologically in the culture medium. In the controls alkaline phosphatase levels showed a continuous increase up to the 7th day which was significantly higher in the presence of GH. There was also a significantly enhanced increase of SM-C/IGF I in the presence of GH during culture in comparison to the controls. Evidently IGF I is produced locally in bone and mediates the effect of GH on bone formation.

Studies on regulation of insulin-like growth factor binding protein (IGFBP)-3 and IGFBP-4 production in human bone cells

1992 ◽  
Vol 127 (6) ◽  
pp. 555-564 ◽  
Author(s):  
Subburaman Mohan ◽  
Donna D Strong ◽  
Uta G Lempert ◽  
Florence Tremollieres ◽  
Jon E Wergedal ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Conditioned Medium ◽  
Cell Cultures ◽  
Bone Cells ◽  
Human Growth ◽  
Bone Cell ◽  
Human Bone ◽  
Human Bone Cell ◽  
Igf I ◽  
Igfbp 3

Previous studies have shown that the actions of IGF-II in bone are determined not only by its concentration, but also by the concentration of IGFBP-4 as well as other IGFBPs. In this study, we sought to determine by Western ligand blotting the effects of growth hormone, IGF-I and IGF-II on the production of IGFBP-3 and IGFBP-4 in TE89 human osteosarcoma cells and in untransformed normal human bone cells derived from rib. Human growth hormone at 10 μg/l decreased the amount of IGFBP-4 but had no effect on the IGFBP-3 level in the conditioned medium of low density cultures of TE89 cells and human bone cells derived from rib. Human growth hormone had no effect on IGFBP-3 or IGFBP-4 levels in the conditioned medium of high density human bone cell cultures. IGF-I and IGF-II, which increased human bone cell proliferation, decreased the level of IGFBP-4 (30% of control at 100 μg/l IGF-I and IGF-II) but increased the level of IGFBP-3 (3–10 fold at 100 μg/l IGF-I and IGF-II) after 48 h of treatment in the conditioned medium of both low and high density TE89 cell cultures. Similar changes in IGFBP-3 and IGFBP-4 levels were also seen in the conditioned medium of human bone cells derived from rib after treatment with IGF-I and IGF-II. Studies to determine the underlying molecular mechanisms by which IGF-II decreased the amount of IGFBP-4 in the conditioned medium revealed that IGF-II decreased the IGFBP-4 mRNA abundance and increased the IGFBP-3 mRNA abundance in human bone cells. Based on the above findings, we conclude that the production of both IGFBP-3 and IGFBP-4 is regulated in bone cells and that local and systemic agents may modulate the responsiveness of bone cells to IGFs by regulated secretion of IGFBP-3 and IGFBP-4.

ORGAN CULTURE OF HUMAN SOMATOTROPHIC PITUITARY ADENOMAS: ULTRASTRUCTURE AND GROWTH HORMONE PRODUCTION

1975 ◽  
Vol 79 (1) ◽  
pp. 217-229 ◽  
Author(s):  
F. Peillon ◽  
M. Gourmelen ◽  
M. Donnadieu ◽  
A. Brandi ◽  
D. Sevaux ◽  
...  
Keyword(s):  
Organ Culture ◽  
Culture Medium ◽  
Secretory Granules ◽  
Electron Microscopic Observation ◽  
Histological Differentiation ◽  
Hormone Production ◽  
Electron Microscopic ◽  
Hormone Synthesis ◽  
Elution Pattern ◽  
Very High

ABSTRACT Ten somatotrophic adenomas removed from acromegalic patients and fragments of the non-tumoural surrounding pituitary were submitted to organ culture for periods of up to one month. Electron microscopic observation shows that these tumours retain their histological differentiation throughout the culture period. The cell morphology of the cultured tumours remains essentially unchanged and in particular the secretory granules keep their initial size (150 and 130 nm). However the granules disappear gradually so that most of the cells look chromophobic by the 4th week of culture, and numerous lysosomes as well as autophagic figures appear at the same time. The hGH concentration in the culture medium has been measured of 4 adenomas. It is very high (10-fold greater than that from non-tumoural pituitary medium) during the first week (range 200–300 μg/ml).It still remains very high in the same experiments until the second week of culture in one experiment (200 μg/ml). After incubation of cultures with 3H-leucine, 3H-hGH is obtained in the medium giving evidence of hormone synthesis by adenoma cells in culture. 3H-hGH represents 40 % of 3H-proteins in the culture medium and gives the same elution pattern as standard hGH on Sephadex G 100 chromatography. A certain degree of correlation is observed between morphological and biological results: the greatest hGH production is obtained from explants which maintain the best histological appearance.

scholarly journals Alkaline phosphatase activity present in serum influences osteogenic differentiation cultures

2021 ◽  
Author(s):  
Sana Ansari ◽  
Keita Ito ◽  
Sandra Hofmann
Keyword(s):  
Alkaline Phosphatase ◽  
Osteogenic Differentiation ◽  
Culture Medium ◽  
Bone Cells ◽  
Mineralization Process ◽  
Matrix Mineralization ◽  
Alp Activity ◽  
Mineral Deposition ◽  
Fetal Bovine ◽  
Mineralized Matrix

Fetal bovine serum (FBS) is a widely used supplement in cell culture medium, despite its known variability in composition which greatly affects cellular function and consequently the outcome of studies. In bone tissue engineering, the deposited mineralized matrix is one of the main outcome parameters, but using different brands of FBS can result in large variations. Alkaline phosphatase (ALP) is present in FBS. Not only is ALP used to judge the osteogenic differentiation of bone cells, it may affect deposition of mineralized matrix. The present study focused on the enzymatic activity of ALP in FBS of different suppliers and its contribution to mineralization in osteogenic differentiation cultures. It was hypothesized that culturing cells in a medium with high intrinsic ALP activity of FBS will lead to higher mineral deposition compared to media with lower ALP activity. The used FBS types were shown to have significant differences in enzymatic ALP activity. Our results indicate that the ALP activity of the medium not only affected the deposited mineralized matrix but also the osteogenic differentiation of cells as measured by a changed cellular ALP activity of human bone marrow derived mesenchymal stromal cells (hBMSC). In media with low inherent ALP activity, the cellular ALP activity was increased and played the major role in the mineralization process; while, in media with high intrinsic ALP activity contribution from the serum, less cellular ALP activity was measured and the ALP activity of the medium also contributed to mineral formation substantially. Our results highlight the diverse effects of ALP activity intrinsic to FBS on osteogenic differentiation and matrix mineralization and how FBS can determine the experimental outcomes, in particular for studies investigating matrix mineralization. Once again, the need to replace FBS with more controlled and known additives is highlighted.

Bone turnover and bone mineral density in young adult patients with panhypopituitarism before and after long-term growth hormone therapy

1995 ◽  
Vol 132 (1) ◽  
pp. 42-46 ◽  
Author(s):  
Rina Balducci ◽  
Vincenzo Toscano ◽  
Anna M Pasquino ◽  
Adele Mangiantini ◽  
Giovanna Municchi ◽  
...  
Keyword(s):  
Bone Mineral Density ◽  
Growth Hormone ◽  
Alkaline Phosphatase ◽  
Bone Turnover ◽  
Bone Mineral ◽  
Serum Levels ◽  
Creatinine Ratio ◽  
Mineral Density ◽  
Urinary Hydroxyproline ◽  
Igf I

Balducci R, Toscano V, Pasquino AM, Mangiantini A, Municchi G, Armenise P, Terracina S, Prossomariti G, Boscherini B. Bone turnover and bone mineral density in young adult partients with panhypopituitarism before and after long-term growth hormone therapy. Eur J Endocrinol 1995;132:42–6. ISSN 0804–4643 We examined the effects of biosynthetic growth hormone (GH) on biochemical indices of bone turnover and on bone mineral density in a group of GH-deficient adults. Thirteen patients (eight males and five females) aged 24 ± 5 years (range 16–35) were studied before and 12 and 24 months after GH treatment (0.1 IU, kg− day−1, 6 days a week). Serum levels of insulin-like growth factor I (IGF-I), calcitonin, parathyroid hormone, alkaline phosphatase, intact osteocalcin, fasting urinary hydroxyproline/creatinine ratio and bone mineral density (BMD), measured at the lumbar spine by dualphoton absorptiometry, were evaluated. After 12 months of treatment, IGF-I, alkaline phosphatase, osteocalcin and the fasting urinary hydroxyproline/creatinine ratio increased significantly. However, after 24 months of therapy, serum levels of osteocalcin decreased to pretreatment values while IGF-I, fasting urinary hydroxyproline/creatinine ratio and alkaline phosphatase remained elevated significantly. No changes were found in parathyroid hormone and calcitonin plasma levels or in BMD either after 12 or 24 months of treatment. These data demonstrate that GH, at the dosage that we used, activates bone turnover during 24 months of therapy in adults with panhypopituitarism, even if a downward trend for osteocalcin became apparent at 24 months. However, this activation in bone turnover was not accompanied by an increase in BMD. We can hypothesize that GH, at the relatively high dosage used, may stimulate osteoclastic activity to a greater extent than osteoblastic activity. It is probable that the dose of GH replacement therapy in adults plays a key role R Balducci, Dipartimento di Sanita Pubblica, Universita "Tor Vergata", Via di Tor Vergata 38, 00173 Roma, Italy

Human Growth Hormone Stimulates Proteinase Activities of Rabbit Bone Cells via IGF-I

2000 ◽  
Vol 268 (3) ◽  
pp. 875-881 ◽  
Author(s):  
Rousselle Anne-Valérie ◽  
Damiens Christelle ◽  
Fortun Yannick ◽  
Passuti Norbert ◽  
Padrines Marc ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Human Growth Hormone ◽  
Bone Cells ◽  
Human Growth ◽  
Igf I ◽  
Rabbit Bone
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