scholarly journals Effects of Chemotherapeutic Agents on the Function of Primary Human Osteoblast-Like Cells Derived from Children

2003 ◽  
Vol 88 (12) ◽  
pp. 6088-6097 ◽  
Author(s):  
J. H. Davies ◽  
B. A. J. Evans ◽  
M. E. M. Jenney ◽  
J. W. Gregory
Keyword(s):  
Type I Collagen ◽  
Cell Function ◽  
Relative Number ◽  
Chemotherapeutic Agents ◽  
Type I ◽  
Human Osteoblast ◽  
Dihydroxyvitamin D3 ◽  
Primary Human Osteoblast ◽  
1,25 Dihydroxyvitamin D3

Abstract Studies in children treated with chemotherapy suggest that chemotherapeutic agents have deleterious effects on bone metabolism. We therefore evaluated the in vitro effects of clinically relevant concentrations of chemotherapeutic agents on the synthesis of type I collagen, alkaline phosphatase (AP) activity, and mineralization by primary human osteoblast-like (HOB) cells derived from children. Because serum 1,25-dihydroxyvitamin D3 concentrations may be reduced during treatment with chemotherapy, the effect of chemotherapeutic agents on HOB cells cultured in the presence or absence of 1,25-dihydroxyvitamin D3 was also evaluated. Type I collagen synthesis was reduced by all agents (P < 0.01) other than methotrexate, whereas the relative AP activity was increased (P < 0.01) by all agents. The relative number of cells staining intensely for AP after culture with agents increased (P < 0.05), and AP mRNA expression was increased (P < 0.01) with vincristine. 1,25-Dihydroxyvitamin D3 ameliorated (P < 0.01) the depletion of HOB cell numbers by chemotherapeutic agents. Furthermore, vincristine and daunorubicin inhibited 1,25-dihydroxyvitamin D3-mediated AP activity (P < 0.01). We conclude that chemotherapeutic agents can adversely affect HOB cell function, and we speculate that this observation may account, in part, for the osteopenia observed during and after treatment of children with chemotherapy.

Creatine supplementation stimulates collagen type I and osteoprotegerin secretion of healthy and osoteopenic primary human osteoblast-like cells in vitro

Bone ◽  
2008 ◽  
Vol 42 ◽  
pp. S21-S22 ◽  
Author(s):  
Isabel Gerber ◽  
Hanswerner Gerber ◽  
Claudio Dora ◽  
Daniel Uebelhart ◽  
Theo Wallimann
Keyword(s):  
Collagen Type ◽  
Creatine Supplementation ◽  
Collagen Type I ◽  
Type I ◽  
Human Osteoblast ◽  
Primary Human Osteoblast

Effect of Adhesion or Collagen Molecules on Cell Attachment, Insulin Secretion, and Glucose Responsiveness in the Cultured Adult Porcine Endocrine Pancreas: A Preliminary Study

2003 ◽  
Vol 12 (4) ◽  
pp. 439-446 ◽  
Author(s):  
Kazuya Edamura ◽  
Koko Nasu ◽  
Yukiko Iwami ◽  
Hiroyuki Ogawa ◽  
Nobuo Sasaki ◽  
...  
Keyword(s):  
Insulin Secretion ◽  
Type I Collagen ◽  
Cell Function ◽  
Cell Attachment ◽  
Control Group ◽  
Type I ◽  
Collagen Group ◽  
Collagen Molecules ◽  
Glucose Responsiveness

The effect of either adhesion or collagen molecules on cell attachment, insulin secretion, and glucose responsiveness was investigated in adult porcine pancreatic endocrine (PE) cells that were cultured for a longer term in vitro. Six different types of molecules—laminin, fibronectin, poly-L-lysine (PLL), type I collagen, gelatin, and Matrigel—were used. Approximately 2.0 × 105 cells per dish of each molecule type were cultured for 4 weeks. In the laminin group, the insulin accumulation was maintained at a significantly higher level than in the control group at 4 weeks of culture, and glucose-stimulated insulin secretion and the insulin-positive rate were also higher than in the control group. In the Matrigel group, islet-like cell clusters were formed, but insulin accumulation rapidly decreased at 3–4 weeks of culture. A large number of PE cells attached tightly and spread in the fibronectin group until the fourth week of culture, but their function was not better than those in the control group. In the PLL and gelatin groups, the PE cell function was not significantly different from that of the control group. In the type I collagen group, insulin secretion was inferior to that of the other groups. The results of this study suggest that laminin is the most suitable extracellular matrix for the long-term culture preservation of PE cells.

scholarly journals 1,25-Dihydroxyvitamin D3 Induces NAD(+)-Dependent 15-Hydroxyprostaglandin Dehydrogenase in Human Neonatal Monocytes

Blood ◽  
1997 ◽  
Vol 89 (6) ◽  
pp. 2105-2112 ◽  
Author(s):  
F. Pichaud ◽  
S. Roux ◽  
J.L. Frendo ◽  
R. Delage-Mourroux ◽  
J. Maclouf ◽  
...  
Keyword(s):  
Mrna Level ◽  
Cyclooxygenase 2 ◽  
Type I ◽  
Blood Monocytes ◽  
Monocytic Differentiation ◽  
Dihydroxyvitamin D3 ◽  
1,25 Dihydroxyvitamin D3 ◽  
Key Enzyme ◽  
Polymerase Chain

Abstract 1,25-Dihydroxyvitamin D3 [1,25-(OH)2D3 ] induces the differentiation of monocytes into macrophage-like cells in vitro. To identify the genes expressed during this process, we performed differential display polymerase chain reaction on RNA extracted from cord blood monocytes (CBMs) treated with 1,25-(OH)2D3 . Treated CBMs expressed type-I 15-hydroxyprostaglandin dehydrogenase (type-I 15-PGDH), the key enzyme of prostaglandin E2 (PGE2 ) catabolism and a 15-PGDH–related mRNA (15-PGDHr). This newly described 15-PGDH–related mRNA was constitutively expressed in adult monocytes. 15-PGDH gene(s) transcription was accompanied by the appearance of the 15-PGDH activity in treated CBMs. In addition, the cyclooxygenase 2 mRNA level was decreased and PGE2 levels in the culture mediums were lowered (50%). Our results stress that 1,25-(OH)2D3 , at least in neonatal monocytes, can exert, directly or indirectly, a dual control on key enzymes of PGE2 metabolism. In conclusion, we suggest that modifications in prostaglandin metabolism, induced by the expression of type-I 15-PGDH and the downregulation of cyclooxygenase 2, could be involved in monocytic differentiation.

scholarly journals Decellularization and Solubilization of Porcine Liver for Use as a Substrate for Porcine Hepatocyte Culture

2017 ◽  
Vol 26 (12) ◽  
pp. 1840-1854 ◽  
Author(s):  
Ramon E. Coronado ◽  
Maria Somaraki-Cormier ◽  
Shanmugasundaram Natesan ◽  
Robert J. Christy ◽  
Joo L. Ong ◽  
...  
Keyword(s):  
Liver Tissue ◽  
Type I Collagen ◽  
Cell Function ◽  
Ammonium Hydroxide ◽  
Human Monocyte ◽  
Comparative Approach ◽  
Type I ◽  
Porcine Liver ◽  
Porcine Hepatocytes

Biologic substrates, prepared by decellularizing and solubilizing tissues, have been of great interest in the tissue engineering field because of the preservation of complex biochemical constituents found in the native extracellular matrix (ECM). The integrity of the ECM is critical for cell behavior, adhesion, migration, differentiation, and proliferation that in turn affect homeostasis and tissue regeneration. Previous studies have shown that various processing methods have a distinctive way of affecting the composition of the decellularized ECM. In this study, we developed a bioactive substrate for hepatocytes in vitro, made of decellularized and solubilized liver tissue. The present work is a comparative approach of 2 different methods. First, we decellularized porcine liver tissue with ammonium hydroxide versus a sodium deoxycholate method, then characterized the decellularized tissue using various methods including double stranded DNA (dsDNA) content, DNA size, immunogenicity, and mass spectrometry. Second, we solubilized the decellularized porcine liver with hydrochloric acid versus acetic acid (AA) and characterized the resultant solubilized tissues using relevant methodologies including protein yield, immunogenicity, and bioactivity. Finally, we isolated primary porcine hepatocytes, cultured, and evaluated their bioactivity on the optimized decellularized–solubilized liver substrate. The decellularized porcine liver ECM processed by the ammonium hydroxide method and solubilized with AA displayed higher ECM integrity, low dsDNA, no evidence of intact nuclei, low human monocyte chemoattraction, and the presence of key molecules typically found in the native liver, a very important element for normal cell function. In addition, primary porcine hepatocytes showed enhanced functionality including albumin and urea production and bile canaliculi formation when cultured on the developed liver substrate compared to type I collagen.

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