Document Detail


Bone cell survival in microgravity: evidence that modeled microgravity increases osteoblast sensitivity to apoptogens.
MedLine Citation:
PMID:  15644346     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Studies were performed to evaluate the effects of modeled microgravity on the induction of osteoblast apoptosis. MC3T3-E1 osteoblast-like cells were cultured in alginate carriers in the NASA-approved high aspect ratio vessel (HARV). This system subjects the cells to a time-averaged gravitational field (vector-averaged gravity) to simulate low gravity conditions. Cells were cultured in the HARV for five days, and then examined for apoptosis. In simulated microgravity, the cells remained vital, although analysis of expressed genes indicated that there was loss of the mature osteoblast phenotype. Additionally, we noted that there was a loss of the mitochondrial membrane potential, a low level of the antiapoptotic protein Bcl-2, as well as Akt protein, and the redox status of the cells was disturbed. All of these parameters indicated that vector-averaged gravity disrupts mitochondrial function, thereby sensitizing osteoblasts to apoptosis. We then used a challenge assay to evaluate the apoptotic sensitivity of the cells subjected to vector-averaged gravity. When challenged with staurosporine, cells subjected to vector-averaged gravity evidenced elevated levels of cell death relative to control cell populations. Another objective of the study was to improve upon conventional carriers by using alginate encapsulation to support cells in the HARV. We have demonstrated that the alginate carrier system affords a more robust system than surface-seeded carriers. This new system has the advantage of shielding cells from mechanical damage and fluid shear stresses on cells in the HARV, permitting carefully controlled studies of the effects of vector-averaged gravity.
Authors:
M A Bucaro; J Fertala; C S Adams; M Steinbeck; P Ayyaswamy; K Mukundakrishnan; I M Shapiro; M V Risbud
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Annals of the New York Academy of Sciences     Volume:  1027     ISSN:  0077-8923     ISO Abbreviation:  Ann. N. Y. Acad. Sci.     Publication Date:  2004 Nov 
Date Detail:
Created Date:  2005-01-12     Completed Date:  2005-02-18     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7506858     Medline TA:  Ann N Y Acad Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  64-73     Citation Subset:  IM; S    
Affiliation:
Department of Orthopaedic Surgery, Thomas Jefferson University, 1015 Walnut Street, Philadelphia, PA 19107, USA.
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MeSH Terms
Descriptor/Qualifier:
3T3 Cells
Alginates / chemistry
Animals
Annexin A5 / pharmacology
Apoptosis
Biophysics / methods
Blotting, Western
Bone and Bones / cytology*
Cell Size
Cell Survival
Flow Cytometry
Glucuronic Acid / chemistry
Gravity, Altered
Hexuronic Acids / chemistry
Humans
Membrane Potentials
Mice
Mitochondria / pathology
Musculoskeletal System
Osteoblasts / metabolism,  pathology*
Phenotype
Proto-Oncogene Proteins c-bcl-2 / metabolism
RNA / chemistry
Reverse Transcriptase Polymerase Chain Reaction
Space Flight
Staurosporine / pharmacology
Stress, Mechanical
Time Factors
Weightlessness*
Weightlessness Simulation
Chemical
Reg. No./Substance:
0/Alginates; 0/Annexin A5; 0/Hexuronic Acids; 0/Proto-Oncogene Proteins c-bcl-2; 576-37-4/Glucuronic Acid; 62996-74-1/Staurosporine; 63231-63-0/RNA; 9005-32-7/alginic acid
Investigator
Investigator/Affiliation:
I M Shapiro / Thomas Jefferson U, Philadelphia, PA

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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