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A Lab-On-A-Chip Platform for Stimulating Osteocyte Mechanotransduction and Analyzing Functional Outcomes of Bone Remodeling
Published on: May 21, 2020
Identification of mechanosensitive genes in osteoblasts by comparative microarray studies using the rotating wall
Mamta J Patel1, Wenbin Liu, Michelle C Sykes
1Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, Georgia 30322, USA.
Journal of Cellular Biochemistry
|January 24, 2007
Summary
Spaceflight microgravity causes bone loss by reducing bone formation. Ground-based simulators like the Rotating Wall Vessel (RWV) alter gene expression in preosteoblast cells, offering insights into bone loss mechanisms during space travel.
Area of Science:
- Space biology
- Cell biology
- Biotechnology
Background:
- Spaceflight-induced bone loss is a significant concern, primarily due to decreased bone formation.
- The exact mechanisms behind reduced bone formation in microgravity remain largely unknown.
- Ground-based simulators, such as the Rotating Wall Vessel (RWV) and Random Positioning Machine (RPM), are crucial for studying space biology on Earth.
Purpose of the Study:
- To investigate the effects of RWV exposure on 2T3 preosteoblast cell differentiation and gene expression.
- To compare RWV-induced gene expression changes with those from RPM and mechanical loading models.
- To identify mechanosensitive genes that may be involved in bone formation regulation during spaceflight.
Main Methods:
- 2T3 preosteoblast cells were exposed to RWV conditions for 3 days.
- Microarray analysis was used to assess global gene expression changes.
- Alkaline phosphatase activity was measured as a marker of differentiation.
- Real-time PCR and Western blots validated specific gene and protein expression changes.
- RWV data were compared with previously published RPM data and external mechanical loading data.
Main Results:
- RWV exposure inhibited alkaline phosphatase activity in 2T3 cells.
- Microarray analysis revealed significant alterations in gene expression: 61 genes downregulated and 45 genes upregulated by over twofold.
- Seven genes, including osteomodulin, runx2, and osteoglycin, were validated via PCR and Western blot.
- 14 mechanosensitive genes showed similar expression trends between RWV and RPM.
- Three genes, including osteoglycin, were downregulated by RWV/RPM but upregulated by mechanical loading of mouse tibiae.
Conclusions:
- RWV exposure significantly impacts preosteoblast differentiation and gene expression profiles.
- A subset of mechanosensitive genes responds similarly to RWV and RPM, but oppositely to mechanical loading.
- These identified mechanosensitive genes offer potential targets for developing countermeasures against bone loss in spaceflight and related pathologies.
