Journal
CELL STRESS & CHAPERONES
Volume 17, Issue 2, Pages 203-214Publisher
SPRINGER
DOI: 10.1007/s12192-011-0300-8
Keywords
Bone healing; Heating; Heat shock protein; Growth factors; Preosteoblasts
Categories
Funding
- National Science Foundation [CBET 0966546]
- Institute for Critical Technologies and Applied Sciences (ICTAS) at Virginia Tech.
- Div Of Chem, Bioeng, Env, & Transp Sys
- Directorate For Engineering [1067654] Funding Source: National Science Foundation
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Conditioning protocols involving mechanical stress independently or with chemical cues such as growth factors (GFs) possess significant potential to enhance bone regeneration. However, utilization of thermal stress conditioning alone or with GFs for bone therapy has been under-investigated. In this study, a preosteoblast cell line (MC3T3-E1) was exposed to treatment with water bath heating (44A degrees C, 4 and 8 min) and osteoinductive GFs (bone morphogenetic protein-2 and transforming growth factor-beta 1) individually or in combination to investigate whether these stimuli could promote induction of bone-related markers, an angiogenic factor, and heat shock proteins (HSPs). Cells remained viable when heating durations were less than 20 min at 40A(0)C, 16 min at 42A(0)C, and 10 min at 44A(0)C. Increasing heating duration at 44A degrees C, promoted gene expression of HSPs, osteocalcin (OCN), and osteopontin (OPN) at 8 h post-heating (PH). Heating in combination with GFs caused the greatest gene induction of osteoprotegerin (OPG; 6.9- and 1.6-fold induction compared to sham-treated and GF only treated groups, respectively) and vascular endothelial growth factor (VEGF; 16.0- and 1.6-fold compared to sham and GF-only treated groups, respectively) at 8 h PH. Both heating and GFs independently suppressed the matrix metalloproteinase-9 (MMP-9) gene. GF treatment caused a more significant decrease in MMP-9 protein secretion to non-detectable levels compared to heating alone at 72 h PH. Secretion of OCN, OPN, and OPG increased with the addition of GFs but diminished with heating as measured by ELISA at 72 h PH. These results suggest that conditioning protocols utilizing heating and GFs individually or in combination can induce HSPs, bone-related proteins, and VEGF while also causing downregulation of osteoclastic activity, potentially providing a promising bone therapeutic strategy.
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