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Response of preosteoblasts to thermal stress conditioning and osteoinductive growth factors

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Cell Stress and Chaperones Aims and scope

Abstract

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 (44°C, 4 and 8 min) and osteoinductive GFs (bone morphogenetic protein-2 and transforming growth factor-β1) 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 40ºC, 16 min at 42ºC, and 10 min at 44ºC. Increasing heating duration at 44°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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Funding sources

The following are the funding sources of this work: National Science Foundation grant CBET 0966546 and Institute for Critical Technologies and Applied Sciences (ICTAS) at Virginia Tech.

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All authors have no conflicts of interest.

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Authors and Affiliations

  1. School of Biomedical Engineering and Sciences, Virginia Tech–Wake Forest University, Virginia Tech, ICTAS Bldg., Stanger Street (MC 0298), Blacksburg, VA, 24061, USA

    Eunna Chung & Marissa Nichole Rylander

  2. Department of Mechanical Engineering, Virginia Tech, Virginia Tech, ICTAS Bldg., Stanger Street (MC 0298), Blacksburg, VA, 24061, USA

    Marissa Nichole Rylander

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  1. Eunna Chung
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  2. Marissa Nichole Rylander
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Correspondence to Marissa Nichole Rylander.

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Chung, E., Rylander, M.N. Response of preosteoblasts to thermal stress conditioning and osteoinductive growth factors. Cell Stress and Chaperones 17, 203–214 (2012). https://doi.org/10.1007/s12192-011-0300-8

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  • DOI: https://doi.org/10.1007/s12192-011-0300-8

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