Downregulation of Heat Shock Protein 70 Impairs Osteogenic and Chondrogenic Differentiation in Human Mesenchymal Stem Cells

Sci Rep. 2018 Jan 11;8(1):553. doi: 10.1038/s41598-017-18541-1.

Abstract

Human mesenchymal stem cells (hMSCs) show promise for bone and cartilage regeneration. Our previous studies demonstrated that hMSCs with periodic mild heating had enhanced osteogenic and chondrogenic differentiation with significantly upregulated heat shock protein 70 (HSP70). However, the role of HSP70 in adult tissue regeneration is not well studied. Here, we revealed an essential regulatory mechanism of HSP70 in osteogenesis and chondrogenesis using adult hMSCs stably transfected with specific shRNAs to knockdown HSP70. Periodic heating at 39 °C was applied to hMSCs for up to 26 days. HSP70 knockdown resulted in significant reductions of alkaline phosphatase activity, calcium deposition, and gene expression of Runx2 and Osterix during osteogenesis. In addition, knockdown of HSP70 led to significant decreases of collagens II and X during chondrogenesis. Thus, downregulation of HSP70 impaired hMSC osteogenic and chondrogenic differentiation as well as the enhancement of these processes by thermal treatment. Taken together, these findings suggest a putative mechanism of thermal-enhanced bone and cartilage formation and underscore the importance of HSP70 in adult bone and cartilage differentiation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase / genetics
  • Alkaline Phosphatase / metabolism
  • Calcium / metabolism
  • Cell Differentiation*
  • Cells, Cultured
  • Chondrocytes / cytology*
  • Chondrocytes / metabolism
  • Core Binding Factor Alpha 1 Subunit / genetics
  • Core Binding Factor Alpha 1 Subunit / metabolism
  • Down-Regulation
  • HEK293 Cells
  • HSP72 Heat-Shock Proteins / genetics*
  • HSP72 Heat-Shock Proteins / metabolism
  • Heat-Shock Response
  • Humans
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / metabolism*
  • Osteoblasts / cytology*
  • Osteoblasts / metabolism
  • Sp7 Transcription Factor / genetics
  • Sp7 Transcription Factor / metabolism

Substances

  • Core Binding Factor Alpha 1 Subunit
  • HSP72 Heat-Shock Proteins
  • RUNX2 protein, human
  • Sp7 Transcription Factor
  • Sp7 protein, human
  • Alkaline Phosphatase
  • Calcium