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Pigment epithelium-derived factor (PEDF) reduced expression and synthesis of SOST/sclerostin in bone explant cultures: implication of PEDF-osteocyte gene regulation in vivo

  • Feng Li
  • Jarrett D. Cain
  • Joyce Tombran-Tink
  • Christopher NiyibiziEmail author
Original Article
  • 27 Downloads

Abstract

Mutations in Serpinf1 gene which encodes pigment epithelium-derived factor (PEDF) lead to osteogenesis imperfecta type VI whose hallmark is defective matrix mineralization. We reported previously that PEDF reduced expression and synthesis of Sost/Sclerostin as well as other osteocytes genes encoding proteins that regulate matrix mineralization [1]. To determine whether PEDF had an effect on osteocyte gene expression in bone, we used bone explant cultures. First, osteocytes were isolated from surgical waste of bone fragments obtained from patients undergoing elective foot surgeries under approved IRB protocol by Penn State College of Medicine IRB committee. Primary osteocytes treated with PEDF reduced expression and synthesis of Sost/Sclerostin and matrix phosphoglycoprotein (MEPE) as well as dentin matrix protein (DMP-1). On the whole, PEDF reduced osteocyte protein synthesis by 50% and by 75% on mRNA levels. For bone explants, following collagenase digestion, bone fragments were incubated in alpha-MEM supplemented with 250 ng/ml of PEDF or BSA. After 7 days of incubation in a medium supplemented with PEDF, analysis of mRNA by PCR and protein by western blotting of encoded osteocyte proteins showed reduced Sclerostin synthesis by 39% and MEPE by 27% when compared to fragments incubated in medium supplemented with BSA. mRNA expression levels of osteocytes in bone fragments treated with PEDF were reduced by 50% for both SOST and MEPE when compared to BSA-treated bone fragments. Taken together, the data indicate that PEDF has an effect on osteocyte gene expression in bone and encourage further studies to examine effect of PEDF on bone formation indices in animal models and its effect on osteocyte gene expression in vivo following PEDF administration.

Keywords

Pigment epithelium derived factor Sclerostin Osteocytes Bone explant culture Osteogenesis imperfecta 

Notes

Acknowledgements

This work was supported by NIH R21AR067473. We would like to thank Ananya Das for the assistance in organizing the manuscript.

Compliance with ethical standards

Conflict of interest

The authors have no conflict of interest to declare.

Supplementary material

774_2018_982_MOESM1_ESM.pdf (177 kb)
BSA and GAPDH expression

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Copyright information

© Springer Japan KK, part of Springer Nature 2019

Authors and Affiliations

  • Feng Li
    • 1
  • Jarrett D. Cain
    • 1
  • Joyce Tombran-Tink
    • 1
    • 2
  • Christopher Niyibizi
    • 1
    • 3
  1. 1.Department of Orthopaedics and Rehabilitation H089Penn State College of MedicineHersheyUSA
  2. 2.Department of Neural and Behavioral SciencesPenn State College of MedicineHersheyUSA
  3. 3.Biochemistry and Molecular BiologyPenn State College of MedicineHersheyUSA

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