Article

Calcified Tissue International

, Volume 87, Issue 4, pp 341-350

Development of a New ELISA for Serum Periostin: Evaluation of Growth-Related Changes and Bisphosphonate Treatment in Mice

  • Sylvain ContiéAffiliated withResearch Unit 664, Institut National de la Santé et de la Recherche MédicaleBiochemical Markers, CCBR-Synarc
  • , Nathalie Voorzanger-RousselotAffiliated withBiochemical Markers, CCBR-Synarc
  • , Judith LitvinAffiliated withDepartment of Anatomy and Cell Biology, Fels Institute for Cancer Research, Temple University School of MedicineCardiovascular Research Center, Temple University School of Medicine
  • , Nicolas BonnetAffiliated withService of Bone Diseases, Department of Rehabilitation and Geriatrics, Geneva University Hospitals and Faculty of Medicine
  • , Serge FerrariAffiliated withService of Bone Diseases, Department of Rehabilitation and Geriatrics, Geneva University Hospitals and Faculty of Medicine
  • , Philippe ClézardinAffiliated withResearch Unit 664, Institut National de la Santé et de la Recherche MédicaleUniversité Claude Bernard Lyon 1
  • , Patrick GarneroAffiliated withResearch Unit 664, Institut National de la Santé et de la Recherche MédicaleCisBio Bioassays Email author 

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Abstract

Periostin is a gamma-carboxyglutamic acid protein preferentially expressed in periosteum and bone mesenchymal stem cells. Lack of a precise assay for measuring circulating levels impairs the investigation of its biological significance. We developed a new ELISA and studied changes of periostin levels both locally at the bone site and systemically in circulating blood during growth and after bisphosphonate-induced inhibition of bone remodeling in the mouse. The ELISA we developed is based on an affinity-purified polyclonal antibody that was raised against the C-terminal sequence of mouse periostin. Reproducibility, repeatability, precision, and accuracy tests met standards of acceptance. Serum periostin and levels of the bone turnover markers osteocalcin, PINP, CTX-I, and TRAP5b were measured in (1) 4-, 6-, 8-, 10-, and 12-week-old wild-type female Balb/c mice and (2) adult ovariectomized female Balb/c mice treated with zoledronic acid or vehicle. Serum periostin decreased during growth and stabilized from 8 weeks and older, its levels correlating with bone turnover markers. Immunohistochemistry in bones from different growth stages showed that periostin localized specifically at the sites of endochondral and intramembranous ossification, especially at the periosteal envelopes. Zoledronic acid induced a marked decrease in bone remodeling markers but did not alter serum periostin levels or periostin immunostaining pattern. The novel ELISA is highly specific and allows accurate and precise measurements of serum periostin levels in mice.

Keywords

Periostin Periosteum ELISA Bone marker Ontogenetic ossification