Development of a new ELISA for serum periostin: evaluation of growth-related changes and bisphosphonate treatment in...

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.