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Novel bioactivity of phosvitin in connective tissue and bone organogenesis revealed by live calvarial bone organ culture models.

Abstract：

:Egg yolk phosvitin is one of the most highly phosphorylated extracellular matrix proteins known in nature with unique physico-chemical properties deemed to be critical during ex-vivo egg embryo development. We have utilized our unique live mouse calvarial bone organ culture models under conditions which dissociates the two bone remodeling stages, viz., resorption by osteoclasts and formation by osteoblasts, to highlight important and to date unknown critical biological functions of egg phosvitin. In our resorption model live bone cultures were grown in the absence of ascorbate and were stimulated by parathyroid hormone (PTH) to undergo rapid osteoclast formation/differentiation with bone resorption. In this resorption model native phosvitin potently inhibited PTH-induced osteoclastic bone resorption with simultaneous new osteoid/bone formation in the absence of ascorbate (vitamin C). These surprising and critical observations were extended using the bone formation model in the absence of ascorbate and in the presence of phosvitin which supported the above results. The results were corroborated by analyses for calcium release or uptake, tartrate-resistant acid phosphatase activity (marker for osteoclasts), alkaline phosphatase activity (marker for osteoblasts), collagen and hydroxyproline composition, and histological and quantitative histomorphometric evaluations. The data revealed that the discovered bioactivity of phosvitin mirrors that of ascorbate during collagen synthesis and the formation of new osteoid/bone. Complementing those studies use of the synthetic collagen peptide analog and cultured calvarial osteoblasts in conjunction with mass spectrometric analysis provided results that augmented the bone organ culture work and confirmed the capacity of phosvitin to stimulate differentiation of osteoblasts, collagen synthesis, hydroxyproline formation, and biomineralization. There are striking implications and interrelationships of this affect that relates to the evolutionary inactivation of the gene of an enzyme L-gulono-γ-lactone oxidase, which is involved in the final step of ascorbate biosynthesis, in many vertebrate species including passeriform birds, reptiles and teleost fish whose egg yolk contain phosvitin. These represent examples of how developing ex-vivo embryos of such species can achieve connective tissue and skeletal system formation in the absence of ascorbate.

journal_name

Dev Biol

journal_title

Developmental biology

authors

Liu J,Czernick D,Lin SC,Alasmari A,Serge D,Salih E

doi

10.1016/j.ydbio.2013.06.005

keywords:

["BSP","CKI","Ca-P","Calvarial bone organ culture","Collagen synthesis","DMEM","Dulbecco's modified Eagle medium","ECM","ER","Egg yolk embryo organogenesis","Evolution","LC–ESI–MS\/MS","MS","Mass spectrometry","OPN","Osteoblasts","Osteoclasts","P-Ser","P-Thr","P-Tyr","PTH","Phosvitin","RANKL","TRAP","bone sialoprotein","calcium phosphate","casein kinase I","endoplasmic reticulum","extracellular matrix","liquid-chromatography–electrospray-ionization–tandem mass spectrometry","mCKII","mass spectrometry","microsomal casein kinase II","osteopontin","parathyroid hormone","phosphoserine","phosphothreonine","phosphotyrosine","receptor activator of nuclear factor-Κβ ligand","tartrate resistant acid phosphatase"]

subject

Has Abstract

pub_date

2013-09-01 00:00:00

pages

256-75

issue

eissn

0012-1606

issn

1095-564X

pii

S0012-1606(13)00320-5

journal_volume

381

pub_type

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Novel bioactivity of phosvitin in connective tissue and bone organogenesis revealed by live calvarial bone organ culture models.