Metabolic expression of intrinsic developmental programs for dentine and enamel biomineralization in serumless,chemically-defined,organotypic culture |
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Authors: | John Evans Pablo Bringas Jr Masanori Nakamura Etsuko Nakamura Valentino Santos Professor Harold C Slavkin |
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Institution: | (1) Laboratory For Developmental Biology, Gerontology Center, University of Southern California, Room 314, 90089-019 Los Angeles, California, USA |
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Abstract: | Summary Biomineralization was investigated using embryonic mouse mandibular first molars (M1) cultured in the presence or absence of fetal calf serum. Metabolic features including cell division and Ca2+ and phosphate incorporation into dentine and enamel extracellular matrices were analyzed. The relative timing and magnitude
of DNA synthesis for serumless cultures was comparable toin vivo controls. Isotopic calcium and phosphate incorporation into the mineral phase of dentine and enamel matrices, in the absence
of serum, fluctuated during development. Molar tooth morphogenesis, cytodifferentiation, and extracellular matrix formation
approximated late crown-stage development in serumless cultures. Von Kossa histochemical staining indicated calcium phosphate
salt formation in serumless cultures. Analysis of anhydrous fixation-prepared enamel and dentine representing serumless cultured
explants indicated that crystal size and orientation were comparable toin vivo enamel and dentine. In contrast, serum-supplemented cultures showed atypical crystal size and orientation. Calcium/phosphorous
(Ca/P) ratio values for serumless cultures after 21 days showed Ca/P enamel values of 2.03 (SD±0.04, p<0.025) and dentine
values of 1.89 (SD±0.01, p<0.025). Electron diffraction patterns of enamel and dentine formed in serumless cultures were principally
those of highly-ordered crystalline hydroxyapatite. Our results suggest that tissue-specific dentine and enamel biomineralization
is regulated by endogenous factors intrinsic to the developmental program of embryonic tooth organs during serumless culture. |
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Keywords: | Enamel Dentine Biomineralization Energy-dispersive X-ray microanalysis |
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