Tooth morphogenesis and differentiation of odontoblasts and ameloblasts depends upon reciprocal epithelial-mesenchymal interactions. These heterotypic tissue interactions appear to mediate both morphogenesis and differentiation. Nutritional requirements for tooth development are not known. The specific function of nutrients for odontoblast and ameloblast differentiation, and for dentine/enamel formation are not known. We have recently developed an organ culture method which supports embryonic tooth morphogenesis and differentiation in a serumless, chemically-defined medium. Under these experimental conditions both dentine and enamel extracellular matrices are formed without mineralization. It is established that 10-20% fetal calf serum (FCS) supplementation provides the humoral requirements for biological mineralization. We propose studies to investigate the nutritional requirements for both dentine and enamel matrix mineralization in vitro in a serumless, chemically-defined medium using mouse molar tooth organs at the cap stage of development. Experiments are designed to determine the molecule(s), macromolecule(s) and ion(s) found in serum which are specifically required for dentine and enamel formation in vitro. Two different strategies are proposed: (1) to isolate from serum those fractions which promote dentine and enamel formation and to then identify the chemical components of these fractions; and (2) to determine the nutrients and their concentration (e.g. vitamins and trace elements) which will optimize dentine and enamel formation in serumless medium. Methods to analyze the effects of nutrient supplementation include metabolic labeling studies for dentine and enamel matrix proteins using immunodetection assays for dentine phosphoprotein and enamel proteins, assays for phosphate, lipids and calcium, and transmission electron microscopy. We propose to determine the nutrient requirements for optimal biological mineralization during in vitro mammalian tooth development.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
5R01DE006425-02
Application #
3219993
Study Section
Oral Biology and Medicine Study Section (OBM)
Project Start
1984-03-01
Project End
1987-02-28
Budget Start
1985-03-01
Budget End
1986-02-28
Support Year
2
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of Southern California
Department
Type
Schools of Dentistry/Oral Hygn
DUNS #
041544081
City
Los Angeles
State
CA
Country
United States
Zip Code
90033
Caton, Javier; Bringas Jr, Pablo; Zeichner-David, Margarita (2005) IGFs increase enamel formation by inducing expression of enamel mineralizing specific genes. Arch Oral Biol 50:123-9
Takahashi, K; Yamane, A; Bringas, P et al. (1998) Induction of amelogenin and ameloblastin by insulin and insulin-like growth factors (IGF-I and IGF-II) during embryonic mouse tooth development in vitro. Connect Tissue Res 38:269-78;discussion 295-303
Slavkin, H C; Diekwisch, T G (1997) Molecular strategies of tooth enamel formation are highly conserved during vertebrate evolution. Ciba Found Symp 205:73-80; discussion 81-4
Slavkin, H C; Diekwisch, T (1996) Evolution in tooth developmental biology: of morphology and molecules. Anat Rec 245:131-50
Hu, J C; Zhang, C; Slavkin, H C (1995) The role of platelet-derived growth factor in the development of mouse molars. Int J Dev Biol 39:939-45
Diekwisch, T G; Berman, B J; Gentner, S et al. (1995) Initial enamel crystals are not spatially associated with mineralized dentine. Cell Tissue Res 279:149-67
Nakamura, M; Bringas Jr, P; Nanci, A et al. (1994) Translocation of enamel proteins from inner enamel epithelia to odontoblasts during mouse tooth development. Anat Rec 238:383-96
Diekwisch, T; David, S; Bringas Jr, P et al. (1993) Antisense inhibition of AMEL translation demonstrates supramolecular controls for enamel HAP crystal growth during embryonic mouse molar development. Development 117:471-82
Slavkin, H C (1993) Rieger syndrome revisited: experimental approaches using pharmacologic and antisense strategies to abrogate EGF and TGF-alpha functions resulting in dysmorphogenesis during embryonic mouse craniofacial morphogenesis. Am J Med Genet 47:689-97;discussion 687-8
Shum, L; Sakakura, Y; Bringas Jr, P et al. (1993) EGF abrogation-induced fusilli-form dysmorphogenesis of Meckel's cartilage during embryonic mouse mandibular morphogenesis in vitro. Development 118:903-17

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