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Title : Identification and Application of Reaggregated Tooth Using Bone Title : Identification and application of reaggregated tooth using bone marrow mesenchymal stem cells Address : Division in Anatomy and Developmental Biology, Department of Oral Biology, Research Center for Orofacial Hard Tissue Regeneration, Oral Science Research Center, College of Dentistry, Brain Korea 21 project for Medical Science, Yonsei Center of Biotechnology, Yonsei University, 134 Shinchon-Dong, Seodaemun-Gu, Seoul, Korea [email protected] Abstract : Tooth bud requires specific and complex epithelial-mesenchymal interactions for development during organogenesis. The results of epithelial-mesenchymal recombination assay showed that molar tooth bud has the potential to develop tooth with reaggregated dental mesenchymal cells at E13.5. We could obtain the reaggregated tooth after recombination between the dissociated dental mesenchymal cells and oral epithelium. According to the number of the dissociated mesenchymal cells, teeth shapes were altered, such as Molar1-like (M1-like), slope- like and the crater-like shapes. The M1-like teeth were examined by H-E staining and in situ hybridization with signaling molecules. Moreover, We tried to produce reaggregated teeth by using bone marrow mesenchymal stem cells (BMMSCs) which have the ability to differentiate the source tissues of bone, cartilage and other mesoderm. BMMSCs committed to differentiate into teeth when BMMSCs were recombined with oral epithelium after reaggregation with the dental mesenchymal cells at certain ratio. These teeth were also examined by H-E staining and in situ hybridization with Shh and Bmp-4. In particular, the M1-like teeth were implanted into mouse alveolar bone to validate the possibility of bioengineered tooth. We could find that this bioengineered tooth, which was produced by using BMMSCs, might be a launching case for tooth development and tissue engineering. Affiliations : Division in Anatomy and Developmental Biology, Department of Oral Biology, Research Center for Orofacial Hard Tissue Regeneration, Oral Science Research Center, College of Dentistry, Brain Korea 21 project for Medical Science, Yonsei Center of Biotechnology, Yonsei University, Korea. Authors : Jinglei Cai, Sung-Won Cho, Eun-Jung Kim, Jae-Young Kim, Han-Sung Jung Department : Division in Anatomy and Developmental Biology, Department of Oral Biology Forename : Jinglei Institute : Research Center for Orofacial Hard Tissue Regeneration, Oral Science Research Center, College of Dentistry, Brain Korea 21 project for Medical Science, Yonsei Center of Biotechnology, Yonsei University Room : Double Submit : Submit Surname : Cai e-mail : [email protected] Title : IGFs Induce Enamel Formation by Increase Expression of Enamel Mineralizing Specific Genes Address : 2250 Alcazar St CSA 103 Los Angeles, Ca 90033 Affiliations : Center for Craniofacial Molecular Biology, University of So. California [email protected] Abstract : Insulin-like growth factors (IGF-I and IGF-II) have been shown to play an important role in growth and differentiation in a number of tissues including mineralizing bone. Little is known about their role in tooth mineralization. Previous work in our laboratory has shown the presence of IGFs ligands, their receptors, and their binding proteins during mouse tooth morphogenesis, and the expression of IGF I coincides with the expression of amelogenin, ameloblastin and enamelin at the late bell and secretory stage. Furthermore, the addition of IGF ligands to tooth organ cultures in vitro results in an increase in the thickness of the enamel layer while at the same time there is a decrease in the dentin layer. The objective of this study is to determine the mechanisms by which IGFs modulate enamel and dentin formation. Mouse first mandibular molars were dissected from E16, E17 and E18 mouse embryos and placed in organ culture in the presence of 100 ng of IGF-1 or IGF-2.! The molars were harvested after 1, 3 and 6 days in culture and either RNA or nuclear proteins were extracted. RNA was subjected to real time RT-PCR and the nuclear extracts were used to determine the activity of transcription factors using a TF/DNA microarrays. Our results show an induction of enamelin, ameloblastin and amelogenin mRNA expression in E17 tooth organs treated with IGF-I for 24hrs. The microarray results showed changes in DNA binding activity for several transcription factors known to be related to tooth development. The DNA binding activity was increased for Pbx-1, NF-1, Yy-1, CREB(1) but decreased for Brn-3 and CBF as compared to the untreated control. These results suggest that IGFs increase enamel formation by the induction of enamelin, ameloblastin and amelogenin gene expression via the transcription factors Pbx-1, NF-1, Yy-1, CREB(1). Studies are underway to determine a possible mechanism for these factors. This study was supported by NIH-NIDCR grant DE! 006425 Authors : Javier Catón, Pablo Bringas Jr. and Margarita Zeichner-David. Department : CCMB Forename : Javier Institute : USC Room : Single Submit : Submit Surname : Caton Title : The Nature and significant regeneration of primary enamel knot Address : Division in Anatomy and Developmental Biology, Department of Oral Biology, Research Center for Orofacial Hard Tissue Regeneration, Oral Science Research Center, College of Dentistry, Brain Korea 21 project for Medical Science, Yonsei Center of Biotechnology, Yonsei University, 134 Shinchon-Dong, Seodaemun-Gu, Seoul, Korea [email protected] Abstract : The development of individual teeth is regulated by epithelial-mesenchymal interactions that are mediated by signals shared with other organs such as skin appendages. Three transient signaling centers in dental epithelium, such as early signaling center, primary enamel knot (PEK) and secondary enamel knots (SEKs), direct the differential growth and subsequent folding of the dental epithelium and also regulate the growth of the tooth shape. The present study shows the origin, some characteristics and inducing mechanism of enamel knots. The center part of dental lamina moved and formed the PEK region and SEKs were not from the PEK cells. After removal of signaling centers, the remained epithelium was recombined with dental mesenchyme. The regeneration possibility of enamel knots was investigated in these recombined tooth germs with two evidences. One is the epithelial cell cluster expressing Shh and Fgf4 in cap stage and the other is the sharp epithelial edge of inn! er dental epithelium between SEKs in bell stage. The PEK could be regenerated at the tooth germ at stage E12 and E14. To find out necessary factors in PEK regeneration, some of gene expressions, such as Lef1, Bmp2 and Bmp4, were investigated in the E16 recombined tooth germ, which could not regenerate the PEK, and were compared with those gene expressions in E14 recombined tooth germ. Also, inter-stage recombination between epithelium and mesenchyme was applied to identify necessary tissue in PEK regeneration. The necessary tissue in regeneration of PEK might be dental epithelium holding important epithelial signals. Also, the position of regenerated PEK was determined by dental epithelium rather than dental mesenchyme. It is concluded that destiny of PEK including origin, regeneration, size and position is determined by dental epithelium rather than dental mesenchyme. Affiliations : Division in Anatomy and Developmental Biology, Department of Oral Biology, Research Center for Orofacial Hard Tissue Regeneration, Oral Science Research Center, College of Dentistry, Brain Korea 21 project for Medical Science, Yonsei Center of Biotechnology, Yonsei University, 134 Shinchon-Dong, Seodaemun-Gu, Seoul, Korea Authors : Sung-Won Cho, Eun-Jung Kim, Heui-Jung Hwang, Jae-Young Kim, Soo-Jin Song, Hitoshi Yamamoto and Han-Sung Jung Department : Division in Anatomy and Developmental Biology, Department of Oral Biology Forename : Sung-Won Institute : Research Center for Orofacial Hard Tissue Regeneration, Oral Science Research Center, College of Dentistry, Brain Korea 21 project for Medical Science, Yonsei Center of Biotechnology, Yonsei University Room : Double Submit : Submit Surname : Cho Title : Dentition pattern, tooth development and amelogenin expression in the lizard, Chalcides viridanus (Scincidae, Squamata) e-mail : [email protected] Abstract : We have studied the formation of the dentition pattern, tooth replacement, tooth morphology and tooth development, with particular attention paid on amelogenesis and amelogenin expression, in a series of the scincid lizard, Chalcides viridanus, from late embryos to 6 year-old specimens, using light, scanning and transmission electron microscopy, in situ hybridization on sections and immunogold labelling. Tooth shape changes during ontogeny. Through successive replacements, the simple, conical shape of the embryonic tooth modifies into bicuspid in the juveniles. The dental lamina of the first teeth branches off from the oral epithelium, while it spreads from the enamel organ of the previous tooth where a replacement tooth is initiated. The cells of the dental lamina proliferate and differentiate into the three layers of the enamel organ: the outer and inner dental epithelium (i.d.e) separated by the stellate reticulum. Next, the i.d.e cells differentiate into amel! oblasts. Facing them mesenchymal cells differentiate into odontoblasts and deposit predentin matrix. Opposite, the enamel matrix is synthesised by the ameloblasts. Mineralization of the dental matrices starts at the top of the tooth, where the ameloblasts stop
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