Biomedical subjects

Thimios A Mitsiadis

Publications and source records attributed to Thimios A Mitsiadis.

13 recordsLinked to original sources

PubMed2006

Waking-up the sleeping beauty: recovery of the ancestral bird odontogenic program.

Recent advances in molecular and developmental genetics have provided tools for understanding evolutionary changes in the nature of the epithelial-mesenchymal interactions regulating the patterned outgrowth of the tooth primordia. Tissue recombination experiments in mice have identified the oral epithelium as providing the instructive information for the initiation of tooth development. Teeth were lost in birds for more than 80 million years ago, but despite their disappearance, a number of gene products and the requisite tissue interactions needed for tooth formation are found in the avian oral region. It is believed that the avian ectomesenchyme has lost the odontogenic capacity, whilst the oral epithelium retains the molecular signaling required to induce odontogenesis. In order to investigate the odontogenic capacity of the neural crest-derived mesenchyme and its potential activation of the avian oral epithelium, we have realized mouse neural tube transplantations to chick embryos to replace the neural crest cells of chick with those of mouse. Teeth are formed in the mouse/chick chimeras, indicating that timing is critical for the acquisition of the odontogenic potential by the epithelium and, furthermore, suggesting that odontogenesis is initially directed by species-specific mesenchymal signals interplaying with common epithelial signals.

Animals↗

PubMed2006

How do genes make teeth to order through development?

This introduction to new patterning theories for the vertebrate dentition outlines the historical concepts to explain graded sequences in tooth shape in mammals (incisors, canines, premolars, molars) which change in evolution in a linked manner, constant for each region. The classic developmental models for shape regulation, known as the 'regional field' and 'dental clone' models, were inspired by the human dentition, where it is known that the last tooth in each series is the one commonly absent. The mouse, as a valuable experimental model, has provided data to test these models and more recently, based on spatial-temporal gene expression data, the 'dental homeobox code' was proposed to specify regions and regulate tooth shape. We have attempted to combine these hypotheses in a new model of the combinatorial homeobox gene expression pattern with the clone and field theories in one of 'co-operative genetic interaction'. This also explains the genetic absence of teeth in humans ascribed to point mutations in mesenchymally expressed genes, which affect tooth number in each series.

Animals↗

PubMed2006

Mesenchymal multipotency of adult human periosteal cells demonstrated by single-cell lineage analysis.

OBJECTIVE: To investigate whether periosteal cells from adult humans have features of multipotent mesenchymal stem cells (MSCs) at the single-cell level. METHODS: Cell populations were enzymatically released from the periosteum of the proximal tibia obtained from adult human donors and then expanded in monolayer. Single-cell-derived clonal populations were obtained by limiting dilution. Culture-expanded periosteal cell populations were tested for their growth potential and for expression of conventional markers of MSCs and were subjected to in vitro assays to investigate their multilineage potential. To assess their multipotency in vivo, periosteal cells were injected into a regenerating mouse tibialis anterior muscle for skeletal myogenesis or were either seeded into an osteoinductive matrix and implanted subcutaneously into nude mice for osteogenesis or implanted in a joint surface defect under a periosteal flap into goats for chondrogenesis. Cell phenotypes were analyzed by histochemistry and immunohistochemistry and by reverse transcription-polymerase chain reaction for the expression of lineage-related marker genes. RESULTS: Regardless of donor age, periosteal cells were clonogenic and could be expanded extensively in monolayer, maintaining linear growth curves over at least 30 population doublings. They displayed long telomeres and expressed markers of MSCs. Under specific conditions, both parental and single-cell-derived clonal cell populations differentiated to the chondrocyte, osteoblast, adipocyte, and skeletal myocyte lineages in vitro and in vivo. CONCLUSION: Our study demonstrates that, regardless of donor age, the adult human periosteum contains cells that, upon enzymatic release and culture expansion, are multipotent MSCs at the single-cell level.

Adult↗

PubMed2006

Activation of WNT and BMP signaling in adult human articular cartilage following mechanical injury.

Acute full thickness joint surface defects can undergo repair, which involves tissue patterning and endochondral bone formation. Molecular signals regulating this process may contribute to the repair outcome, chronic evolution and, eventually, the onset of osteoarthritis. We tested the hypothesis that mechanical injury modulates morphogenetic pathways in adult human articular cartilage explants. Adjacent articular cartilage explants were obtained from preserved areas of the femoral condyles of patients undergoing arthroplasty for osteoarthritis, or from a normal joint of a patient undergoing lower limb amputation. Paired explants were individually maintained in explant culture. From each pair, one explant was mechanically injured and the other left uninjured as a control. Cultures were terminated at different time points for histochemistry, immunohistochemistry and gene expression analysis by reverse transcription real time PCR. Bone morphogenetic protein 2 (BMP-2) mRNA was upregulated in the injured explants. We detected phosphorylation of SMAD-1 and SMAD-5, consistent with activation of the bone morphogenetic protein (BMP) pathway. FRZB-1 mRNA was downregulated in the injured explants, suggesting de-repression of WNT signaling. Accordingly, expression of the canonical WNT target genes Axin-2 and c-JUN was upregulated in the injured explants. Activation of the canonical WNT signaling pathway by LiCl treatment induced upregulation of COL2A1 and Aggrecan mRNA, suggesting an anabolic effect. Phosphorylation of SMAD-1/-5 and downregulation of FRZB were confirmed in vivo in a mouse model of joint surface injury. Taken together, these data show modulation of the BMP and WNT pathways following mechanical injury in vitro and in vivo, which may play a role in the reparative response of the joint surface. These pathways may, therefore, represent potential targets in protocols of biological joint surface defect repair.

Aged↗

PubMed2005

Polymerized bonding agents and the differentiation in vitro of human pulp cells into odontoblast-like cells.

OBJECTIVES: Odontoblasts are highly differentiated post-mitotic cells, which under pathological conditions such as carious lesions and dental injuries may degenerate and be replaced by other pulp cells. We have recently shown that this physiological event can be reproduced in an in vitro assay system, but is highly modified by the presence of unpolymerized resinous monomers. Our hypothesis was that the photopolymerization of the monomers in the bonding agents might abolish these negative effects. The purpose of this study was to evaluate the effects of polymerized dentin bonding agents, through dentin slices, on odontoblast differentiation in vitro. METHODS: Pulp cells were obtained from human third molars. They were used to study the effects of four dentin bonding agents through 0.7 mm dentin slices which served as a barrier between the bonding agents and the culture medium. The media containing the bonding agents' extracts were added at non-toxic concentrations onto the cultured cells. Immunohistochemistry was performed to study the differentiation of pulp fibroblasts into odontoblasts under these conditions by evaluating the expression of several odontoblast specific genes. RESULTS: Pulp fibroblasts cultivated under these conditions synthesized type I collagen, osteonectin, dentin sialoprotein and nestin at the same level as in control cultures. Moreover, pulp cells synthesized a mineralized nodular extracellular matrix. Expression of these proteins was higher in the cells contributing to the nodule formation. In addition, except nestin, all these proteins were expressed in the mineral nodules. SIGNIFICANCE: This work shows the lack of effects of photopolymerized bonding agents, through dentin slices, on cytodifferentiation of secondary odontoblasts.

Adolescent↗

PubMed2004

Role of the Notch signalling pathway in tooth morphogenesis.

Notch receptors are involved in cell fate decisions through the process of lateral inhibition or inductive signalling. Jagged2 belongs to the family of transmembrane proteins that serve as the ligands for Notch receptors. We have analysed the expression of the Jagged2 gene in developing mouse teeth. Jagged2 expression is restricted in inner enamel epithelial cells that give rise to the ameloblasts. We have also examined the role of Jagged2 in tooth development using mutant mice that lack the domain of the Jagged2 protein required for interaction with the Notch receptors (DSL domain). Homozygous mutant mice die after birth, exhibit abnormal tooth morphology and fusions between the palatal and mandibular shelves. These results demonstrate that Notch signalling plays an essential role in tooth development.

Ameloblasts↗

PubMed2003

Development of teeth in chick embryos after mouse neural crest transplantations.

Teeth were lost in birds 70-80 million years ago. Current thinking holds that it is the avian cranial neural crest-derived mesenchyme that has lost odontogenic capacity, whereas the oral epithelium retains the signaling properties required to induce odontogenesis. To investigate the odontogenic capacity of ectomesenchyme, we have used neural tube transplantations from mice to chick embryos to replace the chick neural crest cell populations with mouse neural crest cells. The mouse/chick chimeras obtained show evidence of tooth formation showing that avian oral epithelium is able to induce a nonavian developmental program in mouse neural crest-derived mesenchymal cells.

Animals↗

PubMed2003

Notch2 protein distribution in human teeth under normal and pathological conditions.

Notch signaling is essential for the appropriate differentiation of many cell types during development and, furthermore, is implicated in a variety of human diseases. Previous studies have shown that although the Notch1, -2, and -3 receptors are expressed in developing and injured rodent teeth, Notch2 expression was predominant after a lesion. To pursue the role of the Notch pathway in tooth development and disease, we have analyzed the expression of the Notch2 protein in embryonic and adult wounded human teeth. During the earlier stages of tooth development, the Notch2 protein was expressed in the epithelium, but was absent from proliferating cells of the inner enamel epithelium. At more advanced stages, Notch2 was expressed in the enamel-producing ameloblasts, while it was absent in mesenchyme-derived odontoblasts that synthesize the dentin matrix. Although Notch2 was not expressed in the pulp of adult intact teeth, it was reexpressed during dentin repair processes in odontoblasts and subodontoblastic cells. Transforming growth factor beta-1, which stimulates odontoblast differentiation and hard tissue formation after dental injury, downregulated Notch2 expression in cultured human dental slices, in vitro. These observations are consistent with the notion that Notch signaling is an important element in dental physiological and pathogenic conditions.

Adult↗

PubMed2003

BEN/DM-GRASP/SC1 expression during mouse facial development: differential expression and regulation in molars and incisors.

The cell adhesion molecule BEN/DM-GRASP/SC1 is expressed in a variety of tissues during embryogenesis. Here, we studied the expression pattern of BEN/DM-GRASP/SC1 in different organs involved in facial mouse development, especially in the developing teeth. BEN/DM-GRASP/SC1 was expressed in nose, whisker, gland, and tongue epithelia, as well as in myogenic mesenchyme. In molars, BEN/DM-GRASP/SC1 was firstly expressed in the condensed mesenchyme and thereafter expression was confined to mesenchymal cells of the dental follicle. In contrast, in incisors, transient BEN/DM-GRASP/SC1 expression was restricted to epithelium. In tissue recombination experiments, BEN/DM-GRASP/SC1 expression in mesenchyme was activated by molar, but not incisor epithelium.

Activated-Leukocyte Cell Adhesion Molecule↗

PubMed2003

Role of Islet1 in the patterning of murine dentition.

It is believed that mouse dentition is determined by a prepatterning of the oral epithelium into molar (proximal) and incisor (distal) regions. The LIM homeodomain protein Islet1 (ISL1) is involved in the regulation of differentiation of many cell types and organs. During odontogenesis, we find Islet1 to be exclusively expressed in epithelial cells of the developing incisors but not during molar development. Early expression of Islet1 in presumptive incisor epithelium is coincident with expression of Bmp4, which acts to induce Msx1 expression in the underlying mesenchyme. To define the role of ISL1 in the acquisition of incisor shape, we have analysed regulation of Islet1 expression in mandibular explants. Local application of bone morphogenetic protein 4 (BMP4) in the epithelium of molar territories either by bead implantation or by electroporation stimulated Islet1 expression. Inhibition of BMP signalling with Noggin resulted in a loss of Islet1 expression. Inhibition of Islet1 in distal epithelium resulted in a loss of Bmp4 expression and a corresponding loss of Msx1 expression, indicating that a positive regulatory loop exists between ISL1 and BMP4 in distal epithelium. Ectopic expression of Islet1 in proximal epithelium produces a loss of Barx1 expression in the mesenchyme and resulted in inhibition of molar tooth development. Using epithelial/mesenchymal recombinations we show that at E10.5 Islet1 expression is independent of the underlying mesenchyme whereas at E12.5 when tooth shape specification has passed to the mesenchyme, Islet1 expression requires distal (presumptive incisor) mesenchyme. Islet1 thus plays an important role in regulating distal gene expression during jaw and tooth development.

Animals↗

PubMed2002

Influence of resinous monomers on the differentiation in vitro of human pulp cells into odontoblasts.

Odontoblasts are highly differentiated postmitotic cells, which under pathological conditions such as carious lesions and dental injuries may degenerate and be replaced by other pulp cells. A recent work showed that this physiological event can be reproduced in an in vitro assay system. The purpose of the present study was to evaluate the effects of resinous monomers on odontoblast differentiation in vitro. Pulp cores from extracted human third molars were cultured with beta-glycerophosphate (2 mM) and used to evaluate the effects of TEGDMA, HEMA, UDMA, and Bis-GMA on the differentiation of pulp fibroblasts into odontoblasts. The effect of the monomers was studied by evaluating the expression of several odontoblast specific genes. In the absence of monomers, mineral nodule formation was observed. Pulp cells contributing to the nodule formation synthesized type I collagen, osteonectin, and dentin sialoprotein (DSP). In addition, Fourier transform infrared microspectroscopy showed that the mineral and organic composition of the nodules were characteristic of dentin. When the monomers were added at nontoxic concentrations, the effects of HEMA and Bis-GMA were more evident than that of TEGDMA and UDMA on collagen 1, osteonectin, and DSP expression. However, all monomers significantly decreased DSP expression and completely inhibited the mineral nodule formation.

Acrylates↗

PubMed2002

E- and N-cadherin distribution in developing and functional human teeth under normal and pathological conditions.

Cadherins are calcium-dependent cell adhesion molecules involved in the regulation of various biological processes such as cell recognition, intercellular communication, cell fate, cell polarity, boundary formation, and morphogenesis. Although previous studies have shown E-cadherin expression during rodent or human odontogenesis, there is no equivalent study available on N-cadherin expression in dental tissues. Here we examined and compared the expression patterns of E- and N-cadherins in both embryonic and adult (healthy, injured, carious) human teeth. Both proteins were expressed in the developing teeth during the cap and bell stages. E-cadherin expression in dental epithelium followed an apical-coronal gradient that was opposite to that observed for N-cadherin. E-cadherin was distributed in proliferating cells of the inner and outer enamel epithelia but not in differentiated cells such as ameloblasts, whereas N-cadherin expression was up-regulated in differentiated epithelial cells. By contrast to E-cadherin, N-cadherin was also expressed in mesenchymal cells that differentiate into odontoblasts and produce the hard tissue matrix of dentin. Although N-cadherin was not detected in permanent intact teeth, it was re-expressed during dentin repair processes in odontoblasts surrounding carious or traumatic sites. Similarly, N-cadherin re-expression was seen in vitro, in cultured primary pulp cells that differentiate into odontoblast-like cells. Taken together these results suggest that E- and N-cadherins may play a role during human tooth development and, moreover, indicate that N-cadherin is important for odontoblast function in normal development and under pathological conditions.

Adolescent↗

PubMed2002

In vivo and in vitro expression of connexin 43 in human teeth.

Gap junctions are composed of transmembrane proteins belonging to the connexin family. These proteins permit the exchange of mall regulatory molecules directly between cells for the control of growth, development and differentiation. Although the presence of gap junctions in teeth has been already evidenced, the involved connexins have not yet been identified in human species. Here, we examined the distribution of connexin 43 (Cx43) in embryonic and permanent intact and carious human teeth. During tooth development, Cx43 localized both in epithelial and mesenchymal dental cells, correlated with cytodifferentiation gradients. In adult intact teeth, Cx43 was distributed in odontoblast processes. While Cx43 expression was downregulated in mature intact teeth, Cx43 appeared to be upregulated in odontoblasts facing carious lesions. In cultured pulp cells, Cx43 expression was related to the formation of mineralized nodules. These results indicate that Cx43 expression is developmentally regulated in human dental tissues, and suggest that Cx43 may participate in the processes of dentin formation and pathology.

Adolescent↗