Molecular characterisation of odontoblast during primary, secondary ...

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Introduction communication, which may be involved in regulating the pulp healing process (Magloire, Couble et al. 2004). Figure 8: Histology of first upper bovine molar. 7µm section, Haematoxylin and eosin stained. Bar scale=500µm (A): higher magnification view – bar scale - 250µm Unlike osteocytes, odontoblasts do not become incorporated in the matrix, except for their processes that are embedded in the tubules. This is why dentine must not be considered as an individual tissue but rather as the dentine-pulp complex. The odontoblast processes contain limited organelles (which are believed to be responsible for the later secretion of intra-tubular dentine), but are mostly filled by a dense network of micro-filaments and other micro-tubules. Increasingly, coronal odontoblasts are regarded as being different from those found in the root (Nanci 2003). The coronal odontoblasts are elongated and pyramidal with 32
Introduction an apical nucleus, whereas radicular odontoblasts are more cuboidal, which is potentially an indication of significantly lower cellular activity. This fact is seldom taken into account, but would explain why therapeutic interventions that have been validated at a coronal level (pulp capping, for instance) will often fail when applied to the radicular pulp (pulpotomy). 1.3.2 Final differentiation process of odontoblasts during development. Odontoblasts synthesise the components of the predentine (collagen, glycoproteins and other non-collagenous proteins) and are responsible for its mineralisation. As for the other cells of the dental pulp, the odontoblasts are derived from neural crest mesenchymal cells and their differentiation is under the control of the internal dental epithelium (IDE). Differentiation is initiated at each cusp tip and is carried out according to a specific temporo-spatial pattern. In the mouse, the first differentiated odontoblasts appear on the 18 th day of embryonic development, on the top of the main cusp of the first lower molar. The differentiation process gradually extends to the apical area and leads to the formation of a gradient of differentiation (Ruch, Lesot et al. 1995). 1.3.2.1 From preodontoblast to functional odontoblast In the early stages of development, undifferentiated cells derived from the cranial neural crest migrate to the first branchial arch. The oral epithelium plays a key role in the initial pattern of the tooth, by controlling cellular commitment as well as the morphology of the tooth germs (incisor, molar, etc). Final differentiation of preodontoblasts into odontoblast occurs after a specific number of cell divisions (Ruch, Lesot et al. 1995). All of the cells of the dental papilla have the potential to differentiate into odontoblasts, however, only those in contact with the basal membrane at the interface with the Internal Dental Epithelium (IDE) 33
Page 1 and 2: ECOLE DOCTORALE : Génétique, Cell
Page 3 and 4: ACKNOWLEDGEMENTS I wish to express
Page 5 and 6: Abstract : This research aimed to i
Page 7 and 8: CONTENTS List of Figures ………
Page 9 and 10: 4.5.2 MSX2 and pulp repair ........
Page 11 and 12: Figure 11 : Dentine bridge formatio
Page 13 and 14: minutes. (DMP = dentine matrix prot
Page 15 and 16: lipofectamin ® vector only (MDPC v
Page 17 and 18: List of Tables: Table 1: Primer det
Page 19 and 20: 1.1 Towards a biological approach I
Page 21 and 22: Introduction odontoblast processes
Page 23 and 24: Introduction Secondary dentine: thi
Page 25 and 26: Introduction Understanding the dist
Page 27 and 28: Introduction terms of the communica
Page 29 and 30: Figure 5 : There is a differential
Page 31 and 32: Introduction Movement of the dentin
Page 33: 1.2.3.3 Consequences of new technol
Page 37 and 38: Introduction creates a physical bar
Page 39 and 40: Introduction polarisation and the d
Page 41 and 42: Introduction variations in pressure
Page 43 and 44: Introduction also express neurogeni
Page 45 and 46: 1.4.1 Reactionary dentinogenesis In
Page 47 and 48: Introduction responses, including a
Page 49 and 50: Introduction superficial cells move
Page 51 and 52: Introduction induce dentine bridge
Page 53 and 54: Introduction Because of its highly
Page 55 and 56: Introduction (Hayashi 1982). At 11
Page 57 and 58: Introduction To date, involvement o
Page 59 and 60: Introduction In deeper cavities, wh
Page 61 and 62: Introduction dental papilla cells.
Page 63 and 64: 2 Materials and Methods Material an
Page 65 and 66: Material and methods containing; 12
Page 67 and 68: 2.1.2.3 Microarray analysis Materia
Page 69 and 70: Material and methods 2.2 Part Two:
Page 71 and 72: Material and methods a Modulyo free
Page 73 and 74: Material and methods Figure 16: Pri
Page 75 and 76: Material and methods To date, sever
Page 77 and 78: Material and methods (pH=6.8) with
Page 79 and 80: Material and methods (P), and calci
Page 81 and 82: Material and methods (Sigma, la Ver
Page 83 and 84: Material and methods visualized by
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2.4.2.6 MSX2 over-expression in imm
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Material and methods channel. Data
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Results Amelogenin, IBSP, DCN, Alka
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Results data previously reported fo
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91 MAPK PATHWAY probe set GENE ID b
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Results Alkaline phosphatase and am
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Figure 24: Mouse upper incisor (A)
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Figure 25: Phosphorylated p38 prote
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3.3 MSX2 proteins and dentinogenesi
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Figure 29: First upper molar in +/+
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3.3.1.3 At 21days post natal : Resu
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Results expression, obtained using
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Figure 37: Immunohistochemical anal
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3.3.4 BrdU IHC Results BrdU was inj
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3.3.6 Quantitative RT-PCR (Q-RT-PCR
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Results 3.4 Design and use of a new
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Results Figure 46: Histology at 2 w
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Results cells is also characteristi
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4 Discussion Discussion 4.1 Regulat
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Discussion odontoblast secretory be
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Discussion 4.2 Molecular characteri
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Discussion al. 2004; Magloire, Coub
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Discussion differences. Bone cells
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Discussion we can conclude that the
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Discussion pathway is complex and i
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Discussion odontoblast physiologica
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Discussion dentinogenesis. By perfo
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Discussion cellular and molecular s
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Discussion Notably, our results are
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Discussion contribute to reparative
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Discussion other species (Abedi, To
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Discussion dentinogenesis, some cel
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Discussion morphology. It was not p
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5 Conclusion Discussion Research on
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Discussion Figure 50: 14 months pos
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References during normal and in vit
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References Faraco, I. M., Jr. and R
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References Kidd, E. A., A. Banerjee
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References antigen-expressing cells
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References Smith, A. J. and H. Leso
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References Yamashiro, T., M. Tummer
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References Appendix 1: Full list of
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probe set GENE ID baseline mean (LS
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Appendix 3: Molecular characterizat
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Appendix 5: Evaluation of a new lab
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