Molecular characterisation of odontoblast during primary, secondary ...

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Introduction In summary, primary dentinogenesis occurs during development leading to formation of the crown and root of the tooth, whereas secondary dentine is secreted throughout the life of the tooth, and is responsible for reduction in the size of the pulp chamber, root canals and continued deposition of peritubular dentine (Baume 1980). Tertiary dentine: is secreted in response to external stimuli, such as caries or abrasion, in order to protect the underlying pulp. In the case of moderate stress, which does not result in destruction of odontoblasts, the secreted dentine is termed “reactionary dentine”; when the stress is more intense and odontoblast survival is compromised (i.e., dentine bridge formation at sites of exposure), it is termed “reparative dentine” (Figure 3). Figure 3: The two types of tertiary dentinogenesis. . 22
Introduction Understanding the distinction between the three types of dentine is important, and above all the distinction between secondary (physiological) and tertiary (healing) dentine must be clear, as they are often confused when reported in the literature. 1.2.2 Dentine and bone Notably, dentine and bone are very similar in their composition, but show some structural differences. Many studies have shown that the compositions of these tissues are more similar than previously appreciated. Indeed, certain molecules that were originally considered to be specific to dentine (such as Dentine sialoprotein; DSP) have been shown to be present in alveolar bone (Butler, Brunn et al. 2003; Huang, Sun et al. 2008). It is therefore more difficult to find specific markers to characterise these tissues. Nevertheless, the profiles of the SIBLINGS protein for example, show differences between the two tissues (Huang, Sun et al. 2008). On a structural level, bone and dentine exhibit several common features. Cells of both bone and dentine are of a similar mesodermal embryonic origin, which may explain some of the similarities found in their formation and structures. In bone, secretion of the matrix is performed by osteoblasts; once embedded in the mineralised matrix, they appear to change into quiescent cells called osteocytes which communicate with each other through an extensive network of cell processes embedded in canuliculi. In the dentine-pulp complex, odontoblasts remain on the formative surface of the dentine matrix but have a cell process extending through the dentine with many lateral processes. The similarities between the morphologies of dentinal tubules and osteocyte canuliculi have recently been highlighted (Lu, Xie et al. 2007) and recent molecular data demonstrate striking similarities in the behaviour of odontoblasts and bone cells at different stages of their life cycles (Simon, Smith et al. 2009). 23
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: Introduction Secondary dentine: thi
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 and 34: 1.2.3.3 Consequences of new technol
Page 35 and 36: Introduction an apical nucleus, whe
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
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Material and methods (pH=6.8) with
Page 79 and 80:
Material and methods (P), and calci
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Material and methods (Sigma, la Ver
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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
Page 97 and 98:
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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