1. Dental germ

2. Morphological and biological characteristics of the tooth germ

3. Origin of teeth Teeth have a dual origin: Ectodermal; Mesodermal:
Ectomesenchymal.

4. EPITHELIUM-ECTOMESEMCHYME INTERACTION IN THE TOOTH DEVELOPMENT
Combine tissue type (neural crest cells + mesenchyme) is called ectomesenchyme.
In tooth development, an interaction occurs between oral ectoderm and ectomesenchyme to initiate the process.
Cellular interactions, between epithelium+ectomesenchyme: Formation of dentine, enamel and cementum as well as crown;

5. The processes involved in tooth development
Morphogenesis is stimulated by molecular signals that control:
Cell growth;
Migration;
Cell fate and differentiation.
For every developmental event a complex and intricate cascade of gene expression take place to direct the cells to the right place and onto the proper differentiation pathway;
Many of these pathways all result from epithelial- mesenchymal interactions in which essentially the same molecular mediators are implicated.

6. Embryonic terms to consider Induction
First process to occur during embryogenesis;
Interaction between developing embryonic cells.

7. Morphogenesis May also be called morphodifferentiation;
Development of form and specific tissues;
Results from migration of embryonic cells and inductive interactions between these cells.

8. Patterning
Differentiation
Specification of the embryo through segmentation;
Patterning is both a spatial and temporal event as exemplified by regional development of incisors, canines, premolars, and molars, which occurs at different times and involves the classical processes of induction, competence, and differentiation.
Process of specialization of embryonic cells.

9. interstitial growth Growth deep within a tissue of organ;
As opposed to appositional growth – growth at the periphery through the addition of additional cell layers;

10. Stages of development Initiation; Proliferation; Histodifferentiation;
Morphodifferentiation;
Apposition;
Maturation;
Eruption.

11. Initiation of tooth development
An intriguing question is how dental development is initiated?

12. Iniciation of tooth development
The newly formed stomadeum is lined by a primitive two- or three-cell-thick layered epithelium covering an embryonic connective tissue that, because neural crest cells have migrated in it, is termed ectomesenchyme;
The ectomesenchyme consists of a few spindle-shaped cells separated by a gelatinous ground substance.

13. PHASE 1-initiation phase
Formation of primary epithelial thickening, dental lamina, tooth bud & the cap, bell stage cannot take place in absence of dentally active ectomesenchyme which becomes concentrated in regions of presumptive tooth development;
Inner epithelium of enamel organ induces ectomesenchyme of dental papilla to become orientated towards the intervening basal lamina and differentiate into odontoblasts.

14. Iniciation of tooth development
Thickening of the epithelium.

15. Stages of the initiation
Primary epithelial band;
Vestibular lamina;
Dental lamina.

16. Primary epithelial band
A continuous band of thickened epithelium forms around the mouth in the presumptive upper and lower jaws;
This bands are roughly horseshoe shaped and correspond in position of the future dental arches of the jaws;
Each band of epithelium quickly gives rise to two subdivisions:
The vestibular lamina and
The dental lamina just behind it.

17. The position of the primary epithelial band

18. Initiation of the tooth development

19. Upper epithelial
band
tongue
lower epithelial
band

20. Upper and lower
epithelial bands

21. The reason for epithelial thickening
The formation of these thickened epithelial bands is the result not so much of increased proliferative activity within the epeithelium as of a change in orientation of the mitotic spindle and clevage plane of dividing cells.

22. Sagittal section through the head of an embryo.
A. Primary epithelial bands; B. The same structure at higher magnification; C The change in plane of clevage.

23. Development of the epithelial proliferation
Upper epithelial
band
Lower
Epithelial
band

24. Deepening of the epithelial proliferation

25. Vestibular Lamina
The first subdivision of the primary epithelial band is vestibular lamina;
The vestibule forms as a result of the proliferation of the vestibular lamina into ectomesenchyme;
Its cells rapidly enlarge and then degenerate to form a cleft that becomes the vestibule between the cheek and tooth-bearing area.

26. Dental lamina
The other subdivision of the primary epithelial band is dental lamina;
Dental lamina is located orally to the vestibular lamina;
Within the dental lamina, continued and localized proliferative activity leads to the formation of a series of epithelial outgrowths into the ectomesenchyme at sites corresponding to the positions of the future deciduous teeth.

27. Formation of the vestibule of the vestibular lamina

28. Dental lamina, Vestibular lamina
Vestibular proliferation leads to the formation of the vestibular lamina;
Oral proliferation leads to the formation of the dental lamina.
Vestibular
lamina

29. tongue
Vestibular
lamina
Dental
lamina

30. Next development of dental lamina
Begin localized proliferations;
Early in the medial part - incisors;
Then in the lateral part - canines and molars;
In the upper and lower dental lamina are formed 10 proliferations;
The process is called “the formation of tooth bud”
This is the beginning of the tooth development;

31. Initiation of tooth development
Teeth develop from two typs of cells:
Oral epithelial cells;
Mesenchymal cells from the dental papilla;
Neural crest cells – ectomesenchymal cells;
Epithelial cells are flat on the surface of the epithelium and cylindrical immediately above to the basal lamina;
The underlying ectomesenchymal cells accumulate around the epithelial autgrowths.

32. The flat and cylindrical epithelial cells, basal lamina immediately below, and ectomesenchymal cells
Flat cells
Cylindrical cells

33. Iniciaction of the tooth

34. The controlling signals (BMP-7) in place of the thickened epithelium

35. Proliferation The proliferation is: The proliferation is observed:
Multiplication of cells;
Increases the amount of tooth bud;
The proliferation is observed:
In the epithelium;
In the ectomesenchym.

36. Certain cells of the basal layer begin to proliferate at a more rapid rate then do the adjacent cells.
These proliferating cells contain the entire growth potential of the teeth .

37. Toothe development proceeds in three stages:
Dental bud;
Dental cap;
Dental bell.
These terms are discriptive of the morphology but we have to describe the significant fuctional changes that occur during development, such as morphogenesis and histodifferentiation;
Because development is a continuous process, clear distinction brtween the transition stages is not possible.

38. Dental bud
1. Dental bud;
2. Oral epithelium;
3. Ectomesenchyme.

39. Histodifferentiation
Histological appearance of different cells with:
Different form;
Different size;
Different function;
Than are formed different layers of cells :
In the epithelium;
In the ectomesenchym.

40. Dental Lamina and Vestibular Lamina

41. Morphogenesis with morphodifferentiation
The differentiated cells shall be arranged under different layers;
Each layer acquires a separate function;
Each layer takes part in creating the structures of the tooth, in determining the shape and size of the tooth.

42. Stages in the development of tooth germ

43. Stages in the development of tooth germ

44. Elements of the tooth germ
Enamel organ - epithelial;
Dental papilla:
Мesenchymal;
Ectomesenchymal;
Dental follicle:
Ectomesenchymal.

45. Bud stage

46. Bud stage

47. Bud stage of tooth development
Proliferation of epithelial cells;
Mitotic cell division;
Increasing the cells number;
Increasing the size of the cells;
Still no cells differentiation.

48. Bud stage
The bud stage is represented by the first epithelial incursion into the ectomesenchyme of the jaw;
The epithelial cells show little if any change in shape or function;
The supporting ectomesenchymal cells are packed closely and around the epithelial bud.

49. Formation and growth of tooth bud - activation of mesoderm

50. Ectomesenchylal cells arriving directly from neural crests
Stem cells involved in the formation of the tooth bud;
This is pluripotent cell capable to become virtually any cell.

51. Dental bud At this stage there is no cell differentiation;
There is only cell division and cell mitotic activity;
The cells are identical in shape and size.

52. Dental lamina and dental bud

53. Dental bud The tooth bud now is a ball of cells;
The tooth bud grows rapidly in volume.

54. Bud to Cap Transition
The transition from bud to cap marks the onset of morphological differences between tooth germs that give rise to different types of teeth;
As the epithelial bud continues to proliferate into the ectomesenchyme, cellular density increases immediately adjacent to the epithelial outgrowth;
This process referred to as a condensation of the ectomesenchyme, results from a local grouping of cells that have failed to produce extracellular substance and have thus not separated from each other.

55. Lateral lamina or Gubernaculum dentis
As the tooth bud grows larger, it drags along with it part of the dental lamina;
So from that point on the developing tooth is tethered to the dental lamina by an extension called gubernaculum dentis or lateral lamina.

56. The extension of the dental bud into the ectomesenchyme

57. The extension continues and can be seen condensing ectomesenchyme.

58. Transition from dental bud to dental cap
Transition from dental bud to dental cap. There is condensed ectomesenchyme.

59. Model of mutual stimulation between the epithelial and ectomesenchymal growth factors.
Separated from ectomesenchyme factors (red) stimulated epithelial cells (blue), which in turn cause the release of others in the ectomesenchyme (pink);
Immediately the bone morphogenetic protein (green) inhibits the further release of epithelial factors (blue);
This is the starting signal for the cells differentiation.

60. Crossing the bud stage in cap stage

61. Transition from bud to cap

62. Cap stage

63. Concavity at the top of the tooth bud under the control of mesenchymal factors

64. Processes taking place in cap stage
It is the beginning of cellular differentiation;
Differentiated cells are arranged in layers;
The formative elements of the tooth are already visible:
The epithelial outgrowth, which resemble a cap sitting on a ball of condensed ectomesenchyme, is referred as the enamel organ (it will form the enamel of the tooth);
The ball of condensed ectomesenchymal cells, called dental papilla, forms the dentin and pulp;
The condensed ectomesenchyme limiting the dental papilla and encapsulating the enamel organ – the dental follicle or sac – give rise to the supporting tissues of the tooth.

65. Cap stage – early and late

66. Cup stage
Because the enamel organ sits over the dental papilla like a cap, this stage of the tooth development is known as the cap stage.

67. The concavity of the enamel organ

68. Histodifferentiation of the cells of the enamel organ
The cells that covering the enamel organ are outer dental (or enamel) epithelium;
The cells, which are covering the recess of the enamel organ are inner dental epithelium;
The center of the enamel organ is termed the stella reticulum:
The cells in the center of the enamel organ synthesize and secrete glycosaminoglycans into the extracellular compartment between the epithelial cells;
Glycosaminoglycans are hydrophilic and so pull water into the enamel organ;
The increasing amount of fluid increases the volume of the extracellular compartement of the enamel organ, and the central cells are forced apart;
Because they retain connections with each other through their desmosomal contact, they become star shaped.

70. The layers of the enamel organ in the cap stage
You can see three separate cell layers :
Outer epithelium – a single row of cuboidal cells;
Inner epithelium - cuboid cells adjacent to dental papilla;
Stellate reticulum -star shaped cells.They are the largest volume with large intercellular spaces.

71. Formation of three new structures
Enamel knots – are cluster of nondividing epithelial cells;
Enamel cord – enamel knot is extends between the inner and outer epithelia as the enamel cord;
Enamel niche - spaces between the epithelium of the dental lamina and outer enamel epithelium.

72. Transition from the cap to the bell stage
1. Dental lamina
2. Dental follicle
3. Outer enamel epithelium;
4. Stellate reticulum;
5. Inner enamel epithelium;
6. Dental papilla;
7. Enamel cord - epithelial proliferation, which seemed to divide enamel organ into two parts;
8. Enamel
9. Blood vessels;
10. Enamel niche;
11. Permanent tooth bud
Transition from the cap to the bell stage

73. Enamel knot Enamel knots are clusters of nondividing epithelial cells;
The current view is that the enamel knot represents an organizational center, which orchestrates morphogenesis.

74. Enamel knot
Blue dots are the boundary between epithelial and ectomesenchyme cells;
Red circles and arrows indicate enamel knot - the command center.

75. Enamel knot formation in tooth cap
The primary enamel knot is under the influence of beta- catenin;
The progression of a bud in the cap is done under mesenchymal incentives that activate cells from the tip of the tooth bud to form enamel knot. This is the epithelial signaling center.
The enamel knot is an important regulator of tooth shape;
Induction of the enamel knot is required for the transition from bud to cap;

76. Enamel knot Enamel knot is cluster of nondividing epithelial cells.
If the enamel knot be removed, the tooth is not developing.

77. Enamel knot - the expression of growth factors

78. Enamel knot and enamel cord
Each tooth germ has a single primary enamel knot and enamel cord at the cap stage, and as these disappear, secondary enamel knots appear at the tip of the future cusps in molars.

79. Enamel cord Localized group of cells in the enamel organ;
Enamel cord be located under the outer enamel epithelium;
It directs and binds to enamel knot;
The function of the enamel cord is obviously associated with the enamel knot.

80. Enamel cord
Enamel
knot

81. Enamel niche
This structure is created by the plane of section cutting through a curved lateral lamina so that mesenchyme appears to be surrounded by dental epithelium.

82. Bell stage

83. Bell stage
Continued growth of the tooth germ leads to the next stage – bell stage, so called because the enamel organ comes to resemble a bell as the undersurface of the epithelial cap deepens;
During this stage, the tooth crown assumes its final shape (morphodifferentiation);
The cells that will be making the hard tissues of the crown (ameloblasts and odontoblasts) acquire their distinctive phenotype (histodifferentiation).

84. Enamel organ has four layers of cells
At the periphery of the enamel organ the cells assume a low cuboidal shape and form the outer dental epithelium;
The cells bordering on the dental papilla assume a short columnar shape – inner dental epithelium;
The star-shaped cells forms stellate reticulum;
In the bell stage, some epithelial cells between the inner dental epithelium and the stellate reticulum differentiate into a layer called the stratum intermedium.

86. Bell stage of the enamel organ

87. Bell stage of the enamel organ
Represents enlargement of the overall size of the tooth germ and deepening of its under surface;
Cells at the center secrete an acid mucopolysaccharide into the extracellular space between the epithelial cells covering the germ, (drawing in of the water cause enlargement of germ);
A zone of stretched but interconnected cells (stellate reticulum) produced at center of the germ;
Epithelial cells:
Next to the papilla develop into an enamel-producing layer of cells (inner dental epithelium);
Along leading edge of germ form the outer dental epithelium (dental cuticle).
The transition zone between outer and inner dental epithelia forms the cervical loop.

89. Outer enamel epithelium
This layer cover enamel organ;
The cells are cuboidal shape;
They are connected to dental lamina;
The function of this cell layer is to transport products from dental follicle to enamel organ and to limit the growth impulses of enamel organ.

90. Inner enamel epithelium
In the early bell stage they have short columnar shape;
The cells become tall and columnar;
The cells are located directly above the basement membrane, which separates them from the cells of the dental papilla;
They have highly developed intracellular organelles;
At this stage they receive nutrients from the dental papilla;
Cell nucleus is large and positioned above the basement membrane.

91. Inner epithelium

93. Functions of the inner epithelium
These cells are called "pre ameloblasts”
The functions are:
They are differentiated for protein synthesis;
They are preparing for secretory function;
They stimulate the underlying ectomesenchyme of the dental papilla;
They produce and secrete the enamel;
They determine the shape of the dental crown.

95. Stellate reticulum
The star-shaped cells are conected to each other, to the cells of the outer dental epithelium, and to the stratum intermedium by attachment plaques known as desmosomes;
Their cytoplasm contains all of the usual organelles, but these are distributed sparsely;

96. Stellate reticulum

97. Functions of the stellate reticulum
This layer provides a space for the development of enamel;
It transports nutrients and incentives for the ameloblasts;
In the wide intercellular space they stored the nutrients and water;
There takes place transformation of the different substances.

98. Stratum intermedium
The cells of stratum intermedium are connected to each other and to the cells of the stellate reticulum and inner epithelium also by desmosomes;
There are openings between the cells;
These cells can to be further differentiate into the cells of the neighboring layers;
This is the reason this layer is called the germinative layer.

99. Stratum intermedium

100. Two other important events occur during the bell stage:
First, the dental lamina (and lateral lamina) joining the tooth germ to the oral epithelium breaks up into discret islands of epithelial cells, thus separating the developing tooth from the oral epithelium;
Second, the inner dental epithelium completes its folding, making it possible to recognize the shape of the future crown pattern of the tooth.

101. Epithelial pearls
Fragmentation of the dental lamina results in the formation of discrete clusters of epithelial cells that normally degenerate, but some may persist and are given the name of epithelial pearls;
These may form small cysts (eruption cysts) and delayed eruption;
May give rise to odontoma;
Or may be activated to form supernumerary teeth.

102. Disintegration of the dental lamina

103. Disintegration of the dental lamina at higher magnification

104. Zone of reflexion or cervical loop
The region where the inner and outer dental epithelium meet at the rim of the enamel organ is known as the zone of reflexion or “cervical loop”;
This point is where the cells continue to divide until the tooth crown attains its full size and which, after crown formation, gives rise to epithelial component of root formation;
Later epithelial cells of the cervical loop proliferate to form a double layer of cells known as Hertwig`s epithelial root sheath.

105. Cervical
loops

106. Cervical loop

107. Cervical loop

109. Hertwig's epithelial root sheath
Hertwig's epithelial root sheath (frequently abbreviated as "HERS") is a proliferation of epithelial cells located at the cervical loop of the enamel organ in a developing tooth.
Hertwig's epithelial root sheath initiates the formation of dentin in the root of a tooth by causing the differentiation of odontoblasts from the dental papilla.
The root sheath will further dictate whether the tooth will have single or multiple roots.

110. Functions of the Herwig’s epithelial root sheath
It stimulates the ectomesenchyme of the dental papilla to differentiation;
It simulates the ectomesenchyme of the dental follicle to differentiation;
It contains information on the number, size and shape of the dental root.

111. Formation of the permanent dentition
So far, only the initial development of the decidious (or primary) dentition has been described;
The permanent dentition also arise from the dental lamina;
The tooth germs that gives rise to the permanent incisors, canines, and premolars form as a result of further proliferative activity within the dental lamina at its deepest extremity;
This increased activity leads to the formation of another tooth bud on the lingual aspect of the deciduous tooth germ, which remains dormant for some time;

112. The molars of the permanent dentition
They have not deciduous predecessors, so their tooth germs do not originate in the same way;
The dental lamina burrows posteriorly beneath the lining epithelium of the oral mucosa into the ectomesenchyme;
This backward extention successively form the tooth germs of the first, second , and third molars.

113. Crown Pattern Determination
The cessation of mitotic division within cells of the inner dental epithelium determinise the shape of a tooth;
When the tooth germ is growing rapidly during the cap to bell stage, cell division occurs throughout the inner dental epithelium;
Division ceases at a particular point because the cells are beginning to differentiate and assume their eventual function of producing enamel;
The point at which inner epithelial cell differentiation first occurs represents the site of future cusp development, or the growth center.

114. Summary of crown pattern formation in the internal dental epithelium
The zone of cell devision is indicated by the brekened area in the inner dental epithelium, and the zone where cells have differentiated by the white area.

115. Toothe type determination
The determination of specific tooth types at their correct positions in the jaws is referred to as patterning of the dentition;
Two hypothetical models have been proposed to explain how these different shapes are determined, and evidence exist tu support both:
The first is the field model;
The second is the clone model.

116. Field model
The field model proposes that the factors responsible for tooth shape reside within the ectomesenchyme in distinct but graded field for each tooth family;

117. For example:
There is a "incisal field" responsible for dental shape of the incisors;
It is concentrated in the central incisal area;
The stimulation strongly reduced at the end of the field.

118. Presumptive
incisor mesenchyme
Molar
field
Incisal field

119. Clone model
The clone model proposes that each tooth class is derived from a clone of ectomesenchymal cells programmed by epithelium to produce teeth of a given pattern;

120. Clone theory
The molar clon has induced the dental lamina to begin tooth development. At its posterior border the clone and dental lamina grow posteriorly by means of progress zona.
B. When a clone reaches the critical size, a tooth bud is initiated at its center. A zone of inhibition surrounds the tooth bud.
C. The next tooth bud is not initiated until the progress zone of the clone has escaped its influence.

121. Development of the dental papilla
The dental papilla is separated from the enamel organ by a basement membrane;
The cells of the dental papilla appear as undifferentiated ectomesenchymal cells, having an uncomplicated structures with all the usual organelles;
Blood vessels appear early in the dental papilla, initially in the central region along with nerve fibers associated with these vessels;
The vessels bring nutrition to the rapidly growing organ;
As the papilla grows, smaller vessels are also seen in the periphery of the area, bringing nutrition to the elongating odontoblasts and ameloblasts.

122. Dental
papille

125. Functions of the dental papilla
The ectomesenchyme of the dental papilla forms:
Preodontoblasts in the early bell stage;
Odontoblasts in the late bell stage;
The papilla cells are believed to be significant in furthering enamel organ bud formation into the cap and bell stage;
This cell density is maintained as the enamel organ grows.
Dental papilla gives rise to the dentin and dental pulp;

126. Dental follicle
-The dental follicle is distinguished clearly from the dental papilla in that many more collagen fibrils occupy the extracellular spaces between the follicular fibroblasts; -These generally are oriented circulary around the dental organ and dental papilla; -Clusters of blood vessels are found ramifying around the tooth germ in the dental follicle adjasent to the outer dental epithelium

127. Dental follicle
Dental follicle is dental sac with its enclosed developing tooth.
It gives rise to three important entities: cementoblasts, osteoblasts, and fibroblasts.
Cementoblasts form the cementum;
Osteoblasts form alveolar bone;
Fibroblasts form periodontal ligaments which connect teeth to the alveolar bone;
The dental follicle is formed partly from cells at the base of the dental papilla that flatten and migrate to enclose the whole of tooth germ.

128. Dental
follicle

129. The role of dental follicle in the development of tooth structure
Dental follicle brings nutrition to the enamel organ and dental papilla;
It limits their growth impulses;
After stimulation of the Herwig’s epithelial root sheath dental follicle forms cement, periodontium and alveolar bone.

130. The role of the tooth germ elements in the formation of teeth
Enamel organ:
Forms enamel;
Furthered dental papilla and dental follicle;
Dental papilla:
Forms dentin and dental pulp;
Dental follicle:
Forms cement, periodontium and alveolar bone.

131. Stages of the dental germ development