PHYSICAL AND CHEMICAL PROPERTIES OF ENAMEL AND ITS DEVELOPMENT

Presented By: Isha Gupta. MDS(First Year) Department Of Cons & Endo. Page 1

INTRODUCTION
Enamel is HIGHLY MINERALIZED TISSUE. 96% mineral, 4% water. Inorganic crystalline calcium phosphate
HYDROXYAPPATITE.

Various ions like strontium, magnesium, lead and fluoride are present at some point during Enamel formation.
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 Enamel is very brittle but the underlying dentin provides some resilience. Organized into Rods (prisms) and Interrods (interprismatic substance) Built from closely packed ribbon-like Hydroxyapatite crystals 60 to 70 nm in width and 25 to 30 nm in thickness.

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PHYSICAL CHARACTERISTICS

Enamel is the hardest calcified tissue. Structure and hardness of the Enamel render it brittle. SPECIFIC GRAVITY of Enamel is 2.8 THICKNESS: Enamel attains a maximum thickness of about 2 to 2.5 mm on the cusps , thinning down to almost a knife edge at the neck of the tooth.
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Another physical property of Enamel is its PERMEABILITY. COLOUR CHARACTERICTICS: The color of the Enamelcovered crown ranges from yellowish white to grayish white. The color is determined by differences in TRANSLUCENCY of Enamel which attributed to variations in the degree of calcification and homogeneity of the Enamel.
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CHEMICAL PROPERTIES
Inorganic material (96%) and only a small amount of organic substance and water (4%). Enamel matrix mineralization begins immediately after it is secreted. Enamel primary and secondary mineralization increase mineral content in a relatively smooth curve. Relative space occupied by the organic framework and entire Enamel is almost equal.
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 Chemical analysis of matrix of mature Enamel indicate that proteins can be isolated in several different fractions high % of serine, glutamic acid and glycine. X-ray diffraction studies reveal that molecular structure is typical of group of proteins called CROSS BETA PROTEIN.
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TRANSMISSION ELECTRON MICROGRAPH

ENAMEL CRSYTALS ARE LONG PARALLEL ORGANIZED HYDROXYAPPATIE CRYSTALS ORGANIZED IN BUNDLESWHICH ARE CALLED PRISMS. ROWS OF ENAMEL PRISMS ARE ORGANIZED PERPENDICULAR TO EACH OTHER.

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DEVELOPMENT
EPITHELIUM ENAMEL ORGAN originating from the primitive oral cavity, consists of four layers.
    OUTER ENAMEL EPITHELIUM STELLATE RETICULUM STRATUM INTERMEDIUM INNERENAMEL EPITHELIUM

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 DENTINOENAMEL JUNCTION is the borderline between inner Enamel epithelium and the connective tissues of dental papilla. The Inner Enamel Epithelium reflects onto the Outer Enamel Epithelium at wide basal opening of Enamel organ and forms cervical loop. The different layers of epithelial cells of the Enamel organ are named according to their -MORPHOLOGY -FUNCTION -LOCATION
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OUTER ENAMEL EPITHELIUM

Outer Enamel Epithelium consists of a single layer of cuboid cells. The capillaries in the connective tissues surrounding the Epithelial Enamel organ proliferate and protrude toward it. As the Enamel formation starts capillaries may even indent the stellate reticulum. This increases blood supply to inner Enamel epithelium which ensures a rich metabolism. Cell specialization occurs during Enamel formation for active transport of materials, cells develop villi and cytoplasmic vesicles and large number of mitochondria.
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STELLATE RETICULUM
Middle part of Enamel organ is formed by STELLATE RETICULUM, which has cells that are separated by wide intercellular substance. The cells are star shaped with long processes reaching in all directions from the central body. They are connected with each other and with the cells of Outer Enamel Epithelium and stratum intermedium by DESMOSOMES. The structure of stellate reticulum renders it resistant and elastic.
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STRATUM INTERMEDIUM
The Stratum intermedium are situated between stellate reticulum and inner Enamel epithelium. They are connected to each other and their neighboring cells by desmosomes. The function of the stratum intermedium is not understood ,but it is believed to play a role in production of Enamel itself either through

Control of fluid diffusion into and out of the ameloblasts By actual contribution of necessary formative elements or enzymes.
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INNER ENAMEL EPITHELIUM

Cells of the Inner Enamel Epithelium are derived from the basal cell layer of the oral epithelium. Before Enamel formation begins, these cells assume a columnar form and differentiate into AMELOBLASTS that produce the Enamel matrix. The cell differentiation occurs earlier in the region of the incisal edge or cusps than in the area of the cervical loop.
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CERVICAL LOOP
At the free border of the Enamel Organ the outer and inner Enamel epithelium layers are continuous and reflected into one another as a CERVICAL LOOP. When the crown has been formed, the cells of this portion give rise to Hertwigs epithelial root sheath.

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LIFE CYCLE OF AMELOBLASTS

The life span of the cells of the Inner Enamel Epithelium can be divided into six stages based on their FUNCTION: MORPHOGENIC ORGANIZING FORMATIVE MATURATIVE PROTECTIVE DESMOLYTIC
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MORPHOGENIC STAGE
Before the ameloblasts are fully differentiated to produce Enamel, they interact with adjacent mesenchymal cells and determining shape of the Dentinoenamel junction and the crown. Morphologic changes occurs during this are:  Cells are short and cuboidal or low columnar with large oval nuclei that almost fill the cell body.  Golgi apparatus and the centrioles are located in the proximal end of the cell  Mitochondria are evenly dispersed throughout the cytoplasm.  Terminal bars appear concomitantly with the migration of the mitochondria to the basal region of the cells  The terminal bars represent points of close contact between cells.

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ORGANIZING STAGE
Inner Enamel Epithelium interacts with the adjacent connective tissue cells, which differentiate into ODONTOBLASTS. This stage is characterized by a change in the appearance of the cells of the Inner Enamel Epithelium. Amount of rough endoplasmic reticulum increases significantly and most of mitochondria cluster in proximal region.
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 REVERSAL OF FUNCTIONAL POLARITY of these cells takes place by the migration of the centrioles and Golgi bodies from the proximal ends of the cells into their distal ends. The clear free zone between the Inner Enamel Epithelium and the dental papilla disappears probably because of elongation of the epithelium cells toward the papilla.

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Elongation of epithelial cells toward dental papilla occur

Epithelial cells come in close contact with papilla

Formation of dentin takes place

The first appearance of dentin seems to be a critical phase in the life cycle of the inner Enamel Epithelium AS REVERSAL OF NUTRITIONAL SOURCE takes place.
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 Until cells are in contact with the Dental Papilla, it receives nutrient material from the blood vessels of this tissues. When DENTIN FORMS, it cuts off the ameloblasts from their original source of nourishment, then on they are supplied by the capillaries that surround and may even penetrate the Outer Enamel Epithelium.
REVERSAL OF NUTRITIONAL SOURCE is

characterized by proliferation of capillaries of the dental sac and by reduction and gradual appearance of the stellate reticulum.

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FORMATIVE STAGE
When the first layer of dentin has been formed, ameloblasts enter their formative stage. This mutual interaction between one group of cells and another is one of the fundamental laws of organogenesis and histodifferentiation. The ameloblasts retain approximately the same length and arrangement, during the Enamel matrix formation.
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 Initiation of secretion of Enamel matrix influences the changes in organization and number of cytoplasmic organelles. There is development of blunt cell processes on the ameloblasts surfaces, which penetrate the basal lamina and enters predentin.
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MATURATIVE STAGE
Enamel maturation occurs after most of the thickness of the enamel matrix has been formed in the occlusal or incisal area. Enamel matrix is still progressing at this time in cervical areas. Ameloblasts are slightly reduced in length, with decrease in volume and organic content. During maturation, Ameloblasts display microvilli at their distal extremities, and cytoplasmic vacuoles containing material resembling Enamel matrix are present. These structures indicate an absorptive function these cells.
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PROTECTIVE STAGE
The ameloblasts cease to be arranged in a well defined layer and thus they cannot be differentiated from the cells of the stratum intermedium and Outer Enamel Epithelium. These cell layers then form a stratified epithelial covering of the Enamel, the so called REDUCED ENAMEL EPITHELIUM. The function of this layer is protecting the mature Enamel by separating it from the connective tissue until the tooth erupts. If Connective tissues comes in contact with the Enamel, anomalies, may develop.
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DESMOLYTIC STAGE
Reduced Enamel Epithelium proliferates and seems to induce atrophy of the connective tissue separating it from the oral epithelium, so that fusion of the two epithelia can occur. Epithelial cells elaborate enzymes that are able to destroy connective tissues fibers by Desmolysis. Premature degeneration of the reduced Enamel epithelium may prevent the eruption of a tooth.

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FUNCTIONS OF CELLS OF INNER ENAMEL EPITHELIUM

FUNCTIONS Determining Crown Pattern Of the Tooth. MORPHOLOGIC CHANGES OF CELLS Cells are small, Cuboidal, Centrally placed Nuclei. Morphogenic changes occur within them and forms ameloblasts. Ameloblasts develop conical extension (TOMESS PROCESS) They exhibits cyclical variation with ruffle and smooth ended border against Enamel surface Cells surfaces until the time of tooth eruption

Differentiation Of Odontoblasts.

Secretion of Enamel matrix

Maturation of Enamel phase

Protection of newly formed Enamel

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AMELOGENESIS
On the basis of ultrastructure and composition, two processes are involved in the development of Enamel: ORGANIC MATRIX FORMATION MINERALIZATION AND MATURATION

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AMELOBLAST DIFFERENTIATION
Amelogenesis is two step process involving secretion a distinctive organic matrix which is almost immediately partially mineralized First formed Enamel is partially mineralized when amelogenesis is completed it become fully mineralized as a result of maturation activity on the part of ameloblasts. Inner Enamel Epithelium cells gets differentiated into ameloblasts beginning first at tips of cusp outlines and then sweeping down the slopes.
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ENAMEL MATRIX FORMATION

Next step in process of amelogenesis is synthesis of organic matrix of Enamel. In amelogenesis the term matrix is used not only for organic component but also the inorganic component of first formed Enamel. First formed (partially mineralized) Enamel is referred to simply as DEVELOPING ENAMEL. Its organic component is referred to as ENAMEL PROTEIN.

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ENAMEL PROTEIN

Term AMELOGENIN was coined to describe the nature of Enamel protein which was characterized as having a mol. Weight of 22 to 30 kilodaltons. It is hydrophobic and rich in proline, histidine and glutamine with an ability to flow under pressure. Amelogenin is now known to constitute approximately 90% of Enamel protein in secretory stage.
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 A second Enamel protein of higher mol. Weight 48 to 70 kds which is acidic and rich in glutamic acid and asparatic acid and serine. In particular , Amelogenin contains a 45 aminoacid residue sequence known as TRAP(Tyrosine rich amelogenin protein) As soon as amelogenin is secreted, it undergoes a series of varying and discrete modifications.
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DEVELOPMENT OF TOMES PROCESSES

Surfaces of ameloblasts facing the developing Enamel are not smooth. The projections of the ameloblasts into Enamel matrix have been named TOMES PROCESSES. Tomes processes are partly delineated by incomplete septa, they also contain typical secretions granules as well as rough endoplasmic reticulum and mitochondria

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DISTAL TERMINAL BARS

At the time tomes processes begin to form, terminal bars appear at distal ends of the ameloblasts, separating the tomes processes from the cell proper. Structurally they are localized condensations of cytoplasmic substance closely associated with thickened cell membranes.

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AMELOBLAST COVERING MATURING ENAMEL

Ameloblasts over Maturing Enamel are considerably shorter. These short ameloblasts have a villous surface near the Enamel and the ends of cells are packed with mitochondria Striking difference between Enamel and other mineralized tissues is the organic component as well as water is lost in mineralization. Over 90% of initially secreted protein is lost during Enamel maturation and that which remains forms envelopes around individual crystals
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MINERALIZATION AND MATURATION OF ENAMEL MATRIX

Mineralization of Enamel matrix takes place in four stages FIRST STAGE :

As the matrix segments and the interprismatic substances are laid down their partial mineralization occurs. Chemical analyses indicate that initial influx may amount to 25 to 30% of the eventual total mineral content.
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SECOND STAGE OR MATURATION

Characterized by the gradual completion of mineralization. Maturation starts from the height of the crown and progresses cervically. At each level, maturation seems to begin at the dentinal end of the rods.
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 Thus there is an integration of the two processes:

EACH ROD MATURES FROM THE DEPTH OF THE SURFACE THE SEQUENCE OFMATURING RODS IS FROM CUSPS OR INCISAL EDGE TOWARD THE CERVICAL LINE.

THIRD STAGE Tertiary increase in mineral rebounding from innermost layer out toward the Enamel surface. Surface layer some 15micrometer wide can be distinguished during this phase, it mineralizes more slowly.
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FOURTH STAGE The outer layer mineralizes rapidly and heavily (quaternary mineralization) becoming the most mineralized part of Enamel. Maturation begins before the matrix has reached its full thickness. Thus it is going on in the inner, first formed matrix then on outer, recently formed matrix.

The advancing front is at first parallel to dentin Enamel junction and later to the Outer Enamel surface.
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 At the Ultrastructural level, maturation is characterized by growth of the crystals seen in the primary phase. Enamel is most highly mineralized at its surface with degree of mineralization decreasing toward DEJ until innermost layer is reached.
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